Effect of cultivating conditions on α-galactosidase production by a novel Aspergillus foetidus ZU-G1 strain in solid-state fermentation

PubMed Central

Liu, Cai-qin; Chen, Qi-he; Cheng, Qian-jun; Wang, Jin-ling; He, Guo-qing

2007-01-01

The work is intended to achieve optimum culture conditions of α-galactosidase production by a mutant strain Aspergillus foetidus ZU-G1 in solid-state fermentation (SSF). Certain fermentation parameters involving moisture content, incubation temperature, cultivation period of seed, inoculum volume, initial pH value, layers of pledget, load size of medium and period of cultivation were investigated separately. The optimal cultivating conditions of α-galactosidase production in SSF were 60% initial moisture of medium, 28 °C incubation temperature, 18 h cultivation period of seed, 10% inoculum volume, 5.0~6.0 initial pH of medium, 6 layers of pledget and 10 g dry matter loadage. Under the optimized cultivation conditions, the maximum α-galactosidase production was 2 037.51 U/g dry matter near the 144th hour of fermentation. PMID:17542067

Encapsulation of brewing yeast in alginate/chitosan matrix: lab-scale optimization of lager beer fermentation.

PubMed

Naydenova, Vessela; Badova, Mariyana; Vassilev, Stoyan; Iliev, Vasil; Kaneva, Maria; Kostov, Georgi

2014-03-04

Two mathematical models were developed for studying the effect of main fermentation temperature ( T MF ), immobilized cell mass ( M IC ) and original wort extract (OE) on beer fermentation with alginate-chitosan microcapsules with a liquid core. During the experiments, the investigated parameters were varied in order to find the optimal conditions for beer fermentation with immobilized cells. The basic beer characteristics, i.e. extract, ethanol, biomass concentration, pH and colour, as well as the concentration of aldehydes and vicinal diketones, were measured. The results suggested that the process parameters represented a powerful tool in controlling the fermentation time. Subsequently, the optimized process parameters were used to produce beer in laboratory batch fermentation. The system productivity was also investigated and the data were used for the development of another mathematical model.

Encapsulation of brewing yeast in alginate/chitosan matrix: lab-scale optimization of lager beer fermentation

PubMed Central

Naydenova, Vessela; Badova, Mariyana; Vassilev, Stoyan; Iliev, Vasil; Kaneva, Maria; Kostov, Georgi

2014-01-01

Two mathematical models were developed for studying the effect of main fermentation temperature (T MF), immobilized cell mass (M IC) and original wort extract (OE) on beer fermentation with alginate-chitosan microcapsules with a liquid core. During the experiments, the investigated parameters were varied in order to find the optimal conditions for beer fermentation with immobilized cells. The basic beer characteristics, i.e. extract, ethanol, biomass concentration, pH and colour, as well as the concentration of aldehydes and vicinal diketones, were measured. The results suggested that the process parameters represented a powerful tool in controlling the fermentation time. Subsequently, the optimized process parameters were used to produce beer in laboratory batch fermentation. The system productivity was also investigated and the data were used for the development of another mathematical model. PMID:26019512

Culture Condition Optimization and Pilot Scale Production of the M12 Metalloprotease Myroilysin Produced by the Deep-Sea Bacterium Myroides profundi D25.

PubMed

Shao, Xuan; Ran, Li-Yuan; Liu, Chang; Chen, Xiu-Lan; Zhang, Xi-Ying; Qin, Qi-Long; Zhou, Bai-Cheng; Zhang, Yu-Zhong

2015-06-29

The protease myroilysin is the most abundant protease secreted by marine sedimental bacterium Myroides profundi D25. As a novel elastase of the M12 family, myroilysin has high elastin-degrading activity and strong collagen-swelling ability, suggesting its promising biotechnological potential. Because myroilysin cannot be maturely expressed in Escherichia coli, it is important to be able to improve the production of myroilysin in the wild strain D25. We optimized the culture conditions of strain D25 for protease production by using single factor experiments. Under the optimized conditions, the protease activity of strain D25 reached 1137 ± 53.29 U/mL, i.e., 174% of that before optimization (652 ± 23.78 U/mL). We then conducted small scale fermentations of D25 in a 7.5 L fermentor. The protease activity of strain D25 in small scale fermentations reached 1546.4 ± 82.65 U/mL after parameter optimization. Based on the small scale fermentation results, we further conducted pilot scale fermentations of D25 in a 200 L fermentor, in which the protease production of D25 reached approximately 1100 U/mL. These results indicate that we successfully set up the small and pilot scale fermentation processes of strain D25 for myroilysin production, which should be helpful for the industrial production of myroilysin and the development of its biotechnological potential.

Optimization of fermentation parameters to study the behavior of selected lactic cultures on soy solid state fermentation.

PubMed

Rodríguez de Olmos, A; Bru, E; Garro, M S

2015-03-02

The use of solid fermentation substrate (SSF) has been appreciated by the demand for natural and healthy products. Lactic acid bacteria and bifidobacteria play a leading role in the production of novel functional foods and their behavior is practically unknown in these systems. Soy is an excellent substrate for the production of functional foods for their low cost and nutritional value. The aim of this work was to optimize different parameters involved in solid state fermentation (SSF) using selected lactic cultures to improve soybean substrate as a possible strategy for the elaboration of new soy food with enhanced functional and nutritional properties. Soy flour and selected lactic cultures were used under different conditions to optimize the soy SSF. The measured responses were bacterial growth, free amino acids and β-glucosidase activity, which were analyzed by applying response surface methodology. Based on the proposed statistical model, different fermentation conditions were raised by varying the moisture content (50-80%) of the soy substrate and temperature of incubation (31-43°C). The effect of inoculum amount was also investigated. These studies demonstrated the ability of selected strains (Lactobacillus paracasei subsp. paracasei and Bifidobacterium longum) to grow with strain-dependent behavior on the SSF system. β-Glucosidase activity was evident in both strains and L. paracasei subsp. paracasei was able to increase the free amino acids at the end of fermentation under assayed conditions. The used statistical model has allowed the optimization of fermentation parameters on soy SSF by selected lactic strains. Besides, the possibility to work with lower initial bacterial amounts to obtain results with significant technological impact was demonstrated. Copyright © 2014 Elsevier B.V. All rights reserved.

Optimization of Hyaluronidase Inhibition Activity from Prunus davidiana (Carriere) Franch Fruit Extract Fermented by its Isolated Bacillus subtilis Strain SPF4211.

PubMed

Kim, Won-Baek; Park, So Hae; Koo, Kyoung Yoon; Kim, Bo Ram; Kim, Minji; Lee, Heeseob

2016-09-28

Strain SPF4211, having hyaluronidase (HAase) inhibition activity, was isolated from P. davidiana (Carriere) Franch fruit (PrDF) sugar extract. The phenotypic and biochemical properties based on 16S rDNA sequencing and an API 50 CHB kit suggested that the organism was B. subtilis. To optimize the HAase inhibition activity of PrDF extract by fermentation of strain SPF4211, a central composite design (CCD) was introduced based on three variables: concentration of PrDF extract (X₁: 1-5%), amount of starter culture (X₂: 1-5%), and fermentation time (X₃: 0-7 days). The experimental data were fitted with quadratic regression equations, and the accuracy of the equations was analyzed by ANOVA. The statistical model predicted the highest HAase inhibition activity of 37.936% under the optimal conditions of X₁ = 1%, X₂ = 2.53%, and X₃ = 7 days. The optimized conditions were validated by observation of an actual HAase inhibition activity of 38.367% from extract of PrDF fermented by SPF4211. These results agree well with the predicted model value.

Optimization of culture conditions for gamma-aminobutyric acid production in fermented adzuki bean milk.

PubMed

Song, Hung Yi; Yu, Roch Chui

2018-01-01

γ-Aminobutyric acid (GABA), a nonprotein amino acid, is widely distributed in nature and fulfills several physiological functions. In this study, various lactic acid strains commonly used to produce fermented milk products were inoculated into adzuki bean milk for producing GABA. The high GABA producing strain was selected in further experiment to improve the GABA production utilizing culture medium optimization. The results demonstrated that adzuki bean milk inoculated with Lactobacillus rhamnosus GG increased GABA content from 0.05 mg/mL to 0.44 mg/mL after 36 hours of fermentation, which showed the greatest elevation in this study. Furthermore, the optimal cultural condition to adzuki bean milk inoculated with L. rhamnosus GG to improve the GABA content was performed using response surface methodology. The results showed that GABA content was dependent on the addition of galactose, monosodium glutamate, and pyridoxine with which the increasing ratios of GABA were 23-38%, 24-68%, and 8-36%, respectively. The optimal culture condition for GABA production of adzuki bean milk was found at the content of 1.44% galactose, 2.27% monosodium glutamate, and 0.20% pyridoxine. Under the optimal cultural condition, the amount of GABA produced in the fermented adzuki bean milk was 1.12 mg/mL, which was 22.4-fold higher than that of the unfermented adzuki bean milk (0.05 mg/100 mL). The results suggested that the optimized cultural condition of adzuki bean milk inoculated with L. rhamnosus GG can increase GABA content for consumers as a daily supplement as suggested. Copyright © 2017. Published by Elsevier B.V.

Efficient Accumulation and In Vitro Antitumor Activities of Triterpene Acids from Submerged Batch--Cultured Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes).

PubMed

Wang, Xiao-Ling; Ding, Zhong-Yang; Zhao, Yan; Liu, Gao-Qiang; Zhou, Guo-Ying

2017-01-01

Triterpene acids are among the major bioactive constituents of lucidum. However, submerged fermentation techniques for isolating triterpene acids from G. lucidum have not been optimized for commercial use, and the antitumor activity of the mycelial triterpene acids needs to be further proven. The aim of this work was to optimize the conditions for G. lucidum culture with respect to triterpene acid production, scaling up the process, and examining the in vitro antitumor activity of mycelial triterpene acids. The key conditions (i.e., initial pH, fermentation temperature, and rotation speed) were optimized using response surface methodology, and the in vitro antitumor activity was evaluated using the MTT method. The optimum key fermentation conditions for triterpene acid production were pH 6.0; rotation speed, 161.9 rpm; and temperature, 30.1°C, resulting in a triterpene acid yield of 291.0 mg/L in the validation experiment in a 5-L stirred bioreactor; this yield represented a 70.8% increase in titer compared with the nonoptimized conditions. Furthermore, the optimized conditions were then successfully scaled up to a production scale of 200 L, and a triterpene productivity of 47.9 mg/L/day was achieved, which is, to our knowledge, the highest reported in the large-scale fermentation of G. lucidum. In addition, the mycelial triterpene acids were found to be cytotoxic to the SMMC-7721 and SW620 cell lines in vitro. Chemical analysis showed that the key active triterpene acid compounds, ganoderic acids T and Me, predominated in the extract, at 69.2 and 41.6 mg/g, respectively. Thus, this work develops a simple and feasible batch fermentation technique for the large-scale production of antitumor triterpene acids from G. lucidum.

Genome-wide Fitness Profiles Reveal a Requirement for Autophagy During Yeast Fermentation

PubMed Central

Piggott, Nina; Cook, Michael A.; Tyers, Mike; Measday, Vivien

2011-01-01

The ability of cells to respond to environmental changes and adapt their metabolism enables cell survival under stressful conditions. The budding yeast Saccharomyces cerevisiae (S. cerevisiae) is particularly well adapted to the harsh conditions of anaerobic wine fermentation. However, S. cerevisiae gene function has not been previously systematically interrogated under conditions of industrial fermentation. We performed a genome-wide study of essential and nonessential S. cerevisiae gene requirements during grape juice fermentation to identify deletion strains that are either depleted or enriched within the viable fermentative population. Genes that function in autophagy and ubiquitin-proteasome degradation are required for optimal survival during fermentation, whereas genes that function in ribosome assembly and peroxisome biogenesis impair fitness during fermentation. We also uncover fermentation phenotypes for 139 uncharacterized genes with no previously known cellular function. We demonstrate that autophagy is induced early in wine fermentation in a nitrogen-replete environment, suggesting that autophagy may be triggered by other forms of stress that arise during fermentation. These results provide insights into the complex fermentation process and suggest possible means for improvement of industrial fermentation strains. PMID:22384346

Modelling and Optimization Studies on a Novel Lipase Production by Staphylococcus arlettae through Submerged Fermentation

PubMed Central

Chauhan, Mamta; Chauhan, Rajinder Singh; Garlapati, Vijay Kumar

2013-01-01

Microbial enzymes from extremophilic regions such as hot spring serve as an important source of various stable and valuable industrial enzymes. The present paper encompasses the modeling and optimization approach for production of halophilic, solvent, tolerant, and alkaline lipase from Staphylococcus arlettae through response surface methodology integrated nature inspired genetic algorithm. Response surface model based on central composite design has been developed by considering the individual and interaction effects of fermentation conditions on lipase production through submerged fermentation. The validated input space of response surface model (with R 2 value of 96.6%) has been utilized for optimization through genetic algorithm. An optimum lipase yield of 6.5 U/mL has been obtained using binary coded genetic algorithm predicted conditions of 9.39% inoculum with the oil concentration of 10.285% in 2.99 hrs using pH of 7.32 at 38.8°C. This outcome could contribute to introducing this extremophilic lipase (halophilic, solvent, and tolerant) to industrial biotechnology sector and will be a probable choice for different food, detergent, chemical, and pharmaceutical industries. The present work also demonstrated the feasibility of statistical design tools integration with computational tools for optimization of fermentation conditions for maximum lipase production. PMID:24455210

Optimized Production of Xylitol from Xylose Using a Hyper-Acidophilic Candida tropicalis.

PubMed

Tamburini, Elena; Costa, Stefania; Marchetti, Maria Gabriella; Pedrini, Paola

2015-08-19

The yeast Candida tropicalis DSM 7524 produces xylitol, a natural, low-calorie sweetener, by fermentation of xylose. In order to increase xylitol production rate during the submerged fermentation process, some parameters-substrate (xylose) concentration, pH, aeration rate, temperature and fermentation strategy-have been optimized. The maximum xylitol yield reached at 60-80 g/L initial xylose concentration, pH 5.5 at 37 °C was 83.66% (w/w) on consumed xylose in microaerophilic conditions (kLa = 2·h(-1)). Scaling up on 3 L fermenter, with a fed-batch strategy, the best xylitol yield was 86.84% (w/w), against a 90% of theoretical yield. The hyper-acidophilic behaviour of C. tropicalis makes this strain particularly promising for industrial application, due to the possibility to work in non-sterile conditions.

Optimized Production of Xylitol from Xylose Using a Hyper-Acidophilic Candida tropicalis

PubMed Central

Tamburini, Elena; Costa, Stefania; Marchetti, Maria Gabriella; Pedrini, Paola

2015-01-01

The yeast Candida tropicalis DSM 7524 produces xylitol, a natural, low-calorie sweetener, by fermentation of xylose. In order to increase xylitol production rate during the submerged fermentation process, some parameters-substrate (xylose) concentration, pH, aeration rate, temperature and fermentation strategy-have been optimized. The maximum xylitol yield reached at 60–80 g/L initial xylose concentration, pH 5.5 at 37 °C was 83.66% (w/w) on consumed xylose in microaerophilic conditions (kLa = 2·h−1). Scaling up on 3 L fermenter, with a fed-batch strategy, the best xylitol yield was 86.84% (w/w), against a 90% of theoretical yield. The hyper-acidophilic behaviour of C. tropicalis makes this strain particularly promising for industrial application, due to the possibility to work in non-sterile conditions. PMID:26295411

Screening a strain of Aspergillus niger and optimization of fermentation conditions for degradation of aflatoxin B₁.

PubMed

Zhang, Wei; Xue, Beibei; Li, Mengmeng; Mu, Yang; Chen, Zhihui; Li, Jianping; Shan, Anshan

2014-11-13

Aflatoxin B₁, a type of highly toxic mycotoxin produced by some species belonging to the Aspergillus genus, such as Aspergillus flavus and Aspergillus parasiticus, is widely distributed in feed matrices. Here, coumarin was used as the sole carbon source to screen microorganism strains that were isolated from types of feed ingredients. Only one isolate (ND-1) was able to degrade aflatoxin B₁ after screening. ND-1 isolate, identified as a strain of Aspergillus niger using phylogenetic analysis on the basis of 18S rDNA, could remove 26.3% of aflatoxin B₁ after 48 h of fermentation in nutrient broth (NB). Optimization of fermentation conditions for aflatoxin B₁ degradation by selected Aspergillus niger was also performed. These results showed that 58.2% of aflatoxin B₁ was degraded after 24 h of culture under the optimal fermentation conditions. The aflatoxin B₁ degradation activity of Aspergillus niger supernatant was significantly stronger than cells and cell extracts. Furthermore, effects of temperature, heat treatment, pH, and metal ions on aflatoxin B₁ degradation by the supernatant were examined. Results indicated that aflatoxin B₁ degradation of Aspergillus niger is enzymatic and this process occurs in the extracellular environment.

Production of Laccase by a New Myrothecium verrucaria MD-R-16 Isolated from Pigeon Pea [Cajanus cajan (L.) Millsp.] and its Application on Dye Decolorization.

PubMed

Sun, Jiao; Guo, Na; Niu, Li-Li; Wang, Qing-Fang; Zang, Yu-Ping; Zu, Yuan-Gang; Fu, Yu-Jie

2017-04-23

The present study was conducted to screen a laccase-producing fungal endophyte, optimize fermentation conditions, and evaluate the decolorization ability of the laccase. A new fungal endophyte capable of laccase-producing was firstly isolated from pigeon pea and identified as Myrothecium verrucaria based on a ITS-rRNA sequences analysis. Meanwhile, various fermentation parameters on the laccase production were optimized via response surface methodology (RSM). The optimal fermentation conditions were a fermentation time of five days, temperature 30 °C and pH 6.22. Laccase activity reached 16.52 ± 0.18 U/mL under the above conditions. Furthermore, the laccase showed effective decolorization capability toward synthetic dyes (Congo red, Methyl orange, Methyl red, and Crystal violet) in the presence of the redox mediator ABTS, with more than 70% of dyes decolorizing after 24 h of incubation. Additionally, the activity of laccase was relatively stable with pH (4.5-6.5) and a temperature range of 35-55 °C. Therefore, the high laccase production of the strain and the new fungal laccase could provide a promising alterative approach for industrial and environmental applications.

Modeling of acetate-type fermentation of sugar-containing wastewater under acidic pH conditions.

PubMed

Huang, Liang; Pan, Xin-Rong; Wang, Ya-Zhou; Li, Chen-Xuan; Chen, Chang-Bin; Zhao, Quan-Bao; Mu, Yang; Yu, Han-Qing; Li, Wen-Wei

2018-01-01

In this study, a kinetic model was developed based on Anaerobic Digestion Model No. 1 to provide insights into the directed production of acetate and methane from sugar-containing wastewater under low pH conditions. The model sufficiently described the dynamics of liquid-phase and gaseous products in an anaerobic membrane bioreactor by comprehensively considering the syntrophic bioconversion steps of sucrose hydrolysis, acidogenesis, acetogenesis and methanogenesis under acidic pH conditions. The modeling results revealed a significant pH-dependency of hydrogenotrophic methanogenesis and ethanol-producing processes that govern the sucrose fermentative pathway through changing the hydrogen yield. The reaction thermodynamics of such acetate-type fermentation were evaluated, and the implications for process optimization by adjusting the hydraulic retention time were discussed. This work sheds light on the acid-stimulated acetate-type fermentation process and may lay a foundation for optimization of resource-oriented processes for treatment of food wastewater. Copyright © 2017 Elsevier Ltd. All rights reserved.

Optimization of culture conditions for penicilazaphilone C production by a marine-derived fungus Penicillium sclerotiorum M-22.

PubMed

Zhao, H-G; Wang, M; Lin, Y-Y; Zhou, S-L

2018-03-01

The aim of this study was to optimize the culture conditions of a marine-derived fungus Penicillium sclerotiorum M-22 for the production of penicilazaphilone C (PAC), a novel azaphilonidal derivative exhibiting broad cytotoxic and antibacterial effects. By single factor experiments, the effects to the production of PAC of aged seawater concentration, initial pH values, fermentation time, carbon sources, nitrogen sources and inorganic salt sources were investigated individually. Response surface methodology (RSM) analysis was adopted to investigate the interactions between variables and determine the optimal values for maximum PAC production. Evaluation of the experimental results signified that the optimum conditions for maximum production of PAC (19·85 mg l -1 ) in 250 ml Erlenmeyer flask were fermentation time 24·83 days, pH of 7·00, corn meal concentration of 10·72 g l -1 , yeast extract concentration of 4·58 g l -1 , crude sea salt concentration of 20·59 g l -1 . Production under optimized conditions increased to 1·344-fold comparing to its production prior to optimization. The higher PAC production and the penicilazaphilone C -producing marine fungus would be provide a promising alterative approach for industrial and commercial applications. Penicilazaphilone C (PAC) was a novel azaphilonidal derivative which had exhibited selective cytotoxicity and antibacterial activity. To further enhance production of PAC by optimizing fermentation conditions of Penicillium sclerotiorum M-22 would provide a promising alterative approach for industrial and commercial applications. We used the single factor test to determine the key factors which influence the PAC production. Then through the Response surface methodology and Box-Behnken design to determine the best fermentation condition for maximum production of PAC. Through these experimental designs and analysis will help us improve experimental efficiency and save time and materials. © 2017 The Society for Applied Microbiology.

Cellulase with high β-glucosidase activity by Penicillium oxalicum under solid state fermentation and its use in hydrolysis of cassava residue.

PubMed

Su, Lin-Hui; Zhao, Shuai; Jiang, Sui-Xin; Liao, Xu-Zhong; Duan, Cheng-Jie; Feng, Jia-Xun

2017-02-01

In this study, we investigated cellulase production by Penicillium oxalicum EU2106 under solid-state fermentation (SSF) and its hydrolysis efficiency toward NaOH-H 2 O 2 -pretreated cassava residue (NHCR) produced after bioethanol fermentation. Optimization of SSF cultivation conditions for P. oxalicum EU2106 using a Box-behnken design-based response-surface methodology resulted in maximal cellulase activity of 34.0 ± 2.8 filter-paper units/g dry substrate, exhibiting a ~ twofold increase relative to activities obtained under non-optimized conditions. Furthermore, SSF-derived cellulase converted 94.3 ± 1.5% of NHCR cellulose into glucose within 96 h. Interestingly, P. oxalicum EU2106 produced higher β-glucosidase activity under SSF conditions than that under submerged-state fermentation conditions, resulting in the elimination of cellobiose inhibition during the early stages of NHCR cellulose hydrolysis. Overall, this work provided an alternative for a potential cellulase source and a preferred option for cassava residue biotechnological application.

Optimization of fermentation parameters for production of ethanol from kinnow waste and banana peels by simultaneous saccharification and fermentation.

PubMed

Sharma, Naresh; Kalra, K L; Oberoi, Harinder Singh; Bansal, Sunil

2007-12-01

A study was taken up to evaluate the role of some fermentation parameters like inoculum concentration, temperature, incubation period and agitation time on ethanol production from kinnow waste and banana peels by simultaneous saccharification and fermentation using cellulase and co-culture of Saccharomyces cerevisiae G and Pachysolen tannophilus MTCC 1077. Steam pretreated kinnow waste and banana peels were used as substrate for ethanol production in the ratio 4:6 (kinnow waste: banana peels). Temperature of 30°C, inoculum size of S. cerevisiae G 6% and (v/v) Pachysolen tannophilus MTCC 1077 4% (v/v), incubation period of 48 h and agitation for the first 24 h were found to be best for ethanol production using the combination of two wastes. The pretreated steam exploded biomass after enzymatic saccharification containing 63 gL(-1) reducing sugars was fermented with both hexose and pentose fermenting yeast strains under optimized conditions resulting in ethanol production, yield and fermentation efficiency of 26.84 gL(-1), 0.426 gg (-1) and 83.52 % respectively. This study could establish the effective utilization of kinnow waste and banana peels for bioethanol production using optimized fermentation parameters.

Increase of content and bioactivity of total phenolic compounds from spent coffee grounds through solid state fermentation by Bacillus clausii.

PubMed

Rochín-Medina, Jesús J; Ramírez, Karina; Rangel-Peraza, Jesús G; Bustos-Terrones, Yaneth A

2018-03-01

Spent coffee grounds are waste material generated during coffee beverage preparation. This by-product disposal causes a negative environmental impact, in addition to the loss of a rich source of nutrients and bioactive compounds. A rotating central composition design was used to determine the optimal conditions for the bioactivity of phenolic compounds obtained after the solid state fermentation of spent coffee grounds by Bacillus clausii . To achieve this, temperature and fermentation time were varied according to the experimental design and the total phenolic and flavonoid content, antioxidant activity and antimicrobial activity were determined. Surface response methodology showed that optimum bioprocessing conditions were a temperature of 37 °C and a fermentation time of 39 h. Under these conditions, total phenolic and flavonoid contents increased by 36 and 13%, respectively, in fermented extracts as compared to non-fermented. In addition, the antioxidant activity was increased by 15% and higher antimicrobial activity was observed against Gram positive and negative bacteria. These data demonstrated that bioprocessing optimization of spent coffee grounds using the surface response methodology was an important tool to improve phenolic extraction, which could be used as an antioxidant and antimicrobial agents incorporated into different types of food products.

[Screening and optimization of cholesterol conversion strain].

PubMed

Fan, Dan; Xiong, Bingjian; Pang, Cuiping; Zhu, Xiangdong

2014-10-04

Bacterial strain SE-1 capable of transforming cholesterol was isolated from soil and characterized. The transformation products were identified. Fermentation conditions were optimized for conversion. Cholesterol was used as sole carbon source to isolate strain SE-1. Morphology, physiological and biochemical characteristics of strain SE-1 were studied. 16S rRNA gene was sequenced and subjected to phylogenetic analysis. Fermentation supernatants were extracted with chloroform, the transformation products were analyzed by silica gel thin layer chromatography and Sephadex LH20. Their structures were identified by 1H-NMR and 13C-NMR. Fermentation medium including carbon and nitrogen, methods of adding substrates and fermentation conditions for Strain SE-1 were optimized. Strain SE-1 was a Gram-negative bacterium, exhibiting the highest homologs to Burkholderia cepacia based on the physiological analysis. The sequence analysis of 16S rRNA gene of SE-1 strain and comparison with related Burkholderia show that SE-1 strain was very close to B. cepacia (Genbank No. U96927). The similarity was 99%. The result of silica gel thin layer chromatography shows that strain SE-1 transformed cholesterol to two products, 7beta-hydroxycholesterol and the minor product was 7-oxocholesterol. The optimum culture conditions were: molasses 5%, (NH4 )2SO4 0.3%, 4% of inoculation, pH 7.5 and 36 degrees C. Under the optimum culture condition, the conversion rate reached 34.4% when concentration of cholesterol-Tween 80 was 1 g/L. Cholesterol 7beta-hydroxylation conversion rate under optimal conditions was improved by 20.8%. Strain SE-1 isolated from soil is capable of converting cholesterol at lab-scale.

Optimization of fermentation conditions for alcohol production.

PubMed

Bowman, L; Geiger, E

1984-12-01

The quantitative effects of carbohydrate levels, degree of initial saccharification, glucoamylase dosage, temperature, and fermentation time were investigated using a Box-Wilson central composite design protocol. With Saccharomyces cerevisiae ATCC 4126, it was found that the use of a partially saccharified starch substrate markedly increased yields and attainable alcohol levels. Balancing the degree of initial saccharification with the level of glucoamylase used to complete hydrolysis was found necessary to obtain optimum yields. The temperature optimum was found to be 36 degrees C. The regression equations obtained were used to model the fermentation in order to determine optimum fermentation conditions.

Microbial recycling of glycerol to biodiesel.

PubMed

Yang, Liu; Zhu, Zhi; Wang, Weihua; Lu, Xuefeng

2013-12-01

The sustainable supply of lipids is the bottleneck for current biodiesel production. Here microbial recycling of glycerol, byproduct of biodiesel production to biodiesel in engineered Escherichia coli strains was reported. The KC3 strain with capability of producing fatty acid ethyl esters (FAEEs) from glucose was used as a starting strain to optimize fermentation conditions when using glycerol as sole carbon source. The YL15 strain overexpressing double copies of atfA gene displayed 1.7-fold increase of FAEE productivity compared to the KC3 strain. The titer of FAEE in YL15 strain reached to 813 mg L(-1) in minimum medium using glycerol as sole carbon source under optimized fermentation conditions. The titer of glycerol-based FAEE production can be significantly increased by both genetic modifications and fermentation optimization. Microbial recycling of glycerol to biodiesel expands carbon sources for biodiesel production. Copyright © 2013 Elsevier Ltd. All rights reserved.

Optimization of Fermentation Conditions for the Production of Bacteriocin Fermentate

DTIC Science & Technology

2015-03-30

pentosaceus 43200 were obtained from ATCC. Lactobacillus (L.) plantarum NCDO 955, which was used as an indicator organism, and a strain of pediocin-producing...Development of food grade media for the preparation of Lactobacillus plantarum starter culture. J. Gen. Appl. Microbiol. 52: 349-356. Shurtleff, W...MEDIA FERMENTATION BROTH SAFETY INHIBITION BACTERIOCINS LACTOBACILLUS ANTIMICROBIAL AGENTS

Process optimization and analysis of product inhibition kinetics of crude glycerol fermentation for 1,3-Dihydroxyacetone production.

PubMed

Dikshit, Pritam Kumar; Padhi, Susant Kumar; Moholkar, Vijayanand S

2017-11-01

In present study, statistical optimization of biodiesel-derived crude glycerol fermentation to DHA by immobilized G. oxydans cells over polyurethane foam is reported. Effect of DHA (product) inhibition on crude glycerol fermentation was analyzed using conventional biokinetic models and new model that accounts for both substrate and product inhibition. Optimum values of fermentation parameters were: pH=4.7, temperature=31°C, initial substrate concentration=20g/L. At optimum conditions, DHA yield was 89% (17.83g/L). Effect of product inhibition on fermentation was trivial for DHA concentrations ≤30g/L. At higher concentrations (≥50g/L), kinetics and yield of fermentation showed marked reduction with sharp drop in V max and K S values. Inhibition effect was more pronounced for immobilized cells due to restricted transport of fermentation mixture across polyurethane foam. Retention of fermentation mixture in immobilized matrix resulted in higher localized DHA concentration that possibly enhanced inhibition effect. Copyright © 2017 Elsevier Ltd. All rights reserved.

Camptothecine production by mixed fermentation of two endophytic fungi from Nothapodytes nimmoniana.

PubMed

Bhalkar, Bhumika N; Patil, Swapnil M; Govindwar, Sanjay P

2016-01-01

Two endophytic fungi isolated from the endangered plant Nothapodytes nimmoniana (Grah.) Mabb. were found to effectively synthesize CPT independent of their host plant under submerged fermentation conditions. Molecular characterization of fungi revealed their identity as Colletotrichum fructicola SUK1 (F1) and Corynespora cassiicola SUK2 (F2). Mixed fermentation experiments were carried out to study the effect of microbial signalling between the two fungal species on camptothecine production. Effect of culture parameters on CPT production was studied for both mono-cultures (F1 and F2) separately as well as for the mixed fermentation (F1 + F2). Further the most influencing ones were optimized statistically using response surface methodology. Statistical model based optimized parameters were whey (70 %), agitation rate (110 rpm), temperature (30 °C), and incubation period (7 d) for the mixed fermentation. Monocultures of the two fungal species F1 and F2 yielded CPT up to 33 ± 1.1 mg l(-1)and 69 ± 1.1 mg l(-1), respectively; while their mixed fermentation under the optimized conditions yielded up to 146 ± 0.2 mg l(-1). HPLC and LC-MS/MS techniques were used to analyze the products obtained. Copyright © 2016 British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Process optimization for the preparation of straw feedstuff for rearing yellow mealworms (Tenebrio molitor L.) in BLSS

NASA Astrophysics Data System (ADS)

Li, Leyuan; Liu, lh64. Hong

2012-07-01

It has been confirmed in our previous work that in bioregenerative life support systems, feeding yellow mealworms (Tenebrio molitor L.) using fermented straw has the potential to provide good animal protein for astronauts, meanwhile treating with plant wastes. However, since the nitrogen content in straw is very low, T. molitor larvae can not obtain sufficient nitrogen, which results in a relatively low growth efficiency. In this study, wheat straw powder was mixed with simulated human urine before fermentation. Condition parameters, e.g. urine:straw ratio, moisture content, inoculation dose, fermentation time, fermentation temperature and pH were optimized using Taguchi method. Larval growth rate and average individual mass of mature larva increased significantly in the group of T. molitor larvae fed with feedstuff prepared with the optimized process.

Investigation of vinegar production using a novel shaken repeated batch culture system.

PubMed

Schlepütz, Tino; Büchs, Jochen

2013-01-01

Nowadays, bioprocesses are developed or optimized on small scale. Also, vinegar industry is motivated to reinvestigate the established repeated batch fermentation process. As yet, there is no small-scale culture system for optimizing fermentation conditions for repeated batch bioprocesses. Thus, the aim of this study is to propose a new shaken culture system for parallel repeated batch vinegar fermentation. A new operation mode - the flushing repeated batch - was developed. Parallel repeated batch vinegar production could be established in shaken overflow vessels in a completely automated operation with only one pump per vessel. This flushing repeated batch was first theoretically investigated and then empirically tested. The ethanol concentration was online monitored during repeated batch fermentation by semiconductor gas sensors. It was shown that the switch from one ethanol substrate quality to different ethanol substrate qualities resulted in prolonged lag phases and durations of the first batches. In the subsequent batches the length of the fermentations decreased considerably. This decrease in the respective lag phases indicates an adaptation of the acetic acid bacteria mixed culture to the specific ethanol substrate quality. Consequently, flushing repeated batch fermentations on small scale are valuable for screening fermentation conditions and, thereby, improving industrial-scale bioprocesses such as vinegar production in terms of process robustness, stability, and productivity. Copyright © 2013 American Institute of Chemical Engineers.

Novel fermented chickpea milk with enhanced level of γ-aminobutyric acid and neuroprotective effect on PC12 cells.

PubMed

Li, Wen; Wei, Mingming; Wu, Junjun; Rui, Xin; Dong, Mingsheng

2016-01-01

In this study, novel fermented chickpea milk with high γ -aminobutyric acid (GABA) content and potential neuroprotective activity was developed. Fermentation starter that can produce GABA was selected from 377 strains of lactic acid bacteria isolated from traditional Chinese fermented foods. Among the screened strains, strain M-6 showed the highest GABA-producing capacity in De Man-Rogosa and Sharp (MRS) broth and chickpea milk. M-6 was identified as Lactobacillus plantarum based on Gram staining, API carbohydrate fermentation pattern testing, and 16s rDNA sequencing. The complete gene encoding glutamate decarboxylase was cloned to confirm the presence of the gene in L. plantarum M-6. The fermentation condition was optimized by response surface methodology. Results demonstrated that L. plantarum M-6 produced the highest GABA content of 537.23 mg/L. The optimal condition included an inoculum concentration of 7%, presence of 0.2% (m/v) monosodium glutamate and 55 µ M pyridoxal-5-phosphate, incubation temperature of 39 °C and fermentation time of 48 h . GABA-enriched chickpea milk exerted protective effects on PC12 cells against MnCl2 -induced injury. GABA-enriched chickpea milk improved cell viability and markedly attenuated the release of lactate dehydrogenase compared with the impaired cells.

Effect of fermentation parameters on bio-alcohols production from glycerol using immobilized Clostridium pasteurianum: an optimization study.

PubMed

Khanna, Swati; Goyal, Arun; Moholkar, Vijayanand S

2013-01-01

This article addresses the issue of effect of fermentation parameters for conversion of glycerol (in both pure and crude form) into three value-added products, namely, ethanol, butanol, and 1,3-propanediol (1,3-PDO), by immobilized Clostridium pasteurianum and thereby addresses the statistical optimization of this process. The analysis of effect of different process parameters such as agitation rate, fermentation temperature, medium pH, and initial glycerol concentration indicated that medium pH was the most critical factor for total alcohols production in case of pure glycerol as fermentation substrate. On the other hand, initial glycerol concentration was the most significant factor for fermentation with crude glycerol. An interesting observation was that the optimized set of fermentation parameters was found to be independent of the type of glycerol (either pure or crude) used. At optimum conditions of agitation rate (200 rpm), initial glycerol concentration (25 g/L), fermentation temperature (30°C), and medium pH (7.0), the total alcohols production was almost equal in anaerobic shake flasks and 2-L bioreactor. This essentially means that at optimum process parameters, the scale of operation does not affect the output of the process. The immobilized cells could be reused for multiple cycles for both pure and crude glycerol fermentation.

Influence of dietary fiber on inflammatory bowel disease and colon cancer: importance of fermentation pattern.

PubMed

Rose, Devin J; DeMeo, Mark T; Keshavarzian, Ali; Hamaker, Bruce R

2007-02-01

The benefits of dietary fiber on inflammatory bowel disease may be related to the fermentative production of butyrate in the colon, which appears to decrease the inflammatory response. The benefits of dietary fiber against colon cancer may be related to both fermentative and non-fermentative processes, although poorly fermentable fibers appear more influential. Dietary fiber fermentation profiles are important in determining optimal fibers for colonic health, and may be a function of structure, processing conditions, and other food components. A greater understanding of the relationships between fermentation rate and dietary fiber structure would allow for development of dietary fibers for optimum colonic health.

Monitoring of Lactobacillus fermentation process by using ion chromatography with a series piezoelectric quartz crystal detector.

PubMed

Zhang, J; Xie, Y; Dai, X; Wei, W

2001-03-01

A new method monitoring Lactobacillus fermentation process, which combines ion chromatography (IC) with a series piezoelectric quartz crystal (SPQC) technique, is presented in this paper. Monitoring of the fermentation process was realized by examining the rate of production of lactic acid. An automatic membrane dialyser was used for the pretreatment of the sample in on-line monitoring. A mixture of p-hydroxybenzoic acid and N,N-diethylethanolamine was adopted as mobile phase and its flow rate was 0.8 ml/min. The effects of some fermentation conditions were also discussed in detail. Accordingly, the optimal fermentation conditions were obtained. This method is simple and convenient while the results obtained are accurate and reliable.

Effects of protectant and rehydration conditions on the survival rate and malolactic fermentation efficiency of freeze-dried Lactobacillus plantarum JH287.

PubMed

Lee, Sae-Byuk; Kim, Dong-Hwan; Park, Heui-Dong

2016-09-01

In this study, Lactobacillus plantarum JH287 was used as a malolactic fermentation starter in Campbell Early wine production. L. plantarum JH287 was first lyophilized, and the malolactic fermentation potential of freeze-dried L. plantarum JH287 was investigated. Different protective media and rehydration conditions were tested to improve the survival rate of freeze-dried L. plantarum JH287. Optimal protective medium contained 10 % sorbitol and 10 % skim milk. The optimal rehydration condition was a 1-h rehydration time conducted in the same protective media, and the combination of these two methods produced a survival rate of 86.37 %. In addition, a 77.71 % survival rate was achieved using freeze-dried samples that were stored at 4 °C for 2 months. Freeze-dried L. plantarum JH287 and Saccharomyces cerevisiae Fermivin were used to inoculate the Campbell Early grape must to decrease its malic acid content. Using this mixed-fermentation method, wine showed a decrease in malic acid content after 9 days of fermentation. GC-MS analysis detected 15 volatile ester compounds in the wine. A sensory evaluation showed that the taste and aroma of mix-fermented wine were better than those of the control that had not been inoculated with L. plantarum JH287.

Effect of cultural conditions on antrodin C production by basidiomycete Antrodia camphorata in solid-state fermentation.

PubMed

Xia, Yongjun; Wang, Yuanlong; Zhang, Bobo; Xu, Ganrong; Ai, Lianzhong

2014-01-01

Antrodia camphorata is a medicinal fungus and antrodin C is one of the main bioactive components of A. camphorata in the submerged fermentation (SmF). To optimize the culture conditions, the factors influencing the production of antrodin C by A. camphorata under solid-state fermentation (SSF) were investigated in this study. Different solid substrates and external nitrogen sources were tested for their efficiency in producing antrodin C. The response surface methodology was applied to evaluate the influence of several variables, namely, the concentrations of soybean meal, initial moisture content, and inoculum density on antrodin C production in solid-state fermentation. The experimental results show that the optimum fermentation medium for antrodin C production by A. camphorata was composed of 0.578 g soybean meal, 0.05 g Na2 HPO4 , 0.05 g MgSO4 for 100 g rice, with 51.83% initial moisture content, 22 day culture time, 28 °C culture temperature, and 35.54% inoculum density. At optimized conditions, 6,617.36 ± 92.71 mg kg(-1) yield of antrodin C was achieved. Solid-state fermentation is one good cultural method to improve the production of antrodin C by A. camphorata. © 2014 International Union of Biochemistry and Molecular Biology, Inc.

Optimization of CMCase production from sorghum straw by Aspergillus terreus SUK-1 under solid substrate fermentation using response surface methodology

NASA Astrophysics Data System (ADS)

Tibin, El Mubarak Musa; Al-Shorgani, Najeeb Kaid Naseer; Abuelhassan, Nawal Noureldaim; Hamid, Aidil Abdul; Kalil, Mohd Sahaid; Yusoff, Wan Mohtar Wan

2013-11-01

The cellulase production using sorghum straw as substrate by fungal culture of Aspergillus terreus SUK-1 was investigated in solid substrate fermentation (SSF). The optimum CMCase was achieved by testing most effective fermentation parameters which were: incubation temperature, pH and moisture content using Response Surface Methodology (RSM) based on Central Composite Design (CCD). The carboxymethyl cellulase activity (CMCase) was measured as the defining factor. The results were analysed by analysis of variance (ANOVA) and the regression quadratic model was obtained. The model was found to be significant (p<0.05) and the effect of temperature (25-40°C) and pH (4-7) was found to be not significant on CMCase activity whereas the moisture content was significant in the SSF conditions employed. The high yield of predicted CMCase activity (0.2 U/ml) was obtained under the optimized conditions (temperature 40 □C, pH 5.4 and moisture content of 80%). The model was validated by applying the optimized conditions and it was found that the model was valid.

Statistical optimization of lovastatin production by Omphalotus olearius (DC.) singer in submerged fermentation.

PubMed

Atlı, Burcu; Yamaç, Mustafa; Yıldız, Zeki; Isikhuemhen, Omoanghe S

2016-01-01

In this study, culture conditions were optimized to improve lovastatin production by Omphalotus olearius, isolate OBCC 2002, using statistical experimental designs. The Plackett-Burman design was used to select important variables affecting lovastatin production. Accordingly, glucose, peptone, and agitation speed were determined as the variables that have influence on lovastatin production. In a further experiment, these variables were optimized with a Box-Behnken design and applied in a submerged process; this resulted in 12.51 mg/L lovastatin production on a medium containing glucose (10 g/L), peptone (5 g/L), thiamine (1 mg/L), and NaCl (0.4 g/L) under static conditions. This level of lovastatin production is eight times higher than that produced under unoptimized media and growth conditions by Omphalotus olearius. To the best of our knowledge, this is the first attempt to optimize submerged fermentation process for lovastatin production by Omphalotus olearius.

Temperature and pH Conditions That Prevail during Fermentation of Sausages Are Optimal for Production of the Antilisterial Bacteriocin Sakacin K

PubMed Central

Leroy, Frédéric; de Vuyst, Luc

1999-01-01

Sakacin K is an antilisterial bacteriocin produced by Lactobacillus sake CTC 494, a strain isolated from Spanish dry fermented sausages. The biokinetics of cell growth and bacteriocin production of L. sake CTC 494 in vitro during laboratory fermentations were investigated by making use of MRS broth. The data obtained from the fermentations was used to set up a predictive model to describe the influence of the physical factors temperature and pH on microbial behavior. The model was validated successfully for all components. However, the specific bacteriocin production rate seemed to have an upper limit. Both cell growth and bacteriocin activity were very much influenced by changes in temperature and pH. The production of biomass was closely related to bacteriocin activity, indicating primary metabolite kinetics, but was not the only factor of importance. Acidity dramatically influenced both the production and the inactivation of sakacin K; the optimal pH for cell growth did not correspond to the pH for maximal sakacin K activity. Furthermore, cells grew well at 35°C but no bacteriocin production could be detected at this temperature. L. sake CTC 494 shows special promise for implementation as a novel bacteriocin-producing sausage starter culture with antilisterial properties, considering the fact that the temperature and acidity conditions that prevail during the fermentation process of dry fermented sausages are optimal for the production of sakacin K. PMID:10049850

Characteristic of fermented spinach (Amaranthus spp.) polyphenol by kombucha culture for antioxidant compound

NASA Astrophysics Data System (ADS)

Aspiyanto, Susilowati, Agustine; Iskandar, Jeti M.; Melanie, Hakiki; Maryati, Yati; Lotulung, Puspa D.

2017-01-01

Fermentation on spinach (Amaranthus sp.) vegetable by kombucha culture as an effort to get poliphenol as antioxidant compound had been done. Purification of fermented spinach extract suspension was carried out through microfiltration (MF) membrane (pore size 0.15 µm) fitted in dead-end Stirred Ultrafiltration Cell (SUFC) mode at fixed condition (stirrer rotation 400 rpm, room temperature, pressure 40 psia). Result of the experimental activity showed that long fermentation time increased total acids, total polyphenol and Total Plate Count (TPC), and decreased total solids and reducing sugar in biomass. The optimal fermentation time was reached for 2 weeks with total polyphenol recovery increasing of 92.76 % from before and after fermentation. On this optimal fermentation time, biomass had identified galic acid with relative intensity of 8 %, while as polyphenol monomer was resulted 5 kinds of polyphenol compounds with total intensity 27.97 % and molecular weight (MW) 191.1736, 193.1871 and 194.2170 at T2.5, T2.86 and T3.86. Long fermentation time increased functional properties of polyphenol as antioxidant.

Development of a fermented ice-cream as influenced by in situ exopolysaccharide production: Rheological, molecular, microstructural and sensory characterization.

PubMed

Dertli, Enes; Toker, Omer S; Durak, M Zeki; Yilmaz, Mustafa T; Tatlısu, Nevruz Berna; Sagdic, Osman; Cankurt, Hasan

2016-01-20

This study aimed to investigate the role of in situ exopolysaccharide (EPS) production by EPS(+)Streptococcus thermophilus strains on physicochemical, rheological, molecular, microstructural and sensory properties of ice cream in order to develop a fermented and consequently functional ice-cream in which no stabilizers would be required in ice-cream production. For this purpose, the effect of EPS producing strains (control, strain 1, strain 2 and mixture) and fermentation conditions (fermentation temperature; 32, 37 and 42 °C and time; 2, 3 and 4h) on pH, S. thermophilus count, EPS amount, consistency coefficient (K), and apparent viscosity (η50) were investigated and optimized using single and multiple response optimization tools of response surface methodology. Optimization analyses indicated that functional ice-cream should be fermented with strain 1 or strain mixture at 40-42 °C for 4h in order to produce the most viscous ice-cream with maximum EPS content. Optimization analysis results also revealed that strain specific conditions appeared to be more effective factor on in situ EPS production amount, K and η50 parameters than did fermentation temperature and time. The rheological analysis of the ice-cream produced by EPS(+) strains revealed its high viscous and pseudoplastic non-Newtonian fluid behavior, which demonstrates potential of S. thermophilus EPS as thickening and gelling agent in dairy industry. FTIR analysis proved that the EPS in ice-cream corresponded to a typical EPS, as revealed by the presence of carboxyl, hydroxyl and amide groups with additional α-glycosidic linkages. SEM studies demonstrated that it had a web-like compact microstructure with pores in ice-cream, revealing its application possibility in dairy products to improve their rheological properties. Copyright © 2015. Published by Elsevier Ltd.

Optimization of Exopolysaccharide Production by Lactobacillus delbrueckii subsp. bulgaricus RR Grown in a Semidefined Medium

PubMed Central

Kimmel, Stacy A.; Roberts, Robert F.; Ziegler, Gregory R.

1998-01-01

The optimal fermentation temperature, pH, and Bacto-casitone (Difco Laboratories, Detroit, Mich.) concentration for production of exopolysaccharide by Lactobacillus delbrueckii subsp. bulgaricus RR in a semidefined medium were determined by using response surface methods. The design consisted of 20 experiments, 15 unique combinations, and five replications. All fermentations were conducted in a fermentor with a 2.5-liter working volume and were terminated when 90% of the glucose in the medium had been consumed. The population of L. delbrueckii subsp. bulgaricus RR and exopolysaccharide content were measured at the end of each fermentation. The optimum temperature, pH, and Bacto-casitone concentration for exopolysaccharide production were 38°C, 5, and 30 g/liter, respectively, with a predicted yield of 295 mg of exopolysaccharide/liter. The actual yield under these conditions was 354 mg of exopolysaccharide/liter, which was within the 95% confidence interval (217 to 374 mg of exopolysaccharide/liter). An additional experiment conducted under optimum conditions showed that exopolysaccharide production was growth associated, with a specific production at the endpoint of 101.4 mg/g of dry cells. Finally, to obtain material for further characterization, a 100-liter fermentation was conducted under optimum conditions. Twenty-nine grams of exopolysaccharide was isolated from centrifuged, ultrafiltered fermentation broth by ethanol precipitation. PMID:9464404

Effects of different microbes on fermenting feed for sea cucumber ( Apostichopus japonicus)

NASA Astrophysics Data System (ADS)

Jiang, Yan; Wang, Yingeng; Mai, Kangsen; Zhang, Zheng; Liao, Meijie; Rong, Xiaojun

2015-10-01

The effects of different microbes on fermenting feed for sea cucumber ( Apostichopus japonicus) were compared to select the optimal fermentation strain in this study. Saccharomgces cerevisae, Candida utilis, Bacillus subtilis and Geotrichum candidum were independently added into the experimental compound feed, while only saline was mixed with the control feed. The fermentation treatments were inoculated with 10% seed solution under the condition of 25°C and 70% water content, which lasted for 5 days to elucidate the optimal microbe strain for fermenting effect. Physicochemical indexes and sensorial characteristics were measured per day during the fermentation. The indexes included dry matter recovery (DMR), crude protein (CP), the percentage of amino acid nitrogen to total nitrogen (AA-N/tN), the percentage of ammonia nitrogen to total nitrogen (NH3-N/tN), and the ratio of fermentation strains and vibrios to the total microbes, color, smell and viscosity. The results showed that DMR, CP and AA-N/tN of the S. cerevisae group reached the highest level on day 3, but the ratio of fermentation strain was second to C. utilis group. In addition, its NH3-N/tN and the ratio of vibrios were maintained at low levels, and the sensory evaluation score including smell, color and viscosity was the highest in S. cerevisae group on day 3. Therefore, S. cerevisae could be the optimal strain for the feed fermentation for sea cucumber. This research developed a new production method of fermentation feed for sea cucumber.

Biorefinery of cellulosic primary sludge towards targeted Short Chain Fatty Acids, phosphorus and methane recovery.

PubMed

Crutchik, Dafne; Frison, Nicola; Eusebi, Anna Laura; Fatone, Francesco

2018-06-01

Cellulose from used toilet paper is a major untapped resource embedded in municipal wastewater which recovery and valorization to valuable products can be optimized. Cellulosic primary sludge (CPS) can be separated by upstream dynamic sieving and anaerobically digested to recover methane as much as 4.02 m 3 /capita·year. On the other hand, optimal acidogenic fermenting conditions of CPS allows the production of targeted short-chain fatty acids (SCFAs) as much as 2.92 kg COD/capita·year. Here propionate content can be more than 30% and can optimize the enhanced biological phosphorus removal (EBPR) processes or the higher valuable co-polymer of polyhydroxyalkanoates (PHAs). In this work, first a full set of batch assays were used at three different temperatures (37, 55 and 70 °C) and three different initial pH (8, 9 and 10) to identify the best conditions for optimizing both the total SCFAs and propionate content from CPS fermentation. Then, the optimal conditions were applied in long term to a Sequencing Batch Fermentation Reactor where the highest propionate production (100-120 mg COD/g TVS fed ·d) was obtained at 37 °C and adjusting the feeding pH at 8. This was attributed to the higher hydrolysis efficiency of the cellulosic materials (up to 44%), which increased the selective growth of Propionibacterium acidopropionici in the fermentation broth up to 34%. At the same time, around 88% of the phosphorus released during the acidogenic fermentation was recovered as much as 0.15 kg of struvite per capita·year. Finally, the potential market value was preliminary estimated for the recovered materials that can triple over the conventional scenario of biogas recovery in existing municipal wastewater treatment plants. Copyright © 2018 Elsevier Ltd. All rights reserved.

Optimization of cultural conditions for conversion of glycerol to ethanol by Enterobacter aerogenes S012

PubMed Central

2013-01-01

The aim of this research is to optimize the cultural conditions for the conversion of glycerol to ethanol by Enterobacter aerogenes S012. Taguchi method was used to screen the cultural conditions based on their signal to noise ratio (SN). Temperature (°C), agitation speed (rpm) and time (h) were found to have the highest influence on both glycerol utilization and ethanol production by the organism while pH had the lowest. Full factorial design, statistical analysis, and regression model equation were used to optimize the selected cultural parameters for maximum ethanol production. The result showed that fermentation at 38°C and 200 rpm for 48 h would be ideal for the bacteria to produce maximum amount of ethanol from glycerol. At these optimum conditions, ethanol production, yield and productivity were 25.4 g/l, 0.53 g/l/h, and 1.12 mol/mol-glycerol, repectively. Ethanol production increased to 26.5 g/l while yield and productivity decreased to 1.04 mol/mol-glycerol and 0.37 g/l/h, respectively, after 72 h. Analysis of the fermentation products was performed using HPLC, while anaerobic condition was created by purging the fermentation vessel with nitrogen gas. PMID:23388539

Dextran Utilization During Its Synthesis by Weissella cibaria RBA12 Can Be Overcome by Fed-Batch Fermentation in a Bioreactor.

PubMed

Baruah, Rwivoo; Deka, Barsha; Kashyap, Niharika; Goyal, Arun

2018-01-01

Weissella cibaria RBA12 produced a maximum of 9 mg/ml dextran (with 90% efficiency) using shake flask culture under the optimized concentration of medium components viz. 2% (w/v) of each sucrose, yeast extract, and K 2 HPO 4 after incubation at optimized conditions of 20 °C and 180 rpm for 24 h. The optimized medium and conditions were used for scale-up of dextran production from Weissella cibaria RBA12 in 2.5-l working volume under batch fermentation in a bioreactor that yielded a maximum of 9.3 mg/ml dextran (with 93% efficiency) at 14 h. After 14 h, dextran produced was utilized by the bacterium till 18 h in its stationary phase under sucrose depleted conditions. Dextran utilization was further studied by fed-batch fermentation using sucrose feed. Dextran on production under fed-batch fermentation in bioreactor gave 35.8 mg/ml after 32 h. In fed-batch mode, there was no decrease in dextran concentration as observed in the batch mode. This showed that the utilization of dextran by Weissella cibaria RBA12 is initiated when there is sucrose depletion and therefore the presence of sucrose can possibly overcome the dextran hydrolysis. This is the first report of utilization of dextran, post-sucrose depletion by Weissella sp. studied in bioreactor.

Dilute sulfuric acid fractionation of Korean food waste for ethanol and lactic acid production by yeast.

PubMed

Kim, Yong Seon; Jang, Ji Yeon; Park, Seong Jik; Um, Byung Hwan

2018-04-01

Fermentation of food waste biomass can be used to produce biochemicals such as lactic acid and ethanol in a cost-effective manner. Korean food waste (KFW) dewatered by a screw press contains 23.1% glucan on a dry basis and is a potential raw material for the production of ethanol and lactic acid through fermentation. This study was conducted to optimize the dilute acid fractionation conditions for KFW fermentation with respect to the H 2 SO 4 concentration (0-0.8% w/v), temperature (130-190 °C), and residence time (1-128 min) using response surface methodology. Dilute sulfuric acid fractionation was carried out using a 30-mL stainless steel reactor under conditions, and then the dilute acid fractionation was scaled-up in 1-L and 7-L stainless steel reactors under the optimal conditions. The hydrolysate was concentrated, liquid-liquid extracted and neutralized for lactic acid and ethanol production. The highest concentration of glucose obtained from the KFW was 26.4 g/L using fractionation with 0.37% w/v H 2 SO 4 at 156 °C for 123.6 min. Using recombinant Saccharomyces cerevisiae containing a codon-optimized lactate dehydrogenase, the yield of lactic acid and ethanol was 77% of the theoretical yield for 17.4 g/L of fermentable sugar at pH 5.5. Additionally, the yield of ethanol produced by Issatchenkia orientalis was 89% of the theoretical yield for 25 g/L of fermentable sugar at pH 3. Copyright © 2018 Elsevier Ltd. All rights reserved.

High-level production of β-1,3-1,4-glucanase by Rhizomucor miehei under solid-state fermentation and its potential application in the brewing industry.

PubMed

Yang, S Q; Xiong, H; Yang, H Y; Yan, Q J; Jiang, Z Q

2015-01-01

To improve the β-1,3-1,4-glucanase production by Rhizomucor miehei under solid-state fermentation (SSF) for industrial application. The fermentation conditions for β-1,3-1,4-glucanase production by R. miehei CAU432 under SSF were optimized using a 'one-factor-at-a-time' method. Under the optimized fermentation conditions, viz. oatmeal (0·45-0·9 mm) as sole carbon source, 5% (w/w) peptone as sole nitrogen source, initial moisture of 80% (w/w), initial culture pH of 5·0, incubation temperature of 50°C and incubation time of 6 days, the highest β-1,3-1,4-glucanase activity of 20,025 U g(-1) dry substrate was achieved, which represents the highest yield for β-1,3-1,4-glucanase production ever reported. The crude enzyme was extracted and purified to homogeneity with a purification fold of 4·6 and a recovery yield of 9·0%. The addition of the purified β-1,3-1,4-glucanase in mash obviously reduced its filtration time (24·6%) and viscosity (2·61%). The optimal fermentation conditions for maximal β-1,3-1,4-glucanase production under SSF was obtained, and the enzyme was suitable for application in the malting process. The high production yield and excellent capability of the enzyme may enable it great potential in industries, especially in brewing industry. © 2014 The Society for Applied Microbiology.

Fermentation performance optimization in an ectopic fermentation system.

PubMed

Yang, Xiaotong; Geng, Bing; Zhu, Changxiong; Li, Hongna; He, Buwei; Guo, Hui

2018-07-01

Ectopic fermentation systems (EFSs) were developed for wastewater treatment. Previous studies have investigated the ability of thermophilic bacteria to improve fermentation performance in EFS. Continuing this research, we evaluated EFS performance using principle component analysis and investigated the addition of different proportions of cow dung. Viable bacteria communities were clustered and identified using BOX-AIR-based repetitive extragenic palindromic-PCR and 16S rDNA analysis. The results revealed optimal conditions for the padding were maize straw inoculated with thermophilic bacteria. Adding 20% cow dung yielded the best pH values (6.94-8.56), higher temperatures, increased wastewater absorption, improved litter quality, and greater microbial quantities. The viable bacteria groups were enriched by the addition of thermophilic consortium, and exogenous strains G21, G14, G4-1, and CR-15 were detected in fermentation process. The proportion of Bacillus species in treatment groups reached 70.37% after fermentation, demonstrating that thermophilic bacteria, especially Bacillus, have an important role in EFS, supporting previous predictions. Copyright © 2018 Elsevier Ltd. All rights reserved.

Optimization of culture condition for ACEI and GABA production by lactic acid bacteria.

PubMed

Tung, Yi-Ting; Lee, Bao-Hong; Liu, Chin-Feng; Pan, Tzu-Ming

2011-01-01

Gamma-aminobutyric acid (GABA) and angiotensin-converting enzyme inhibitor (ACEI) are compounds which can influence hypertension. The goal of this study is to optimize the culture condition for GABA and ACEI production by Lactobacillus plantarum NTU 102 fermented skim milk. In this study, we used 3-factor-3-level Box-Behnken design combining with response surface methodology, where the 3 factors represent the concentration of skim milk, the concentration of monosodium glutamate, and culture temperature. Best conditions for GABA and ACEI production differed. The results indicated that L. plantarum NTU 102 produced the highest combined levels of GABA and ACEI at 37 °C, in milk having 8% to 12% nonfat solids supplemented with 0.6% to 1% MSG. Agitation of the medium during fermentation had no effect on GABA or ACEI production but extended incubation (up to 6 d) increases levels of the bioactive compounds. L. plantarum NTU 102 fermented products may be a potential functional food source for regulating hypertension. © 2011 Institute of Food Technologists®

Strain-Dependent Transcriptome Signatures for Robustness in Lactococcus lactis

PubMed Central

Dijkstra, Annereinou R.; Alkema, Wynand; Starrenburg, Marjo J. C.; van Hijum, Sacha A. F. T.; Bron, Peter A.

2016-01-01

Recently, we demonstrated that fermentation conditions have a strong impact on subsequent survival of Lactococcus lactis strain MG1363 during heat and oxidative stress, two important parameters during spray drying. Moreover, employment of a transcriptome-phenotype matching approach revealed groups of genes associated with robustness towards heat and/or oxidative stress. To investigate if other strains have similar or distinct transcriptome signatures for robustness, we applied an identical transcriptome-robustness phenotype matching approach on the L. lactis strains IL1403, KF147 and SK11, which have previously been demonstrated to display highly diverse robustness phenotypes. These strains were subjected to an identical fermentation regime as was performed earlier for strain MG1363 and consisted of twelve conditions, varying in the level of salt and/or oxygen, as well as fermentation temperature and pH. In the exponential phase of growth, cells were harvested for transcriptome analysis and assessment of heat and oxidative stress survival phenotypes. The variation in fermentation conditions resulted in differences in heat and oxidative stress survival of up to five 10-log units. Effects of the fermentation conditions on stress survival of the L. lactis strains were typically strain-dependent, although the fermentation conditions had mainly similar effects on the growth characteristics of the different strains. By association of the transcriptomes and robustness phenotypes highly strain-specific transcriptome signatures for robustness towards heat and oxidative stress were identified, indicating that multiple mechanisms exist to increase robustness and, as a consequence, robustness of each strain requires individual optimization. However, a relatively small overlap in the transcriptome responses of the strains was also identified and this generic transcriptome signature included genes previously associated with stress (ctsR and lplL) and novel genes, including nanE and genes encoding transport proteins. The transcript levels of these genes can function as indicators of robustness and could aid in selection of fermentation parameters, potentially resulting in more optimal robustness during spray drying. PMID:27973578

Fermentation process using specific oxygen uptake rates as a process control

DOEpatents

Van Hoek, Pim; Aristidou, Aristos; Rush, Brian J.

2016-08-30

Specific oxygen uptake (OUR) is used as a process control parameter in fermentation processes. OUR is determined during at least the production phase of a fermentation process, and process parameters are adjusted to maintain the OUR within desired ranges. The invention is particularly applicable when the fermentation is conducted using a microorganism having a natural PDC pathway that has been disrupted so that it no longer functions. Microorganisms of this sort often produce poorly under strictly anaerobic conditions. Microaeration controlled by monitoring OUR allows the performance of the microorganism to be optimized.

Fermentation process using specific oxygen uptake rates as a process control

DOEpatents

Van Hoek, Pim [Minnetonka, MN; Aristidou, Aristos [Maple Grove, MN; Rush, Brian [Minneapolis, MN

2011-05-10

Specific oxygen uptake (OUR) is used as a process control parameter in fermentation processes. OUR is determined during at least the production phase of a fermentation process, and process parameters are adjusted to maintain the OUR within desired ranges. The invention is particularly applicable when the fermentation is conducted using a microorganism having a natural PDC pathway that has been disrupted so that it no longer functions. Microorganisms of this sort often produce poorly under strictly anaerobic conditions. Microaeration controlled by monitoring OUR allows the performance of the microorganism to be optimized.

Fermentation process using specific oxygen uptake rates as a process control

DOEpatents

Hoek, Van; Pim, Aristidou [Minnetonka, MN; Aristos, Rush [Maple Grove, MN; Brian, [Minneapolis, MN

2007-06-19

Specific oxygen uptake (OUR) is used as a process control parameter in fermentation processes. OUR is determined during at least the production phase of a fermentation process, and process parameters are adjusted to maintain the OUR within desired ranges. The invention is particularly applicable when the fermentation is conducted using a microorganism having a natural PDC pathway that has been disrupted so that it no longer functions. Microorganisms of this sort often produce poorly under strictly anaerobic conditions. Microaeration controlled by monitoring OUR allows the performance of the microorganism to be optimized.

Fermentation process using specific oxygen uptake rates as a process control

DOEpatents

Van Hoek, Pim; Aristidou, Aristos; Rush, Brian

2014-09-09

Specific oxygen uptake (OUR) is used as a process control parameter in fermentation processes. OUR is determined during at least the production phase of a fermentation process, and process parameters are adjusted to maintain the OUR within desired ranges. The invention is particularly applicable when the fermentation is conducted using a microorganism having a natural PDC pathway that has been disrupted so that it no longer functions. Microorganisms of this sort often produce poorly under strictly anaerobic conditions. Microaeration controlled by monitoring OUR allows the performance of the microorganism to be optimized.

Kinetic modeling of simultaneous saccharification and fermentation of corn starch for ethanol production.

PubMed

Białas, Wojciech; Czerniak, Adrian; Szymanowska-Powałowska, Daria

2014-01-01

Fuel ethanol production, using a simultaneous saccharification and fermentation process (SSF) of native starch from corn flour, has been performed using Saccharomyces cerevisiae and a granular starch hydrolyzing enzyme. The quantitative effects of mash concentration, enzyme dose and pH were investigated with the use of a Box-Wilson central composite design protocol. Proceeding from results obtained in optimal fermentation conditions, a kinetics model relating the utilization rates of starch and glucose as well as the production rates of ethanol and biomass was tested. Moreover, scanning electron microscopy (SEM) was applied to investigate corn starch granule surface after the SFF process. A maximum ethanol concentration of 110.36 g/l was obtained for native corn starch using a mash concentration of 25%, which resulted in ethanol yield of 85.71%. The optimal conditions for the above yield were found with an enzyme dose of 2.05 ml/kg and pH of 5.0. These results indicate that by using a central composite design, it is possible to determine optimal values of the fermentation parameters for maximum ethanol production. The investigated kinetics model can be used to describe SSF process conducted with granular starch hydrolyzing enzymes. The SEM micrographs reveal randomly distributed holes on the surface of granules.

Astaxanthin preparation by fermentation of esters from Haematococcus pluvialis algal extracts with Stenotrophomonas species.

PubMed

Dong, Hao; Li, Xuemin; Xue, Changhu; Mao, Xiangzhao

2016-05-01

Natural astaxanthin (Ax) is an additive that is widely used because of its beneficial biochemical functions. However, the methods used to produce free Ax have drawbacks. Chemical saponification methods produce several by-products, and lipase-catalyzed hydrolysis methods are not cost effective. In this study, a bacterial strain of Stenotrophomonas sp. was selected to enzymatically catalyze the saponification of Ax esters to produce free all-trans-Ax. Through single-factor experiments and a Box-Behnken design, the optimal fermentation conditions were determined as follows: a seed culture age of 37.79 h, an inoculum concentration of 5.92%, and an initial broth pH of 6.80. Under these conditions, a fermentation curve was drawn, and the optimal fermentation time was shown to be 60 h. At 60 h, the degradation rate of the Ax esters was 98.08%, and the yield of free all-trans-Ax was 50.130 μg/mL. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:649-656, 2016. © 2016 American Institute of Chemical Engineers.

Effect of dilute alkaline pretreatment on the conversion of different parts of corn stalk to fermentable sugars and its application in acetone-butanol-ethanol fermentation.

PubMed

Cai, Di; Li, Ping; Luo, Zhangfeng; Qin, Peiyong; Chen, Changjing; Wang, Yong; Wang, Zheng; Tan, Tianwei

2016-07-01

To investigate the effect of dilute alkaline pretreatment on different parts of biomass, corn stalk was separated into flower, leaf, cob, husk and stem, which were treated by NaOH in range of temperature and chemical loading. The NaOH-pretreated solid was then enzymatic hydrolysis and used as the substrate for batch acetone-butanol-ethanol (ABE) fermentation. The results demonstrated the five parts of corn stalk could be used as potential feedstock separately, with vivid performances in solvents production. Under the optimized conditions towards high product titer, 7.5g/L, 7.6g/L, 9.4g/L, 7g/L and 7.6g/L of butanol was obtained in the fermentation broth of flower, leaf, cob, husk and stem hydrolysate, respectively. Under the optimized conditions towards high product yield, 143.7g/kg, 126.3g/kg, 169.1g/kg, 107.7g/kg and 116.4g/kg of ABE solvent were generated, respectively. Copyright © 2016 Elsevier Ltd. All rights reserved.

Cellulase production through solid-state tray fermentation, and its use for bioethanol from sorghum stover.

PubMed

Idris, Ayman Salih Omer; Pandey, Ashok; Rao, S S; Sukumaran, Rajeev K

2017-10-01

The production of cellulase by Trichoderma reesei RUT C-30 under solid-state fermentation (SSF) on wheat bran and cellulose was optimized employing a two stage statistical design of experiments. Optimization of process parameters resulted in a 3.2-fold increase in CMCase production to 959.53IU/gDS. The process was evaluated at pilot scale in tray fermenters and yielded 457IU/gDS using the lab conditions and indicating possibility for further improvement. The cellulase could effectively hydrolyze alkali pretreated sorghum stover and addition of Aspergillus niger β-glucosidase improved the hydrolytic efficiency 174%, indicating the potential to use this blend for effective saccharification of sorghum stover biomass. The enzymatic hydrolysate of sorghum stover was fermented to ethanol with ∼80% efficiency. Copyright © 2017 Elsevier Ltd. All rights reserved.

Optimization of High Solids Dilute Acid Hydrolysis of Spent Coffee Ground at Mild Temperature for Enzymatic Saccharification and Microbial Oil Fermentation.

PubMed

Wang, Hui-Min David; Cheng, Yu-Shen; Huang, Chi-Hao; Huang, Chia-Wei

2016-10-01

Soluble coffee, being one of the world's most popular consuming drinks, produces a considerable amount of spent coffee ground (SCG) along with its production. The SCG could function as a potential lignocellulosic feedstock for production of bioproducts. The objective of this study is to investigate the possible optimal condition of dilute acid hydrolysis (DAH) at high solids and mild temperature condition to release the reducing sugars from SCG. The optimal condition was found to be 5.3 % (w/w) sulfuric acid concentration and 118 min reaction time. Under the optimal condition, the mean yield of reducing sugars from enzymatic saccharification of defatted SCG acid hydrolysate was 563 mg/g. The SCG hydrolysate was then successfully applied to culture Lipomyces starkeyi for microbial oil fermentation without showing any inhibition. The results suggested that dilute acid hydrolysis followed by enzymatic saccharification has the great potential to convert SCG carbohydrates to reducing sugars. This study is useful for the further developing of biorefinery using SCG as feedstock at a large scale.

Reduction of product-related species during the fermentation and purification of a recombinant IL-1 receptor antagonist at the laboratory and pilot scale.

PubMed

Schirmer, Emily B; Golden, Kathryn; Xu, Jin; Milling, Jesse; Murillo, Alec; Lowden, Patricia; Mulagapati, Srihariraju; Hou, Jinzhao; Kovalchin, Joseph T; Masci, Allyson; Collins, Kathryn; Zarbis-Papastoitsis, Gregory

2013-08-01

Through a parallel approach of tracking product quality through fermentation and purification development, a robust process was designed to reduce the levels of product-related species. Three biochemically similar product-related species were identified as byproducts of host-cell enzymatic activity. To modulate intracellular proteolytic activity, key fermentation parameters (temperature, pH, trace metals, EDTA levels, and carbon source) were evaluated through bioreactor optimization, while balancing negative effects on growth, productivity, and oxygen demand. The purification process was based on three non-affinity steps and resolved product-related species by exploiting small charge differences. Using statistical design of experiments for elution conditions, a high-resolution cation exchange capture column was optimized for resolution and recovery. Further reduction of product-related species was achieved by evaluating a matrix of conditions for a ceramic hydroxyapatite column. The optimized fermentation process was transferred from the 2-L laboratory scale to the 100-L pilot scale and the purification process was scaled accordingly to process the fermentation harvest. The laboratory- and pilot-scale processes resulted in similar process recoveries of 60 and 65%, respectively, and in a product that was of equal quality and purity to that of small-scale development preparations. The parallel approach for up- and downstream development was paramount in achieving a robust and scalable clinical process. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Solid state fermentation and production of rifamycin SV using Amycolatopsis mediterranei.

PubMed

Nagavalli, M; Ponamgi, S P D; Girijashankar, V; Venkateswar Rao, L

2015-01-01

Production of Rifamycin SV from cheaper agro-industrial by-products using mutant strain of Amycolatopsis mediterranei OVA5-E7 in solid state fermentation (SSF) was optimized. Among the agro-based substrates used, ragi bran was found suitable for maximizing the yield of Rifamycin SV (1310 mg 100 g(-1) ds). The yield can be further enhanced to 19·7 g Kg(-1) of dry substrate by supplementing the substrate with deoiled cotton cake (10% w/w) using optimized fermentation parameters such as maintaining 80% moisture, pH 7·0, 30°C incubation temperature, inoculum 25% v/w and carrying the solid state fermenting for 9 days. Manipulating these seven specifications, the end product yield achieved in our experimentation was 20 g of Rifamycin SV Kg(-1) ds. Eventually, an overall 5-fold improvement in Rifamycin SV production was achieved. Antibiotics such as rifamycin are broad-spectrum antimicrobial drugs used in large-scale worldwide as human medicine towards controlling diseases. Amycolatopsis mediterranei strain which produces this antibiotic was earlier used in submerged fermentation yielded lower amounts of rifamycin. By employing cheaper agro-industrial by-products, we produced upto 20 g rifamycin SV per Kg dry substrate used under optimized solid state fermentation conditions. Keeping in view, the role of rifamycin in meeting the medical demands of world's increasing population; we successfully used an improved strain on cheaper substrates with optimized fermentation parameters and achieved a 5-fold improvement in rifamycin SV production. © 2014 The Society for Applied Microbiology.

Current progress on truffle submerged fermentation: a promising alternative to its fruiting bodies.

PubMed

Tang, Ya-Jie; Liu, Rui-Sang; Li, Hong-Mei

2015-03-01

Truffle (Tuber spp.), also known as "underground gold," is popular in various cuisines because of its unique and characteristic aroma. Currently, truffle fruiting bodies are mostly obtained from nature and semi-artificial cultivation. However, the former source is scarce, and the latter is time-consuming, usually taking 4 to 12 years before harvest of the fruiting body. The truffle submerged fermentation process was first developed in Tang's lab as an alternative to its fruiting bodies. To the best of our knowledge, most reports of truffle submerged fermentation come from Tang's group. This review examines the current state of the truffle submerged fermentation process. First, the strategy to optimize the truffle submerged fermentation process is summarized; the final conditions yielded not only the highest reported truffle biomass but also the highest production of extracellular and intracellular polysaccharides. Second, the comparison of metabolites produced by truffle fermentation and fruiting bodies is presented, and the former were superior to the latter. Third, metabolites (i.e., volatile organic compounds, equivalent umami concentration, and sterol) derived from truffle fermentation could be regulated by fermentation process optimization. These findings indicated that submerged fermentation of truffles can be used for commercial production of biomass and metabolites as a promising alternative to generating its fruiting bodies in bioreactor.

A Novel Technique that Enables Efficient Conduct of Simultaneous Isomerization and Fermentation (SIF) of Xylose

NASA Astrophysics Data System (ADS)

Rao, Kripa; Chelikani, Silpa; Relue, Patricia; Varanasi, Sasidhar

Of the sugars recovered from lignocellulose, D-glucose can be readily converted into ethanol by baker's or brewer's yeast (Saccharomyces cerevisiae). However, xylose that is obtained by the hydrolysis of the hemicellulosic portion is not fermentable by the same species of yeasts. Xylose fermentation by native yeasts can be achieved via isomerization of xylose to its ketose isomer, xylulose. Isomerization with exogenous xylose isomerase (XI) occurs optimally at a pH of 7-8, whereas subsequent fermentation of xylulose to ethanol occurs at a pH of 4-5. We present a novel scheme for efficient isomerization of xylose to xylulose at conditions suitable for the fermentation by using an immobilized enzyme system capable of sustaining two different pH microenvironments in a single vessel. The proof-of-concept of the two-enzyme pellet is presented, showing conversion of xylose to xylulose even when the immobilized enzyme pellets are suspended in a bulk solution whose pH is sub-optimal for XI activity. The co-immobilized enzyme pellets may prove extremely valuable in effectively conducting "simultaneous isomerization and fermentation" (SIF) of xylose. To help further shift the equilibrium in favor of xylulose formation, sodium tetraborate (borax) was added to the isomerization solution. Binding of tetrahydroxyborate ions to xylulose effectively reduces the concentration of xylulose and leads to increased xylose isomerization. The formation of tetrahydroxyborate ions and the enhancement in xylulose production resulting from the complexation was studied at two different bulk pH values. The addition of 0.05 M borax to the isomerization solution containing our co-immobilized enzyme pellets resulted in xylose to xylulose conversion as high as 86% under pH conditions that are suboptimal for XI activity. These initial findings, which can be optimized for industrial conditions, have significant potential for increasing the yield of ethanol from xylose in an SIF approach.

Submerged fermentation of Lactobacillus rhamnosus YS9 for γ-aminobutyric acid (GABA) production

PubMed Central

Lin, Qian

2013-01-01

γ-Aminobutyric acid (GABA) is a major inhibitory neurotransmitter in central nervous system, and its application in drugs and functional foods has attracted great attention. To enhance production of γ-aminobutyric acid, Lactobacillus rhamnosus YS9, a strain isolated from Chinese traditional fermented food pickled vegetable, was grown under submerged fermentation. Its cultivation conditions were investigated. When culture pH condition was adjusted to the optimal pH of glutamate decarboxylase activity, culture of Lb. rhamnosus YS9 in medium supplemented with 200 mM of monosodium glutamate and 200 μM of pyridoxal phosphate (PLP), produced 187 mM of GABA. PMID:24159304

Metabolomics-based optimal koji fermentation for tyrosinase inhibition supplemented with Astragalus radix.

PubMed

Kim, Ah Jin; Choi, Jung Nam; Kim, Jiyoung; Yeo, Soo Hwan; Choi, Ji Ho; Lee, Choong Hwan

2012-01-01

The present study was focused on improving the quality of rice koji by fermentation with a selected Aspergillus oryzae strain and a plant Astragalus radix. A. oryzae KCCM 60345 was used as main inoculant and the Astragalus radix was added as supplement in rice koji preparation. LC-MS based metabolite analysis and tyrosinase inhibitory activities were studied for different time periods. A. oryzae KCCM 60345 fermented rice koji supplemented with Astragalus showed higher tyrosinase inhibition activity at 4 d of fermentation and metabolite analysis with PCA and PLS-DA indicated differences in kojic acid, calycosin-7-O-β-D-glucoside, ononin, calycosin, and formononetin as compared with other forms of rice koji fermentation. By correlation analysis between metabolites and tyrosinase inhibitory activity, calycosin and kojic acid were identified as major tyrosinase inhibitors. Based on these results, we concluded that A. oryzae KCCM 60345 supplemented with Astragalus radix is useful for whitening effects, and we identified optimal conditions for rice koji preparation.

Mechanisms and kinetics of cellulose fermentation for protein production

NASA Technical Reports Server (NTRS)

Dunlap, C. A.

1971-01-01

The development of a process (and ancillary processing and analytical techniques) to produce bacterial single-cell protein of good nutritional quality from waste cellulose is discussed. A fermentation pilot plant and laboratory were developed and have been in operation for about two years. Single-cell protein (SCP) can be produced from sugarcane bagasse--a typical agricultural cellulosic waste. The optimization and understanding of this process and its controlling variables are examined. Both batch and continuous fermentation runs have been made under controlled conditions in the 535 liter pilot plant vessel and in the laboratory 14-liter fermenters.

Simultaneous saccharification and ethanol fermentation of oxalic acid pretreated corncob assessed with response surface methodology

Treesearch

Jae-Won Lee; Rita C.L.B. Rodrigues; Thomas W. Jeffries

2009-01-01

Response surface methodology was used to evaluate optimal time, temperature and oxalic acid concentration for simultaneous saccharification and fermentation (SSF) of corncob particles by Pichia stipitis CBS 6054. Fifteen different conditions for pretreatment were examined in a 23 full factorial design with six axial points. Temperatures ranged from 132 to 180º...

Antitumour activity of 3-nitropropionic acid from Phomopsis sp. and optimization of fermentation conditions.

PubMed

Lu, F T; Ma, D C; Yan, W; Guo, J; Bai, L H

2015-08-01

In this study, 3-nitropropionic acid (3-NPA) was separated and purified from endophytic fungi belonging to Phomopsis sp. and its cytotoxicity was determined by MTT assay. Treatment with 3-NPA for 24 h resulted in a dose-dependent apoptosis in MCF-7 cells. Through quantitative detection of the genes that are closely related to the Bcl-2 signalling pathway, there was an increased expression of p53 and Bax and a decreased expression of Bcl-2, which indicated apoptosis in these cells. Meanwhile, the overexpression of PARA (poly ADP-ribose polymerase) and apoptosis inducing factor (AIF) also suggested that 3-NPA induced cellular apoptosis through a caspase-3-independent pathway in caspase-3-deficient MCF-7 cells. The fermentation condition was also improved to produce more 3-NPA: glucose as a carbon source and yeast extract as a nitrogen source, fermentation for 8 days at 32°C and a solution environment of pH 5·0. Under these conditions, the yield of 3-NPA was increased to 529 mg l(-1) compared with 410 mg l(-1) under traditional fermentation conditions. 3-Nitropropionic acid is a mitochondrial inhibitor and has some useful bioactivities such as antibacterial activity. In this paper we found that 3-NPA also has obvious cytotoxicity, so we studied its antitumour activity and tried to determine the antitumour molecular mechanism, opening a new perspective for potential antitumour prodrug development. As 3-NPA is often obtained from natural products with a low yield, in order to overcome the disadvantage of an endophytic fungi source of 3-NPA, we optimized the fermentation conditions for 3-NPA in Phomopsis sp. to obtain the maximum production of 3-NPA. © 2015 The Society for Applied Microbiology.

Utilization of cellulosic materials through enzyamtic hydrolysis. I. Fermentation of hydrolysate to ethanol and single-cell protein.

PubMed

Cysewski, G R; Wilke, C R

1976-09-01

Ethanol fermentation studies were conducted with Saccharomyces cerevisiae ATCC "4126, to determine the optimal conditions of oxygen tension and feed sugar concentration. In long-term continuous culture maximum ethanol production was found to occur at 0.07 mmHg oxygen tension and 10% glucose feed concentration. Preliminary process design and cost studies are developed for industrial scale fermentations to produce ethanol and torula yeast from sugars obtained by enzymatic hydrolysis of newsprint.

Improved fermentation performance to produce bioethanol from Gelidium amansii using Pichia stipitis adapted to galactose.

PubMed

Sukwong, Pailin; Ra, Chae Hun; Sunwoo, In Yung; Tantratian, Sumate; Jeong, Gwi-Taek; Kim, Sung-Koo

2018-03-23

This study employed a statistical method to obtain optimal hyper thermal acid hydrolysis conditions using Gelidium amansii (red seaweed) as a source of biomass. The optimal hyper thermal acid hydrolysis using G. amansii as biomass was determined as 12% (w/v) slurry content, 358.3 mM H 2 SO 4 , and temperature of 142.6 °C for 11 min. After hyper thermal acid hydrolysis, enzymatic saccharification was carried out. The total monosaccharide concentration was 45.1 g/L, 72.2% of the theoretical value of the total fermentable monosaccharides of 62.4 g/L based on 120 g dry weight/L in the G. amansii slurry. To increase ethanol production, 3.8 g/L 5-hydroxymethylfurfural (HMF) in the hydrolysate was removed by treatment with 3.5% (w/v) activated carbon for 2 min and fermented with Pichia stipitis adapted to high galactose concentrations via separate hydrolysis and fermentation. With complete HMF removal and the use of P. stipitis adapted to high galactose concentrations, 22 g/L ethanol was produced (yield 0.50). Fermentation with total HMF removal and yeast adapted to high galactose concentrations increased the fermentation performance and decreased the fermentation time from 96 to 36 h compared to traditional fermentation.

Open fermentative production of fuel ethanol from food waste by an acid-tolerant mutant strain of Zymomonas mobilis.

PubMed

Ma, Kedong; Ruan, Zhiyong; Shui, Zongxia; Wang, Yanwei; Hu, Guoquan; He, Mingxiong

2016-03-01

The aim of present study was to develop a process for open ethanol fermentation from food waste using an acid-tolerant mutant of Zymomonas mobilis (ZMA7-2). The mutant showed strong tolerance to acid condition of food waste hydrolysate and high ethanol production performance. By optimizing fermentation parameters, ethanol fermentation with initial glucose concentration of 200 g/L, pH value around 4.0, inoculum size of 10% and without nutrient addition was considered as best conditions. Moreover, the potential of bench scales fermentation and cell reusability was also examined. The fermentation in bench scales (44 h) was faster than flask scale (48 h), and the maximum ethanol concentration and ethanol yield (99.78 g/L, 0.50 g/g) higher than that of flask scale (98.31 g/L, 0.49 g/g). In addition, the stable cell growth and ethanol production profile in five cycles successive fermentation was observed, indicating the mutant was suitable for industrial ethanol production. Copyright © 2015 Elsevier Ltd. All rights reserved.

Statistical culture-based strategies to enhance chlamydospore production by Trichoderma harzianum SH2303 in liquid fermentation.

PubMed

Li, Ya-Qian; Song, Kai; Li, Ya-Chai; Chen, Jie

2016-08-01

Trichoderma-based formulations are applied as commercial biocontrol agents for soil-borne plant pathogens. Chlamydospores are active propagules in Trichoderma spp., but their production is currently limited due to a lack of optimal liquid fermentation technology. In this study, we explored response surface methodologies for optimizing fermentation technology in Trichoderma SH2303. Our initial studies, using the Plackett-Burman design, identified cornmeal, glycerol, and initial pH levels as the most significant factors (P<0.05) for enhancing the production of chlamydospores. Subsequently, we applied the Box-Behnken design to study the interactions between, and optimal levels of, a number of factors in chlamydospore production. These statistically predicted results indicated that the highest number of chlamydospores (3.6×10(8) spores/ml) would be obtained under the following condition: corn flour 62.86 g/L, glycerol 7.54 ml/L, pH 4.17, and 6-d incubation in liquid fermentation. We validated these predicted values via three repeated experiments using the optimal culture and achieved maximum chlamydospores of 4.5×10(8) spores/ml, which approximately a 8-fold increase in the number of chlamydospores produced by T. harzianum SH2303 compared with that before optimization. These optimized values could help make chlamydospore production cost-efficient in the future development of novel biocontrol agents.

Statistical culture-based strategies to enhance chlamydospore production by Trichoderma harzianum SH2303 in liquid fermentation*

PubMed Central

Li, Ya-qian; Song, Kai; Li, Ya-chai; Chen, Jie

2016-01-01

Trichoderma-based formulations are applied as commercial biocontrol agents for soil-borne plant pathogens. Chlamydospores are active propagules in Trichoderma spp., but their production is currently limited due to a lack of optimal liquid fermentation technology. In this study, we explored response surface methodologies for optimizing fermentation technology in Trichoderma SH2303. Our initial studies, using the Plackett-Burman design, identified cornmeal, glycerol, and initial pH levels as the most significant factors (P<0.05) for enhancing the production of chlamydospores. Subsequently, we applied the Box-Behnken design to study the interactions between, and optimal levels of, a number of factors in chlamydospore production. These statistically predicted results indicated that the highest number of chlamydospores (3.6×108 spores/ml) would be obtained under the following condition: corn flour 62.86 g/L, glycerol 7.54 ml/L, pH 4.17, and 6-d incubation in liquid fermentation. We validated these predicted values via three repeated experiments using the optimal culture and achieved maximum chlamydospores of 4.5×108 spores/ml, which approximately a 8-fold increase in the number of chlamydospores produced by T. harzianum SH2303 compared with that before optimization. These optimized values could help make chlamydospore production cost-efficient in the future development of novel biocontrol agents. PMID:27487807

Bioconversion of shrimp waste Penaeus merguiensis using lactic acid fermentation: An alternative procedure for chemical extraction of chitin and chitosan.

PubMed

Sedaghat, Fatemeh; Yousefzadi, Morteza; Toiserkani, Hojjat; Najafipour, Sohrab

2017-11-01

Chitin extraction from shrimp wastes by biological treatment, using the Pseudomonas aeruginosa was a positive and simple method. In order to look for the optimal conditions, the wastes were incubated at 30°C and 100rpm in different glucose (0%, 10%, 15% and 20%) and inoculation (10%, 15% and 20%) concentrations for 4 and 6days. At the end of fermentation, Protease activity was investigated at different temperatures and temperature 50°C was considered as the optimum. The results obtained also showed a direct relationship between the concentration of different parameters and deproteinization and demineralization rates, so that the optimal conditions were 20% glucose, 20% inoculation and 6days fermentation. These conditions led to 82% demineralization, 92% deproteinization and chitin yield of 47%. Then, chitin was converted to chitosan using microwave, autoclave and traditional methods. The highest yield (87%) was obtained with autoclave method. At the end, the chitin and chitosan were characterized by elemental analysis and FTIR. Copyright © 2017 Elsevier B.V. All rights reserved.

Lactic acid production from submerged fermentation of broken rice using undefined mixed culture.

PubMed

Nunes, Luiza Varela; de Barros Correa, Fabiane Fernanda; de Oliva Neto, Pedro; Mayer, Cassia Roberta Malacrida; Escaramboni, Bruna; Campioni, Tania Sila; de Barros, Natan Roberto; Herculano, Rondinelli Donizetti; Fernández Núñez, Eutimio Gustavo

2017-04-01

The present work aimed to characterize and optimize the submerged fermentation of broken rice for lactic acid (LA) production using undefined mixed culture from dewatered activated sludge. A microorganism with amylolytic activity, which also produces LA, Lactobacillus amylovorus, was used as a control to assess the extent of mixed culture on LA yield. Three level full factorial designs were performed to optimize and define the influence of fermentation temperature (20-50 °C), gelatinization time (30-60 min) and broken rice concentration in culture medium (40-80 g L -1 ) on LA production in pure and undefined mixed culture. LA production in mixed culture (9.76 g L -1 ) increased in sixfold respect to pure culture in optimal assessed experimental conditions. The optimal conditions for maximizing LA yield in mixed culture bioprocess were 31 °C temperature, 45 min gelatinization time and 79 g L -1 broken rice concentration in culture medium. This study demonstrated the positive effect of undefined mixed culture from dewatered activated sludge to produce LA from culture medium formulated with broken rice. In addition, this work establishes the basis for an efficient and low-cost bioprocess to manufacture LA from this booming agro-industrial by-product.

Iso-migrastatin Titer Improvement in the Engineered Streptomyces lividans SB11002 Strain by Optimization of Fermentation Conditions

PubMed Central

Wu, Xueyun; Yang, Dong; Zhu, Xiangcheng; Feng, Zhiyang; Lv, Zhengbin; Zhang, Yaozhou; Shen, Ben; Xu, Zhinan

2011-01-01

The heterologous production of iso-migrastatin (iso-MGS) was successfully demonstrated in an engineered S. lividans SB11002 strain, which was derived from S. lividans K4–114, following introduction of pBS11001, which harbored the entire mgs biosynthetic gene cluster. However, under similar fermentation conditions, the iso-MGS titer in the engineered strain was significantly lower than that in the native producer - Streptomyces platensis NRRL 18993. To circumvent the problem of low iso-MGS titers and to expand the utility of this heterologous system for iso-MGS biosynthesis and engineering, systematic optimization of the fermentation medium was carried out. The effects of major components in the cultivation medium, including carbon, organic and inorganic nitrogen sources, were investigated using a single factor optimization method. As a result, sucrose and yeast extract were determined to be the best carbon and organic nitrogen sources, resulting in optimized iso-MGS production. Conversely, all other inorganic nitrogen sources evaluated produced various levels of inhibition of iso-MGS production. The final optimized R2YE production medium produced iso-MGS with a titer of 86.5 mg/L, about 3.6-fold higher than that in the original R2YE medium, and 1.5 fold higher than that found within the native S. platensis NRRL 18993 producer. PMID:21625393

[Isolation and fermentation condition of milk-clotting enzyme producing strain from glutinous rice wine].

PubMed

Cheng, Qiaoling; Bai, Xiaojia; Wang, Yanping

2008-06-01

Glutinous rice wine is a traditional food in south of China and it can coagulate milk. It has been proved that its function of coagulating milk is because of the presence of milk-clotting enzyme produced by microorganisms in glutinous rice wine. The aim of this work is to isolate milk-clotting enzyme producing strain from glutinous rice wine and study the fermentation condition. We screened out four bacteria and fungus by gradient dilution. It was proved that mold played the most important role in the production of milk-clotting enzyme. This is further confirmed by casein plate method. The optimization of fermentation conditions revealed that two times concentrated potato medium supplemented with 5% glucose without additional nitrogen was better for production of the enzyme. The enzyme activity was increased 144% under the conditions established.

Expanding a dynamic flux balance model of yeast fermentation to genome-scale

PubMed Central

2011-01-01

Background Yeast is considered to be a workhorse of the biotechnology industry for the production of many value-added chemicals, alcoholic beverages and biofuels. Optimization of the fermentation is a challenging task that greatly benefits from dynamic models able to accurately describe and predict the fermentation profile and resulting products under different genetic and environmental conditions. In this article, we developed and validated a genome-scale dynamic flux balance model, using experimentally determined kinetic constraints. Results Appropriate equations for maintenance, biomass composition, anaerobic metabolism and nutrient uptake are key to improve model performance, especially for predicting glycerol and ethanol synthesis. Prediction profiles of synthesis and consumption of the main metabolites involved in alcoholic fermentation closely agreed with experimental data obtained from numerous lab and industrial fermentations under different environmental conditions. Finally, fermentation simulations of genetically engineered yeasts closely reproduced previously reported experimental results regarding final concentrations of the main fermentation products such as ethanol and glycerol. Conclusion A useful tool to describe, understand and predict metabolite production in batch yeast cultures was developed. The resulting model, if used wisely, could help to search for new metabolic engineering strategies to manage ethanol content in batch fermentations. PMID:21595919

Utilization of deoiled Jatropha curcas seed cake for production of xylanase from thermophilic Scytalidium thermophilum.

PubMed

Joshi, Chetna; Khare, S K

2011-01-01

Jatropha curcas is a major biodiesel crop. Large amount of deoiled cake is generated as by-product during biodiesel production from its seeds. Deoiled J. curcas seed cake was assessed as substrate for the production of xylanase from thermophilic fungus Scytalidium thermophilum by solid-state fermentation. The seed cake was efficiently utilized by S. thermophilum for its growth during which it produced good amount of heat stable extracellular xylanase. The solid-state fermentation conditions were optimized for maximum xylanase production. Under the optimized conditions viz. deoiled seed cake supplemented with 1% oat-spelt xylan, adjusted to pH 9.0, moisture content 1:3 w/v, inoculated with 1×10(6) spores per 5 g cake and incubated at 45 °C, 1455 U xylanase/g deoiled seed cake was obtained. The xylanase was useful in biobleaching of paper pulp. Solid-state fermentation of deoiled cake appears a potentially viable approach for its effective utilization. Copyright © 2010 Elsevier Ltd. All rights reserved.

Statistical optimization of process parameters for the production of tannase by Aspergillus flavus under submerged fermentation.

PubMed

Mohan, S K; Viruthagiri, T; Arunkumar, C

2014-04-01

Production of tannase by Aspergillus flavus (MTCC 3783) using tamarind seed powder as substrate was studied in submerged fermentation. Plackett-Burman design was applied for the screening of 12 medium nutrients. From the results, the significant nutrients were identified as tannic acid, magnesium sulfate, ferrous sulfate and ammonium sulfate. Further the optimization of process parameters was carried out using response surface methodology (RSM). RSM has been applied for designing of experiments to evaluate the interactive effects through a full 31 factorial design. The optimum conditions were tannic acid concentration, 3.22 %; fermentation period, 96 h; temperature, 35.1 °C; and pH 5.4. Higher value of the regression coefficient (R 2  = 0.9638) indicates excellent evaluation of experimental data by second-order polynomial regression model. The RSM revealed that a maximum tannase production of 139.3 U/ml was obtained at the optimum conditions.

Overexpression and characterization of laccase from Trametes versicolor in Pichia pastoris.

PubMed

Li, Q; Pei, J; Zhao, L; Xie, J; Cao, F; Wang, G

2014-01-01

A laccase-encoding gene of Trametes versicolor, lccA, was cloned and expressed in Pichia pastoris X33. The lccA gene consists ofa 1560 bp open reading frame encoding 519 amino acids, which was classified into family copper blue oxidase. To improve the expression level of recombinant laccase in P. pastoris, conditions of the fermentation were optimized by the single factor experiments. The optimal fermentation conditions for the laccase production in shake flask cultivation using BMGY medium were obtained: the optimal initial pH 7.0, the presence of 0.5 mM Cu2+, 0.6% methanol added into the culture every 24 h. The laccase activity was up to 11.972 U/L under optimal conditions after 16 days of induction in a medium with 4% peptone. After 100 h of large scale production in 5 L fermenter the enzyme activity reached 18.123 U/L. The recombinant laccase was purified by ultrafiltration and (NH4)2SO4 precipitation showing a single band on SDS-PAGE, which had a molecular mass of 58 kDa. The optimum pH and temperature for the laccase were pH 2.0 and 50 degrees C with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as a substrate. The recombinant laccase was stable over a pH range of 2.0-7.0. The K(m) and the V(max) value of LccA were 0.43 mM and 82.3 U/mg for ABTS, respectively.

Direct concentration and viability measurement of yeast in corn mash using a novel imaging cytometry method.

PubMed

Chan, Leo L; Lyettefi, Emily J; Pirani, Alnoor; Smith, Tim; Qiu, Jean; Lin, Bo

2011-08-01

Worldwide awareness of fossil-fuel depletion and global warming has been increasing over the last 30 years. Numerous countries, including the USA and Brazil, have introduced large-scale industrial fermentation facilities for bioethanol, biobutanol, or biodiesel production. Most of these biofuel facilities perform fermentation using standard baker's yeasts that ferment sugar present in corn mash, sugar cane, or other glucose media. In research and development in the biofuel industry, selection of yeast strains (for higher ethanol tolerance) and fermentation conditions (yeast concentration, temperature, pH, nutrients, etc.) can be studied to optimize fermentation performance. Yeast viability measurement is needed to identify higher ethanol-tolerant yeast strains, which may prolong the fermentation cycle and increase biofuel output. In addition, yeast concentration may be optimized to improve fermentation performance. Therefore, it is important to develop a simple method for concentration and viability measurement of fermenting yeast. In this work, we demonstrate an imaging cytometry method for concentration and viability measurements of yeast in corn mash directly from operating fermenters. It employs an automated cell counter, a dilution buffer, and staining solution from Nexcelom Bioscience to perform enumeration. The proposed method enables specific fluorescence detection of viable and nonviable yeasts, which can generate precise results for concentration and viability of yeast in corn mash. This method can provide an essential tool for research and development in the biofuel industry and may be incorporated into manufacturing to monitor yeast concentration and viability efficiently during the fermentation process.

Optimization of fermentation conditions for green pigment production from Bacillus cereus M¹ 16 (MTCC 5521) and its pharmacological application.

PubMed

Banerjee, D; Mondal, A; Gupta, M; Guha, A K; Ray, L

2014-01-01

Optimal culture conditions for the production of green pigment was investigated. The optimal culture condition for the production of an extracellular green pigment by growing Bacillus cereus M(1) 16 (MTCC 5521) in a complex medium containing (g l(-1) ) Peptone-4.0, Beef Extract-9.0, NaCl-7.0, MgSO4 .7H2 O-1.0 and KH2 PO4 -5.0 was as follows pH-7.0 at 30°C for 72 h in a 5 l fermenter. Aeration rate and agitator speed had no effect on the pigment production. Thin layer chromatogram of the pigment extracted from the fermented broth with chloroform on silica gel GF254 using ethyl acetate and hexane (1 : 1) as solvent showed three fractions. The major fraction (C3 ) was separated out and identified as 9-methyl-1, 4, 5, 8-tetra-azaphenanthrene. Acute toxicity test revealed the nontoxic nature upto a dose of 2000 mg kg(-1) , b.wt., of mice. MTT assay showed the cytotoxic nature in HL60 cells having an IC50 of 2.47 mmol. So, this biopigment may have application in food, textile colorant and pharmaceutical industry. This study demonstrated the optimum production of a biopigment (9-methyl-1, 4, 5, 8-tetra-azaphenanthrene) by fermentation of a complex medium with Bacillus cereus M(1) 16 (MTCC 5521) in submerged fermentation. This is the first investigation of toxicity and cytotoxicity activities of this biopigment. The study showed that the purified pigment had no toxicity to healthy albino mice but a high cytotoxicity activity in HL60 cancer cell line in vitro. The biopigment had further displayed dyeing capability to both solidified agar and cotton cloth. Therefore, it may represent a nontoxic and natural alternative to chemical dyes and pigments. © 2013 The Society for Applied Microbiology.

Optimization of fermentation conditions for the production of curcumin by engineered Escherichia coli.

PubMed

Couto, Márcia R; Rodrigues, Joana L; Rodrigues, Lígia R

2017-08-01

Curcumin is a plant secondary metabolite with outstanding therapeutic effects. Therefore, there is a great interest in developing new strategies to produce this high-value compound in a cheaper and environmentally friendly way. Curcumin heterologous production in Escherichia coli using artificial biosynthetic pathways was previously demonstrated using synthetic biology approaches. However, the culturing conditions to produce this compound were not optimized and so far only a two-step fermentation process involving the exchange of culture medium allowed high concentrations of curcumin to be obtained, which limits its production at an industrial scale. In this study, the culturing conditions to produce curcumin were evaluated and optimized. In addition, it was concluded that E. coli BL21 allows higher concentrations of curcumin to be produced than E. coli K-12 strains. Different isopropyl β-d-thiogalactopyranoside concentrations, time of protein expression induction and substrate type and concentration were also evaluated. The highest curcumin production obtained was 959.3 µM (95.93% of per cent yield), which was 3.1-fold higher than the highest concentration previously reported. This concentration was obtained using a two-stage fermentation with lysogeny broth (LB) and M9. Moreover, terrific broth was also demonstrated to be a very interesting alternative medium to produce curcumin because it also led to high concentrations (817.7 µM). The use of this single fermentation medium represents an advantage at industrial scale and, although the final production is lower than that obtained with the LB-M9 combination, it leads to a significantly higher production of curcumin in the first 24 h of fermentation. This study allowed obtaining the highest concentrations of curcumin reported so far in a heterologous organism and is of interest for all of those working with the heterologous production of curcuminoids, other complex polyphenolic compounds or plant secondary metabolites. © 2017 The Author(s).

Quantitative analysis of pyoluteorin in anti-fungal fermentation liquor of Pseudomonas species by capillary zone electrophoresis with UV-vis detector.

PubMed

Wang, Qiu-Ling; Zhang, Xue-Hong; Fan, Liu-Yin; Zhang, Wei; Xu, Yu-Qian; Hu, Hong-Bo; Cao, Cheng-Xi

2005-11-05

This paper investigated potential utility of capillary zone electrophoresis (CZE) for very succinct but robust quantitative analysis of pyoluteorin (Plt) in anti-fungal fermentation liquor of Pseudomonas species. The experimental conditions for the separation and quantification of Plt were optimized at first. The optimized conditions are: 80 mmol/L pH 8.40 Gly-NaOH buffer, 51 cm total length (42 cm effective) and 75 microm I.D. capillary, 230 nm wavelength, 25 kV, 13 mbar 10s pressure sample injection and 24 degrees C air-cooling. Under the optimized conditions, the migration times of Plt and the internal standard phenobarbital are 2.09 and 2.49 min, respectively, the linear response of Plt concentration ranges from 5.0 to 1000 microg/mL with high correlation coefficient (r=0.99977, n=9), the limits of detection (LOD) and quantification (LOQ) for Plt are 0.66 and 2.2 microg/mL, the precision values (expressed as R.S.D.) of intra- and inter-day are 1.19-1.94% and 1.55-6.21%, respectively, the recoveries of Plt at three concentration levels of 750, 250 and 50 microg/mL range from 90.31% to 97.85% and to 98.96%, respectively. The developed method can be well used for the quantification of Plt in the fermentation liquor.

Fermentation Process and Metabolic Flux of Ethanol Production from the Detoxified Hydrolyzate of Cassava Residue

PubMed Central

Li, Xingjiang; Deng, Yongdong; Yang, Ying; Wei, Zhaojun; Cheng, Jieshun; Cao, Lili; Mu, Dongdong; Luo, Shuizhong; Zheng, Zhi; Jiang, Shaotong; Wu, Xuefeng

2017-01-01

With the growth of the world population, energy problems are becoming increasingly severe; therefore, sustainable energy sources have gained enormous importance. With respect to ethanol fuel production, biomass is gradually replacing grain as the main raw material. In this study, we explored the fermentation of five- and six-carbon sugars, the main biomass degradation products, into alcohol. We conducted mutagenic screening specifically for Candida tropicalis CICC1779 to obtain a strain that effectively used xylose (Candida tropicalis CICC1779-Dyd). By subsequently studying fermentation conditions under different initial liquid volume oxygen transfer coefficients (kLα), and coupling control of the aeration rate and agitation speed under optimal conditions, the optimal dissolved oxygen change curve was obtained. In addition, we constructed metabolic flow charts and equations to obtain a better understanding of the fermentation mechanism and to improve the ethanol yield. In our experiment, the ethanol production of the wild type stain was 17.58 g·L−1 at a kLα of 120. The highest ethanol yield of the mutagenic strains was 24.85 g·L−1. The ethanol yield increased to 26.56 g·L−1 when the dissolved oxygen content was optimized, and the conversion of sugar into alcohol reached 0.447 g·g−1 glucose (the theoretical titer of yeast-metabolized xylose was 0.46 g ethanol/g xylose and the glucose ethanol fermentation titer was 0.51 g ethanol/g glucose). Finally, the detected activity of xylose reductase and xylose dehydrogenase was higher in the mutant strain than in the original, which indicated that the mutant strain (CICC1779-Dyd) could effectively utilize xylose for metabolism. PMID:28878755

Evaluation of a kinetic model for computer simulation of growth and fermentation by Scheffersomyces (Pichia) stipitis fed D-xylose.

PubMed

Slininger, P J; Dien, B S; Lomont, J M; Bothast, R J; Ladisch, M R; Okos, M R

2014-08-01

Scheffersomyces (formerly Pichia) stipitis is a potential biocatalyst for converting lignocelluloses to ethanol because the yeast natively ferments xylose. An unstructured kinetic model based upon a system of linear differential equations has been formulated that describes growth and ethanol production as functions of ethanol, oxygen, and xylose concentrations for both growth and fermentation stages. The model was validated for various growth conditions including batch, cell recycle, batch with in situ ethanol removal and fed-batch. The model provides a summary of basic physiological yeast properties and is an important tool for simulating and optimizing various culture conditions and evaluating various bioreactor designs for ethanol production. © 2014 Wiley Periodicals, Inc.

Optimization of culture conditions and bench-scale production of L-asparaginase by submerged fermentation of Aspergillus terreus MTCC 1782.

PubMed

Gurunathan, Baskar; Sahadevan, Renganathan

2012-07-01

Optimization of culture conditions for L-asparaginase production by submerged fermentation of Aspergillus terreus MTCC 1782 was studied using a 3-level central composite design of response surface methodology and artificial neural network linked genetic algorithm. The artificial neural network linked genetic algorithm was found to be more efficient than response surface methodology. The experimental L-asparaginase activity of 43.29 IU/ml was obtained at the optimum culture conditions of temperature 35 degrees C, initial pH 6.3, inoculum size 1% (v/v), agitation rate 140 rpm, and incubation time 58.5 h of the artificial neural network linked genetic algorithm, which was close to the predicted activity of 44.38 IU/ml. Characteristics of L-asparaginase production by A. terreus MTCC 1782 were studied in a 3 L bench-scale bioreactor.

Strain improvement by combined UV mutagenesis and ribosome engineering and subsequent fermentation optimization for enhanced 6'-deoxy-bleomycin Z production.

PubMed

Zhu, Xiangcheng; Kong, Jieqian; Yang, Hu; Huang, Rong; Huang, Yong; Yang, Dong; Shen, Ben; Duan, Yanwen

2018-02-01

The bleomycins (BLMs) are important clinical drugs extensively used in combination chemotherapy for the treatment of various cancers. Dose-dependent lung toxicity and the development of drug resistance have restricted their wide applications. 6'-Deoxy-BLM Z, a recently engineered BLM analogue with improved antitumor activity, has the potential to be developed into the next-generation BLM anticancer drug. However, its low titer in the recombinant strain Streptomyces flavoviridis SB9026 has hampered current efforts, which require sufficient compound, to pursue preclinical studies and subsequent clinical development. Here, we report the strain improvement by combined UV mutagenesis and ribosome engineering, as well as the fermentation optimization, for enhanced 6'-deoxy-BLM production. A high producer, named S. flavoviridis G-4F12, was successfully isolated, producing 6'-deoxy-BLM at above 70 mg/L under the optimized fermentation conditions, representing a sevenfold increase in comparison with that of the original producer. These findings demonstrated the effectiveness of combined empirical breeding methods in strain improvement and set the stage for sustainable production of 6'-deoxy-BLM via pilot-scale microbial fermentation.

Co-fermentation of carbon sources by Enterobacter aerogenes ATCC 29007 to enhance the production of bioethanol.

PubMed

Thapa, Laxmi Prasad; Lee, Sang Jun; Yang, Xiao Guang; Yoo, Hah Young; Kim, Sung Bong; Park, Chulhwan; Kim, Seung Wook

2014-06-01

We investigated the enhancement of bioethanol production in Enterobacter aerogenes ATCC 29007 by co-fermentation of carbon sources such as glycerol, glucose, galactose, sucrose, fructose, xylose, starch, mannitol and citric acid. Biofuel production increases with increasing growth rate of microorganisms; that is why we investigated the optimal growth rate of E. aerogenes ATCC 29007, using mixtures of different carbon sources with glycerol. E. aerogenes ATCC 29007 was incubated in media containing each carbon source and glycerol; growth rate and bioethanol production improved in all cases compared to those in medium containing glycerol alone. The growth rate and bioethanol production were highest with mannitol. Fermentation was carried out at 37 °C for 18 h, pH 7, using 50 mL defined production medium in 100 mL serum bottles at 200 rpm. Bioethanol production under optimized conditions in medium containing 16 g/L mannitol and 20 g/L glycerol increased sixfold (32.10 g/L) than that containing glycerol alone (5.23 g/L) as the carbon source in anaerobic conditions. Similarly, bioethanol production using free cells in continuous co-fermentation also improved (27.28 g/L) when 90.37 % of 16 g/L mannitol and 67.15 % of 20 g/L glycerol were used. Although naturally existing or engineered microorganisms can ferment mixed sugars sequentially, the preferential utilization of glucose to non-glucose sugars often results in lower overall yield and productivity of ethanol. Here, we present new findings in E. aerogenes ATCC 29007 that can be used to improve bioethanol production by simultaneous co-fermentation of glycerol and mannitol.

Artificial neural network - Genetic algorithm to optimize wheat germ fermentation condition: Application to the production of two anti-tumor benzoquinones.

PubMed

Zheng, Zi-Yi; Guo, Xiao-Na; Zhu, Ke-Xue; Peng, Wei; Zhou, Hui-Ming

2017-07-15

Methoxy-ρ-benzoquinone (MBQ) and 2, 6-dimethoxy-ρ-benzoquinone (DMBQ) are two potential anticancer compounds in fermented wheat germ. In present study, modeling and optimization of added macronutrients, microelements, vitamins for producing MBQ and DMBQ was investigated using artificial neural network (ANN) combined with genetic algorithm (GA). A configuration of 16-11-1 ANN model with Levenberg-Marquardt training algorithm was applied for modeling the complicated nonlinear interactions among 16 nutrients in fermentation process. Under the guidance of optimized scheme, the total contents of MBQ and DMBQ was improved by 117% compared with that in the control group. Further, by evaluating the relative importance of each nutrient in terms of the two benzoquinones' yield, macronutrients and microelements were found to have a greater influence than most of vitamins. It was also observed that a number of interactions between nutrients affected the yield of MBQ and DMBQ remarkably. Copyright © 2017 Elsevier Ltd. All rights reserved.

Study on fermentation kinetics and extraction process of rhamnolipid production by papermaking wastewater

NASA Astrophysics Data System (ADS)

Yu, Keer

2018-01-01

Paper mill wastewater (PMW) is the outlet water generated during pulp and papermaking process in the paper industry. Fermentation by wastewater can lower the cost of production as well as alleviate the pressure of wastewater treatment. Rhamnolipids find broad placations as natural surfactants. This paper studied the rhamnolipids fermentation by employing Pseudomonas aeruginosa isolated by the laboratory, and determined to use wastewater which filtered by medium speed filter paper and strain Z2, the culture conditions were optimized, based on the flask shaking fermentation. On the basis of 5L tank fermentation, batch fermentation was carried out, the yield of fermentation reached 7.067g/L and the fermentation kinetics model of cell growth, product formation and substrate consumption was established by using origin software, and the fermentation process could be simulated well. And studied on the extraction process of rhamnolipids, through fermentation dynamic equation analysis can predict the in fill material yield can be further improved. Research on the extraction process of rhamnolipid simplifies the operation of extraction, and lays the foundation for the industrial extraction.

Exploring the influence of culture conditions on kefir's anticancer properties.

PubMed

Hatmal, Ma'mon M; Nuirat, Abeer; Zihlif, Malek A; Taha, Mutasem O

2018-05-01

Cancer is a major health problem in many parts of the world. Conventional anticancer treatments are painful, expensive, and unsafe. Therefore, demand is increasing for cancer treatments preferentially in the form of functional foods or nutritional supplements. Kefir, a traditional fermented milk dairy product, has significant antimutagenic and antitumor properties. This research addresses the hypothesis that kefir's anticancer properties are affected by fermentation conditions. Initially, kefir extracts prepared under standard conditions were screened against 7 cancer cell lines using the tetrazolium dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide colorimetric assay. Colon cancer and chronic myelogenous leukemia cells were found to be most susceptible to kefir extracts. Subsequently, a factorial design was implemented to assess the effects of 3 fermentation times (24, 48, and 72 h), 3 kefir-to-milk ratios (2, 5, and 10% wt/vol), and 3 fermentation temperatures (4, 25, and 40°C) on kefir's anticancer properties. Remarkably, exploration of the fermentation conditions allowed the anticancer properties of kefir to be enhanced by 5- to 8-fold against susceptible cell lines. Overall, these results demonstrate the possibility of optimizing the anticancer properties of kefir as a functional food in cancer therapy. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Medium optimization for ε-poly-L-lysine production by Streptomyces diastatochromogenes using response surface methodology.

PubMed

Guo, F; Zheng, H; Cheng, Y; Song, S; Zheng, Z; Jia, S

2018-02-01

Poly-ε-L-lysine is a natural homo-polyamide of L-lysine with excellent antimicrobial properties, which can be used as a novel preservative and has a wide range of applications. In this paper, the fermentation medium for ε-PL production by Streptomyces diastatochromogenes 6#-7 was optimized by Response Surface Methodology. The results of Plackett-Burman design showed that glucose, yeast extract and (NH 4 ) 2 SO 4 were the major influencing factors in ε-PL production of S. diastatochromogenes 6#-7. The optimal concentrations of glucose, yeast extract and (NH 4 ) 2 SO 4 were determined to be 60, 7·5 and 7·5 g l -1 according to Box-Behnken experiment and regression analysis, respectively. Under the optimized conditions, the ε-PL yield in shake-flask fermentation was 0·948 ± 0·030 g l -1 , which was in good agreement with the predicted value of 0·970 g l -1 . The yield was improved by 43·1% from that with the initial medium. In 5 l jar-fermenter the ε-PL yield reached 25·5 g l -1 , which was increased by 56·4% from the original medium. In addition, the fermentation time was reduced from 174 to 120 h. Medium optimization is a very practical and valuable tool for fermentation industry to improve product yield and minimize by-products as well as reduce overall manufacturing costs. The response surface methodology is not new, but it is still a very effective method in medium optimization research. This study used ε-polylysine fermentation as an example to demonstrate how the product yield can be significantly increased by medium optimization through surface response methodology. Similar approach can be used in other microbial fermentations such as in pharmaceutical, food, agricultural and energy industries. As an example, ε-polylysine is one of a few newly approved natural food-grade antimicrobials for food and beverages preservations. Yield improvement is economically beneficial to not only ε-polylysine manufacturers but also to their users and consumers due to lower costs and price. © 2017 The Society for Applied Microbiology.

Manufacture of TATB and TNT from Biosynthesized Phloroglucinols

DTIC Science & Technology

2010-07-01

the microbial synthesis of mono-O-methylphloroglucinols, phloroglucinol O-methyl transferase (POMT) from Rosa chinensis var. spontanea has been...successfully de novo synthesized in codon-optimized form for expression in E. coli, which is the host currently used for microbial synthesis of...efforts had been made in both strain development and optimizing fermentation conditions for microbial phloroglucinol synthesis . Under optimized resin

Cellulase production in a new mutant strain of Penicillium decumbens ML-017 by solid state fermentation with rice bran.

PubMed

Liu, Yun-Tao; Luo, Ze-Yu; Long, Chuan-Nan; Wang, Hai-Dong; Long, Min-Nan; Hu, Zhong

2011-10-01

To produce cellulolytic enzyme efficiently, Penicillium decumbens strain L-06 was used to prepare mutants with ethyl methane sulfonate (EMS) and UV-irradiation. A mutant strain ML-017 is shown to have a higher cellulase activity than others. Box-Behnken's design (BBD) and response surface methodology (RSM) were adopted to optimize the conditions of cellulase (filter paper activity, FPA) production in strain ML-017 by solid-state fermentation (SSF) with rice bran as the substrate. And the result shows that the initial pH, moisture content and culture temperature all have significant effect on the production of cellulase. The optimized condition shall be initial pH 5.7, moisture content 72% and culture temperature 30°C. The maximum cellulase (FPA) production was obtained under the optimized condition, which is 5.76 IU g(-1), increased by 44.12% to its original strain. It corresponded well with the calculated results (5.15 IU g(-1)) by model prediction. The result shows that both BBD and RSM are the cellulase optimization methods with good prospects. Copyright © 2011 Elsevier B.V. All rights reserved.

Physico-chemical characterization of natural fermentation process of Conservolea and Kalamàta table olives and developement of a protocol for the pre-selection of fermentation starters.

PubMed

Bleve, Gianluca; Tufariello, Maria; Durante, Miriana; Grieco, Francesco; Ramires, Francesca Anna; Mita, Giovanni; Tasioula-Margari, Maria; Logrieco, Antonio Francesco

2015-04-01

Table olives are one of the most important traditional fermented vegetables in Europe and their world consumption is constantly increasing. Conservolea and Kalamàta are the most important table olives Greek varieties. In the Greek system, the final product is obtained by spontaneous fermentations, without any chemical debittering treatment. This natural fermentation process is not predictable and strongly influenced by the physical-chemical conditions and by the presence of microorganisms contaminating the olives. Natural fermentations of Conservolea and Kalamàta cultivars black olives were studied in order to determine microbiological, biochemical and chemical evolution during the process. Following the process conditions generally used by producers, in both cultivars, yeasts were detected throughout the fermentation, whereas lactic acid bacteria (LAB) appeared in the last staged of the process. A new optimized specific protocol was developed to select autochthonous yeast and LAB isolates that can be good candidates as starters. These microorganisms were pre-selected for their ability to adapt to model brines, to have beta-glucosidase activity, not to produce biogenic amines. Chemical compounds deriving by microbiological activities and associated to the three different phases (30, 90 and 180 days) of the fermentation process were identified and were proposed as chemical descriptors to follow the fermentation progress. Copyright © 2014 Elsevier Ltd. All rights reserved.

Comparative Transcriptomic Analysis Reveals Similarities and Dissimilarities in Saccharomyces cerevisiae Wine Strains Response to Nitrogen Availability

PubMed Central

Barbosa, Catarina; García-Martínez, José; Pérez-Ortín, José E.; Mendes-Ferreira, Ana

2015-01-01

Nitrogen levels in grape-juices are of major importance in winemaking ensuring adequate yeast growth and fermentation performance. Here we used a comparative transcriptome analysis to uncover wine yeasts responses to nitrogen availability during fermentation. Gene expression was assessed in three genetically and phenotypically divergent commercial wine strains (CEG, VL1 and QA23), under low (67 mg/L) and high nitrogen (670 mg/L) regimes, at three time points during fermentation (12h, 24h and 96h). Two-way ANOVA analysis of each fermentation condition led to the identification of genes whose expression was dependent on strain, fermentation stage and on the interaction of both factors. The high fermenter yeast strain QA23 was more clearly distinct from the other two strains, by differential expression of genes involved in flocculation, mitochondrial functions, energy generation and protein folding and stabilization. For all strains, higher transcriptional variability due to fermentation stage was seen in the high nitrogen fermentations. A positive correlation between maximum fermentation rate and the expression of genes involved in stress response was observed. The finding of common genes correlated with both fermentation activity and nitrogen up-take underlies the role of nitrogen on yeast fermentative fitness. The comparative analysis of genes differentially expressed between both fermentation conditions at 12h, where the main difference was the level of nitrogen available, showed the highest variability amongst strains revealing strain-specific responses. Nevertheless, we were able to identify a small set of genes whose expression profiles can quantitatively assess the common response of the yeast strains to varying nitrogen conditions. The use of three contrasting yeast strains in gene expression analysis prompts the identification of more reliable, accurate and reproducible biomarkers that will facilitate the diagnosis of deficiency of this nutrient in the grape-musts and the development of strategies to optimize yeast performance in industrial fermentations. PMID:25884705

Lignin degradation in corn stalk by combined method of H2O2 hydrolysis and Aspergillus oryzae CGMCC5992 liquid-state fermentation.

PubMed

Zhang, Zhicai; Xia, Lili; Wang, Feng; Lv, Peng; Zhu, Maxiaoqi; Li, Jinhua; Chen, Keping

2015-01-01

Lignin peroxidase (LiP) is the primary enzyme responsible for lignin degradation. In our previous work, in order to shorten the pretreatment time and increase the lignin degradation, we have pretreated the corn stalk (CS) using a combination of Aspergillus oryzae CGMCC 5992 solid-state fermentation and H2O2 treatment. In the present study, one-factor-at-a-time design and response surface design were applied to optimize the nutritional constituents for LiP production in liquid-state fermentation by A. oryzae CGMCC 5992 and the conditions for CS degradation by A. oryzae CGMCC 5992. The optimal medium included CS of 30 g/L, glucose of 4.6 g/L, sodium nitrate of 1.2 g/L, corn steep liquor of 1 g/L, yeast extract of 1.2 g/L, and vitamin B1 of 0.15 g/L. Under these optimal conditions, the LiP production reached its maximum of 652.34 U/L. The optimal condition for CS degradation included CS of 20 g, A. oryzae CGMCC 5992 broth of 50 mL, 1.5 % H2O2 solution of 80 mL, H2O2 flow rate of 0.4 mL/min, water volume of 240 mL (water/material ratio of 12:1), hydrolysis temperature of 39 °C, and hydrolysis time of 8 h. Before hydrolysis, CS and water were pretreated at 113 °C for 11 min. Under these optimal conditions, the sugar yield reached its maximum of 46.28 %. Our newly developed method had great advantages in pretreatment of CS due to its quickness, convenience, safety, no special equipment and high sugar yield.Graphical abstractThe schematic diagram of corn straw hydrolysis.

Optimization of polyhydroxybutyrate production by Bacillus sp. CFR 256 with corn steep liquor as a nitrogen source.

PubMed

Vijayendra, S V N; Rastogi, N K; Shamala, T R; Anil Kumar, P K; Kshama, L; Joshi, G J

2007-06-01

Polyhydroxyalkanotes (PHAs), the eco-friendly biopolymers produced by many bacteria, are gaining importance in curtailing the environmental pollution by replacing the non-biodegradable plastics derived from petroleum. The present study was carried out to economize the polyhydroxybutyrate (PHB) production by optimizing the fermentation medium using corn steep liquor (CSL), a by-product of starch processing industry, as a cheap nitrogen source, by Bacillus sp. CFR 256. Response surface methodology (RSM) was used to optimize the fermentation medium using the variables such as corn steep liquor (5-25 g l(-1)), Na(2)HPO(4) 2H(2)O (2.2-6.2 g l(-1)), KH(2)PO(4) (0.5-2.5 g l(-1)), sucrose (5-55 g l(-1)) and inoculum concentration (1-25 ml l(-1)). Central composite rotatable design (CCRD) experiments were carried out to study the complex interactions of the variables.The optimum conditions for maximum PHB production were (g l(-1)): CSL-25, Na(2)HPO(4) 2H(2)O-2.2, KH(2)PO(4) - 0.5, sucrose - 55 and inoculum - 10 (ml l(-1)). After 72 h of fermentation, the amount of PHA produced was 8.20 g l(-1) (51.20% of dry cell biomass). It is the first report on optimization of fermentation medium using CSL as a nitrogen source, for PHB production by Bacillus sp.

Assessment of the yeast species composition of cocoa bean fermentations in different cocoa-producing regions using denaturing gradient gel electrophoresis.

PubMed

Papalexandratou, Zoi; De Vuyst, Luc

2011-11-01

The yeast species composition of 12 cocoa bean fermentations carried out in Brazil, Ecuador, Ivory Coast and Malaysia was investigated culture-independently. Denaturing gradient gel electrophoresis of 26S rRNA gene fragments, obtained through polymerase chain reaction with universal eukaryotic primers, was carried out with two different commercial apparatus (the DCode and CBS systems). In general, this molecular method allowed a rapid monitoring of the yeast species prevailing during fermentation. Under similar and optimal denaturing gradient gel electrophoresis conditions, the CBS system allowed a better separated band pattern than the DCode system and an unambiguous detection of the prevailing species present in the fermentation samples. The most frequent yeast species were Hanseniaspora sp., followed by Pichia kudriavzevii and Saccharomyces cerevisiae, independent of the origin of the cocoa. This indicates a restricted yeast species composition of the cocoa bean fermentation process. Exceptionally, the Ivorian cocoa bean box fermentation samples showed a wider yeast species composition, with Hyphopichia burtonii and Meyerozyma caribbica among the main representatives. Yeasts were not detected in the samples when the temperature inside the fermenting cocoa pulp-bean mass reached values higher than 45 °C or under early acetic acid production conditions. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

No-Cook Process for Ethanol Production Using Indian Broken Rice and Pearl Millet

PubMed Central

Gohel, Vipul; Duan, Gang

2012-01-01

No-cook process using granular starch hydrolyzing enzyme (GSHE) was evaluated for Indian broken rice and pearl millet. One-factor-at-a-time optimization method was used in ethanol production to identify optimum concentration of GSHE, under yeast fermentation conditions using broken rice and pearl millet as fermentation feedstocks. An acid fungal protease at a concentration of 0.2 kg per metric ton of grain was used along with various dosages of GSHE under yeast fermentation conditions to degrade the grain proteins into free amino nitrogen for yeast growth. To measure the efficacy of GSHE to hydrolyze no-cook broken rice and pearl millet, the chemical composition, fermentation efficiency, and ethanol recovery were determined. In both feedstocks, fermentation efficiency and ethanol recovery obtained through single-step no-cook process were higher than conventional multistep high-temperature process, currently considered the ideal industrial process. Furthermore, the no-cook process can directly impact energy consumption through steam saving and reducing the water cooling capacity needs, compared to conventional high-temperature process. PMID:22518148

Whole slurry saccharification and fermentation of maleic acid-pretreated rice straw for ethanol production.

PubMed

Jung, Young Hoon; Park, Hyun Min; Kim, Kyoung Heon

2015-09-01

We evaluated the feasibility of whole slurry (pretreated lignocellulose) saccharification and fermentation for producing ethanol from maleic acid-pretreated rice straw. The optimized conditions for pretreatment were to treat rice straw at a high temperature (190 °C) with 1 % (w/v) maleic acid for a short duration (3 min ramping to 190 °C and 3 min holding at 190 °C). Enzymatic digestibility (based on theoretical glucose yield) of cellulose in the pretreated rice straw was 91.5 %. Whole slurry saccharification and fermentation of pretreated rice straw resulted in 83.2 % final yield of ethanol based on the initial quantity of glucan in untreated rice straw. These findings indicate that maleic acid pretreatment results in a high yield of ethanol from fermentation of whole slurry even without conditioning or detoxification of the slurry. Additionally, the separation of solids and liquid is not required; therefore, the economics of cellulosic ethanol fuel production are significantly improved. We also demonstrated whole slurry saccharification and fermentation of pretreated lignocellulose, which has rarely been reported.

Immobilized anaerobic fermentation for bio-fuel production by Clostridium co-culture.

PubMed

Xu, Lei; Tschirner, Ulrike

2014-08-01

Clostridium thermocellum/Clostridium thermolacticum co-culture fermentation has been shown to be a promising way of producing ethanol from several carbohydrates. In this research, immobilization techniques using sodium alginate and alkali pretreatment were successfully applied on this co-culture to improve the bio-ethanol fermentation performance during consolidated bio-processing (CBP). The ethanol yield obtained increased by over 60 % (as a percentage of the theoretical maximum) as compared to free cell fermentation. For cellobiose under optimized conditions, the ethanol yields were approaching about 85 % of the theoretical efficiency. To examine the feasibility of this immobilization co-culture on lignocellulosic biomass conversion, untreated and pretreated aspen biomasses were also used for fermentation experiments. The immobilized co-culture shows clear benefits in bio-ethanol production in the CBP process using pretreated aspen. With a 3-h, 9 % NaOH pretreatment, the aspen powder fermentation yields approached 78 % of the maximum theoretical efficiency, which is almost twice the yield of the untreated aspen fermentation.

Biological Hydrogen Production: Simultaneous Saccharification and Fermentation With Nitrogen and Phosphorus Removal from Wastewater Effluent

DTIC Science & Technology

2010-01-01

requiring thermochemical pretreatment , aswould typically be required with lignocellulosic feedstocks. Therefore it offers a readily-processed and...Standards and Technology. The pH of the reactors was controlled throughout all fermentations by the automatic addition of 0.1 N NaOH . Total organic...nutrients. The optimized conditions developed with paper as a substrate may also convey to the use of a similar process with lignocellulosic biomass

Cultivation, LD(50) determination and experimental model of Streptococcus suis serotype 2 strain HA9801.

PubMed

Zhao, Zhanzhong; Wang, Jian; Liu, Peihong; Zhang, Suhua; Gong, Jianpei; Huang, Xiqin; Li, Bin; Xue, Feiqun

2009-04-01

The effects of nutritional components and submerged culture conditions on colony-forming unit (CFU) counts by Streptococcus suis serotype 2 strain HA9801 in flask culture was investigated, and the optimal medium and cultivation conditions was confirmed by using a 50l bioreactor. The LD(50) values of HA9801 in pigs before and after fermentation were 1.8 x 10(7)CFU, which indicated that the virulence of HA9801 was very stable in the fermentation process. In addition, an experimental model that closely mimics naturally occurring disease in conventional pigs was established.

Enhanced Production of Gamma-Aminobutyric Acid by Optimizing Culture Conditions of Lactobacillus brevis HYE1 Isolated from Kimchi, a Korean Fermented Food.

PubMed

Lim, Hee Seon; Cha, In-Tae; Roh, Seong Woon; Shin, Hae-Hun; Seo, Myung-Ji

2017-03-28

This study evaluated the effects of culture conditions, including carbon and nitrogen sources, L-monosodium glutamate (MSG), and initial pH, on gamma-aminobutyric acid (GABA) production by Lactobacillus brevis HYE1 isolated from kimchi, a Korean traditional fermented food. L. brevis HYE1 was screened by the production analysis of GABA and genetic analysis of the glutamate decarboxylase gene, resulting in 14.64 mM GABA after 48 h of cultivation in MRS medium containing 1% (w/v) MSG. In order to increase GABA production by L. brevis HYE1, the effects of carbon and nitrogen sources on GABA production were preliminarily investigated via one-factor-at-a-time optimization strategy. As the results, 2% maltose and 3% tryptone were determined to produce 17.93 mM GABA in modified MRS medium with 1% (w/v) MSG. In addition, the optimal MSG concentration and initial pH were determined to be 1% and 5.0, respectively, resulting in production of 18.97 mM GABA. Thereafter, response surface methodology (RSM) was applied to determine the optimal conditions of the above four factors. The results indicate that pH was the most significant factor for GABA production. The optimal culture conditions for maximum GABA production were also determined to be 2.14% (w/v) maltose, 4.01% (w/v) tryptone, 2.38% (w/v) MSG, and an initial pH of 4.74. In these conditions, GABA production by L. brevis HYE1 was predicted to be 21.44 mM using the RSM model. The experiment was performed under these optimized conditions, resulting in GABA production of 18.76 mM. These results show that the predicted and experimental values of GABA production are in good agreement.

Time-dependent fermentation control strategies for enhancing synthesis of marine bacteriocin 1701 using artificial neural network and genetic algorithm.

PubMed

Peng, Jiansheng; Meng, Fanmei; Ai, Yuncan

2013-06-01

The artificial neural network (ANN) and genetic algorithm (GA) were combined to optimize the fermentation process for enhancing production of marine bacteriocin 1701 in a 5-L-stirred-tank. Fermentation time, pH value, dissolved oxygen level, temperature and turbidity were used to construct a "5-10-1" ANN topology to identify the nonlinear relationship between fermentation parameters and the antibiotic effects (shown as in inhibition diameters) of bacteriocin 1701. The predicted values by the trained ANN model were coincided with the observed ones (the coefficient of R(2) was greater than 0.95). As the fermentation time was brought in as one of the ANN input nodes, fermentation parameters could be optimized by stages through GA, and an optimal fermentation process control trajectory was created. The production of marine bacteriocin 1701 was significantly improved by 26% under the guidance of fermentation control trajectory that was optimized by using of combined ANN-GA method. Copyright © 2013 Elsevier Ltd. All rights reserved.

Optimization of erythritol and glycerol accumulation in conidia of Beauveria bassiana by solid-state fermentation, using response surface methodology.

PubMed

Tarocco, Federico; Lecuona, Roberto E; Couto, Alicia S; Arcas, Jorge A

2005-09-01

Entomopathogenic fungi are widely produced for use as mycoinsecticides. Therefore, improvement of the shelf life of fungal propagules under good and adverse conditions should be a pre-requisite of their production. In order to improve conidial physiology as well as mycoinsecticide efficiency, culture conditions may be varied. The Doehlert design was used to generate response surfaces with an estimation of the parameters of the quadratic model allowing the study of three different factors at a different number of levels. This experimental design was applied to optimize water activity (aw), pH, and fermentation time for Beauveria bassiana conidial production and accumulation of polyols in solid-state fermentation. Thus, it was possible to identify the region in the experimental range in which the optimum values of these parameters were simultaneously achieved. Maximal conidia production was achieved at pH 5-6 and aw=0.999. Under these conditions, polyol accumulation was 3 mg erythritol/g conidia and 29.6 mg glycerol/g conidia. However, maximal polyol accumulation was achieved at pH 4.5 and aw 0.950; erythritol production increased 33-fold and glycerol production 4.5-fold. Under these conditions conidia production was 1,000 times lower. The possibilities of increasing the quality of the biocontrol agent without neglecting yield are discussed.

Production, optimization and characterization of lactic acid by Lactobacillus delbrueckii NCIM 2025 from utilizing agro-industrial byproduct (cane molasses).

PubMed

Srivastava, Abhinay Kumar; Tripathi, Abhishek Dutt; Jha, Alok; Poonia, Amrita; Sharma, Nitya

2015-06-01

In the present work Lactobacillus delbrueckii was used to utilize agro-industrial byproduct (cane molasses) for lactic acid production under submerged fermentation process. Screening of LAB was done by Fourier transform infra red spectroscopy (FTIR). Effect of different amino acids (DL-Phenylalanine, L-Lysine and DL-Aspartic acid) on the fermentation process was done by high performance liquid chromatography (HPLC). Central composite rotatable design (CCRD) was used to optimize the levels of three parameters viz. tween 80, amino acid and cane molasses concentration during fermentative production of lactic acid. Under optimum condition lactic acid production was enhanced from 55.89 g/L to 84.50 g/L. Further, validation showed 81.50 g/L lactic acid production. Scale up was done on 7.5 L fermentor. Productivity was found to be 3.40 g/L/h which was higher than previous studies with reduced fermentation time from 24 h to 12 h. Further characterization of lactic acid was done by FTIR.

Mycelium and polysaccharide production of Agaricus blazei Murrill by submerged fermentation.

PubMed

Lin, Jr-Hui; Yang, Shang-Shyng

2006-04-01

Over the last decade, Agaricus blazei Murrill has been studied and developed as a novel functional food in Japan, Korea, China, and Taiwan. Due to the low yields, the fruiting bodies of A. blazei Murrill are relatively expensive, and a cheap and stable source of A. blazei Murrill mycelium for commercial purposes is highly desirable. Culture media and conditions were investigated with a view to reducing the cost and improving the mycelium and polysaccharide production of A. blazei Murrill by submerged fermentation. Thirty six isolates of A. blazei Murrill were isolated from 22 fruiting bodies produced in Taiwan, and 16 of them could be successfully cultivated on mannitol-egg yolk-polymyxin medium. The isolates were identified by species-specific polymerase chain reaction (PCR) and optimized for the culture media and conditions by submerged fermentation for mycelium and polysaccharide production. Some properties of polysaccharide extract were also investigated. All of the PCR products with species-specific primers showed high identity and matched the internal transcribed spacer 1 sequences of A. blazei Murrill. The phylogenic tree of A. blazei Murrill isolates generated from random amplified polymorphic DNAs arranged all samples into 3 groups and 2 independent cases. The optimal culture media of mycelium production in submerged fermentation were 5% malt extract, 0.1% yeast extract, and 0.5% peptone at pH 6.0, while the optimal culture conditions were 200 mL medium in 500 mL Hinton flask, shaking at 90 rpm for 3 days and then shifting to 105 rpm for 5 days at 27 degrees C. Each liter of A. blazei Murrill M72 yielded 10.83 +/- 0.24 g dried mycelia weight and each liter of A. blazei Murrill M152 produced 0.251 +/- 0.004 g crude polysaccharide (3.03 +/- 0.05% of dried mycelia weight). Crude polysaccharide of A. blazei Murrill M162 contained 82.27-99.14% of total sugar and less than 1.63% of protein; it had 4 major molecular weight components (274.1, 32.7, 7.5, and 2.1 kDa, respectively), with the 2.1 kDa portion possibly a beta-(1,3)-glucan. These results show that selection of media and conditions can be employed in order to improve the mycelium and polysaccharide production of A. blazei Murrill M72 or M152 by submerged fermentation. Mycelia and polysaccharide production of A. blazei Murrill with submerged fermentation is potentially feasible.

The roles of xylan and lignin in oxalic acid pretreated corncob during separate enzymatic hydrolysis and ethanol fermentation

Treesearch

Jae-Won Lee; Rita C.L.B. Rodrigues; Hyun Joo Kim; In-Gyu Choi; Thomas W. Jeffries

2010-01-01

High yields of hemicellulosic and cellulosic sugars are critical in obtaining economical conversion of agricultural residues to ethanol. To optimize pretreatment conditions, we evaluated oxalic acid loading rates, treatment temperatures and times in a 23 full factorial design. Response-surface analysis revealed an optimal oxalic acid pretreatment...

Use of sourdough fermentation and pseudo-cereals and leguminous flours for the making of a functional bread enriched of gamma-aminobutyric acid (GABA).

PubMed

Coda, Rossana; Rizzello, Carlo Giuseppe; Gobbetti, Marco

2010-02-28

Lactobacillus plantarum C48 and Lactococcus lactis subsp. lactis PU1, previously selected for the biosynthesis of gamma-aminobutyric acid (GABA), were used for sourdough fermentation of cereal, pseudo-cereal and leguminous flours. Chickpea, amaranth, quinoa and buckwheat were the flours most suitable to be enriched of GABA. The parameters of sourdough fermentation were optimized. Addition of 0.1mM pyridoxal phosphate, dough yield of 160, inoculum of 5 x 10(7)CFU/g of starter bacteria and fermentation for 24h at 30 degrees C were found to be the optimal conditions. A blend of buckwheat, amaranth, chickpea and quinoa flours (ratio 1:1:5.3:1) was selected and fermented with baker's yeast (non-conventional flour bread, NCB) or with Lb. plantarum C48 sourdough (non-conventional flour sourdough bread, NCSB) and compared to baker's yeast started wheat flour bread (WFB). NCSB had the highest concentration of free amino acids and GABA (ca. 4467 and 504 mg/kg, respectively). The concentration of phenolic compounds and antioxidant activity of NCSB bread was the highest, as well as the rate of in vitro starch hydrolysis was the lowest. Texture analysis showed that sourdough fermentation enhances several characteristics of NCSB with respect to NCB, thus approaching the features of WFB. Sensory analysis showed that sourdough fermentation allowed to get good palatability and overall taste appreciation. (c) 2009 Elsevier B.V. All rights reserved.

Optimal Design of Biomass Utilization System for Rural Area Includes Technical and Economic Dimensions

NASA Astrophysics Data System (ADS)

Morioka, Yasuki; Nakata, Toshihiko

In order to design optimal biomass utilization system for rural area, OMNIBUS (The Optimization Model for Neo-Integrated Biomass Utilization System) has been developed. OMNIBUS can derive the optimal system configuration to meet different objective function, such as current account balance, amount of biomass energy supply, and CO2 emission. Most of biomass resources in a focused region e.g. wood biomass, livestock biomass, and crop residues are considered in the model. Conversion technologies considered are energy utilization technologies e.g. direct combustion and methane fermentation, and material utilization technologies e.g. composting and carbonization. Case study in Miyakojima, Okinawa prefecture, has been carried out for several objective functions and constraint conditions. Considering economics of the utilization system as a priority requirement, composting and combustion heat utilization are mainly chosen in the optimal system configuration. However gasification power plant and methane fermentation are included in optimal solutions, only when both biomass energy utilization and CO2 reduction have been set as higher priorities. External benefit of CO2 reduction has large impacts on the system configuration. Provided marginal external benefit of more than 50,000 JPY/t-C, external benefit becomes greater than the revenue from electricity and compost etc. Considering technological learning in the future, expensive technologies such as gasification power plant and methane fermentation will have economic feasibility as well as market competitiveness.

Improvement of ε-poly-L-lysine production through seed stage development based on in situ pH monitoring.

PubMed

Sun, Qi-Xing; Chen, Xu-Sheng; Ren, Xi-Dong; Mao, Zhong-Gui

2015-01-01

Nissin, natamycin, and ε-poly-L-lysine (ε-PL) are three safe, microbial-produced food preservatives used today in the food industry. However, current industrial production of ε-PL is only performed in several countries. In order to realize large-scale ε-PL production by fermentation, the effects of seed stage on cell growth and ε-PL production were investigated by monitoring of pH in situ in a 5-L laboratory-scale fermenter. A significant increase in ε-PL production in fed-batch fermentation by Streptomyces sp. M-Z18 was achieved, at 48.9 g/L, through the optimization of several factors associated with seed stage, including spore pretreatment, inoculum age, and inoculum level. Compared with conventional fermentation approaches using 24-h-old shake-flask seed broth as inoculum, the maximum ε-PL concentration and productivity were enhanced by 32.3 and 36.6 %, respectively. The effect of optimized inoculum conditions on ε-PL production on a large scale was evaluated using a 50-L pilot-scale fermenter, attaining a maximum ε-PL production of 36.22 g/L in fed-batch fermentation, constituting the first report of ε-PL production at pilot scale. These results will be helpful for efficient ε-PL production by Streptomyces at pilot and plant scales.

Liberation of fermentable sugars from soybean hull biomass using ionic liquid 1-butyl-3-methylimidazolium acetate and their bioconversion to ethanol.

PubMed

da Cunha-Pereira, Fernanda; Rech, Rosane; Záchia Ayub, Marco Antônio; Pinheiro Dillon, Aldo; Dupont, Jairton

2016-03-01

Optimized hydrolysis of lignocellulosic waste biomass is essential to achieve the liberation of sugars to be used in fermentation process. Ionic liquids (ILs), a new class of solvents, have been tested in the pretreatment of cellulosic materials to improve the subsequent enzymatic hydrolysis of the biomass. Optimized application of ILs on biomass is important to advance the use of this technology. In this research, we investigated the effects of using 1-butyl-3-methylimidazolium acetate ([bmim][Ac]) on the decomposition of soybean hull, an abundant cellulosic industrial waste. Reaction aspects of temperature, incubation time, IL concentration, and solid load were optimized before carrying out the enzymatic hydrolysis of this residue to liberate fermentable glucose. Optimal conditions were found to be 75°C, 165 min incubation time, 57% (mass fraction) of [bmim][Ac], and 12.5% solid loading. Pretreated soybean hull lost its crystallinity, which eased enzymatic hydrolysis, confirmed by Fourier Transform Infrared analysis. The enzymatic hydrolysis of the biomass using an enzyme complex from Penicillium echinulatum liberated 92% of glucose from the cellulose matrix. The hydrolysate was free of any toxic compounds, such as hydroxymethylfurfural and furfural. The obtained hydrolysate was tested for fermentation using Candida shehatae HM 52.2, which was able to convert glucose to ethanol at yields of 0.31. These results suggest the possible use of ILs for the pretreatment of some lignocellulosic waste materials, avoiding the formation of toxic compounds, to be used in second-generation ethanol production and other fermentation processes. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 32:312-320, 2016. © 2015 American Institute of Chemical Engineers.

Recovery of fermented inulin fiber by lactic acid bacteria (LAB) from inulin hydrolysate using fungi inulinase enzymes of Scopulariopsis sp.-CBS1 and class of Deuteromycetes-CBS4 as cholesterol binder

NASA Astrophysics Data System (ADS)

Susilowati, Agustine; Melanie, Hakiki; Maryati, Yati; Aspiyanto

2017-01-01

Fermentation of Lactobacillus Acid Bacteria (LAB) which are mixtures of Lactobacillus acidophilus, Bifidobacteriumbifidum, Lactobacillus bulgaricus and Streptococcus thermophillus on hydrolysate as a result of inulin hydrolysis using inulinase enzymes obtained from endophytic fungi ofScopulariopsis sp.-CBS1 (inulin hydrolysate of S) and Class of Deuteromycetes-CBS4 (inulin hydrolysate of D) generate potential fermented inulin fiber as cholesterol binder. Fermentation process was conducted under concentrations of inulin hydrolysate 50% (w/v), LAB 15% (v/v) and skim milk 12.5% (w/v) at room temperature and 40°C for 0, 12, 24, 36 and 48 hours, respectively. Result of experimental work showed that longer time of LAB fermentation increased total acids, TPC and CBC at pH 2, but decreased total sugar, reducing, IDF, SDF, CBC pH 2 and CBC pH 7. Based on Cholesterol Binding Capacity (CBC), optimization of fermentation process on inulin hydrolysate of S was achieved by combining treatment at 40°C for 24 hours resulted in CBC pH 2 of 19.11 mg/g TDF and inulin hydrolysate of D was achieved by fermentation at 40 °C for 48 hours resulted in CBC pH 2 of 24.28 mg/g TDF. Inulin hydrolysate of class of Deutrymecetes CBS4 fermented by LAB had better functional property as cholesterol binder than that inulin hydrolysate of S fermented by LAB. This is due to cholesterol binder and cholesterol derivatives as a result of degradation of LAB on digestive system (stomach) when compared to higher colon under optimal process condition.

Bacteriocinogenic LAB Strains for Fermented Meat Preservation: Perspectives, Challenges, and Limitations.

PubMed

Favaro, Lorenzo; Todorov, Svetoslav Dimitrov

2017-12-01

Over the last decades, much research has focused on lactic acid bacteria (LAB) bacteriocins because of their potential as biopreservatives and their action against the growth of spoilage microbes. Meat and fermented meat products are prone to microbial contamination, causing health risks, as well as economic losses in the meat industry. The use of bacteriocin-producing LAB starter or protective cultures is suitable for fermented meats. However, although bacteriocins can be produced during meat processing, their levels are usually much lower than those achieved during in vitro fermentations under optimal environmental conditions. Thus, the direct addition of a bacteriocin food additive would be desirable. Moreover, safety and technological characteristics of the bacteriocinogenic LAB must be considered before their widespread applications. This review describes the perspectives and challenges toward the complete disclosure of new bacteriocins as effective preservatives in the production of safe and "healthy" fermented meat products.

Fermentation optimization for the production of bioactive polysaccharides from Cordyceps sinensis fungus UM01.

PubMed

Wang, Lan-Ying; Cheong, Kit-Leong; Wu, Ding-Tao; Meng, Lan-Zhen; Zhao, Jing; Li, Shao-Ping

2015-08-01

The optimal fermentation conditions and medium for the production of bioactive polysaccharides from the mycelium of Cordyceps sinensis fungus UM01 were investigated by using orthogonal design and high performance size exclusion chromatography coupled with multi-angel laser light scattering and refractive index detector (HPSEC-MALLS-RID). Results showed that the optimal temperature, initial pH, rotation speed, medium capacity (ratio of medium volume to the volume of flask bottle) and inoculums volume for the mycelium growth were 15 °C, pH 6.0, 150 rpm, 2/5 (v/v), and 3% (v/v), respectively. Furthermore, bioactive polysaccharides from the mycelium of C. sinensis fungus UM01 were determined as polysaccharide fractions with the molecular weight above 10 kDa. The optimal fermentation medium was determined as a composition of glucose 30.0 g/L, sucrose 30.0 g/L, KH2PO4 1.0 g/L, CaCl2 0.5 g/L, yeast extract 3.0 g/L, and MgCl2 0.1g/L according to the maximum amount of the bioactive polysaccharides (486.16±19.60 mg/L) measured by HPSEC-MALLS/RID. Results are helpful to establish an efficient and controllable fermentation process for the industrial production of bioactive polysaccharides from C. sinensis UM01, and beneficial to develop a unique health and functional product in future. Copyright © 2015 Elsevier B.V. All rights reserved.

Conversion of food waste into biofertilizer for the biocontrol of root knot nematode by Paecilomyces lilacinus.

PubMed

Yu, Zhen; Zhang, You-Chi; Zhang, Xiang; Wang, Yin

2015-01-01

The feasibility of converting food waste into nematocidal biofertilizer by nematophagous fungus Paecilomyces lilacinus (P. lilacinus) was investigated. The culture conditions of P. lilacinus were optimized through response surface methodology. Results showed that fermentation time, the amount of food waste, initial pH and temperature were most important factors for P. lilacinus production. The P. lilacinus production under optimized conditions was 10(9.6 ± 0.3) conidia mL⁻¹. After fermentation, the chemical oxygen demand concentration of food waste was efficiently decreased by 81.92%. Moreover, the property evaluation of the resultant food waste as biofertilizer indicates its high quality with reference to the standard released by the Chinese Ministry of Agriculture. The protease activity and nematocidal ability of P. lilacinus cultured by food waste were 10.8% and 27% higher than those by potato dextrose agar, respectively.

Screening of Functional Rhizopus stolonifer for Alcohol Fermentation and Production of High Quality Korean Traditional Rice Wine

PubMed Central

Song, Jung-Hwa; Kim, Jae-Ho; Ahn, Byung-Hak

2010-01-01

Different strains of mold were screened for the production of high quality Korean traditional rice wine with anti-hypertension and good acceptability. We isolated 867 nuruk mold strains and selected 24 for further study based on measurement of amylase activity. Among them, mold No. 17 showed high ethanol production upon fermentation with Saccharomyces cerevisiae as well as anti-hypertensive properties. The No. 17 strain was therefore selected as the functional mold and later identified as Rhizopus stolonifer based on molecular biological characteristics. Optimal fermentation conditions for the brewing of anti-hypertensive traditional rice wine comprised the addition of R. stolonifer No. 17 koji at a concentration of 35 sp/g and a fermentation period of 10 days at 25℃ using S. cerevisiae. PMID:23956639

Screening of Functional Rhizopus stolonifer for Alcohol Fermentation and Production of High Quality Korean Traditional Rice Wine.

PubMed

Song, Jung-Hwa; Kim, Jae-Ho; Ahn, Byung-Hak; Lee, Jong-Soo

2010-06-01

Different strains of mold were screened for the production of high quality Korean traditional rice wine with anti-hypertension and good acceptability. We isolated 867 nuruk mold strains and selected 24 for further study based on measurement of amylase activity. Among them, mold No. 17 showed high ethanol production upon fermentation with Saccharomyces cerevisiae as well as anti-hypertensive properties. The No. 17 strain was therefore selected as the functional mold and later identified as Rhizopus stolonifer based on molecular biological characteristics. Optimal fermentation conditions for the brewing of anti-hypertensive traditional rice wine comprised the addition of R. stolonifer No. 17 koji at a concentration of 35 sp/g and a fermentation period of 10 days at 25℃ using S. cerevisiae.

Optimization of lipids production by Cryptococcus laurentii 11 using cheese whey with molasses.

PubMed

Castanha, Rodrigo Fernandes; Mariano, Adriano Pinto; de Morais, Lilia Aparecida Salgado; Scramin, Shirlei; Monteiro, Regina Teresa Rosim

2014-01-01

This study aimed the optimization of culture condition and composition for production of Cryptococcus laurentii 11 biomass and lipids in cheese whey medium supplemented with sugarcane molasses. The optimization of pH, fermentation time, and molasses concentration according to a full factorial statistical experimental design was followed by a Plackett-Burman experimental design, which was used to determine whether the supplementation of the culture medium by yeast extract and inorganic salts could provide a further enhancement of lipids production. The following conditions and composition of the culture medium were found to optimize biomass and lipids production: 360 h fermentation, 6.5 pH and supplementation of (g L(-1)): 50 molasses, 0.5 yeast extract, 4 KH2PO4, 1 Na2HPO4, 0.75 MgSO4 · 7H2O and 0.002 ZnSO4 · H2O. Additional supplementation with inorganic salts and yeast extract was essential to optimize the production, in terms of product concentration and productivity, of neutral lipids by C. laurentii 11. Under this optimized condition, the production of total lipids increased by 133% in relation to control experiment (from 1.27 to 2.96 g L(-1)). The total lipids indicated a predominant (86%) presence of neutral lipids with high content of 16- and 18-carbon-chain saturated and monosaturated fatty acids. This class of lipids is considered especially suitable for the production of biodiesel.

Probiotic potential of noni juice fermented with lactic acid bacteria and bifidobacteria.

PubMed

Wang, Chung-Yi; Ng, Chang-Chai; Su, Hsuan; Tzeng, Wen-Sheng; Shyu, Yuan-Tay

2009-01-01

The present study assesses the feasibility of noni as a raw substrate for the production of probiotic noni juice by lactic acid bacteria (Lactobacilluscasei and Lactobacillus plantarum) and bifidobacteria (Bifidobacteriumlongum). Changes in pH, acidity, sugar content, cell survival and antioxidant properties during fermentation were monitored. All tested strains grew well on noni juice, reaching nearly 10⁹ colony-forming units/ml after 48 h fermentation. L.casei produced less lactic acid than B.longum and L. plantarum. After 4 weeks of cold storage at 4°C, B.longum and L. plantarum survived under low-pH conditions in fermented noni juice. In contrast, L.casei exhibited no cell viability after 3 weeks. Moreover, noni juice fermented with B.longum had a high antioxidant capacity that did not differ significantly (P <0.05) from that of lactic acid bacteria. Finally, we found that B.longum and L. plantarum are optimal probiotics for fermentation with noni juice.

Solid state fermentation of Trichoderma viride for enhancement phenolic content, antioxidant and antimicrobial activities in ginger.

PubMed

Saleh, Rashad M; Kabli, Saleh A; Al-Garni, Saleh M; Al-Ghamdi, Maryam A; Abdel-Aty, Azza M; Mohamed, Saleh A

2018-05-04

The phenolic content of methanolic and water extracts of ginger fermented by Trichoderma spp. during solid state fermentation (SSF) was detected as compared with unfermented ginger. The total phenolic content of fermented ginger increased several times. The highest phenolic content of ginger was detected after SSF by T. viride. The optimal physiological conditions for the maximum production of the phenolic content and β-glucosidase activity of fermented ginger by T. viride were detected at day 7 incubation, pH 6.0, 30°C and 30% moisture. There are consistent between the maximum production of β-glucosidase and phenolic content. The SSF of ginger by T. viride greatly enhanced the antioxidant potency of phenolic compounds by using DPPH and ABTS assays. Potent antibacterial activity was appeared by phenolic compounds of fermented ginger against all the tested human-pathogenic bacteria. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Detoxification of acidic catalyzed hydrolysate of Kappaphycus alvarezii (cottonii).

PubMed

Meinita, Maria Dyah Nur; Hong, Yong-Ki; Jeong, Gwi-Taek

2012-01-01

Red seaweed, Kappaphycus alvarezii, holds great promise for use in biofuel production due to its high carbohydrate content. In this study, we investigated the effect of fermentation inhibitors to the K. alvarezii hydrolysate on cell growth and ethanol fermentation. In addition, detoxification of fermentation inhibitors was performed to decrease the fermentation inhibitory effect. 5-Hydroxymethylfurfural and levulinic acid, which are liberated from acidic hydrolysis, was also observed in the hydrolysate of K. alvarezii. These compounds inhibited ethanol fermentation. In order to remove these inhibitors, activated charcoal and calcium hydroxide were introduced. The efficiency of activated charcoals was examined and over-liming was used to remove the inhibitors. Activated charcoal was found to be more effective than calcium hydroxide to remove the inhibitors. Detoxification by activated charcoal strongly improved the fermentability of dilute acid hydrolysate in the production of bioethanol from K. alvarezii with Saccharomyces cerevisiae. The optimal detoxifying conditions were found to be below an activated charcoal concentration of 5%.

Steam pretreatment of Saccharum officinarum L. bagasse by adding of impregnating agents for advanced bioethanol production.

PubMed

Verardi, A; Blasi, A; De Bari, I; Calabrò, V

2016-12-01

The main byproduct of the sugarcane industry, Saccharum officinarum L. bagasse (sugarcane bagasse, SCB), is widely used as lignocellulose biomass for bio-ethanol (EtOH) production. In this research study, SCB was pretreated by steam explosion (SE) method using two different impregnating agents: sulfur dioxide (SD) and hydrogen peroxide (HP). As matter of fact, the use of impregnating agents improves the performance of SE method, increasing the concentrations of fermentable sugars after enzymatic saccharification, and decreasing the inhibitor compounds produced during the steam pretreatment step. The aim of this study was to investigate and compare the use of the two impregnating agents in various SE-conditions in order to optimize pretreatment parameters. For every pretreatment condition, it has been evaluated: concentration of fermentable sugars, glucose and xylose yields, and the effects of the inhibitor compounds on enzymatic hydrolysis step. The obtained results allow to improve the efficiency of the whole process of bio-EtOH synthesis enhancing the amount of fermentable sugars produced and the eco-sustainability of the whole process. Indeed, the optimization of steam pretreatment leads to a reduction of energy requirements and to a lower environmental impact. Copyright © 2015 Elsevier Inc. All rights reserved.

Fermentation optimization of goat milk with Lactobacillus acidophilus and Bifidobacterium bifidum by Box-Behnken design.

PubMed

Shu, Guowei; Bao, Chunju; Chen, He; Wang, Changfeng; Yang, Hui

2016-01-01

Goat milk is only limited to the processing of goat milk powder and liquid milk, the products are mainly about milk powder and a few of them are made as milk tablet. Therefore, the study of probiotic goat milk will have great significance in the full use of goats and the development of the goat milk industry in China. The effect of fermentation temperature (35°C, 37°C, 39°C), strain ratio (1:1:1, 2:1:1, 3:1:1) and inoculum size (4%, 5%, 6%) on viable counts of L. acidophilus and B. bifidum, total bacteria and sensory value during fermentation process of L. acidophilus and B. bifidum goat yogurt (AB-goat yogurt) was investigated. The optimum fermentation conditions for AB-goat yogurt were: fermentation temperature 38°C, the strain ratio 2:1:1, inoculum size 6%. Under the optimum conditions, the viable counts of B. bifidum, L. acidophilus, total bacteria and sensory value reached (4.30 ±0.11)×107  cfu/mL, (1.39 ±0.09)×108  cfu/mL, (1.82±0.06)×109  cfu/mL and 7.90 ±0.14, respectively. The fermentation temperature, the strain ratio and inoculum size had a significant effect on the fermentation of AB-goat yogurt and these results are beneficial for developing AB-goat yogurt.

Saccharomyces cerevisiae and metabolic activators: HXT3 gene expression and fructose/glucose discrepancy in sluggish fermentation conditions.

PubMed

Díaz-Hellín, Patricia; Naranjo, Victoria; Úbeda, Juan; Briones, Ana

2016-12-01

When exposed to mixtures of glucose and fructose, as occurs during the fermentation of grape juice into wine, Saccharomyces cerevisiae uses these sugars at different rates. Moreover, glucose and fructose are transported by the same hexose transporters (HXT), which present a greater affinity for glucose, so that late in fermentation, fructose becomes the predominant sugar. Only a few commercial fermentation activators are available to optimally solve the problems this entails. The aim of this study was to investigate the relation between HXT3 gene expression and fructose/glucose discrepancy in two different media inoculated with a commercial wine strain of S. cerevisiae in the presence of three metabolic activators. Fermentation kinetics, vitality and major metabolites were also measured. Rehydration with ergosterol improved the area under the curve and the growth rate (µ max ) in both studied media. Also, the fructose/glucose discrepancy values were improved with all activator treatments, highlighting rehydration in the presence of ascorbic acid. The yeast rehydration process was demonstrated to influence HXT3 expression under the studied conditions. Tetrahydrofolic acid treatment greatly influenced HXT3 gene expression, especially on the 12th day of the fermentation process. To a lesser extent, ergosterol and ascorbic acid also improved this parameter.

Bio-transformation of agri-food wastes by newly isolated Neurospora crassa and Lactobacillus plantarum for egg production.

PubMed

Liu, P; Li, J; Deng, Z

2016-03-01

Using bio-transferred feedstuff was a cost-effective approach to improve egg quality and production; particularly, the nutritive diet came from agri-food wastes. In this study, optimization of fermentation conditions and co-cultivation of Neurospora crassa with Lactobacillus plantarum was performed in a simple bioreactor. The optimized fermentation of beer lees substrates through N. crassa led to the hydrolysis rates of crude fiber increasing to 43.27%. Compared to that of using N. crassa alone, the combination of N. crassa and L. plantarum enhanced the content of amino acids (13,120 to 18,032 mg/100 g) on oil-tea seed cake substrates particularly. When hens were fed 10% fermented oil-tea seedcake substrate, the ratio of feed to egg decreased from 3.1 to 2.6, egg production ratio increased from 65.71 to 80.10%, and color of vitelline (Roche) increased from 8.20 to 10.20. Fifteen kinds of carotenoids were identified by HPLC in fermented oil-tea seed cake substrates. The results of this study highlighted that the mixed-fermentation by N. crassa and L. plantarum may be an effective way to convert agri-food wastes into high-valued biomass products, which could have a positive effect on hens and their eggs. © 2016 Poultry Science Association Inc.

Systematic engineering of TCA cycle for optimal production of a four-carbon platform chemical 4-hydroxybutyric acid in Escherichia coli.

PubMed

Choi, Sol; Kim, Hyun Uk; Kim, Tae Yong; Lee, Sang Yup

2016-11-01

To address climate change and environmental problems, it is becoming increasingly important to establish biorefineries for the production of chemicals from renewable non-food biomass. Here we report the development of Escherichia coli strains capable of overproducing a four-carbon platform chemical 4-hybroxybutyric acid (4-HB). Because 4-HB production is significantly affected by aeration level, genome-scale metabolic model-based engineering strategies were designed under aerobic and microaerobic conditions with emphasis on oxidative/reductive TCA branches and glyoxylate shunt. Several different metabolic engineering strategies were employed to develop strains suitable for fermentation both under aerobic and microaerobic conditions. It was found that microaerobic condition was more efficient than aerobic condition in achieving higher titer and productivity of 4-HB. The final engineered strain produced 103.4g/L of 4-HB by microaerobic fed-batch fermentation using glycerol. The aeration-dependent optimization strategy of TCA cycle will be useful for developing microbial strains producing other reduced derivative chemicals of TCA cycle intermediates. Copyright © 2016 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

Saccharomyces and non-Saccharomyces Competition during Microvinification under Different Sugar and Nitrogen Conditions

PubMed Central

Lleixà, Jessica; Manzano, Maria; Mas, Albert; Portillo, María del C.

2016-01-01

The inoculation of wines with autochthonous yeast allows obtaining complex wines with a peculiar microbial footprint characteristic from a wine region. Mixed inoculation of non-Saccharomyces yeasts and S. cerevisiae is of interest for the wine industry for technological and sensory reasons. However, the interactions between these yeasts are not well understood, especially those regarding the availability of nutrients. The aim of the present study was to analyze the effect of nitrogen and sugar concentration on the evolution of mixed yeast populations on controlled laboratory-scale fermentations monitored by density, plate culturing, PCR-DGGE and sugar and nitrogen consumption. Furthermore, the effect of the time of inoculation of Saccharomyces cerevisiae respect the initial co-inoculation of three non-Saccharomyces yeasts was evaluated over the evolution of fermentation. Our results have shown that S. cerevisiae inoculation during the first 48 h conferred a stabilizing effect over the fermentations with non-Saccharomyces strains tested and, generally, reduced yeast diversity at the end of the fermentation. On the other hand, nitrogen limitation increased the time of fermentation and also the proportion of non-Saccharomyces yeasts at mid and final fermentation. High sugar concentration resulted in different proportions of the inoculated yeast depending on the time of S. cerevisiae inoculation. This work emphasizes the importance of the concentration of nutrients on the evolution of mixed fermentations and points to the optimal conditions for a stable fermentation in which the inoculated yeasts survived until the end. PMID:27994585

Camptothecin-producing endophytic fungus Trichoderma atroviride LY357: isolation, identification, and fermentation conditions optimization for camptothecin production.

PubMed

Pu, Xiang; Qu, Xixing; Chen, Fei; Bao, Jinku; Zhang, Guolin; Luo, Yinggang

2013-11-01

Camptothecin (CPT), the third largest anticancer drug, is produced mainly by Camptotheca acuminata and Nothapodytes foetida. CPT itself is the starting material for clinical CPT-type drugs, but the plant-derived CPT cannot support the heavy demand from the global market. Research efforts have been made to identify novel sources for CPT. In this study, three CPT-producing endophytic fungi, Aspergillus sp. LY341, Aspergillus sp. LY355, and Trichoderma atroviride LY357, were isolated and identified from C. acuminata. Most CPT produced by these fungi was found in the fermentation broth, and their corresponding CPT yields were 7.93, 42.92, and 197.82 μg l(-1), respectively. The CPT-producing capability of LY341 and LY355 was completely lost after repeat subculturing. A substantial decrease of CPT production was also observed in the second generation of LY357. However, a stable and sustainable production of CPT was found from the second generation through the eighth generation of LY357. The fermentation medium, time, pH, temperature, and agitation rate were optimized for CPT production. Methyl jasmonate and XAD16 were proven to be an optimum elicitor and adsorbent resin, respectively, in view of that CPT yield was increased 3.4- and 11-fold through their use. A 50- to 75-fold increase of CPT yield was obtained when the optimized fermentation conditions, elicitor, and adsorbent resin were combined and applied to the culture of the seventh and eighth generations of LY357, and the highest CPT yield was 142.15 μg l(-1). The CPT-producing T. atroviride LY357 paves a potential to uncover the mysteries of CPT biosynthesis.

Production and characterization of bioactive metabolites from piezotolerant deep sea fungus Nigrospora sp. in submerged fermentation.

PubMed

Arumugam, G K; Srinivasan, S K; Joshi, G; Gopal, D; Ramalingam, K

2015-01-01

To produce and characterize bioactive metabolites from piezotolerant marine fungus Nigrospora sp. in submerged fermentation. A distinct marine strain, Nigrospora sp. NIOT has been isolated from a depth of 800 m at the Arabian Sea. The 18S rRNA and internal transcribed spacers (ITS) analysis demonstrates its close association with the genus Nigrospora. Effect of pH, temperature, salinity, carbon source and amino acids was studied to optimize the fermentation conditions. Optimal mycelia growth and secondary metabolites production were observed at 6·0-8·0 pH, 20-30°C temperature, 7·5% salinity, sucrose as carbon source and tryptophan as amino acid source. The extracellular secondary metabolites exhibited high antimicrobial activities against both gram-positive and gram-negative pathogenic bacteria with minimal inhibitory concentration (MIC) values higher than 30 μg ml(-1). Strongest cytotoxicity was observed in all cell lines tested, GI50 (growth inhibition by 50%) was calculated to be 1·35, 3·2, 0·13 and 0·35 μg ml(-1) against U937, MCF-7, A673 and Jurkat, respectively. Fourier transform infrared spectroscopy (FTIR) and gas chromatography-mass spectrometry (GC-MS) analyses of secondary metabolites confirmed the production of antimicrobial and anticancer substances. A piezotolerant fungus Nigrospora sp. NIOT isolated from deep sea environment was successfully cultured under submerged fermentation. The secondary metabolites produced from this organism showed potent antimicrobial and anticancer activities with immediate application to cosmetics and pharmaceutical industries. This is the first study exploring Nigrospora sp. from 800 m in marine environment. This deep sea fungus under optimized culture conditions effectively produced bioactive secondary metabolites such as griseofulvin, spirobenzofuran and pyrone derivatives at higher concentrations. © 2014 The Society for Applied Microbiology.

Optimization of the liquid biofertilizer production in batch fermentation with by-product from MSG

NASA Astrophysics Data System (ADS)

Namfon, Panjanapongchai; Ratchanok, Sahaworarak; Chalida, Daengbussade

2017-03-01

The long term use of chemical fertilizers destroyed the friability of soil which obviously decreased quantity and quality of crops and especially affect microorganisms living in soils. The bio-fertilizer with microbial consortium is an environmental friendly alternative to solve this bottleneck due to harboring soil microorganisms such as Bacillus sp., Micrococcus sp., Pseudomonas sp., Staphylococcus sp. and Deinococcus sp. produced with natural by-product or waste from industries that is alternative and sustainable such as nutrient-rich (by-product) from Mono Sodium Glutamate (MSG) for producing liquid biofertilizer by batch fermentation. In this work, the concentration of reducing sugar from substrate as main carbon source was evaluated in shake flask with mixed cultures. The optimal conditions were studied comparing with two levels of reducing sugar concentration (10, 20 g/L) and inoculums concentration (10, 20 %v/v) with using (2×2) full factorial design. The results indicated that the by-product from monosodium glutamate is feasible for fermentation and inoculums concentration is mainly influenced the batch fermentation process. Moreover, the combined 20 g/L and 10%v/v were considerably concluded as an optimal condition, of which the concentration of vegetative cells and spores attained at 8.29×109 CFU/mL and 1.97×105 CFU/mL, respectively. Their spores cell yields from reducing sugar (Yx/s) were obtained at 1.22×106 and 3.34×105 CFU/g were markedly different. In conclusion, the liquid Biofertilizer was produced satisfactorily at 20 g/L reducing sugar and 10% v/v inoculums in shake flask culture. Moreover, these results suggested that the by-product from monosodium glutamate is feasible for low-cost substrate in economical scale and environmental-friendly.

Production of Candida utilis on slop by-product of fermentation industries.

PubMed

Foda, M S; El-Naggar, M R; Haroun, B M

1976-01-01

The slop (vinas) liquor, a major by-product a alcohol fermentation industries, has been used as growth medium for production of the torula yeast, Candida utilis. Supplementation of the slop with 0.2% ammonium sulphate and 1% to 5% molasses improved the cell yield significantly. The crude slop gave better results than the diluted or centrifuged liquors. Under optimal conditions, more than 15 grams of yeast were obtained on dry weight basis. The application feasibility of these results is presented.

Application of the Response Surface Methodology to Optimize the Fermentation Parameters for Enhanced Docosahexaenoic Acid (DHA) Production by Thraustochytrium sp. ATCC 26185.

PubMed

Wu, Kang; Ding, Lijian; Zhu, Peng; Li, Shuang; He, Shan

2018-04-22

The aim of this study was to determine the cumulative effect of fermentation parameters and enhance the production of docosahexaenoic acid (DHA) by Thraustochytrium sp. ATCC 26185 using response surface methodology (RSM). Among the eight variables screened for effects of fermentation parameters on DHA production by Plackett-Burman design (PBD), the initial pH, inoculum volume, and fermentation volume were found to be most significant. The Box-Behnken design was applied to derive a statistical model for optimizing these three fermentation parameters for DHA production. The optimal parameters for maximum DHA production were initial pH: 6.89, inoculum volume: 4.16%, and fermentation volume: 140.47 mL, respectively. The maximum yield of DHA production was 1.68 g/L, which was in agreement with predicted values. An increase in DHA production was achieved by optimizing the initial pH, fermentation, and inoculum volume parameters. This optimization strategy led to a significant increase in the amount of DHA produced, from 1.16 g/L to 1.68 g/L. Thraustochytrium sp. ATCC 26185 is a promising resource for microbial DHA production due to the high-level yield of DHA that it produces, and the capacity for large-scale fermentation of this organism.

Manufacture and characterization of a yogurt-like beverage made with oat flakes fermented by selected lactic acid bacteria.

PubMed

Luana, Nionelli; Rossana, Coda; Curiel, José Antonio; Kaisa, Poutanen; Marco, Gobbetti; Rizzello, Carlo Giuseppe

2014-08-18

This study aimed at investigating the suitability of oat flakes for making functional beverages. Different technological options were assayed, including the amount of flakes, the inoculum of the starter and the addition of enzyme preparations. The beverage containing 25% (wt/wt) of oat flakes and fermented with L. plantarum LP09 was considered optimal on the basis of sensory and technological properties. The enzyme addition favored the growth of the starter, shortened the time needed to reach pH4.2 to ca. 8h, and favored a decrease of the quotient of fermentation. Fermentation increased the polyphenols availability and the antioxidant activity (25 and 70% higher, respectively) and decreased the hydrolysis index in vitro. Sensory analyses showed that fermented oat flakes beverage had the typical features of a yogurt-like beverage, enhancing the overall intensity of odor and flavor compared to the non-fermented control. Selection of proper processing and fermentation condition allowed the obtainment of a beverage with better nutritional and sensory properties. Copyright © 2014 Elsevier B.V. All rights reserved.

Optimisation of lactic acid fermentation for improved vinegar flavour during rosy vinegar brewing.

PubMed

Jiang, Yujian; Guo, Jianna; Li, Yudong; Lin, Sen; Wang, Li; Li, Jianrong

2010-06-01

Rosy vinegar is a well-known traditional Chinese product whose flavour is affected by its lactic acid content. In this study, Lactobacillus bacteria were employed to increase the content of lactic acid during the ethanol fermentation stage. The optimised fermentation parameters were determined as an inoculation amount of 3% (v/v), a temperature of 30 degrees C and an initial pH value of 4.0. Fermentation under these optimal conditions resulted in an alcohol degree of 6.2% (v/v), a total acidity of 49.5 g L(-1) and a lactic acid content of 4.14 g L(-1). The content of lactic acid (4.14 g L(-1)), which approached the level achieved by solid state fermentation, was 3.56-fold higher than that in vinegar fermented without lactic acid bacteria (1.16 g L(-1)). The results indicate that mixed fermentation with Lactobacillus plantarum and Saccharomyces cerevisiae strains greatly increases the lactic acid content and improves the flavour of rosy vinegar. Copyright (c) 2010 Society of Chemical Industry.

Dynamics of the microbial community during continuous methane fermentation in continuously stirred tank reactors.

PubMed

Tang, Yue-Qin; Shigematsu, Toru; Morimura, Shigeru; Kida, Kenji

2015-04-01

Methane fermentation is an attractive technology for the treatment of organic wastes and wastewaters. However, the process is difficult to control, and treatment rates and digestion efficiency require further optimization. Understanding the microbiology mechanisms of methane fermentation is of fundamental importance to improving this process. In this review, we summarize the dynamics of microbial communities in methane fermentation chemostats that are operated using completely stirred tank reactors (CSTRs). Each chemostat was supplied with one substrate as the sole carbon source. The substrates include acetate, propionate, butyrate, long-chain fatty acids, glycerol, protein, glucose, and starch. These carbon sources are general substrates and intermediates of methane fermentation. The factors that affect the structure of the microbial community are discussed. The carbon source, the final product, and the operation conditions appear to be the main factors that affect methane fermentation and determine the structure of the microbial community. Understanding the structure of the microbial community during methane fermentation will guide the design and operation of practical wastewater treatments. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Enhanced mannan-derived fermentable sugars of palm kernel cake by mannanase-catalyzed hydrolysis for production of biobutanol.

PubMed

Shukor, Hafiza; Abdeshahian, Peyman; Al-Shorgani, Najeeb Kaid Nasser; Hamid, Aidil Abdul; Rahman, Norliza A; Kalil, Mohd Sahaid

2016-10-01

Catalytic depolymerization of mannan composition of palm kernel cake (PKC) by mannanase was optimized to enhance the release of mannan-derived monomeric sugars for further application in acetone-butanol-ethanol (ABE) fermentation. Efficiency of enzymatic hydrolysis of PKC was studied by evaluating effects of PKC concentration, mannanase loading, hydrolysis pH value, reaction temperature and hydrolysis time on production of fermentable sugars using one-way analysis of variance (ANOVA). The ANOVA results revealed that all factors studied had highly significant effects on total sugar liberated (P<0.01). The optimum conditions for PKC hydrolysis were 20% (w/v) PKC concentration, 5% (w/w) mannanase loading, hydrolysis pH 4.5, 45°C temperature and 72h hydrolysis time. Enzymatic experiments in optimum conditions revealed total fermentable sugars of 71.54±2.54g/L were produced including 67.47±2.51g/L mannose and 2.94±0.03g/L glucose. ABE fermentation of sugar hydrolysate by Clostridium saccharoperbutylacetonicum N1-4 resulted in 3.27±1.003g/L biobutanol. Copyright © 2016 Elsevier Ltd. All rights reserved.

Demonstration-Scale High-Cell-Density Fermentation of Pichia pastoris.

PubMed

Liu, Wan-Cang; Zhu, Ping

2018-01-01

Pichia pastoris has been one of the most successful heterologous overexpression systems in generating proteins for large-scale production through high-cell-density fermentation. However, optimizing conditions of the large-scale high-cell-density fermentation for biochemistry and industrialization is usually a laborious and time-consuming process. Furthermore, it is often difficult to produce authentic proteins in large quantities, which is a major obstacle for functional and structural features analysis and industrial application. For these reasons, we have developed a protocol for efficient demonstration-scale high-cell-density fermentation of P. pastoris, which employs a new methanol-feeding strategy-biomass-stat strategy and a strategy of increased air pressure instead of pure oxygen supplement. The protocol included three typical stages of glycerol batch fermentation (initial culture phase), glycerol fed-batch fermentation (biomass accumulation phase), and methanol fed-batch fermentation (induction phase), which allows direct online-monitoring of fermentation conditions, including broth pH, temperature, DO, anti-foam generation, and feeding of glycerol and methanol. Using this protocol, production of the recombinant β-xylosidase of Lentinula edodes origin in 1000-L scale fermentation can be up to ~900 mg/L or 9.4 mg/g cells (dry cell weight, intracellular expression), with the specific production rate and average specific production of 0.1 mg/g/h and 0.081 mg/g/h, respectively. The methodology described in this protocol can be easily transferred to other systems, and eligible to scale up for a large number of proteins used in either the scientific studies or commercial purposes.

Optimization of Postharvest Conditions To Produce Chocolate Aroma from Jackfruit Seeds.

PubMed

Spada, Fernanda Papa; Zerbeto, Lais Masson; Ragazi, Gabriel Bernardes Cabreira; Gutierrez, Érika Maria Roel; Souza, Miriam Coelho; Parker, Jane K; Canniatti-Brazaca, Solange Guidolin

2017-02-15

Jackfruit seeds are an underutilized waste in many tropical countries. This work demonstrates the potential of roasted jackfruit seeds to develop chocolate aroma. Twenty-seven different roasted jackfruit seed flours were produced from local jackfruit by acidifying or fermenting the seeds prior to drying and then roasting under different time/temperature combinations. The chocolate aroma of groups of four flours were ranked by a sensory panel (n = 162), and response surface methodology was used to identify optimum conditions. The results indicated a significant and positive influence of fermentation and acidification on the production of chocolate aroma. SPME/GC-MS of the flours showed that important aroma compounds such as 2,3-diethyl-5-methylpyrazine and 2-phenylethyl acetate were substantially higher in the fermented product and that the more severe roasting conditions produced 2-3 times more 2,3-diethyl-5-methylpyrazine, but less 3-methylbutanal. Moisture, a w , pH, luminosity, and color were also monitored to ensure that these properties were similar to those of cocoa powder or cocoa substitutes.

Alteration of white-rot basidiomycetes cellulase and xylanase activities in the submerged co-cultivation and optimization of enzyme production by Irpex lacteus and Schizophyllum commune.

PubMed

Metreveli, Eka; Kachlishvili, Eva; Singer, Steven W; Elisashvili, Vladimir

2017-10-01

Mono and dual cultures of four white-rot basidiomycete species were evaluated for cellulase and xylanase activity under submerged fermentation conditions. Co-cultivation of Pycnoporus coccineus or Trametes hirsuta with Schizophyllum commune displayed antagonistic interactions resulting in the decrease of endoglucanase and total cellulase activities. In contrast, increases in cellulase and xylanase activity were revealed through the compatible interactions of Irpex lacteus with S. commune. Co-cultivation conditions were optimized for maximum enzyme production by I. lacteus and S. commune, the best producers of cellulase/xylanase and β-glucosidase, respectively. An optimized medium for the target enzyme production by the mixed culture was established in a laboratory fermenter yielding 7U/mL total cellulase, 142U/mL endoglucanase, 104U/mL xylanase, and 5.2U/mL β-glucosidase. The dual culture approach resulted in an enzymatic mixture with 11% improved lignocellulose saccharification potential compared to enzymes from a monoculture of I. lacteus. Copyright © 2017 Elsevier Ltd. All rights reserved.

Laccase production by Coriolopsis caperata RCK2011: Optimization under solid state fermentation by Taguchi DOE methodology

PubMed Central

Nandal, Preeti; Ravella, Sreenivas Rao; Kuhad, Ramesh Chander

2013-01-01

Laccase production by Coriolopsis caperata RCK2011 under solid state fermentation was optimized following Taguchi design of experiment. An orthogonal array layout of L18 (21 × 37) was constructed using Qualitek-4 software with eight most influensive factors on laccase production. At individual level pH contributed higher influence, whereas, corn steep liquor (CSL) accounted for more than 50% of the severity index with biotin and KH2PO4 at the interactive level. The optimum conditions derived were; temperature 30°C, pH 5.0, wheat bran 5.0 g, inoculum size 0.5 ml (fungal cell mass = 0.015 g dry wt.), biotin 0.5% w/v, KH2PO4 0.013% w/v, CSL 0.1% v/v and 0.5 mM xylidine as an inducer. The validation experiments using optimized conditions confirmed an improvement in enzyme production by 58.01%. The laccase production to the level of 1623.55 Ugds−1 indicates that the fungus C. caperata RCK2011 has the commercial potential for laccase. PMID:23463372

Optimization of γ-aminobutyric acid production by Lactobacillus plantarum Taj-Apis362 from honeybees.

PubMed

Tajabadi, Naser; Ebrahimpour, Afshin; Baradaran, Ali; Rahim, Raha Abdul; Mahyudin, Nor Ainy; Manap, Mohd Yazid Abdul; Bakar, Fatimah Abu; Saari, Nazamid

2015-04-15

Dominant strains of lactic acid bacteria (LAB) isolated from honey bees were evaluated for their γ-aminobutyric acid (GABA)-producing ability. Out of 24 strains, strain Taj-Apis362 showed the highest GABA-producing ability (1.76 mM) in MRS broth containing 50 mM initial glutamic acid cultured for 60 h. Effects of fermentation parameters, including initial glutamic acid level, culture temperature, initial pH and incubation time on GABA production were investigated via a single parameter optimization strategy. The optimal fermentation condition for GABA production was modeled using response surface methodology (RSM). The results showed that the culture temperature was the most significant factor for GABA production. The optimum conditions for maximum GABA production by Lactobacillus plantarum Taj-Apis362 were an initial glutamic acid concentration of 497.97 mM, culture temperature of 36 °C, initial pH of 5.31 and incubation time of 60 h, which produced 7.15 mM of GABA. The value is comparable with the predicted value of 7.21 mM.

Phenolic profile and antioxidant activity of extracts prepared from fermented heat-stabilized defatted rice bran.

PubMed

Webber, Daniel M; Hettiarachchy, Navam S; Li, Ruiqi; Horax, Ronny; Theivendran, Sivarooban

2014-11-01

Heat-stabilized, defatted rice bran (HDRB) serves as a potential source of phenolic compounds which have numerous purported health benefits. An estimated 70% of phenolics present in rice bran are esterified to the arabinoxylan residues of the cell walls. Release of such compounds could provide a value-added application for HDRB. The objective of this study was to extract and quantify phenolics from HDRB using fermentation technology. Out of 8 organisms selected for rice bran fermentation, Bacillus subtilis subspecies subtilis had the maximum phenolic release of 26.8 mg ferulic acid equivalents (FAE) per gram HDRB. Response surface methodology was used to further optimize the release of rice bran phenolics. An optimum of 28.6 mg FAE/g rice bran was predicted at 168 h, 0.01% inoculation level, and 100 mg HDRB/mL. Fermentation of HDRB for 96 h with B. subtilis subspecies subtilis resulted in a significant increase in phenolic yield, phenolic concentration, and radical scavenging capacity. Fermented rice bran had 4.86 mg gentistic acid, 1.38 mg caffeic acid, 6.03 mg syringic acid, 19.02 mg (-)-epicatechin, 4.08 mg p-courmaric acid, 4.64 mg ferulic acid, 10.04 mg sinapic acid, and 17.59 mg benzoic acid per 100 g fermented extract compared to 0.65 mg p-courmaric acid and 0.36 mg ferulic acid per 100 g nonfermented extract. The high phenolic content and antioxidant activity of fermented HDRB extract indicates that rice bran fermentation under optimized condition is a potential means of meeting the demand for an effective and affordable antioxidant. © 2014 Institute of Food Technologists®

Optimization to the Culture Conditions for Phellinus Production with Regression Analysis and Gene-Set Based Genetic Algorithm

PubMed Central

Li, Zhongwei; Xin, Yuezhen; Wang, Xun; Sun, Beibei; Xia, Shengyu; Li, Hui

2016-01-01

Phellinus is a kind of fungus and is known as one of the elemental components in drugs to avoid cancers. With the purpose of finding optimized culture conditions for Phellinus production in the laboratory, plenty of experiments focusing on single factor were operated and large scale of experimental data were generated. In this work, we use the data collected from experiments for regression analysis, and then a mathematical model of predicting Phellinus production is achieved. Subsequently, a gene-set based genetic algorithm is developed to optimize the values of parameters involved in culture conditions, including inoculum size, PH value, initial liquid volume, temperature, seed age, fermentation time, and rotation speed. These optimized values of the parameters have accordance with biological experimental results, which indicate that our method has a good predictability for culture conditions optimization. PMID:27610365

Effect of phosphoric acid pretreatment of corncobs on the fermentability of Clostridium beijerinckii TISTR 1461 for biobutanol production.

PubMed

Boonsombuti, Akarin; Luengnaruemitchai, Apanee; Wongkasemjit, Sujitra

2015-01-01

Corncobs pretreated with H2SO4, HNO3, and H3PO4 were compared to evaluate the fermentation ability of Clostridium beijerinckii TISTR 1461 to produce biobutanol via acetone-butanol-ethanol (ABE) fermentation. It was found that the hydrolysate from H3PO4 pretreatment could be used as a substrate without any inhibitor removal methods. However, in terms of sugar yield, it gave the lowest total sugars in both pretreatment and enzymatic hydrolysis. Response surface methodology was applied to optimize enzymatic hydrolysis of the pretreated corncobs. The optimized conditions reduced the consumption of enzymes and hydrolysis time to 7.68 FPU/g biomass and 63.88 hr, respectively, and yielded 51.82 g/L reducing sugars. The Celluclast 1.5 L and Novozyme 188 enzyme ratio were varied to maximize the hydrolyzed sugars. The ABE fermentation, using substrate from phosphoric acid pretreatment of corncobs, with 10 g/L glucose supplementation produced 11.64 g/L of total ABE, which was close to the control experiment using synthetic medium. This study showed that corncobs pretreated with phosphoric acid could potentially be used as a substrate without using a detoxification process.

Garcina cambogia leaf and seawater for tannase production by marine Aspergillus awamori BTMFW032 under slurry state fermentation.

PubMed

Beena, S P; Basheer, Soorej M; Bhat, Sarita G; Chandrasekaran, M

2011-12-01

Garcinia gummi-gutta (syn. G. cambogia, G. quaesita), known to have medicinal properties, was evaluated as a substrate and inducer for tannase production by a marine Aspergillus awamori BTMFW032, under slurry state fermentation using Czapekdox-minimal medium and sea water as the cultivation medium. Among the various natural tannin substrates evaluated, Garcinia leaf supported maximal tannase production. The cultivation conditions and components of the cultivation medium were optimized employing response surface methodology. The experimental results were fitted to a second-order polynomial model at a 92.2% level of significance (p < 0.0001). The maximal tannase activity was obtained in a slurry state medium containing 26.6%, w/v, Garcinia leaf, supplemented with 0.1% tannic acid as inducer. The optimum values of pH, temperature and inoculum concentration obtained were 5.0, 40 degrees C and 3%, respectively. A Box-Behnken model study of the fermentation conditions was carried out, and the best production of tannase was registered at 40 degrees C without agitation. Optimization strategy employing response surface methodology led to nearly 3-fold increase in the enzyme production from 26.2 U/mL obtained in unoptimized medium to 75.2 Units/mL in Box Behnken design, within 18 h of fermentation. It was observed that sea water could support maximal tannase production by A. awamori compared with other media suggesting that the sea water salts could have played an inducer role in expression of tannase encoding genes. To the best of our knowledge, this is the first report on production of tannase, an industrially important enzyme, utilizing Garcinia leaf as substrate under slurry state fermentation by marine A. awamori and sea water as the cultivation medium.

Biological hydrogen production by dark fermentation: challenges and prospects towards scaled-up production.

PubMed

RenNanqi; GuoWanqian; LiuBingfeng; CaoGuangli; DingJie

2011-06-01

Among different technologies of hydrogen production, bio-hydrogen production exhibits perhaps the greatest potential to replace fossil fuels. Based on recent research on dark fermentative hydrogen production, this article reviews the following aspects towards scaled-up application of this technology: bioreactor development and parameter optimization, process modeling and simulation, exploitation of cheaper raw materials and combining dark-fermentation with photo-fermentation. Bioreactors are necessary for dark-fermentation hydrogen production, so the design of reactor type and optimization of parameters are essential. Process modeling and simulation can help engineers design and optimize large-scale systems and operations. Use of cheaper raw materials will surely accelerate the pace of scaled-up production of biological hydrogen. And finally, combining dark-fermentation with photo-fermentation holds considerable promise, and has successfully achieved maximum overall hydrogen yield from a single substrate. Future development of bio-hydrogen production will also be discussed. Copyright © 2011 Elsevier Ltd. All rights reserved.

Characterization of thermophilic fungal community associated with pile fermentation of Pu-erh tea.

PubMed

Zhang, Wei; Yang, Ruijuan; Fang, Wenjun; Yan, Liang; Lu, Jun; Sheng, Jun; Lv, Jie

2016-06-16

This study aimed to characterize the thermophilic fungi in pile-fermentation process of Pu-erh tea. Physicochemical analyses showed that the high temperature and low pH provided optimal conditions for propagation of fungi. A number of fungi, including Blastobotrys adeninivorans, Thermomyces lanuginosus, Rasamsonia emersonii, Aspergillus fumigatus, Rhizomucor pusillus, Rasamsonia byssochlamydoides, Rasamsonia cylindrospora, Aspergillus tubingensis, Aspergillus niger, Candida tropicalis and Fusarium graminearum were isolated as thermophilic fungi under combination of high temperature and acid culture conditions from Pu-erh tea pile-fermentation. The fungal communities were analyzed by PCR-DGGE. Results revealed that those fungi are closely related to Debaryomyces hansenii, Cladosporium cladosporioides, A. tubingensis, R. emersonii, R. pusillus, A. fumigatus and A. niger, and the last four presented as dominant species in the pile process. These four preponderant thermophilic fungi reached the order of magnitude of 10(7), 10(7), 10(7) and 10(6)copies/g dry tea, respectively, measured by real-time quantitative PCR (q-PCR). The results indicate that the thermophilic fungi play an important role in Pu-erh tea pile fermentation. Copyright © 2016 Elsevier B.V. All rights reserved.

Economics of membrane occupancy and respiro-fermentation

PubMed Central

Zhuang, Kai; Vemuri, Goutham N; Mahadevan, Radhakrishnan

2011-01-01

The simultaneous utilization of efficient respiration and inefficient fermentation even in the presence of abundant oxygen is a puzzling phenomenon commonly observed in bacteria, yeasts, and cancer cells. Despite extensive research, the biochemical basis for this phenomenon remains obscure. We hypothesize that the outcome of a competition for membrane space between glucose transporters and respiratory chain (which we refer to as economics of membrane occupancy) proteins influences respiration and fermentation. By incorporating a sole constraint based on this concept in the genome-scale metabolic model of Escherichia coli, we were able to simulate respiro-fermentation. Further analysis of the impact of this constraint revealed differential utilization of the cytochromes and faster glucose uptake under anaerobic conditions than under aerobic conditions. Based on these simulations, we propose that bacterial cells manage the composition of their cytoplasmic membrane to maintain optimal ATP production by switching between oxidative and substrate-level phosphorylation. These results suggest that the membrane occupancy constraint may be a fundamental governing constraint of cellular metabolism and physiology, and establishes a direct link between cell morphology and physiology. PMID:21694717

A Statistical Approach for Optimization of Simultaneous Production of β-Glucosidase and Endoglucanase by Rhizopus oryzae from Solid-State Fermentation of Water Hyacinth Using Central Composite Design

PubMed Central

Karmakar, Moumita; Ray, Rina Rani

2011-01-01

The production cost of β-glucosidase and endoglucanase could be reduced by using water hyacinth, an aquatic weed, as the sole carbon source and using cost-efficient fermentation strategies like solid-state fermentation (SSF). In the present study, the effect of different production conditions on the yield of β-glucosidase and endoglucanase by Rhizopus oryzae MTCC 9642 from water hyacinth was investigated systematically using response surface methodology. A Central composite experimental design was applied to optimize the impact of three variables, namely, substrate concentration, pH, and temperature, on enzyme production. The optimal level of each parameter for maximum enzyme production by the fungus was determined. Highest activity of endoglucanase of 495 U/mL was achieved at a substrate concentration of 1.23%, pH 7.29, and temperature 29.93°C whereas maximum β-glucosidase activity of 137.32 U/ml was achieved at a substrate concentration of 1.25%, pH 6.66, and temperature 32.09°C. There was a direct correlation between the levels of enzymatic activities and the substrate concentration of water hyacinth as carbon source. PMID:21687577

Metabolomic and proteomic analysis of D-lactate-producing Lactobacillus delbrueckii under various fermentation conditions.

PubMed

Liang, Shaoxiong; Gao, Dacheng; Liu, Huanhuan; Wang, Cheng; Wen, Jianping

2018-05-28

As an important feedstock monomer for the production of biodegradable stereo-complex poly-lactic acid polymer, D-lactate has attracted much attention. To improve D-lactate production by microorganisms such as Lactobacillus delbrueckii, various fermentation conditions were performed, such as the employment of anaerobic fermentation, the utilization of more suitable neutralizing agents, and exploitation of alternative nitrogen sources. The highest D-lactate titer could reach 133 g/L under the optimally combined fermentation condition, increased by 70.5% compared with the control. To decipher the potential mechanisms of D-lactate overproduction, the time-series response of intracellular metabolism to different fermentation conditions was investigated by GC-MS and LC-MS/MS-based metabolomic analysis. Then the metabolomic datasets were subjected to weighted correlation network analysis (WGCNA), and nine distinct metabolic modules and eight hub metabolites were identified to be specifically associated with D-lactate production. Moreover, a quantitative iTRAQ-LC-MS/MS proteomic approach was employed to further analyze the change of intracellular metabolism under the combined fermentation condition, identifying 97 up-regulated and 42 down-regulated proteins compared with the control. The in-depth analysis elucidated how the key factors exerted influence on D-lactate biosynthesis. The results revealed that glycolysis and pentose phosphate pathways, transport of glucose, amino acids and peptides, amino acid metabolism, peptide hydrolysis, synthesis of nucleotides and proteins, and cell division were all strengthened, while ATP consumption for exporting proton, cell damage, metabolic burden caused by stress response, and bypass of pyruvate were decreased under the combined condition. These might be the main reasons for significantly improved D-lactate production. These findings provide the first omics view of cell growth and D-lactate overproduction in L. delbrueckii, which can be a theoretical basis for further improving the production of D-lactate.

Genome-wide study of the adaptation of Saccharomyces cerevisiae to the early stages of wine fermentation.

PubMed

Novo, Maite; Mangado, Ana; Quirós, Manuel; Morales, Pilar; Salvadó, Zoel; Gonzalez, Ramon

2013-01-01

This work was designed to identify yeast cellular functions specifically affected by the stress factors predominating during the early stages of wine fermentation, and genes required for optimal growth under these conditions. The main experimental method was quantitative fitness analysis by means of competition experiments in continuous culture of whole genome barcoded yeast knockout collections. This methodology allowed the identification of haploinsufficient genes, and homozygous deletions resulting in growth impairment in synthetic must. However, genes identified as haploproficient, or homozygous deletions resulting in fitness advantage, were of little predictive power concerning optimal growth in this medium. The relevance of these functions for enological performance of yeast was assessed in batch cultures with single strains. Previous studies addressing yeast adaptation to winemaking conditions by quantitative fitness analysis were not specifically focused on the proliferative stages. In some instances our results highlight the importance of genes not previously linked to winemaking. In other cases they are complementary to those reported in previous studies concerning, for example, the relevance of some genes involved in vacuolar, peroxisomal, or ribosomal functions. Our results indicate that adaptation to the quickly changing growth conditions during grape must fermentation require the function of different gene sets in different moments of the process. Transport processes and glucose signaling seem to be negatively affected by the stress factors encountered by yeast in synthetic must. Vacuolar activity is important for continued growth during the transition to stationary phase. Finally, reduced biogenesis of peroxisomes also seems to be advantageous. However, in contrast to what was described for later stages, reduced protein synthesis is not advantageous for the early (proliferative) stages of the fermentation process. Finally, we found adenine and lysine to be in short supply for yeast growth in some natural grape musts.

Enhancement of gamma-aminobutyric acid (GABA) levels using an autochthonous Lactobacillus futsaii CS3 as starter culture in Thai fermented shrimp (Kung-Som).

PubMed

Sanchart, Chatthaphisuth; Rattanaporn, Onnicha; Haltrich, Dietmar; Phukpattaranont, Pimpimol; Maneerat, Suppasil

2017-08-01

Gamma-aminobutyric acid (GABA) is a non-proteinogenic amino acid, which has a variety of well-characterized beneficial physiological functions. In order to improve GABA levels and the fermentation process of Thai fermented shrimp (Kung-Som), autochthonous Lactobacillus futsaii CS3 was inoculated as a starter culture into Kung-Som, and its effects on the quality of Kung-Som were studied. The optimal conditions for GABA production in Kung-Som as obtained by response surface methodology (RSM) using a central composite design (CCD) were an inoculum size of roughly 10 7 CFU/g (X 1 ) of L. futsaii cells together with the addition of 0.5% (w/w) monosodium glutamate (MSG) (X 2 ), resulting in maximum GABA levels of 10,500 mg per kg fresh product. Under these optimized conditions, the experimental GABA content of Kung-Som with an added starter culture was up to four times higher than that of the control (without starter culture) or commercial Kung-Som products (10,120 mg/kg product). Kung-Som produced by inoculation with L. futsaii CS3 but without addition of MSG showed a considerably increased GABA content of 7790 mg/kg compared to the control. Fermentation time was reduced to less than 1 week for these samples compared to the control batches, which took up to 19 days. Polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) revealed that L. futsaii CS3 remained prominently throughout the Kung-Som fermentation, and that lactic acid bacteria (LAB) rapidly dominated the total microflora because of this inoculation with L. futsaii CS3. Kung-Som samples with starter culture were accepted as well as commercial ones by 30 panelists (p > 0.05). In conclusion, L. futsaii CS3 is a good starter culture for GABA production, resulting in, improved microbiological safety as well as reduced fermentation time.

Comparative evaluation of extracellular β-D-fructofuranosidase in submerged and solid-state fermentation produced by newly identified Bacillus subtilis strain.

PubMed

Lincoln, Lynette; More, Sunil S

2018-04-17

To screen and identify a potential extracellular β-D-fructofuranosidase or invertase producing bacterium from soil, and comparatively evaluate the enzyme biosynthesis under submerged and solid-state fermentation. Extracellular invertase producing bacteria were screened from soil. Identification of the potent bacterium was performed based on microscopic examinations and 16S rDNA molecular sequencing. Bacillus subtilis LYN12 invertase secretion was surplus with wheat bran humidified with molasses medium (70%), with elevated activity at 48 h and 37 °C under solid-state fermentation, whereas under submerged conditions increased activity was observed at 24 h and 45 °C in the molasses medium. The study revealed a simple fermentative medium for elevated production of extracellular invertase from a fast growing Bacillus strain. Bacterial invertases are scarce and limited reports are available. By far, this is the first report on the comparative analysis of optimization of extracellular invertase synthesis from Bacillus subtilis strain by submerged and solid-state fermentation. The use of agricultural residues increased yields resulting in development of a cost-effective and stable approach. Bacillus subtilis LYN12 invertase possesses excellent fermenting capability to utilize agro-industrial residues under submerged and solid-state conditions. This could be a beneficial candidate in food and beverage processing industries. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Ant colony system algorithm for the optimization of beer fermentation control.

PubMed

Xiao, Jie; Zhou, Ze-Kui; Zhang, Guang-Xin

2004-12-01

Beer fermentation is a dynamic process that must be guided along a temperature profile to obtain the desired results. Ant colony system algorithm was applied to optimize the kinetic model of this process. During a fixed period of fermentation time, a series of different temperature profiles of the mixture were constructed. An optimal one was chosen at last. Optimal temperature profile maximized the final ethanol production and minimized the byproducts concentration and spoilage risk. The satisfactory results obtained did not require much computation effort.

Studying the rapid bioconversion of lignocellulosic sugars into ethanol using high cell density fermentations with cell recycle

PubMed Central

2014-01-01

Background The Rapid Bioconversion with Integrated recycle Technology (RaBIT) process reduces capital costs, processing times, and biocatalyst cost for biochemical conversion of cellulosic biomass to biofuels by reducing total bioprocessing time (enzymatic hydrolysis plus fermentation) to 48 h, increasing biofuel productivity (g/L/h) twofold, and recycling biocatalysts (enzymes and microbes) to the next cycle. To achieve these results, RaBIT utilizes 24-h high cell density fermentations along with cell recycling to solve the slow/incomplete xylose fermentation issue, which is critical for lignocellulosic biofuel fermentations. Previous studies utilizing similar fermentation conditions showed a decrease in xylose consumption when recycling cells into the next fermentation cycle. Eliminating this decrease is critical for RaBIT process effectiveness for high cycle counts. Results Nine different engineered microbial strains (including Saccharomyces cerevisiae strains, Scheffersomyces (Pichia) stipitis strains, Zymomonas mobilis 8b, and Escherichia coli KO11) were tested under RaBIT platform fermentations to determine their suitability for this platform. Fermentation conditions were then optimized for S. cerevisiae GLBRCY128. Three different nutrient sources (corn steep liquor, yeast extract, and wheat germ) were evaluated to improve xylose consumption by recycled cells. Capacitance readings were used to accurately measure viable cell mass profiles over five cycles. Conclusion The results showed that not all strains are capable of effectively performing the RaBIT process. Acceptable performance is largely correlated to the specific xylose consumption rate. Corn steep liquor was found to reduce the deleterious impacts of cell recycle and improve specific xylose consumption rates. The viable cell mass profiles indicated that reduction in specific xylose consumption rate, not a drop in viable cell mass, was the main cause for decreasing xylose consumption. PMID:24847379

Effective conversion of maize straw wastes into bio-hydrogen by two-stage process integrating H2 fermentation and MECs.

PubMed

Li, Yan-Hong; Bai, Yan-Xia; Pan, Chun-Mei; Li, Wei-Wei; Zheng, Hui-Qin; Zhang, Jing-Nan; Fan, Yao-Ting; Hou, Hong-Wei

2015-12-01

The enhanced H2 production from maize straw had been achieved through the two-stage process of integrating H2 fermentation and microbial electrolysis cells (MECs) in the present work. Several key parameters affecting hydrolysis of maize straw through subcritical H2O were optimized by orthogonal design for saccharification of maize straw followed by H2 production through H2 fermentation. The maximum reducing sugar (RS) content of maize straw reached 469.7 mg/g-TS under the optimal hydrolysis condition with subcritical H2O combining with dilute HCl of 0.3% at 230 °C. The maximum H2 yield, H2 production rate, and H2 content was 115.1 mL/g-TVS, 2.6 mL/g-TVS/h, and 48.9% by H2 fermentation, respectively. In addition, the effluent from H2 fermentation was used as feedstock of MECs for additional H2 production. The maximum H2 yield of 1060 mL/g-COD appeared at an applied voltage of 0.8 V, and total COD removal reached about 35%. The overall H2 yield from maize straw reached 318.5 mL/g-TVS through two-stage processes. The structural characterization of maize straw was also carefully investigated by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD) spectra.

Expression of food-grade phytase in Lactococcus lactis from optimized conditions in milk broth.

PubMed

Miao, Yuzhi; Xu, Hui; Fei, Baojin; Qiao, Dairong; Cao, Yi

2013-07-01

The major objective of this study was to engineer lactic acid bacteria to produce the enzyme phytase from a gene native to Bacillus subtilis GYPB04. The phytase gene (phyC) of B. subtilis GYPB04 was cloned into the plasmid pMG36e for expression in Lactococcus lactis. The enzyme activity in L. lactis cultured in GM17 broth was 20.25 U/mL at 36°C. The expressed phytase was characterized as active in a pH range of 2.0-9.0 at a temperature range of 20-80°C, with an optimum pH of 5.5-6.5 and temperature of 60°C. When cultured in food-grade milk broth, the transformed L. lactis grew to an OD(600 nm) value of 1.05 and had a phytase yield of 13.58 U/mL. In same broth under optimized conditions for cell growth and phytase production, the transformant reached an OD(600 nm) value of 1.68 and a phytase yield of 42.12 U/mL, representing approximately 1.6-fold and 3.1-fold increases, respectively, compared to growth in natural milk broth. Fermentation was scaled to 5 L under optimized conditions, and product analysis revealed a final OD(600 nm) value of 1.89 and an extracellular enzyme activity of 24.23 U/mL. The results of this study may be used in the dairy fermentation industry for the development of functional, healthy yogurts and other fermented dairy foods that provide both active phytase and viable probiotics to the consumer. Copyright © 2013 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Continuous beer fermentation using immobilized yeast cell bioreactor systems.

PubMed

Brányik, Tomás; Vicente, António A; Dostálek, Pavel; Teixeira, José A

2005-01-01

Traditional beer fermentation and maturation processes use open fermentation and lager tanks. Although these vessels had previously been considered indispensable, during the past decades they were in many breweries replaced by large production units (cylindroconical tanks). These have proved to be successful, both providing operating advantages and ensuring the quality of the final beer. Another promising contemporary technology, namely, continuous beer fermentation using immobilized brewing yeast, by contrast, has found only a limited number of industrial applications. Continuous fermentation systems based on immobilized cell technology, albeit initially successful, were condemned to failure for several reasons. These include engineering problems (excess biomass and problems with CO(2) removal, optimization of operating conditions, clogging and channeling of the reactor), unbalanced beer flavor (altered cell physiology, cell aging), and unrealized cost advantages (carrier price, complex and unstable operation). However, recent development in reactor design and understanding of immobilized cell physiology, together with application of novel carrier materials, could provide a new stimulus to both research and application of this promising technology.

Using millet as substrate for efficient production of monacolin K by solid-state fermentation of Monascus ruber.

PubMed

Zhang, Bo-Bo; Xing, Hong-Bo; Jiang, Bing-Jie; Chen, Lei; Xu, Gan-Rong; Jiang, Yun; Zhang, Da-Yong

2018-03-01

In this study, various grains such as rice, millet, corn, barley and wheat were used as raw materials for monacolin K production by solid-state fermentation of Monascus ruber. Among these substrates, millet was found to be the best one for monacolin K production, by which the yield reached 7.12 mg/g. For enhanced monacolin K production, the effects of fermentation time, charge amount, initial moisture content and inoculum volume were systematically investigated in the solid-state fermentation of M. ruber. Moreover, complementary carbon source and nitrogen source were added for further improving the production of monacolin K. Results showed that the maximum production of monacolin K (19.81 mg/g) could be obtained at the optimal conditions. Compared with the traditional red mold rice, using millet as substrate is promising for high production of monacolin K in the solid-state fermentation of M. ruber. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Effect of the combined physical and chemical treatments with microbial fermentation on corn straw degradation.

PubMed

Guo, Hongwei; Chang, Juan; Yin, Qingqiang; Wang, Ping; Lu, Min; Wang, Xiao; Dang, Xiaowei

2013-11-01

In order to improve corn straw degradation, steam explosion, sodium hydroxide soaking and Aspergillus oryzae fermentation were used. The optimal sodium hydroxide pretreatment condition for lignin degradation was obtained. The degradation rates of hemicellulose, cellulose and lignin were 54.68%, 17.76% and 33.14% for the exploded straw (P<0.05); 67.92%, 2.44% (P>0.05) and 76.54% for the alkali-treated straw (P<0.05); 75.98%, 39.93% and 77.88% for the exploded and alkali-treated straw (P<0.05), respectively. The following microbial fermentation could degrade hemicellulose and cellulose further (P<0.05). Cellulase, amylase and protease activities produced during microbial fermentation in the pretreated corn straw were lower than that in the untreated one (P<0.05); however, glucose content was increased by microbial fermentation (P<0.05). It can be concluded that the combined treatments of steam explosion, sodium hydroxide and microbial fermentation will be a good method for straw degradation. Copyright © 2013 Elsevier Ltd. All rights reserved.

Comprehensive study of the evolution of gas-liquid partitioning of aroma compounds during wine alcoholic fermentation.

PubMed

Morakul, Sumallika; Athes, Violaine; Mouret, Jean-Roch; Sablayrolles, Jean-Marie

2010-09-22

Calculating the gas-liquid partitioning of aromatic molecules during winemaking fermentation is essential to minimize the loss of aroma and to optimize the fermentation conditions. In this study, the effect of the main fermentation parameters on the partition coefficients (ki) of higher alcohols (2-methylpropan-1-ol and 3-methyl butan-1-ol) and esters (ethyl acetate, 3-methyl-1-butyl acetate, and 2-ethyl hexanoate) was assessed. The values of ki were first determined in synthetic media simulating must and wine. They varied considerably with both the hydrophobicity of the compound and the composition of the medium. Then, the effect of temperature on ki was quantified. The absence of any effect of gas composition was also established by replacing air with CO2. Finally, the impact of CO2 stripping was assessed by running specific fermentations in which the rate of CO2 production was kept constant by perfusion with assimilable nitrogen. These fermentations showed that in contrast to temperature and must composition, CO2 stripping did not change the gas-liquid partitioning of higher alcohols and esters.

Optimization of Cellulase Production from Bacteria Isolated from Soil

PubMed Central

Sethi, Sonia; Datta, Aparna; Gupta, B. Lal; Gupta, Saksham

2013-01-01

Cellulase-producing bacteria were isolated from soil and identified as Pseudomonas fluorescens, Bacillus subtilIs, E. coli, and Serratia marcescens. Optimization of the fermentation medium for maximum cellulase production was carried out. The culture conditions like pH, temperature, carbon sources, and nitrogen sources were optimized. The optimum conditions found for cellulase production were 40°C at pH 10 with glucose as carbon source and ammonium sulphate as nitrogen source, and coconut cake stimulates the production of cellulase. Among bacteria, Pseudomonas fluorescens is the best cellulase producer among the four followed by Bacillus subtilis, E. coli, and Serratia marscens. PMID:25937986

Bioprocess optimization for production of thermoalkali-stable protease from Bacillus subtilis K-1 under solid-state fermentation.

PubMed

Singh, Satbir; Bajaj, Bijender Kumar

2016-10-02

Cost-effective production of proteases, which are robust enough to function under harsh process conditions, is always sought after due to their wide industrial application spectra. Solid-state production of enzymes using agro-industrial wastes as substrates is an environment-friendly approach, and it has several advantages such as high productivity, cost-effectiveness, being less labor-intensive, and less effluent production, among others. In the current study, different agro-wastes were employed for thermoalkali-stable protease production from Bacillus subtilis K-1 under solid-state fermentation. Agricultural residues such as cotton seed cake supported maximum protease production (728 U ml(-1)), which was followed by gram husk (714 U ml(-1)), mustard cake (680 U ml(-1)), and soybean meal (653 U ml(-1)). Plackett-Burman design of experiment showed that peptone, moisture content, temperature, phosphates, and inoculum size were the significant variables that influenced the protease production. Furthermore, statistical optimization of three variables, namely peptone, moisture content, and incubation temperature, by response surface methodology resulted in 40% enhanced protease production as compared to that under unoptimized conditions (from initial 728 to 1020 U ml(-1)). Thus, solid-state fermentation coupled with design of experiment tools represents a cost-effective strategy for production of industrial enzymes.

Dietary supplementation with bovine lactoferrampin-lactoferricin produced by Pichia pastoris fed-batch fermentation affects intestinal microflora in weaned piglets.

PubMed

Tang, Xiang-Shan; Shao, Hua; Li, Tie-Jun; Tang, Zhi-Ru; Huang, Rui-Ling; Wang, Sheng-Ping; Kong, Xiang-Feng; Wu, Xin; Yin, Yu-Long

2012-10-01

This work is aimed at investigating the effects of recombinant bovine lactoferrampin-lactoferricin (LFA-LFC) instead of chlortetracycline on intestinal microflora in weaned piglets. The high cost of peptide production from either native digestion or chemical synthesis limits the clinical application of antimicrobial peptides. The expression of recombinant peptides in yeast may be an effective alternative. In the current study, recombinant LFA-LFC was produced via fed-batch fermentation in recombinant strain Pichia pastoris (KM71) XS10. Uniform design U6(6(4)) was used to optimize the fermentation conditions. The target peptide purified via cation-exchange and size-exclusion chromatography was added into the dietary of weaned piglets. After 21 days, the Lactobacilli, Bifidobacteria, and Enterobacteria in the chyme of the gut were quantified using real-time polymerase chain reaction. The results showed that approximately 82 mg of LFA-LFC was secreted into 1 L of medium under optimized conditions. Moreover, purified peptide showed strong antimicrobial activities against all the tested microorganisms. Compared with the control group, the LFA-LFC group increased the amount of Lactobacilli and Bifidobacteria (P<0.05) in the chyme of the stomach, duodenum, jejunum, ileum, colon, and caecum. These results show that dietary supplementation with LFA-LFC can affect intestinal microflora in weaned piglets.

Microbiological Transformation of Terpenes

PubMed Central

Prema, B. R.; Bhattacharyya, P. K.

1962-01-01

Several strains of fungi were tested for their ability to metabolize α-pinene in shake cultures. A strain of Aspergillus niger showing marked efficiency in this respect was selected for further studies. The optimal conditions for fermentation were established with respect to substrate concentration, time, and temperature. From the fermentation products three major metabolites of α-pinene were isolated: a ketone, C10H14O, identified as d-verbenone; an alcohol, C10H16O, identified as d-cis-verbenol; and a crystalline diol, C10H18O2, characterized as d-trans-sobrerol. PMID:16349625

Improved Production and Antitumor Properties of Triterpene Acids from Submerged Culture of Ganoderma lingzhi.

PubMed

Wang, Xiao-Ling; Ding, Zhong-Yang; Liu, Gao-Qiang; Yang, Hailong; Zhou, Guo-Ying

2016-10-20

Triterpene acids (TAs) are the major bioactive constituents in the medicinal fungus Ganoderma lingzhi . However, fermentative production of TAs has not been optimized for commercial use, and whether the TAs isolated from G. lingzhi submerged culture mycelia possess antitumor activity needs to be further proven. In this study, enhanced TA yield and productivity were attained with G. lingzhi using response surface methodology. The interactions of three variables were studied using a Box-Benhnken design, namely initial pH, dissolved oxygen (DO) and fermentation temperature. The optimum conditions were an initial pH of 5.9, 20.0% DO and 28.6 °C. These conditions resulted in a TA yield of 308.1 mg/L in a 5-L stirred bioreactor. Furthermore, the optimized conditions were then successfully scaled up to a production scale of 200 L, and maximum TA production and productivity of 295.3 mg/L and 49.2 mg/L/day were achieved, which represented 80.9% and 111.5% increases, respectively, compared with the non-optimized conditions. Additionally, the triterpene acid extract (TAE) from G. lingzhi mycelia was found to be cytotoxic to the SMMC-7721 and SW620 cell lines in vitro, and the TAE exhibited dose-dependent antitumor activity against the solid tumor sarcoma 180 in vivo. Chemical analysis revealed that the key active triterpene compounds, ganoderic acid T and ganoderic acid Me, predominated in the extract.

Thermophilic Gram-Positive Biocatalysts for Biomass Conversion to Ethanol

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shanmugam, K.T.; Ingram, L.O.; Maupin-Furlow, J.A.

2003-12-01

Production of energy from renewable sources is receiving increased attention due to the finite nature of fossil fuels and the environmental impact associated with the continued large scale use of fossil energy sources. Biomass, a CO2-neutral abundant resource, is an attractive alternate source of energy. Biomass-derived sugars, such as glucose, xylose, and other minor sugars, can be readily fermented to fuel ethanol and commodity chemicals. Extracellular cellulases produced by fungi are commercially developed for depolymerization of cellulose in biomass to glucose for fermentation by appropriate biocatalysts in a simultaneous saccharification and fermentation (SSF) process. Due to the differences in themore » optimum conditions for the activity of the fungal cellulases and the growth and fermentation characteristics of the current industrial biocatalysts, SSF of cellulose is envisioned at conditions that are not optimal for the fungal cellulase activity leading to higher than required cost of cellulase in SSF. We have isolated bacterial biocatalysts whose growth and fermentation requirements match the optimum conditions for commercial fungal cellulase activity (pH 5.0 and 50 deg. C). These isolates fermented both glucose and xylose, major components of cellulose and hemicellulose, respectively, to L(+)-lactic acid. Xylose was metabolized through the pentose-phosphate pathway by these organisms as evidenced by the fermentation profile and analysis of the fermentation products of 13C1-xylose by NMR. As expected for the metabolism of xylose by the pentose-phosphate pathway, 13C-lactate accounted for more than 90% of the total 13C-labeled products. All three strains fermented crystalline cellulose to lactic acid with the addition of fungal cellulase (Spezyme CE) (SSF) at an optimum of about 10 FPU/g cellulose. These isolates also fermented cellulose and sugar cane bagasse hemicellulose acid hydrolysate simultaneously. Based on fatty acid profile and 16S rRNA sequence, these isolates cluster with Bacillus coagulans although B. coagulans type strain, ATCC 7050, failed to utilize xylose as a carbon source. For successful production of ethanol from pyruvate, both pyruvate decarboxylase (PDC) and alcohol dehydrogenase (AHD) need to be produced at optimal levels in these biocatalysts. A plasmid containing the S. ventriculi pdc gene and the adh gene from geobacillus stearothermophilus was constructed using plasmid pWH1520 that was successfully used for expression of pdc in B. megaterium. The resulting portable ethanol (PET) plasmid, pJAM423, was transformed into B. megaterium. After xylose induction, a significant fraction of cell cytoplasm was composed of the S. ventriculi PDC and G. stearothermophilus ADH proteins. In preliminary experiments, the amount of ethanol produced by b. megaterium with plasmid pJAM423 was about twice (20 mM) of the bacterium without the plasmid. These results show that the PET operon is functional in B. megaterium but high level ethanol production needs further genetic and metabolic engineering. A genetic transfer system for the second generation biocatalysts needs to be developed for transferring the plasmid pJAM423 and its derivatives for engineering these organisms for ethanol production from biomass derived sugars and cellulose to ethanol. One of the new biocatalysts, strain P4-102B was found to be transformable with plasmids and the method for introducing plasmid pJAM423 into this strain and expression of the encoded DNA is being optimized. These new second generation biocatalysts have the potential to reduce the cost of SSF by minimizing the amount of fungal cellulases, a significant cost component in the use of biomass as a renewable resource for production of fuels and chemicals.« less

Trehalose accumulation from corn starch by Saccharomycopsis fibuligera A11 during 2-l fermentation and trehalose purification.

PubMed

Chi, Zhe; Wang, Ji-Ming; Chi, Zhen-Ming; Ye, Fang

2010-01-01

In this study, corn starch was used as the substrate for cell growth and trehalose accumulation by Saccharomycopsis fibuligera A11. Effect of different aeration rates, agitation speeds, and concentrations of corn starch on direct conversion of corn starch to trehalose by S. fibuligera A11 were examined using a Biostat B2 2-l fermentor. We found that the optimal conditions for direct conversion of corn starch to trehalose by this yeast strain were that agitation speed was 200 rpm, aeration rate was 4.0 l/min, concentration of corn starch was 2.0% (w/v), initial pH was 5.5, fermentation temperature was 30 degrees C. Under these conditions, over 22.9 g of trehalose per 100 g of cell dry weight was accumulated in the yeast cells, cell mass was 15.2 g/l of the fermentation medium, 0.12% (w/v) of reducing sugar, and 0.21% (w/v) of total sugar were left in the fermented medium within 48 h of the fermentation. It was found that trehalose in the yeast cells could be efficiently extracted by the hot distilled water (80 degrees C). After isolation and purification, the crystal trehalose was obtained from the extract of the cells.

Production of Fumaric Acid in 20-Liter Fermentors

PubMed Central

Rhodes, R. A.; Lagoda, A. A.; Misenheimer, T. J.; Smith, M. L.; Anderson, R. F.; Jackson, R. W.

1962-01-01

The conditions necessary for the production of fumaric acid in 20-liter fermentors by fermentation of glucose with Rhizopus arrhizus strain NRRL 2582 were determined. Continuous neutralization of fumaric acid was necessary for optimal yields. Yields of the calcium salt were in excess of 65 g of fumaric acid from 100 g of sugar consumed during fermentation of sugar concentrations of 10 to 16%. Conditions established for calcium fumarate production include a simple mineral salts medium, 0.5 v:v:min aeration rate, 300 rev/min agitation rate in a baffled tank, 33 C incubation temperature, CaCO3 to neutralize the acid formed, and a 4 to 5% (v/v) vegetative inoculum. A suitable procedure and medium for the preparation of a vigorous vegetative inoculum were established. The tendency for calcium fumarate fermentations to foam excessively was controlled with a proper antifoam agent added prior to sterilization of the medium and again at daily intervals during fermentation. The production of soluble sodium or potassium fumarates was inhibited when the concentration of fumarates reached 3.5 to 4.0%. No means of overcoming this inhibition was found. Starches and certain other grain-derived carbohydrates were fermented to form calcium fumarate in flask experiments with approximately the same efficiency as was glucose. PMID:16349614

Study of the role of anaerobic metabolism in succinate production by Enterobacter aerogenes.

PubMed

Tajima, Yoshinori; Kaida, Kenichi; Hayakawa, Atsushi; Fukui, Keita; Nishio, Yousuke; Hashiguchi, Kenichi; Fudou, Ryosuke; Matsui, Kazuhiko; Usuda, Yoshihiro; Sode, Koji

2014-09-01

Succinate is a core biochemical building block; optimizing succinate production from biomass by microbial fermentation is a focus of basic and applied biotechnology research. Lowering pH in anaerobic succinate fermentation culture is a cost-effective and environmentally friendly approach to reducing the use of sub-raw materials such as alkali, which are needed for neutralization. To evaluate the potential of bacteria-based succinate fermentation under weak acidic (pH <6.2) and anaerobic conditions, we characterized the anaerobic metabolism of Enterobacter aerogenes AJ110637, which rapidly assimilates glucose at pH 5.0. Based on the profile of anaerobic products, we constructed single-gene knockout mutants to eliminate the main anaerobic metabolic pathways involved in NADH re-oxidation. These single-gene knockout studies showed that the ethanol synthesis pathway serves as the dominant NADH re-oxidation pathway in this organism. To generate a metabolically engineered strain for succinate production, we eliminated ethanol formation and introduced a heterogeneous carboxylation enzyme, yielding E. aerogenes strain ΔadhE/PCK. The strain produced succinate from glucose with a 60.5% yield (grams of succinate produced per gram of glucose consumed) at pH <6.2 and anaerobic conditions. Thus, we showed the potential of bacteria-based succinate fermentation under weak acidic conditions.

Optimum design and operation of primary sludge fermentation schemes for volatile fatty acids production.

PubMed

Chanona, J; Ribes, J; Seco, A; Ferrer, J

2006-01-01

This paper presents a model-knowledge based algorithm for optimising the primary sludge fermentation process design and operation. This is a recently used method to obtain the volatile fatty acids (VFA), needed to improve biological nutrient removal processes, directly from the raw wastewater. The proposed algorithm consists in a heuristic reasoning algorithm based on the expert knowledge of the process. Only effluent VFA and the sludge blanket height (SBH) have to be set as design criteria, and the optimisation algorithm obtains the minimum return sludge and waste sludge flow rates which fulfil those design criteria. A pilot plant fed with municipal raw wastewater was operated in order to obtain experimental results supporting the developed algorithm groundwork. The experimental results indicate that when SBH was increased, higher solids retention time was obtained in the settler and VFA production increased. Higher recirculation flow-rates resulted in higher VFA production too. Finally, the developed algorithm has been tested by simulating different design conditions with very good results. It has been able to find the optimal operation conditions in all cases on which preset design conditions could be achieved. Furthermore, this is a general algorithm that can be applied to any fermentation-elutriation scheme with or without fermentation reactor.

Production of Mannitol from a High Concentration of Glucose by Candida parapsilosis SK26.001.

PubMed

Meng, Qing; Zhang, Tao; Wei, Wenting; Mu, Wanmeng; Miao, Ming

2017-01-01

A novel strain, SK26.001, which can produce mannitol from a high concentration of glucose without the addition of fructose, was isolated from sugarcane juice. This strain was identified as Candida parapsilosis based on 18S ribosomal RNA (rRNA) sequence analysis and the morphological and physiological-biochemical characteristics of the strain. Under optimized fermentation conditions, the mannitol concentration in shake flasks reached 68.5 g/L. When batch fermentation was performed, the fed glucose was completely consumed after 72 h, resulting in a final mannitol concentration of 80.3 g/L. Fed-batch fermentation was then performed with glucose feed. During the fed-batch process, ammonia water was added to maintain the pH at 4.0. The mannitol concentration in the fermenter reached 97.1 g/L after 120 h, with a total glucose consumption of 284 g/L.

Producing Acetic Acid of Acetobacter pasteurianus by Fermentation Characteristics and Metabolic Flux Analysis.

PubMed

Wu, Xuefeng; Yao, Hongli; Liu, Qing; Zheng, Zhi; Cao, Lili; Mu, Dongdong; Wang, Hualin; Jiang, Shaotong; Li, Xingjiang

2018-03-19

The acetic acid bacterium Acetobacter pasteurianus plays an important role in acetic acid fermentation, which involves oxidation of ethanol to acetic acid through the ethanol respiratory chain under specific conditions. In order to obtain more suitable bacteria for the acetic acid industry, A. pasteurianus JST-S screened in this laboratory was compared with A. pasteurianus CICC 20001, a current industrial strain in China, to determine optimal fermentation parameters under different environmental stresses. The maximum total acid content of A. pasteurianus JST-S was 57.14 ± 1.09 g/L, whereas that of A. pasteurianus CICC 20001 reached 48.24 ± 1.15 g/L in a 15-L stir stank. Metabolic flux analysis was also performed to compare the reaction byproducts. Our findings revealed the potential value of the strain in improvement of industrial vinegar fermentation.

Co-generation of microbial lipid and bio-butanol from corn cob bagasse in an environmentally friendly biorefinery process.

PubMed

Cai, Di; Dong, Zhongshi; Wang, Yong; Chen, Changjing; Li, Ping; Qin, Peiyong; Wang, Zheng; Tan, Tianwei

2016-09-01

Biorefinery process of corn cob bagasse was investigated by integrating microbial lipid and ABE fermentation. The effects of NaOH concentration on the fermentations performance were evaluated. The black liquor after pretreatment was used as substrate for microbial lipid fermentation, while the enzymatic hydrolysates of the bagasse were used for ABE fermentation. The results demonstrated that under the optimized condition, the cellulose and hemicellulose in raw material could be effectively utilized. Approximate 87.7% of the polysaccharides were converted into valuable biobased products (∼175.7g/kg of ABE along with ∼36.6g/kg of lipid). At the same time, almost half of the initial COD (∼48.9%) in the black liquor could be degraded. The environmentally friendly biorefinery process showed promising in maximizing the utilization of biomass for future biofuels production. Copyright © 2016 Elsevier Ltd. All rights reserved.

Optimum production and characterization of an acid protease from marine yeast Metschnikowia reukaufii W6b

NASA Astrophysics Data System (ADS)

Li, Jing; Peng, Ying; Wang, Xianghong; Chi, Zhenming

2010-12-01

The marine yeast strain W6b isolated from sediment of the South China Sea was found to produce a cell-bound acid protease. The crude acid protease produced by this marine yeast showed the highest activity at pH 3.5 and 40 °C. The optimal pH and temperature for the crude acid protease were in agreement with those for acid protease produced by the terrestrial yeasts. The optimal medium of the acid protease production was seawater containing 1.0% glucose, 1.5% casein, and 0.5% yeast extract, and the optimal cultivation conditions of the acid protease production were pH 4.0, a temperature of 25 °C and a shaking speed of 140 rmin-1. Under the optimal conditions, 72.5 UmL-1 of acid protease activity could be obtained in cell suspension within 48 h of fermentation at shake flask level. The acid protease production was induced by high-molecular-weight nitrogen sources and repressed by low-molecular-weight nitrogen sources. Skimmed-milk-clotting test showed that the crude acid protease from the cell suspension of the yeast W6b had high skimmed milk coagulability. The acid protease produced by M. reukaufii W6b may have highly potential applications in cheese, food and fermentation industries.

Influence of cultivating conditions on the alpha-galactosidase biosynthesis from a novel strain of Penicillium sp. in solid-state fermentation.

PubMed

Wang, C L; Li, D F; Lu, W Q; Wang, Y H; Lai, C H

2004-01-01

The work is intended to achieve optimum culture conditions of alpha-galactosidase production by a mutant strain Penicillium sp. in solid-state fermentation (SSF). Certain fermentation parameters involving incubation temperature, moisture content, initial pH value, inoculum and load size of medium, and incubation time were investigated separately. The optimal temperature and moisture level for alpha-galactosidase biosynthesis was found to be 30 degrees C and 50%, respectively. The range of pH 5.5-6.5 was favourable. About 40-50 g of medium in 250-ml flask and inoculum over 1.0 x 10(6) spores were suitable for enzyme production. Seventy-five hours of incubation was enough for maximum alpha-galactosidase production. Substrate as wheat bran supplemented with soyabean meal and beet pulp markedly improved the enzyme yield in trays. Under optimum culture conditions, the alpha-galactosidase activity from Penicillium sp. MAFIC-6 indicated 185.2 U g(-1) in tray of SSF. The process on alpha-galactosidase production in laboratory scale may have a potentiality of scaling-up.

Enhanced extraction of butyric acid under high-pressure CO2 conditions to integrate chemical catalysis for value-added chemicals and biofuels.

PubMed

Chun, Jaesung; Choi, Okkyoung; Sang, Byoung-In

2018-01-01

Extractive fermentation with the removal of carboxylic acid requires low pH conditions because acids are better partitioned into the solvent phase at low pH values. However, this requirement conflicts with the optimal near-neutral pH conditions for microbial growth. CO 2 pressurization was used, instead of the addition of chemicals, to decrease pH for the extraction of butyric acid, a fermentation product of Clostridium tyrobutyricum , and butyl butyrate was selected as an extractant. CO 2 pressurization (50 bar) improved the extraction efficiency of butyric acid from a solution at pH 6, yielding a distribution coefficient ( D ) 0.42. In situ removal of butyric acid during fermentation increased the production of butyric acid by up to 4.10 g/L h, an almost twofold increase over control without the use of an extraction process. In situ extraction of butyric acid using temporal CO 2 pressurization may be applied to an integrated downstream catalytic process for upgrading butyric acid to value-added chemicals in an organic solvent.

Metabolic Flux Analysis during the Exponential Growth Phase of Saccharomyces cerevisiae in Wine Fermentations

PubMed Central

Quirós, Manuel; Martínez-Moreno, Rubén; Albiol, Joan; Morales, Pilar; Vázquez-Lima, Felícitas; Barreiro-Vázquez, Antonio; Ferrer, Pau; Gonzalez, Ramon

2013-01-01

As a consequence of the increase in global average temperature, grapes with the adequate phenolic and aromatic maturity tend to be overripe by the time of harvest, resulting in increased sugar concentrations and imbalanced C/N ratios in fermenting musts. This fact sets obvious additional hurdles in the challenge of obtaining wines with reduced alcohols levels, a new trend in consumer demands. It would therefore be interesting to understand Saccharomyces cerevisiae physiology during the fermentation of must with these altered characteristics. The present study aims to determine the distribution of metabolic fluxes during the yeast exponential growth phase, when both carbon and nitrogen sources are in excess, using continuous cultures. Two different sugar concentrations were studied under two different winemaking temperature conditions. Although consumption and production rates for key metabolites were severely affected by the different experimental conditions studied, the general distribution of fluxes in central carbon metabolism was basically conserved in all cases. It was also observed that temperature and sugar concentration exerted a higher effect on the pentose phosphate pathway and glycerol formation than on glycolysis and ethanol production. Additionally, nitrogen uptake, both quantitatively and qualitatively, was strongly influenced by environmental conditions. This work provides the most complete stoichiometric model used for Metabolic Flux Analysis of S. cerevisiae in wine fermentations employed so far, including the synthesis and release of relevant aroma compounds and could be used in the design of optimal nitrogen supplementation of wine fermentations. PMID:23967264

Oxygen Response of the Wine Yeast Saccharomyces cerevisiae EC1118 Grown under Carbon-Sufficient, Nitrogen-Limited Enological Conditions

PubMed Central

Aceituno, Felipe F.; Orellana, Marcelo; Torres, Jorge; Mendoza, Sebastián; Slater, Alex W.; Melo, Francisco

2012-01-01

Discrete additions of oxygen play a critical role in alcoholic fermentation. However, few studies have quantitated the fate of dissolved oxygen and its impact on wine yeast cell physiology under enological conditions. We simulated the range of dissolved oxygen concentrations that occur after a pump-over during the winemaking process by sparging nitrogen-limited continuous cultures with oxygen-nitrogen gaseous mixtures. When the dissolved oxygen concentration increased from 1.2 to 2.7 μM, yeast cells changed from a fully fermentative to a mixed respirofermentative metabolism. This transition is characterized by a switch in the operation of the tricarboxylic acid cycle (TCA) and an activation of NADH shuttling from the cytosol to mitochondria. Nevertheless, fermentative ethanol production remained the major cytosolic NADH sink under all oxygen conditions, suggesting that the limitation of mitochondrial NADH reoxidation is the major cause of the Crabtree effect. This is reinforced by the induction of several key respiratory genes by oxygen, despite the high sugar concentration, indicating that oxygen overrides glucose repression. Genes associated with other processes, such as proline uptake, cell wall remodeling, and oxidative stress, were also significantly affected by oxygen. The results of this study indicate that respiration is responsible for a substantial part of the oxygen response in yeast cells during alcoholic fermentation. This information will facilitate the development of temporal oxygen addition strategies to optimize yeast performance in industrial fermentations. PMID:23001663

Oxygen response of the wine yeast Saccharomyces cerevisiae EC1118 grown under carbon-sufficient, nitrogen-limited enological conditions.

PubMed

Aceituno, Felipe F; Orellana, Marcelo; Torres, Jorge; Mendoza, Sebastián; Slater, Alex W; Melo, Francisco; Agosin, Eduardo

2012-12-01

Discrete additions of oxygen play a critical role in alcoholic fermentation. However, few studies have quantitated the fate of dissolved oxygen and its impact on wine yeast cell physiology under enological conditions. We simulated the range of dissolved oxygen concentrations that occur after a pump-over during the winemaking process by sparging nitrogen-limited continuous cultures with oxygen-nitrogen gaseous mixtures. When the dissolved oxygen concentration increased from 1.2 to 2.7 μM, yeast cells changed from a fully fermentative to a mixed respirofermentative metabolism. This transition is characterized by a switch in the operation of the tricarboxylic acid cycle (TCA) and an activation of NADH shuttling from the cytosol to mitochondria. Nevertheless, fermentative ethanol production remained the major cytosolic NADH sink under all oxygen conditions, suggesting that the limitation of mitochondrial NADH reoxidation is the major cause of the Crabtree effect. This is reinforced by the induction of several key respiratory genes by oxygen, despite the high sugar concentration, indicating that oxygen overrides glucose repression. Genes associated with other processes, such as proline uptake, cell wall remodeling, and oxidative stress, were also significantly affected by oxygen. The results of this study indicate that respiration is responsible for a substantial part of the oxygen response in yeast cells during alcoholic fermentation. This information will facilitate the development of temporal oxygen addition strategies to optimize yeast performance in industrial fermentations.

Improved cellulase production by Botryosphaeria rhodina from OPEFB at low level moisture condition through statistical optimization.

PubMed

Bahrin, E K; Ibrahim, M F; Abd Razak, M N; Abd-Aziz, S; Shah, U K Md; Alitheen, N; Salleh, M Md

2012-01-01

The response surface method was applied in this study to improve cellulase production from oil palm empty fruit bunch (OPEFB) by Botryosphaeria rhodina. An experimental design based on a two-level factorial was employed to screen the significant environmental factors for cellulase production. The locally isolated fungus Botryosphaeria rhodina was cultivated on OPEFB under solid-state fermentation (SSF). From the analysis of variance (ANOVA), the initial moisture content, amount of substrate, and initial pH of nutrient supplied in the SSF system significantly influenced cellulase production. Then the optimization of the variables was done using the response surface method according to central composite design (CCD). Botryosphaeria rhodina exhibited its best performance with a high predicted value of FPase enzyme production (17.95 U/g) when the initial moisture content was at 24.32%, initial pH of nutrient was 5.96, and 3.98 g of substrate was present. The statistical optimization from actual experiment resulted in a significant increment of FPase production from 3.26 to 17.91 U/g (5.49-fold). High cellulase production at low moisture content is a very rare condition for fungi cultured in solid-state fermentation.

New Protocol Based on UHPLC-MS/MS for Quantitation of Metabolites in Xylose-Fermenting Yeasts

NASA Astrophysics Data System (ADS)

Campos, Christiane Gonçalves; Veras, Henrique César Teixeira; de Aquino Ribeiro, José Antônio; Costa, Patrícia Pinto Kalil Gonçalves; Araújo, Katiúscia Pereira; Rodrigues, Clenilson Martins; de Almeida, João Ricardo Moreira; Abdelnur, Patrícia Verardi

2017-12-01

Xylose fermentation is a bottleneck in second-generation ethanol production. As such, a comprehensive understanding of xylose metabolism in naturally xylose-fermenting yeasts is essential for prospection and construction of recombinant yeast strains. The objective of the current study was to establish a reliable metabolomics protocol for quantification of key metabolites of xylose catabolism pathways in yeast, and to apply this protocol to Spathaspora arborariae. Ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) was used to quantify metabolites, and afterwards, sample preparation was optimized to examine yeast intracellular metabolites. S. arborariae was cultivated using xylose as a carbon source under aerobic and oxygen-limited conditions. Ion pair chromatography (IPC) and hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) were shown to efficiently quantify 14 and 5 metabolites, respectively, in a more rapid chromatographic protocol than previously described. Thirteen and eleven metabolites were quantified in S. arborariae under aerobic and oxygen-limited conditions, respectively. This targeted metabolomics protocol is shown here to quantify a total of 19 metabolites, including sugars, phosphates, coenzymes, monosaccharides, and alcohols, from xylose catabolism pathways (glycolysis, pentose phosphate pathway, and tricarboxylic acid cycle) in yeast. Furthermore, to our knowledge, this is the first time that intracellular metabolites have been quantified in S. arborariae after xylose consumption. The results indicated that fine control of oxygen levels during fermentation is necessary to optimize ethanol production by S. arborariae. The protocol presented here may be applied to other yeast species and could support yeast genetic engineering to improve second generation ethanol production. [Figure not available: see fulltext.

Biodegradation of toxic chemicals by Pleurotus eryngii in submerged fermentation and solid-state fermentation.

PubMed

Chang, Bea-Ven; Chang, Yi-Ming

2016-04-01

The toxic chemicals bisphenol A (BPA), bisphenol F (BPF), nonylphenol (NP), and tetrabromobisphenol A (TBBPA) are endocrine-disrupting chemicals that have consequently drawn much concern regarding their effect on the environment. The objectives of this study were to investigate the degradation of BPA, BPF, NP, and TBBPA by enzymes from Pleurotus eryngii in submerged fermentation (SmF) and solid-state fermentation (SSF), and also to assess the removal of toxic chemicals in spent mushroom compost (SMC). BPA and BPF were analyzed by high-performance liquid chromatography; NP and TBBPA were analyzed by gas chromatography. NP degradation was enhanced by adding CuSO4 (1 mM), MnSO4 (0.5 mM), gallic acid (1 mM), tartaric acid (20 mM), citric acid (20 mM), guaiacol (1 mM), or 2,2'-azino-bis- (3-ethylbenzothiazoline-6-sulfonic acid; 1 mM), with the last yielding a higher NP degradation rate than the other additives from SmF. The optimal conditions for enzyme activity from SSF were a sawdust/wheat bran ratio of 1:4 and a moisture content of 5 mL/g. The enzyme activities were higher with sawdust/wheat bran than with sawdust/rice bran. The optimal conditions for the extraction of enzyme from SMC required using sodium acetate buffer (pH 5.0, solid/solution ratio 1:5), and extraction over 3 hours. The removal rates of toxic chemicals by P. eryngii, in descending order of magnitude, were SSF > SmF > SMC. The removal rates were BPF > BPA > NP > TBBPA. Copyright © 2014. Published by Elsevier B.V.

Improved production of tannase by Klebsiella pneumoniae using Indian gooseberry leaves under submerged fermentation using Taguchi approach.

PubMed

Kumar, Mukesh; Singh, Amrinder; Beniwal, Vikas; Salar, Raj Kumar

2016-12-01

Tannase (tannin acyl hydrolase E.C 3.1.1.20) is an inducible, largely extracellular enzyme that causes the hydrolysis of ester and depside bonds present in various substrates. Large scale industrial application of this enzyme is very limited owing to its high production costs. In the present study, cost effective production of tannase by Klebsiella pneumoniae KP715242 was studied under submerged fermentation using different tannin rich agro-residues like Indian gooseberry leaves (Phyllanthus emblica), Black plum leaves (Syzygium cumini), Eucalyptus leaves (Eucalyptus glogus) and Babul leaves (Acacia nilotica). Among all agro-residues, Indian gooseberry leaves were found to be the best substrate for tannase production under submerged fermentation. Sequential optimization approach using Taguchi orthogonal array screening and response surface methodology was adopted to optimize the fermentation variables in order to enhance the enzyme production. Eleven medium components were screened primarily by Taguchi orthogonal array design to identify the most contributing factors towards the enzyme production. The four most significant contributing variables affecting tannase production were found to be pH (23.62 %), tannin extract (20.70 %), temperature (20.33 %) and incubation time (14.99 %). These factors were further optimized with central composite design using response surface methodology. Maximum tannase production was observed at 5.52 pH, 39.72 °C temperature, 91.82 h of incubation time and 2.17 % tannin content. The enzyme activity was enhanced by 1.26 fold under these optimized conditions. The present study emphasizes the use of agro-residues as a potential substrate with an aim to lower down the input costs for tannase production so that the enzyme could be used proficiently for commercial purposes.

Optimization of Medium Composition for the Production of Neomycin by Streptomyces fradiae NCIM 2418 in Solid State Fermentation

PubMed Central

Vastrad, B. M.; Neelagund, S. E.

2014-01-01

Neomycin production of Streptomyces fradiae NCIM 2418 was optimized by using response surface methodology (RSM), which is powerful mathematical approach comprehensively applied in the optimization of solid state fermentation processes. In the first step of optimization, with Placket-Burman design, ammonium chloride, sodium nitrate, L-histidine, and ammonium nitrate were established to be the crucial nutritional factors affecting neomycin production significantly. In the second step, a 24 full factorial central composite design and RSM were applied to determine the optimal concentration of significant variable. A second-order polynomial was determined by the multiple regression analysis of the experimental data. The optimum values for the important nutrients for the maximum were obtained as follows: ammonium chloride 2.00%, sodium nitrate 1.50%, L-histidine 0.250%, and ammonium nitrate 0.250% with a predicted value of maximum neomycin production of 20,000 g kg−1 dry coconut oil cake. Under the optimal condition, the practical neomycin production was 19,642 g kg−1 dry coconut oil cake. The determination coefficient (R 2) was 0.9232, which ensures an acceptable admissibility of the model. PMID:25009746

Optimization of cultivation conditions of fermented shaggy ink cap culinary-medicinal mushroom, Coprinus comatus (O.Mull.:Fr.) Pers. (higher Basidiomycetes) rich in Vanadium.

PubMed

Zhang, Chunjing; Qi, Xiaodan; Shi, Yan; Sun, Yan; Li, Shuyan; Gao, Xiulan; Yu, Haitao

2012-01-01

The present paper is mainly aimed at optimization of cultivation conditions of fermented mushrooms of Coprinus comatus rich in vanadium (CCRV). Initial screening of effects of carbon source, temperature, pH, and inoculum size were done by using a one-factor-at-a-time method. The results obtained in that study showed that the optimal medium composition was 30 g glucose/Lin YEPG medium, initial pH 6.0, inoculum volume 10%, and incubation time 120 h. Then the medium was subjected to screening of the most significant parameters using the L9 orthogonal array to solve multivariable equations simultaneously. The results obtained in this study showed that the optimal medium composition was 0.4% V and 30 g glucose/Lin YEPG medium, initial pH 5.0, inoculum volume 15%, and incubation time 120 h. At this medium composition, the mycelial biomass and V content were 7.18 ± 0.24 g/L and 3786.0 ± 17 μg/g, respectively. The anti-diabetic potential of CCRV produced with the optimal level was tested in alloxan-induced diabetes. After the mice were administered (i.g.) with CCRV, the level of blood sugar in the CCRV group was very close to that of the control group. These findings suggested that CCRV produced with the optimal level is useful in the control of diabetes mellitus.

Deletion of FPS1, Encoding Aquaglyceroporin Fps1p, Improves Xylose Fermentation by Engineered Saccharomyces cerevisiae

PubMed Central

Wei, Na; Xu, Haiqing; Kim, Soo Rin

2013-01-01

Accumulation of xylitol in xylose fermentation with engineered Saccharomyces cerevisiae presents a major problem that hampers economically feasible production of biofuels from cellulosic plant biomass. In particular, substantial production of xylitol due to unbalanced redox cofactor usage by xylose reductase (XR) and xylitol dehydrogenase (XDH) leads to low yields of ethanol. While previous research focused on manipulating intracellular enzymatic reactions to improve xylose metabolism, this study demonstrated a new strategy to reduce xylitol formation and increase carbon flux toward target products by controlling the process of xylitol secretion. Using xylitol-producing S. cerevisiae strains expressing XR only, we determined the role of aquaglyceroporin Fps1p in xylitol export by characterizing extracellular and intracellular xylitol. In addition, when FPS1 was deleted in a poorly xylose-fermenting strain with unbalanced XR and XDH activities, the xylitol yield was decreased by 71% and the ethanol yield was substantially increased by nearly four times. Experiments with our optimized xylose-fermenting strain also showed that FPS1 deletion reduced xylitol production by 21% to 30% and increased ethanol yields by 3% to 10% under various fermentation conditions. Deletion of FPS1 decreased the xylose consumption rate under anaerobic conditions, but the effect was not significant in fermentation at high cell density. Deletion of FPS1 resulted in higher intracellular xylitol concentrations but did not significantly change the intracellular NAD+/NADH ratio in xylose-fermenting strains. The results demonstrate that Fps1p is involved in xylitol export in S. cerevisiae and present a new gene deletion target, FPS1, and a mechanism different from those previously reported to engineer yeast for improved xylose fermentation. PMID:23475614

Enhanced saturated fatty acids accumulation in cultures of newly-isolated strains of Schizochytrium sp. and Thraustochytriidae sp. for large-scale biodiesel production.

PubMed

Wang, Qiuzhen; Sen, Biswarup; Liu, Xianhua; He, Yaodong; Xie, Yunxuan; Wang, Guangyi

2018-08-01

Heterotrophic marine protists (Thraustochytrids) have received increasingly global attention as a renewable, sustainable and alternative source of biodiesel because of their high ability of saturated fatty acids (SFAs) accumulation. Yet, the influence of extrinsic factors (nutrients and environmental conditions) on thraustochytrid culture and optimal conditions for high SFAs production are poorly described. In the present study, two different thraustochytrid strains, Schizochytrium sp. PKU#Mn4 and Thraustochytriidae sp. PKU#Mn16 were studied for their growth and SFAs production profiles under various conditions (carbon, nitrogen, temperature, pH, KH 2 PO 4 , salinity, and agitation speed). Of the culture conditions, substrates (C and N) source and conc., temperature, and agitation speed significantly influenced the cell growth and SFAs production of both strains. Although both the strains were capable of growth and SFAs production in the broad range of culture conditions, their physiological responses to KH 2 PO 4 , pH, and salinity were dissimilar. Under their optimal batch culture conditions, peak SFAs productions of 3.3g/L and 2.2g/L with 62% and 49% SFAs contents (relative to total fatty acids) were achieved, respectively. The results of 5-L fed-batch fermentation under optimal conditions showed a nearly 4.5-fold increase in SFAs production (i.e., 7.5g/L) by both strains compared to unoptimized conditions. Of the two strains, the quality of biodiesel produced from the fatty acids of PKU#Mn4 met the biodiesel standard defined by ASTM6751. This study, to the knowledge of the authors, is the first comprehensive report of optimal fermentation conditions demonstrating enhanced SFAs production by strains belonging to two different thraustochytrid genera and provides the basis for large-scale biodiesel production. Copyright © 2018. Published by Elsevier B.V.

Statistical optimization of recycled-paper enzymatic hydrolysis for simultaneous saccharification and fermentation via central composite design.

PubMed

Liu, Qing; Cheng, Ke-ke; Zhang, Jian-an; Li, Jin-ping; Wang, Ge-hua

2010-01-01

A central composite design of the response surface methodology (RSM) was employed to study the effects of temperature, enzyme concentration, and stirring rate on recycled-paper enzymatic hydrolysis. Among the three variables, temperature and enzyme concentration significantly affected the conversion efficiency of substrate, whereas stirring rate was not effective. A quadratic polynomial equation was obtained for enzymatic hydrolysis by multiple regression analysis using RSM. The results of validation experiments were coincident with the predicted model. The optimum conditions for enzymatic hydrolysis were temperature, enzyme concentration, and stirring rate of 43.1 degrees C, 20 FPU g(-1) substrate, and 145 rpm, respectively. In the subsequent simultaneous saccharification and fermentation (SSF) experiment under the optimum conditions, the highest 28.7 g ethanol l(-1) was reached in the fed-batch SSF when 5% (w/v) substrate concentration was used initially, and another 5% added after 12 h fermentation. This ethanol output corresponded to 77.7% of the theoretical yield based on the glucose content in the raw material.

Total solids content: a key parameter of metabolic pathways in dry anaerobic digestion

PubMed Central

2013-01-01

Background In solid-state anaerobic digestion (AD) bioprocesses, hydrolytic and acidogenic microbial metabolisms have not yet been clarified. Since these stages are particularly important for the establishment of the biological reaction, better knowledge could optimize the process performances by process parameters adjustment. Results This study demonstrated the effect of total solids (TS) content on microbial fermentation of wheat straw with six different TS contents ranging from wet to dry conditions (10 to 33% TS). Three groups of metabolic behaviors were distinguished based on wheat straw conversion rates with 2,200, 1,600, and 1,400 mmol.kgVS-1 of fermentative products under wet (10 and 14% TS), dry (19 to 28% TS), and highly dry (28 to 33% TS) conditions, respectively. Furthermore, both wet and dry fermentations showed acetic and butyric acid metabolisms, whereas a mainly butyric acid metabolism occurred in highly dry fermentation. Conclusion Substrate conversion was reduced with no changes of the metabolic pathways until a clear limit at 28% TS content, which corresponded to the threshold value of free water content of wheat straw. This study suggested that metabolic pathways present a limit of TS content for high-solid AD. PMID:24261971

Direct and efficient xylitol production from xylan by Saccharomyces cerevisiae through transcriptional level and fermentation processing optimizations.

PubMed

Li, Zhe; Qu, Hongnan; Li, Chun; Zhou, Xiaohong

2013-12-01

In this study, four engineered Saccharomyces cerevisiae carrying xylanase, β-xylosidase and xylose reductase genes by different transcriptional regulations were constructed to directly convert xylan to xylitol. According to the results, the high-copy number plasmid required a rigid selection for promoter characteristics, on the contrast, the selection of promoters could be more flexible for low-copy number plasmid. For cell growth and xylitol production, glucose and galactose were found more efficient than other sugars. The semi-aerobic condition and feeding of co-substrates were taken to improve the yield of xylitol. It was found that the strain containing high-copy number plasmid had the highest xylitol yield, but it was sensitive to the change of fermentation. However, the strain carrying low-copy number plasmid was more adaptable to different processes. By optimization of the transcriptional regulation and fermentation processes, the xylitol concentration could be increased of 1.7 folds and the yield was 0.71 g xylitol/g xylan. Copyright © 2013 Elsevier Ltd. All rights reserved.

Production of cellulose by Aspergillus niger under submerged and solid state fermentation using coir waste as a substrate

PubMed Central

Mrudula, Soma; Murugammal, Rangasamy

2011-01-01

Aspergillus niger was used for cellulase production in submerged (SmF) and solid state fermentation (SSF). The maximum production of cellulase was obtained after 72 h of incubation in SSF and 96 h in Smf. The CMCase and FPase activities recorded in SSF were 8.89 and 3.56 U per g of dry mycelial bran (DBM), respectively. Where as in Smf the CMase & FPase activities were found to be 3.29 and 2.3 U per ml culture broth, respectively. The productivity of extracellular cellulase in SSF was 14.6 fold higher than in SmF. The physical and nutritional parameters of fermentation like pH, temperature, substrate, carbon and nitrogen sources were optimized. The optimal conditions for maximum biosynthesis of cellulase by A. niger were shown to be at pH 6, temperature 30 °C. The additives like lactose, peptone and coir waste as substrate increased the productivity both in SmF and SSF. The moisture ratio of 1:2 (w/v) was observed for optimum production of cellulase in SSF. PMID:24031730

Fermentation Conditions that Affect Clavulanic Acid Production in Streptomyces clavuligerus: A Systematic Review.

PubMed

Ser, Hooi-Leng; Law, Jodi Woan-Fei; Chaiyakunapruk, Nathorn; Jacob, Sabrina Anne; Palanisamy, Uma Devi; Chan, Kok-Gan; Goh, Bey-Hing; Lee, Learn-Han

2016-01-01

The β-lactamase inhibitor, clavulanic acid is frequently used in combination with β-lactam antibiotics to treat a wide spectrum of infectious diseases. Clavulanic acid prevents drug resistance by pathogens against these β-lactam antibiotics by preventing the degradation of the β-lactam ring, thus ensuring eradication of these harmful microorganisms from the host. This systematic review provides an overview on the fermentation conditions that affect the production of clavulanic acid in the firstly described producer, Streptomyces clavuligerus. A thorough search was conducted using predefined terms in several electronic databases (PubMed, Medline, ScienceDirect, EBSCO), from database inception to June 30th 2015. Studies must involve wild-type Streptomyces clavuligerus, and full texts needed to be available. A total of 29 eligible articles were identified. Based on the literature, several factors were identified that could affect the production of clavulanic acid in S. clavuligerus. The addition of glycerol or other vegetable oils (e.g., olive oil, corn oil) could potentially affect clavulanic acid production. Furthermore, some amino acids such as arginine and ornithine, could serve as potential precursors to increase clavulanic acid yield. The comparison of different fermentation systems revealed that fed-batch fermentation yields higher amounts of clavulanic acid as compared to batch fermentation, probably due to the maintenance of substrates and constant monitoring of certain entities (such as pH, oxygen availability, etc.). Overall, these findings provide vital knowledge and insight that could assist media optimization and fermentation design for clavulanic acid production in S. clavuligerus.

Strategies for Fermentation Medium Optimization: An In-Depth Review

PubMed Central

Singh, Vineeta; Haque, Shafiul; Niwas, Ram; Srivastava, Akansha; Pasupuleti, Mukesh; Tripathi, C. K. M.

2017-01-01

Optimization of production medium is required to maximize the metabolite yield. This can be achieved by using a wide range of techniques from classical “one-factor-at-a-time” to modern statistical and mathematical techniques, viz. artificial neural network (ANN), genetic algorithm (GA) etc. Every technique comes with its own advantages and disadvantages, and despite drawbacks some techniques are applied to obtain best results. Use of various optimization techniques in combination also provides the desirable results. In this article an attempt has been made to review the currently used media optimization techniques applied during fermentation process of metabolite production. Comparative analysis of the merits and demerits of various conventional as well as modern optimization techniques have been done and logical selection basis for the designing of fermentation medium has been given in the present review. Overall, this review will provide the rationale for the selection of suitable optimization technique for media designing employed during the fermentation process of metabolite production. PMID:28111566

Optimization of Culture Medium for the Growth of Candida sp. and Blastobotrys sp. as Starter Culture in Fermentation of Cocoa Beans (Theobroma cacao) Using Response Surface Methodology (RSM).

PubMed

Mahazar, N H; Zakuan, Z; Norhayati, H; MeorHussin, A S; Rukayadi, Y

2017-01-01

Inoculation of starter culture in cocoa bean fermentation produces consistent, predictable and high quality of fermented cocoa beans. It is important to produce healthy inoculum in cocoa bean fermentation for better fermented products. Inoculum could minimize the length of the lag phase in fermentation. The purpose of this study was to optimize the component of culture medium for the maximum cultivation of Candida sp. and Blastobotrys sp. Molasses and yeast extract were chosen as medium composition and Response Surface Methodology (RSM) was then employed to optimize the molasses and yeast extract. Maximum growth of Candida sp. (7.63 log CFU mL-1) and Blastobotrys sp. (8.30 log CFU mL-1) were obtained from the fermentation. Optimum culture media for the growth of Candida sp., consist of 10% (w/v) molasses and 2% (w/v) yeast extract, while for Blastobotrys sp., were 1.94% (w/v) molasses and 2% (w/v) yeast extract. This study shows that culture medium consists of molasses and yeast extract were able to produce maximum growth of Candida sp. and Blastobotrys sp., as a starter culture for cocoa bean fermentation.

Optimization of wastewater microalgae saccharification using dilute acid hydrolysis for acetone, butanol, and ethanol fermentation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Castro, Yessica; Ellis, Joshua T.; Miller, Charles D.

2015-02-01

Exploring and developing sustainable and efficient technologies for biofuel production are crucial for averting global consequences associated with fuel shortages and climate change. Optimization of sugar liberation from wastewater algae through acid hydrolysis was determined for subsequent fermentation to acetone, butanol, and ethanol (ABE) by Clostridium saccharoperbutylacetonicum N1-4. Acid concentration, retention time, and temperature were evaluated to determine optimal hydrolysis conditions by assessing the sugar and ABE yield as well as the associated costs. Sulfuric acid concentrations ranging from 0-1.5 M, retention times of 40-120 min, and temperatures from 23°C- 90°C were combined to form a full factorial experiment. Acidmore » hydrolysis pretreatment of 10% dried wastewater microalgae using 1.0 M sulfuric acid for 120 min at 80-90°C was found to be the optimal parameters, with a sugar yield of 166.1 g for kg of dry algae, concentrations of 5.23 g/L of total ABE, and 3.74 g/L of butanol at a rate of USD $12.83 per kg of butanol.« less

Chitooligomers preparation by chitosanase produced under solid state fermentation using shrimp by-products as substrate.

PubMed

Nidheesh, T; Pal, Gaurav Kumar; Suresh, P V

2015-05-05

Solid state fermentation (SSF) conditions were statistically optimized for the production of chitosanase by Purpureocillium lilacinum CFRNT12 using shrimp by-products as substrate. Central composite design and response surface methodology were applied to evaluate the effect of variables and their optimization. Incubation temperature, incubation time, concentration of inoculum and yeast extract were found to influence the chitosanase production significantly. The R(2) value of 0.94 indicates the aptness of the model. The level of variables for optimal production of chitosanase was 32 ± 1°C temperature, 96 h incubation, 10.5% (w/v) inoculum, 1.05% (w/w) yeast extract and 65% (w/w) moisture content. The chitosanase production was found to increase from 2.34 ± 0.07 to 41.78 ± 0.73 units/g initial dry substrate after optimization. The crude chitosanase produced 4.43 mM of chitooligomers as exclusive end product from colloidal chitosan hydrolysis. These results indicate the potential of P. lilacinum CFRNT12 for the chitosanase production employing cost effective SSF using shrimp by-products. Copyright © 2014 Elsevier Ltd. All rights reserved.

Recombinant host cells and media for ethanol production

DOEpatents

Wood, Brent E; Ingram, Lonnie O; Yomano, Lorraine P; York, Sean W

2014-02-18

Disclosed are recombinant host cells suitable for degrading an oligosaccharide that have been optimized for growth and production of high yields of ethanol, and methods of making and using these cells. The invention further provides minimal media comprising urea-like compounds for economical production of ethanol by recombinant microorganisms. Recombinant host cells in accordance with the invention are modified by gene mutation to eliminate genes responsible for the production of unwanted products other than ethanol, thereby increasing the yield of ethanol produced from the oligosaccharides, relative to unmutated parent strains. The new and improved strains of recombinant bacteria are capable of superior ethanol productivity and yield when grown under conditions suitable for fermentation in minimal growth media containing inexpensive reagents. Systems optimized for ethanol production combine a selected optimized minimal medium with a recombinant host cell optimized for use in the selected medium. Preferred systems are suitable for efficient ethanol production by simultaneous saccharification and fermentation (SSF) using lignocellulose as an oligosaccharide source. The invention also provides novel isolated polynucleotide sequences, polypeptide sequences, vectors and antibodies.

[Transformation of baicalin and wogonoside through liquid fermentation with Bacillus natto].

PubMed

Long, Hou-ning; Zhang, Shuo; Yao, Lei; Zhang, Min; Wang, Peng-jiao; Meng, Xiao-xia; Gao, Xiu; Zhang, Rong-ping

2015-12-01

This experiment aimed to explore and research the process of preparing baicalein and wogonin through liquid fermentation with Bacillus natto. Active enzymes of produced by B. natto was used for the biological transformation of baclin and wogonoside, in order to increase the content of the haicalein and wogonin in the scutellaria. With the content of the baicalein and wogonin as evaluating indexes, the effects of carbon source, nitrogen source, the types and suitable concentration of inorganic salt, medium pH, granularities of medical materials, liquid volume in flask, shaking speed, liquid-to-solid ratio, fermentation time on the fermentation process were studied. The optimal process conditions for liquid fermentation of scutellaria were 1.0% of peptone, 0.05% of NaCl, pH at 6, the granularities of medical materials of the scutellaria screened through 40-mesh sifter, 33% of liquid, shaker incubator speed at 200 r x min(-1), liquid-to-solid ratio of 5:1, temperature at 37 degrees C, fermentation for 6 days, baclin's conversion rate at 97.6% and wogonoside's conversion rate at 97% in the scutellaria. According to the verification test, the process was stable and feasible, and could provide data reference for the industrial production.

Effects of Fermentation Temperature on Key Aroma Compounds and Sensory Properties of Apple Wine.

PubMed

Peng, Bangzhu; Li, Fuling; Cui, Lu; Guo, Yaodong

2015-12-01

Fermentation temperature strongly affects yeast metabolism during apple wine making and thus aromatic and quality profiles. In this study, the temperature effect during apple wine making on both the key aroma compounds and sensory properties of apple wine were investigated. The concentration of nine key aroma compounds (ethyl acetate, isobutyl acetate, isopentylacetate, ethyl caprylate, ethyl 4-hydroxybutanoate, isobutylalcohol, isopentylalcohol, 3-methylthio-1-propanol, and benzeneethanol) in apple wine significantly increased with the increase of fermentation temperature from 17 to 20 °C, and then eight out of the nine key aroma compounds with an exception of ethyl 4-hydroxybutanoate, decreased when the temperature goes up 20 to 26 °C. Sensory analysis showed that the apple wine fermented at 20 °C had the highest acceptance for consumers. Fermentation at the temperature of 20 °C was therefore considered to be the most suitable condition using the selected yeast strain (Saccharomyces cerevisiae AP05) for apple wine making. Changes in the fermentation temperature can considerably affect the production of key aroma compounds and sensory profiles of apple wine. These results could help apple wine producers make better quality production for consumers at the optimal fermentation temperature. © 2015 Institute of Food Technologists®

The impact of oxygen on the final alcohol content of wine fermented by a mixed starter culture.

PubMed

Morales, Pilar; Rojas, Virginia; Quirós, Manuel; Gonzalez, Ramon

2015-05-01

We have developed a wine fermentation procedure that takes advantage of the metabolic features of a previously characterized Metschnikowia pulcherrima strain in order to reduce ethanol production. It involves the use of M. pulcherrima/Saccharomyces cerevisiae mixed cultures, controlled oxygenation conditions during the first 48 h of fermentation, and anaerobic conditions thereafter. The influence of different oxygenation regimes and initial inoculum composition on yeast physiology and final ethanol content was studied. The impact of oxygenation on yeast physiology goes beyond the first aerated step and influences yields and survival rates during the anaerobic stage. The activity of M. pulcherrima in mixed oxygenated cultures resulted in a clear reduction in ethanol yield, as compared to S. cerevisiae. Despite relatively low initial cell numbers, S. cerevisiae always predominated in mixed cultures by the end of the fermentation process. Strain replacement was faster under low oxygenation levels. M. pulcherrima confers an additional advantage in terms of dissolved oxygen, which drops to zero after a few hours of culture, even under highly aerated conditions, and this holds true for mixed cultures. Alcohol reduction values about 3.7 % (v/v) were obtained for mixed cultures under high aeration, but they were associated to unacceptable volatile acidity levels. In contrast, under optimized conditions, only 0.35 g/L acetic acid was produced, for an alcohol reduction of 2.2 % (v/v), and almost null dissolved oxygen during the process.

Actinobacillus succinogenes ATCC 55618 Fermentation Medium Optimization for the Production of Succinic Acid by Response Surface Methodology

PubMed Central

Zhu, Li-Wen; Wang, Cheng-Cheng; Liu, Rui-Sang; Li, Hong-Mei; Wan, Duan-Ji; Tang, Ya-Jie

2012-01-01

As a potential intermediary feedstock, succinic acid takes an important place in bulk chemical productions. For the first time, a method combining Plackett-Burman design (PBD), steepest ascent method (SA), and Box-Behnken design (BBD) was developed to optimize Actinobacillus succinogenes ATCC 55618 fermentation medium. First, glucose, yeast extract, and MgCO3 were identified to be key medium components by PBD. Second, preliminary optimization was run by SA method to access the optimal region of the key medium components. Finally, the responses, that is, the production of succinic acid, were optimized simultaneously by using BBD, and the optimal concentration was located to be 84.6 g L−1 of glucose, 14.5 g L−1 of yeast extract, and 64.7 g L−1 of MgCO3. Verification experiment indicated that the maximal succinic acid production of 52.7 ± 0.8 g L−1 was obtained under the identified optimal conditions. The result agreed with the predicted value well. Compared with that of the basic medium, the production of succinic acid and yield of succinic acid against glucose were enhanced by 67.3% and 111.1%, respectively. The results obtained in this study may be useful for the industrial commercial production of succinic acid. PMID:23093852

Novel stepwise pH control strategy to improve short chain fatty acid production from sludge anaerobic fermentation.

PubMed

Zhao, Jianwei; Wang, Dongbo; Liu, Yiwen; Ngo, Huu Hao; Guo, Wenshan; Yang, Qi; Li, Xiaoming

2018-02-01

This study reports an innovative strategy known as stepwise pH fermentation, developed to enhance the production of short chain volatile fatty acids (SCFA) from waste activated sludge (WAS) anaerobic fermentation. Experimental results confirmed the optimal pH for WAS disruption and acidification was 11 and 9, respectively, and corresponding optimal time was, respectively, 5 d and 2 d. In this scenario, the optimal SCFA yield was 2356 mg chemical oxygen demand (COD)/L, which was much higher than that derived from alkaline fermentation system. Investigation of the mechanism indicated that pH 11 could accelerate the disruption of WAS and inhibit the activities of methanogens; furthermore, pH 9 was beneficial to the activity of acid-producing bacteria, resulting in more SCFA production. Stepwise pH fermentation integrated with sodium chloride (NaCl) present in WAS had synergistic impacts on WAS anaerobic fermentation. Copyright © 2017 Elsevier Ltd. All rights reserved.

Biochemical Characterization of Extracellular Cellulase from Tuber maculatum Mycelium Produced Under Submerged Fermentation.

PubMed

Bedade, Dattatray K; Singhal, Rekha S; Turunen, Ossi; Deska, Jan; Shamekh, Salem

2017-02-01

Interaction of truffle mycelium with the host plant involves the excretion of extracellular enzymes. The ability of Tuber maculatum mycelium to produce an extracellular cellulase during submerged fermentation was demonstrated for the first time. T. maculatum mycelia were isolated and tested for extracellular cellulase production at variable pH on solid agar medium, and the highest activity was observed at pH 7.0. Furthermore, T. maculatum was subjected to submerged fermentation in basal salt medium for cellulase production. Under optimized conditions using sodium carboxymethyl cellulose (0.5 % w/v) as carbon source and an initial pH of 7.0, the enzyme production yielded 1.70 U/mL of cellulase in the cell-free supernatant after 7 days of incubation time. The optimum of the obtained cellulase's activity was at pH 5.0 and a temperature of 50 °C. The enzyme showed good thermostability at 50 °C by retaining 99 % of its maximal activity over an incubation time of 100 min. The cellulase activity was inhibited by Fe 2+ and slightly activated by Mn 2+ and Cu 2+ at 1 mM concentration. The results indicated that truffle mycelium is utilizing cellulosic energy source in the root system, and the optimal conditions are those existing in the acidic Finnish soil.

Evaluation of improved γ-aminobutyric acid production in yogurt using Lactobacillus plantarum NDC75017.

PubMed

Shan, Y; Man, C X; Han, X; Li, L; Guo, Y; Deng, Y; Li, T; Zhang, L W; Jiang, Y J

2015-04-01

Most γ-aminobutyric acid (GABA)-producing microorganisms are lactic acid bacteria (LAB), but the yield of GABA is limited in most of these GABA-producing strains. In this study, the production of GABA was carried out by using Lactobacillus plantarum NDC75017, a strain screened from traditional fermented dairy products in China. Concentrations of substrate (l-monosodium glutamate, L-MSG) and coenzyme (pyridoxal-5-phosphate, PLP) of glutamate decarboxylase (GAD) and culture temperature were investigated to evaluate their effects on GABA yield of Lb. plantarum NDC75017. The results indicated that GABA production was related to GAD activity and biomass of Lb. plantarum NDC75017. Response surface methodology was used to optimize conditions of GABA production. The optimal factors for GABA production were L-MSG at 80 mM, PLP at 18 μM, and a culture temperature of 36 °C. Under these conditions, production of GABA was maximized at 314.56 mg/100 g. Addition of Lb. plantarum NDC75017 to a commercial starter culture led to higher GABA production in fermented yogurt. Flavor and texture of the prepared yogurt and the control yogurt did not differ significantly. Thus, Lb. plantarum NDC75017 has good potential for manufacture of GABA-enriched fermented milk products. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Improving the fermentation production of the individual key triterpene ganoderic acid me by the medicinal fungus Ganoderma lucidum in submerged culture.

PubMed

Liu, Gao-Qiang; Wang, Xiao-Ling; Han, Wen-Jun; Lin, Qin-Lu

2012-10-24

Enhanced ganoderic acid Me (GA-Me, an important anti-tumor triterpene) yield was attained with the medicinal fungus Ganoderma lucidum using response surface methodology (RSM). Interactions were studied with three variables, viz. glucose, peptone and culture time using a Central Composite Design (CCD). The CCD contains a total of 20 experiments with the first 14 experiments organized in a fractional factorial design, with the experimental trails from 15 to 20 involving the replications of the central points. A polynomial model, describing the relationships between the yield of GA-Me and the three factors in a second-order equation, was developed. The model predicted the maximum GA-Me yield of 11.9 mg·L−1 for glucose, peptone, culture time values of 44.4 g·L−1, 5.0 g·L−1, 437.1 h, respectively, and a maximum GA-Me yield of 12.4 mg·L−1 was obtained in the validation experiment, which represented a 129.6% increase in titre compared to that of the non-optimized conditions. In addition, 11.4 mg·L−1 of GA-Me was obtained in a 30-L agitated fermenter under the optimized conditions, suggesting the submerged culture conditions optimized in the present study were also suitable for GA-Me production on a large scale.

Optimization of dilute sulfuric acid pretreatment to maximize combined sugar yield from sugarcane bagasse for ethanol production.

PubMed

Benjamin, Y; Cheng, H; Görgens, J F

2014-01-01

Increasing fermentable sugar yields per gram of biomass depends strongly on optimal selection of varieties and optimization of pretreatment conditions. In this study, dilute acid pretreatment of bagasse from six varieties of sugarcane was investigated in connection with enzymatic hydrolysis for maximum combined sugar yield (CSY). The CSY from the varieties were also compared with the results from industrial bagasse. The results revealed considerable differences in CSY between the varieties. Up to 22.7 % differences in CSY at the optimal conditions was observed. The combined sugar yield difference between the best performing variety and the industrial bagasse was 34.1 %. High ratio of carbohydrates to lignin and low ash content favored the release of sugar from the substrates. At mild pretreatment conditions, the differences in bioconversion efficiency between varieties were greater than at severe condition. This observation suggests that under less severe conditions the glucose recovery was largely determined by chemical composition of biomass. The results from this study support the possibility of increasing sugar yields or improving the conversion efficiency when pretreatment optimization is performed on varieties with improved properties.

Recovering folic acid and its identification on mixed pastes of tempeh and fermented vegetable as natural source of folic acid

NASA Astrophysics Data System (ADS)

Susilowati, Agustine; Aspiyanto, Maryati, Yati; Melanie, Hakiki; Lotulung, Puspa D.

2017-11-01

Mixing between tempeh and both fermented broccoli (Brassica oleracea) and spinach (Amaranthus sp.) were conducted to achieve mixed pastes as natural source of folic acid for 'smart food'. Mixing was performed on soy, mung bean, and kidney bean tempehs with both fermented broccoli and spinach at ratio of 1 : 1, 1 : 2, 1 : 3, 1 : 4, 1 : 5 and 1 : 6, respectively. Result of experimental activity showed that pulverizing ratio becoming more and more low will decrease total solids, soluble protein and N-Amino, but fluctuates on folic acid in mixed paste. Based on folic acid equivalent and the best fermented vegetable efficiency, optimization condition was reached in paste with combination between mung beans tempeh and fermented spinach at ratio of 1 : 2 by increasing folic acid concentration of 83.18 % (0.83 times), dissolved protein 432.29 % (4.32 times) and N-amino 55.36 % (0,55 times). While, it is occurred a lowering total solids 22.16 % (0.22 times) when compared with folic acid, soluble protein, N-amino, and total solids on initial materials of mung bean tempeh. In this condition, it is achieved folic acid monomer with molecular weight (MW) 148.14 Da. with relative intensity 100 %, and glutamic acid monomer 443.50 Da.with relative intensity 0.07 %.

Consolidated bioprocessing strategy for ethanol production from Jerusalem artichoke tubers by Kluyveromyces marxianus under high gravity conditions.

PubMed

Yuan, W J; Chang, B L; Ren, J G; Liu, J P; Bai, F W; Li, Y Y

2012-01-01

Developing an innovative process for ethanol fermentation from Jerusalem artichoke tubers under very high gravity (VHG) conditions. A consolidated bioprocessing (CBP) strategy that integrated inulinase production, saccharification of inulin contained in Jerusalem artichoke tubers and ethanol production from sugars released from inulin by the enzyme was developed with the inulinase-producing yeast Kluyveromyces marxianus Y179 and fed-batch operation. The impact of inoculum age, aeration, the supplementation of pectinase and nutrients on the ethanol fermentation performance of the CBP system was studied. Although inulinase activities increased with the extension of the seed incubation time, its contribution to ethanol production was negligible because vigorously growing yeast cells harvested earlier carried out ethanol fermentation more efficiently. Thus, the overnight incubation that has been practised in ethanol production from starch-based feedstocks is recommended. Aeration facilitated the fermentation process, but compromised ethanol yield because of the negative Crabtree effect of the species, and increases the risk of contamination under industrial conditions. Therefore, nonaeration conditions are preferred for the CBP system. Pectinase supplementation reduced viscosity of the fermentation broth and improved ethanol production performance, particularly under high gravity conditions, but the enzyme cost should be carefully balanced. Medium optimization was performed, and ethanol concentration as high as 94·2 g l(-1) was achieved when 0·15 g l(-1) K(2) HPO(4) was supplemented, which presents a significant progress in ethanol production from Jerusalem artichoke tubers. A CBP system using K. marxianus is suitable for efficient ethanol production from Jerusalem artichoke tubers under VHG conditions. Jerusalem artichoke tubers are an alternative to grain-based feedstocks for ethanol production. The high ethanol concentration achieved using K. marxianus with the CBP system not only saves energy consumption for ethanol distillation, but also significantly reduces the amount of waste distillage discharged from the distillation system. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

Enzymatic hydrolysis of pretreated Alfa fibers (Stipa tenacissima) using β-d-glucosidase and xylanase of Talaromyces thermophilus from solid-state fermentation.

PubMed

Mallek-Fakhfakh, Hanen; Fakhfakh, Jawhar; Walha, Kamel; Hassairi, Hajer; Gargouri, Ali; Belghith, Hafedh

2017-10-01

This work aims at realizing an optimal hydrolysis of pretreated Alfa fibers (Stipa tenacissima) through the use of enzymes produced from Talaromyces thermophilus AX4, namely β-d-glucosidase and xylanase, by a solid state fermentation process of an agro-industrial waste (wheat bran supplemented with lactose). The carbon source was firstly selected and the optimal values of three other parameters were determined: substrate loading (10g), moisture content (85%) and production time (10days); which led to an optimized enzymatic juice. The outcome was then supplemented with cellulases of T. reesei and used to optimize the enzymatic saccharification of alkali-pretreated Alfa fibers (PAF). The maximum saccharification yield of 83.23% was achieved under optimized conditions (substrate concentration 3.7% (w/v), time 144h and enzyme loading of 0.8 FPU, 15U CMCase, 60U β-d-glucosidase and 125U xylanase).The structural modification of PAF due to enzymatic saccharification was supported by the changes of morphologic and chemical composition observed through macroscopic representation, FTIR and X-Ray analysis. Copyright © 2017 Elsevier B.V. All rights reserved.

Different methodologies for sustainability of optimization techniques used in submerged and solid state fermentation.

PubMed

Ashok, Anup; Kumar, Devarai Santhosh

2017-10-01

Optimization techniques are considered as a part of nature's way of adjusting to the changes happening around it. There are different factors that establish the optimum working condition or the production of any value-added product. A model is accepted for a particular process after its sustainability has been verified on a statistical and analytical level. Optimization techniques can be divided into categories as statistical, nature inspired and artificial neural network each with its own benefits and usage in particular cases. A brief introduction about subcategories of different techniques that are available and their computational effectivity will be discussed. The main focus of the study revolves around the applicability of these techniques to any particular operation such as submerged fermentation (SmF) and solid state fermentation (SSF), their ability to produce secondary metabolites and the usefulness in the laboratory and industrial level. Primary studies to determine the enzyme activity of different microorganisms such as bacteria, fungi and yeast will also be discussed. l-Asparaginase, the most commonly used drugs in the treatment of acute lymphoblastic leukemia (ALL) shall be considered as an example, a short discussion on models used in the production by the processes of SmF and SSF will be discussed to understand the optimization techniques that are being dealt. It is expected that this discussion would help in determining the proper technique that can be used in running any optimization process for different purposes, and would help in making these processes less time-consuming with better output.

Understanding short-chain fatty acids accumulation enhanced in waste activated sludge alkaline fermentation: kinetics and microbiology.

PubMed

Zhang, Peng; Chen, Yinguang; Zhou, Qi; Zheng, Xiong; Zhu, Xiaoyu; Zhao, Yuxiao

2010-12-15

Most of the studies on sewage sludge treatment in literature were conducted for methane generation under acidic or near neutral pH conditions. It was reported in our previous studies that the accumulation of short-chain fatty acids (SCFAs), the preferred carbon source of biological wastewater nutrient removal, was significantly enhanced when sludge was fermented under alkaline conditions, but the optimal pH was temperature-dependent (pH 10 at ambient temperature, pH 9 at mesophilic, and pH 8 at thermophilic), and the maximal SCFAs yields were in the following order: thermophilic pH 8 > mesophilic pH 9 > ambient pH 10 > ambient uncontrolled pH. In this study the kinetic and microbiological features of waste activated sludge fermented in the range of pH 7-10 were investigated to understand the mechanism of remarkably high SCFAs accumulation under alkaline conditions. The developed sludge alkaline fermentation model could be applied to predicate the experimental data in either batch or semicontinuous sludge alkaline fermentation tests, and the relationships among alkaline pH, kinetic parameters, and SCFAs were discussed. Further analyses with fluorescence in situ hybridization (FISH) and PCR-based 16S rRNA gene clone library indicated that both the ratio of bacteria to archaea and the fraction of SCFAs producer accounting for bacteria were in the sequence of thermophilic pH 8 > mesophilic pH 9 > ambient pH 10 > ambient uncontrolled pH, which was in correspondence with the observed order of maximal SCFAs yields.

Deep Eutectic Solvents pretreatment of agro-industrial food waste.

PubMed

Procentese, Alessandra; Raganati, Francesca; Olivieri, Giuseppe; Russo, Maria Elena; Rehmann, Lars; Marzocchella, Antonio

2018-01-01

Waste biomass from agro-food industries are a reliable and readily exploitable resource. From the circular economy point of view, direct residues from these industries exploited for production of fuel/chemicals is a winning issue, because it reduces the environmental/cost impact and improves the eco-sustainability of productions. The present paper reports recent results of deep eutectic solvent (DES) pretreatment on a selected group of the agro-industrial food wastes (AFWs) produced in Europe. In particular, apple residues, potato peels, coffee silverskin, and brewer's spent grains were pretreated with two DESs, (choline chloride-glycerol and choline chloride-ethylene glycol) for fermentable sugar production. Pretreated biomass was enzymatic digested by commercial enzymes to produce fermentable sugars. Operating conditions of the DES pretreatment were changed in wide intervals. The solid to solvent ratio ranged between 1:8 and 1:32, and the temperature between 60 and 150 °C. The DES reaction time was set at 3 h. Optimal operating conditions were: 3 h pretreatment with choline chloride-glycerol at 1:16 biomass to solvent ratio and 115 °C. Moreover, to assess the expected European amount of fermentable sugars from the investigated AFWs, a market analysis was carried out. The overall sugar production was about 217 kt yr -1 , whose main fraction was from the hydrolysis of BSGs pretreated with choline chloride-glycerol DES at the optimal conditions. The reported results boost deep investigation on lignocellulosic biomass using DES. This investigated new class of solvents is easy to prepare, biodegradable and cheaper than ionic liquid. Moreover, they reported good results in terms of sugars' release at mild operating conditions (time, temperature and pressure).

Bacillus coagulans MA-13: a promising thermophilic and cellulolytic strain for the production of lactic acid from lignocellulosic hydrolysate.

PubMed

Aulitto, Martina; Fusco, Salvatore; Bartolucci, Simonetta; Franzén, Carl Johan; Contursi, Patrizia

2017-01-01

The transition from a petroleum-based economy towards more sustainable bioprocesses for the production of fuels and chemicals (circular economy) is necessary to alleviate the impact of anthropic activities on the global ecosystem. Lignocellulosic biomass-derived sugars are suitable alternative feedstocks that can be fermented or biochemically converted to value-added products. An example is lactic acid, which is an essential chemical for the production of polylactic acid, a biodegradable bioplastic. However, lactic acid is still mainly produced by Lactobacillus species via fermentation of starch-containing materials, the use of which competes with the supply of food and feed. A thermophilic and cellulolytic lactic acid producer was isolated from bean processing waste and was identified as a new strain of Bacillus coagulans , named MA-13. This bacterium fermented lignocellulose-derived sugars to lactic acid at 55 °C and pH 5.5. Moreover, it was found to be a robust strain able to tolerate high concentrations of hydrolysate obtained from wheat straw pre-treated by acid-catalysed (pre-)hydrolysis and steam explosion, especially when cultivated in controlled bioreactor conditions. Indeed, unlike what was observed in microscale cultivations (complete growth inhibition at hydrolysate concentrations above 50%), B. coagulans MA-13 was able to grow and ferment in 95% hydrolysate-containing bioreactor fermentations. This bacterium was also found to secrete soluble thermophilic cellulases, which could be produced at low temperature (37 °C), still retaining an optimal operational activity at 50 °C. The above-mentioned features make B. coagulans MA-13 an appealing starting point for future development of a consolidated bioprocess for production of lactic acid from lignocellulosic biomass, after further strain development by genetic and evolutionary engineering. Its optimal temperature and pH of growth match with the operational conditions of fungal enzymes hitherto employed for the depolymerisation of lignocellulosic biomasses to fermentable sugars. Moreover, the robustness of B. coagulans MA-13 is a desirable trait, given the presence of microbial growth inhibitors in the pre-treated biomass hydrolysate.

Aroma-active ester profile of ale beer produced under different fermentation and nutritional conditions.

PubMed

Hiralal, Lettisha; Olaniran, Ademola O; Pillay, Balakrishna

2014-01-01

A broad range of aroma-active esters produced during fermentation are vital for the complex flavour of beer. This study assessed the influence of fermentation temperature, pH, and wort nutritional supplements on the production of yeast-derived ester compounds and the overall fermentation performance. The best fermentation performance was achieved when wort was supplemented with 0.75 g/l l-leucine resulting in highest reducing sugar and FAN (free amino nitrogen) utilization and ethanol production. At optimum fermentation pH of 5, 38.27% reducing sugars and 35.28% FAN was utilized resulting in 4.07% (v/v) ethanol. Wort supplemented with zinc sulphate (0.12 g/l) resulted in 5.01% ethanol (v/v) production and 54.32% reducing sugar utilization. Increase in fermentation temperature from 18°C to room temperature (± 22.5°C) resulted in 17.03% increased ethanol production and 14.42% and 62.82% increase in total acetate ester concentration and total ethyl ester concentration, respectively. Supplementation of worth with 0.12 g/l ZnSO4 resulted in 2.46-fold increase in both isoamyl acetate and ethyl decanoate concentration, while a 7.05-fold and 1.96-fold increase in the concentration of isoamyl acetate and ethyl decanoate, respectively was obtained upon 0.75 g/l l-leucine supplementation. Wort supplemented with l-leucine (0.75 g/l) yielded the highest beer foam head stability with a rating of 2.67, while highest yeast viability was achieved when wort was supplemented with 0.12 g/l zinc sulphate. Results from this study suggest that supplementing wort with essential nutrients required for yeast growth and optimizing the fermentation conditions could be an effective way of improving fermentation performance and controlling aroma-active esters in beer. Copyright © 2013 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Modeling and verification of process parameters for the production of tannase by Aspergillus oryzae under submerged fermentation using agro-wastes.

PubMed

Varadharajan, Venkatramanan; Vadivel, Sudhan Shanmuga; Ramaswamy, Arulvel; Sundharamurthy, Venkatesaprabhu; Chandrasekar, Priyadharshini

2017-01-01

Tannase production by Aspergillus oryzae using various agro-wastes as substrates by submerged fermentation was studied in this research. The microbe was isolated from degrading corn kernel obtained from the corn fields at Tiruchengode, India. The microbial identification was done using 18S rRNA gene analysis. The agro-wastes chosen for the study were pomegranate rind, Cassia auriculata flower, black gram husk, and tea dust. The process parameters chosen for optimization study were substrate concentration, pH, temperature, and incubation period. During one variable at a time optimization, the pomegranate rind extract produced maximum tannase activity of 138.12 IU/mL and it was chosen as the best substrate for further experiments. The quadratic model was found to be the effective model for prediction of tannase production by A. oryzae. The optimized conditions predicted by response surface methodology (RSM) with genetic algorithm (GA) were 1.996% substrate concentration, pH of 4.89, temperature of 34.91 °C, and an incubation time of 70.65 H with maximum tannase activity of 138.363 IU/mL. The confirmatory experiment under optimized conditions showed tannase activity of 139.22 IU/mL. Hence, RSM-GA pair was successfully used in this study to optimize the process parameters required for the production of tannase using pomegranate rind. © 2015 International Union of Biochemistry and Molecular Biology, Inc.

Optimization of enzymatic hydrolysis and fermentation conditions for improved bioethanol production from potato peel residues.

PubMed

Ben Taher, Imen; Fickers, Patrick; Chniti, Sofien; Hassouna, Mnasser

2017-03-01

The aim of this work was the optimization of the enzyme hydrolysis of potato peel residues (PPR) for bioethanol production. The process included a pretreatment step followed by an enzyme hydrolysis using crude enzyme system composed of cellulase, amylase and hemicellulase, produced by a mixed culture of Aspergillus niger and Trichoderma reesei. Hydrothermal, alkali and acid pretreatments were considered with regards to the enhancement of enzyme hydrolysis of potato peel residues. The obtained results showed that hydrothermal pretreatment lead to a higher enzyme hydrolysis yield compared to both acid and alkali pretreatments. Enzyme hydrolysis was also optimized for parameters such as temperature, pH, substrate loading and surfactant loading using a response surface methodology. Under optimized conditions, 77 g L -1 of reducing sugars were obtained. Yeast fermentation of the released reducing sugars led to an ethanol titer of 30 g L -1 after supplementation of the culture medium with ammonium sulfate. Moreover, a comparative study between acid and enzyme hydrolysis of potato peel residues was investigated. Results showed that enzyme hydrolysis offers higher yield of bioethanol production than acid hydrolysis. These results highlight the potential of second generation bioethanol production from potato peel residues treated with onsite produced hydrolytic enzymes. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:397-406, 2017. © 2017 American Institute of Chemical Engineers.

Optimizing fermentation process miscanthus-to-ethanol biorefinery scale under uncertain conditions

NASA Astrophysics Data System (ADS)

Bomberg, Matthew; Sanchez, Daniel L.; Lipman, Timothy E.

2014-05-01

Ethanol produced from cellulosic feedstocks has garnered significant interest for greenhouse gas abatement and energy security promotion. One outstanding question in the development of a mature cellulosic ethanol industry is the optimal scale of biorefining activities. This question is important for companies and entrepreneurs seeking to construct and operate cellulosic ethanol biorefineries as it determines the size of investment needed and the amount of feedstock for which they must contract. The question also has important implications for the nature and location of lifecycle environmental impacts from cellulosic ethanol. We use an optimization framework similar to previous studies, but add richer details by treating many of these critical parameters as random variables and incorporating a stochastic sub-model for land conversion. We then use Monte Carlo simulation to obtain a probability distribution for the optimal scale of a biorefinery using a fermentation process and miscanthus feedstock. We find a bimodal distribution with a high peak at around 10-30 MMgal yr-1 (representing circumstances where a relatively low percentage of farmers elect to participate in miscanthus cultivation) and a lower and flatter peak between 150 and 250 MMgal yr-1 (representing more typically assumed land-conversion conditions). This distribution leads to useful insights; in particular, the asymmetry of the distribution—with significantly more mass on the low side—indicates that developers of cellulosic ethanol biorefineries may wish to exercise caution in scale-up.

Potency of microfiltration membrane process in purifying broccoli (Brassica oleracea L.) fermented by lactic acid bacteria (LAB) as functional food

NASA Astrophysics Data System (ADS)

Susilowati, Agustine; Aspiyanto, Maryati, Yati; Melanie, Hakiki; Lotulung, Puspa D.

2017-01-01

Purifying broccoli (Brassica oleracea L.) fermented by Lactic Acid Bacteria (LAB) using mixture of L. bulgaricus, S. thermopillus, L. acidophillusand Bifidobacteriumbifidum and fructooligosaccharides (FOS) as carbon source have been performed to recover biomass concentrate for probiotic and antioxidant. Purification of fermented broccoli was conducted through microfiltration (MF) membrane of 0.15 µm at stirrer rotation speed 400 rpm, room temperature and pressure 40 psia for 30 minutes. Fermented broccoli produced via fermentation process with fermentation time 0 (initial) and 48 hours, and LAB concentration 10% and 20% (v/v) represented as biomass of A, B, C and D. The experimental result showed that based on selectivity of total organic acids, separating optimization was achieved at biomass D (fermentation time 48 hours and mixed LAB culture concentration 20%). Concentrate composition produced in this condition were total acids 6.04%, total solids 24.31%, total polyphenol 0.0252%, reducing sugar 68.25 mg/mL, total sugars 30.89 mg/mL, and dissolved protein 28.54 mg/mL with pH 3.94. In this condition, recovery of biomass concentrate of D for total acids 5.64 folds, total solids 1.82 folds, total polyphenol 3.03 folds, reducing sugar 1.16 folds, total sugars 1.19 folds, and dissolved protein 0.67 folds compared with feed (initial process). Identification of monomer of biomass concentrate D as polyphenol derivatives at T2,01 and T3.01 gave monomer with molecular weight (MW) 192.78 Dalton (Da.), and monomer with MW 191.08, 191.49 and 192.07 Da., while lactic acid derivatives showed MW 251.13, 251.6 and 252.14, and monomer with MW 250.63, 252.14 and 254.22 Da.

Protein enrichment, cellulase production and in vitro digestion improvement of pangolagrass with solid state fermentation.

PubMed

Hu, Chan-Chin; Liu, Li-Yun; Yang, Shang-Shyng

2012-02-01

Pangolagrass, Digitaria decumbens Stent, is a major grass for cow feeding, and may be a good substrate for protein enrichment. To improve the quality of pangolagrass for animal feeding, cellulolytic microbes were isolated from various sources and cultivated with solid state fermentation to enhance the protein content, cellulase production and in vitro digestion. The microbes, culture conditions and culture media were studied. Cellulolytic microbes were isolated from pangolagrass and its extracts, and composts. Pangolagrass supplemented with nitrogen and minerals was used to cultivate the cellulolytic microbes with solid state fermentation. The optimal conditions for protein enrichment and cellulase activity were pangolagrass substrate at initial moisture 65-70%, initial pH 6.0-8.0, supplementation with 2.5% (NH(4))(2)SO(4), 2.5% KH(2)PO(4) and K(2)HPO(4) mixture (2:1, w/w) and 0.3% MgSO(4).7H(2)O and cultivated at 30(o)C for 6 days. The protein content of fermented pangolagrass increased from 5.97-6.28% to 7.09-16.96% and the in vitro digestion improved from 4.11-4.38% to 6.08-19.89% with the inoculation of cellulolytic microbes by solid state fermentation. Each 1 g of dried substrate yielded Avicelase 0.93-3.76 U, carboxymethylcellulase 1.39-4.98 U and β-glucosidase 1.20-6.01 U. The isolate Myceliophthora lutea CL3 was the strain found to be the best at improving the quality of pangolagrass for animal feeding with solid state fermentation. Solid state fermentation of pangolagrass inoculated with appropriate microbes is a feasible process to enrich protein content, increase in vitro digestibility and improve the quality for animal feeding. Copyright © 2011. Published by Elsevier B.V.

Enhanced xylose fermentation by engineered yeast expressing NADH oxidase through high cell density inoculums.

PubMed

Zhang, Guo-Chang; Turner, Timothy L; Jin, Yong-Su

2017-03-01

Accumulation of reduced byproducts such as glycerol and xylitol during xylose fermentation by engineered Saccharomyces cerevisiae hampers the economic production of biofuels and chemicals from cellulosic hydrolysates. In particular, engineered S. cerevisiae expressing NADPH-linked xylose reductase (XR) and NAD + -linked xylitol dehydrogenase (XDH) produces substantial amounts of the reduced byproducts under anaerobic conditions due to the cofactor difference of XR and XDH. While the additional expression of a water-forming NADH oxidase (NoxE) from Lactococcus lactis in engineered S. cerevisiae with the XR/XDH pathway led to reduced glycerol and xylitol production and increased ethanol yields from xylose, volumetric ethanol productivities by the engineered yeast decreased because of growth defects from the overexpression of noxE. In this study, we introduced noxE into an engineered yeast strain (SR8) exhibiting near-optimal xylose fermentation capacity. To overcome the growth defect caused by the overexpression of noxE, we used a high cell density inoculum for xylose fermentation by the SR8 expressing noxE. The resulting strain, SR8N, not only showed a higher ethanol yield and lower byproduct yields, but also exhibited a high ethanol productivity during xylose fermentation. As noxE overexpression elicits a negligible growth defect on glucose conditions, the beneficial effects of noxE overexpression were substantial when a mixture of glucose and xylose was used. Consumption of glucose led to rapid cell growth and therefore enhanced the subsequent xylose fermentation. As a result, the SR8N strain produced more ethanol and fewer byproducts from a mixture of glucose and xylose than the parental SR8 strain without noxE overexpression. Our results suggest that the growth defects from noxE overexpression can be overcome in the case of fermenting lignocellulose-derived sugars such as glucose and xylose.

High solid simultaneous saccharification and fermentation of wet oxidized corn stover to ethanol.

PubMed

Varga, Enikõ; Klinke, Helene B; Réczey, Kati; Thomsen, Anne Belinda

2004-12-05

In this study ethanol was produced from corn stover pretreated by alkaline and acidic wet oxidation (WO) (195 degrees C, 15 min, 12 bar oxygen) followed by nonisothermal simultaneous saccharification and fermentation (SSF). In the first step of the SSF, small amounts of cellulases were added at 50 degrees C, the optimal temperature of enzymes, in order to obtain better mixing condition due to some liquefaction. In the second step more cellulases were added in combination with dried baker's yeast (Saccharomyces cerevisiae) at 30 degrees C. The phenols (0.4-0.5 g/L) and carboxylic acids (4.6-5.9 g/L) were present in the hemicellulose rich hydrolyzate at subinhibitory levels, thus no detoxification was needed prior to SSF of the whole slurry. Based on the cellulose available in the WO corn stover 83% of the theoretical ethanol yield was obtained under optimized SSF conditions. This was achieved with a substrate concentration of 12% dry matter (DM) acidic WO corn stover at 30 FPU/g DM (43.5 FPU/g cellulose) enzyme loading. Even with 20 and 15 FPU/g DM (corresponding to 29 and 22 FPU/g cellulose) enzyme loading, ethanol yields of 76 and 73%, respectively, were obtained. After 120 h of SSF the highest ethanol concentration of 52 g/L (6 vol.%) was achieved, which exceeds the technical and economical limit of the industrial-scale alcohol distillation. The SSF results showed that the cellulose in pretreated corn stover can be efficiently fermented to ethanol with up to 15% DM concentration. A further increase of substrate concentration reduced the ethanol yield significant as a result of insufficient mass transfer. It was also shown that the fermentation could be followed with an easy monitoring system based on the weight loss of the produced CO2.

Production of 2,3-butanediol by a low-acid producing Klebsiella oxytoca NBRF4.

PubMed

Han, Sung-Hyuk; Lee, Jung-Eun; Park, Kyungmoon; Park, Yong-Cheol

2013-01-25

2,3-Butanediol (2,3-BDO) is a value-added chemical with great potential for the industrial production of synthetic rubber, plastic and solvent. For microbial production of 2,3-BDO, in this study, Klebsiella oxytoca NBRF4 was constructed by chemical mutation and screening against NaBr, NaBrO(3) and fluoroacetate. Among metabolic enzymes involved in the production of lactate, acetate and 2,3-BDO, K. oxytoca NBRF4 possessed 1.2 times lower specific activities of lactate dehydrogenase and phosphotransacetylase, and 22% higher specific acetoin reductase activity than the K. oxytoca ATCC43863 control strain. A series of batch fermentations in a defined medium and application of a statistical tool of response surface method led to the determination of optimal culture conditions: 10% dissolved oxygen level, pH 4.3 and 38°C. The actual results of batch fermentation at the optimal conditions using 44 g/L glucose were coincident with the predetermined values: 14.4 g/L 2,3-BDO concentration, 0.32 g/g yield. To increase 2,3-BDO titer, fed-batch fermentation of K. oxytoca NBRF4 was performed by an intermittent feeding of 800 g/L glucose to control its concentration around 5-20 g/L in the culture broth. Finally, 34.2g/L 2,3-BDO concentration and 0.35 g/g yield were obtained without organic acid production in 70 hours of the fed-batch culture, which were 2.4 and 1.2 times higher than those of the batch fermentation using 44 g/L glucose. Copyright © 2012 Elsevier B.V. All rights reserved.

An alternative feedstock of corn meal for industrial fuel ethanol production: delignified corncob residue.

PubMed

Lei, Cheng; Zhang, Jian; Xiao, Lin; Bao, Jie

2014-09-01

Delignified corncob residue is an industrial solid waste from xylose production using corncob as feedstock. In this study, delignified corncob residue was used as the feedstock of ethanol production by simultaneous saccharification and fermentation (SSF) and the optimal fermentation performance was investigated under various operation conditions. The ethanol titer and yield reached 75.07 g/L and 89.38%, respectively, using a regular industrial yeast strain at moderate cellulase dosage and high solids loading. A uniform SSF temperature of 37°C at both prehydrolysis and SSF stages was tested. The fermentation performance and cost of delignified corncob residue and corn meal was compared as feedstock of ethanol fermentation. The result shows that the delignified corncob residue is competitive to corn meal as ethanol production feedstock. The study gives a typical case to demonstrate the potential of intensively processed lignocellulose as the alternative feedstock of corn meal for industrial fuel ethanol production. Copyright © 2014 Elsevier Ltd. All rights reserved.

Optimization study of ethanolic fermentation from oil palm trunk, rubberwood and mixed hardwood hydrolysates using Saccharomyces cerevisiae.

PubMed

Chin, K L; H'ng, P S; Wong, L J; Tey, B T; Paridah, M T

2010-05-01

Ethanolic fermentation using Saccharomyces cerevisiae was carried out on three types of hydrolysates produced from lignocelulosic biomass which are commonly found in Malaysia such as oil palm trunk, rubberwood and mixed hardwood. The effect of fermentation temperature and pH of hydrolysate was evaluated to optimize the fermentation efficiency which defined as maximum ethanol yield in minimum fermentation time. The fermentation process using different temperature of 25 degrees Celsius, 30 degrees Celsius and 40 degrees Celsius were performed on the prepared fermentation medium adjusted to pH 4, pH 6 and pH 7, respectively. Results showed that the fermentation time was significantly reduced with the increase of temperature but an adverse reduction in ethanol yield was observed using temperature of 40 degrees Celsius. As the pH of hydrolysate became more acidic, the ethanol yield increased. Optimum fermentation efficiency for ethanolic fermentation of lignocellulosic hydrolysates using S. cerevisiae can be obtained using 33.2 degrees Celsius and pH 5.3. Copyright 2009 Elsevier Ltd. All rights reserved.

Optimization of the marinating conditions of cassava fish (Pseudotolithus sp.) fillet for Lanhouin production through application of Doehlert experimental design.

PubMed

Kindossi, Janvier Mêlégnonfan; Anihouvi, Victor Bienvenu; Vieira-Dalodé, Générose; Akissoé, Noël Houédougbé; Hounhouigan, Djidjoho Joseph

2016-03-01

Lanhouin is a traditional fermented salted fish made from the spontaneous and uncontrolled fermentation of whole salted cassava fish (Pseudotolithus senegalensis) mainly produced in the coastal regions of West Africa. The combined effects of NaCl, citric acid concentration, and marination time on the physicochemical and microbiological characteristics of the fish fillet used for Lanhouin production were studied using a Doehlert experimental design with the objective of preserving its quality and safety. The marination time has significant effects on total viable and lactic acid bacteria counts, and NaCl content of the marinated fish fillet while the pH was significantly affected by citric acid concentration and marination duration with high regression coefficient R (2) of 0.83. The experiment showed that the best conditions for marination process of fish fillet were salt ratio 10 g/100 g, acid citric concentration 2.5 g/100 g, and marination time 6 h. These optimum marinating conditions obtained present the best quality of marinated flesh fish leading to the safety of the final fermented product. This pretreatment is necessary in Lanhouin production processes to ensure its safety quality.

Periodic peristalsis increasing acetone-butanol-ethanol productivity during simultaneous saccharification and fermentation of steam-exploded corn straw.

PubMed

Li, Jingwen; Wang, Lan; Chen, Hongzhang

2016-11-01

The acetone-butanol-ethanol (ABE) fermentation of lignocellulose at high solids content has recently attracted extensive attention. However, the productivity of high solids ABE fermentation of lignocellulose is typically low in traditional processes due to the lack of efficient intensifying methods. In the present study, periodic peristalsis, a novel intensifying method, was applied to improve ABE production by the simultaneous saccharification and fermentation (SSF) of steam-exploded corn straw using Clostridium acetobutylicum ATCC824. The ABE concentration and the ABE productivity of SSF at a solids content of 17.5% (w/w) with periodic peristalsis were 17.1 g/L and 0.20 g/(L h), respectively, which were higher than those obtained under static conditions (15.2 g/L and 0.14 g/(L h)). The initial sugar conversion rate over the first 12 h with periodic peristalsis was 4.67 g/(L h) at 10 FPU/g cellulase dosage and 15% (w/w) solids content, an increase of 49.7% compared with the static conditions. With periodic peristalsis, the period of batch fermentation was shortened from 108 h to 84 h. The optimal operating regime was a low frequency (6 h -1 ) of periodic peristalsis in the acid-production phase (0-48 h) of SSF. Therefore, periodic peristalsis should be an effective intensifying method to increase the productivity of ABE fermentation at high solids content. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Utilization of methanol in crude glycerol to assist lipid production in non-sterilized fermentation from Trichosporon oleaginosus.

PubMed

Chen, Jiaxin; Zhang, Xiaolei; Tyagi, Rajeshwar Dayal; Drogui, Patrick

2018-04-01

In this work, methanol in crude glycerol solution was used to assist the lipid production with oleaginous yeast Trichosporon oleaginosus cultivated under non-sterilized conditions. The investigated methanol concentration was 0%, 1.4%, 2.2%, 3.3% and 4.4% (w/v). The results showed that methanol played a significant role in the non-sterilized fermentation for lipid production. The optimal methanol concentration was around 1.4% (w/v) in which the growth of T. oleaginosus was promoted and overcame that of the contaminants. The non-sterilized fed-batch fermentation with initial methanol concentration of 1.4% (w/v) was then performed and high biomass production (43.39 g/L) and lipid production (20.42 g/L) were achieved. Copyright © 2018 Elsevier Ltd. All rights reserved.

Production, statistical optimization and application of endoglucanase from Rhizopus stolonifer utilizing coffee husk.

PubMed

Navya, P N; Pushpa, S Murthy

2013-08-01

Coffee cherry husk (CH) is one of the major by-products obtained from coffee processing industry and accounts to 43 ± 5.9% of cellulose. Screening of fungal organism for cellulase production was carried out and the potential organism was identified as Rhizopus stolonifer by internal transcribed spacer's (ITS)-5.8S rDNA analysis. A systematic study with response surface methodology (RSM) based on CCRD was used to study the interactions among the variables such as pH (3-7), moisture (40-80%) and progression duration (72-168 h) of the fermentation process to maximize the enzyme production. Under the optimized cultivation condition, R. stolonifer synthesized 22,109 U/gds. Model validations at optimum operating conditions showed excellent agreement between the experimental results and the predicted responses with a confidence level of 95%. Endoglucanase thus produced was utilized for ethanol production by simultaneous saccharification and fermentation and maximum of 65.5 g/L of ethanol was obtained. This fungal cellulase has also reported to be efficient detergent additives and promising for commercial use. The present study demonstrates coffee husk as a significant bioprocess substrate. Statistical optimization with major parameters for cellulase production can be highly applicable for industrial scale. Furthermore, value addition to coffee husk with sustainable waste management leading to environment conservation can be achieved.

Optimization of a low-cost hyperosmotic medium and establishing the fermentation kinetics of erythritol production by Yarrowia lipolytica from crude glycerol.

PubMed

Yang, Li-Bo; Zhan, Xiao-Bei; Zhu, Li; Gao, Min-Jie; Lin, Chi-Chung

2016-05-18

The production of erythritol by Yarrowia lipolytica from low-cost substitutable substrates for high yield was investigated. Crude glycerol, urea, and NaCl related to osmotic pressure were the most significant factors affecting erythritol production. An artificial neural network model and genetic algorithm were used to search the optimal composition of the significant factors and locate the resulting erythritol yield. Medium with 232.39 g/L crude glycerol, 1.57 g/L urea, and 31.03 g/L NaCl led to predictive maximum erythritol concentration of 110.7 g/L. The erythritol concentration improved from 50.4 g/L to 109.2 g/L with the optimized medium, which was reproducible. Erythritol fermentation kinetics were investigated in a batch system. Multistep fermentation kinetic models with hyperosmotic inhibitory effects were developed. The resulting mathematical equations provided a good description of temporal variations such as microbial growth (X), substrate consumption (S), and product formation (P) in erythritol fermentation. The accordingly derived model is the first reported model for fermentative erythritol production from glycerol, providing useful information to optimize the growth of Y. lipolytica and contributing visual description for the erythritol fermentation process under high osmotic pressure, as well as improvement of productivity and efficiency.

Optimization of submerged fermentation medium for citrinin-free monascin production by Monascus.

PubMed

Chen, Di; Xue, Yuan; Chen, Mianhua; Li, Zhenjing; Wang, Changlu

2016-11-16

Microbial fermentation of citrinin-free Monascus pigments is in favor in the development of food industry. This study investigated the influences of carbon source, nitrogen source, and mineral salts on the cell growth, monascin (MS), and citrinin (CT) production in Monascus M9. A culture medium composition was established for maximizing the production of citrinin-free MS in submerged culture, as follows: 50 g/L Japonica rice powder, 20 g/L NH 4 NO 3 , 3 g/L NaNO 3 , 1.5 g/L KH 2 PO 4 , 1 g/L MgSO 4  · 7H 2 O, 0.2 g/L MnSO 4 . Under these conditions, no CT was detectable by high performance liquid chromatography. The yield of MS reached 14.11 mg/g, improving approximately 30% compared with before optimization.

Production of phytase by Mucor racemosus in solid-state fermentation.

PubMed

Bogar, Barbara; Szakacs, George; Pandey, Ashok; Abdulhameed, Sabu; Linden, James C; Tengerdy, Robert P

2003-01-01

Phytase production was studied by three Mucor and eight Rhizopus strains by solid-state fermentation (SSF) on three commonly used natural feed ingredients (canola meal, coconut oil cake, wheat bran). Mucor racemosus NRRL 1994 (ATCC 46129) gave the highest yield (14.5 IU/g dry matter phytase activity) on coconut oil cake. Optimizing the supplementation of coconut oil cake with glucose, casein and (NH(4))(2)SO(4), phytase production in solid-state fermentation was increased to 26 IU/g dry matter (DM). Optimization was carried out by Plackett-Burman and central composite experimental designs. Using the optimized medium phytase, alpha-amylase and lipase production of Mucor racemosus NRRL 1994 was compared in solid-state fermentation and in shake flask (SF) fermentation. SSF yielded higher phytase activity than did SF based on mass of initial substrate. Because this particular isolate is a food-grade fungus that has been used for sufu fermentation in China, the whole SSF material (crude enzyme, in situ enzyme) may be used directly in animal feed rations with enhanced cost efficiency.

Saccharomyces cerevisiae and S. kudriavzevii Synthetic Wine Fermentation Performance Dissected by Predictive Modeling.

PubMed

Henriques, David; Alonso-Del-Real, Javier; Querol, Amparo; Balsa-Canto, Eva

2018-01-01

Wineries face unprecedented challenges due to new market demands and climate change effects on wine quality. New yeast starters including non-conventional Saccharomyces species, such as S. kudriavzevii , may contribute to deal with some of these challenges. The design of new fermentations using non-conventional yeasts requires an improved understanding of the physiology and metabolism of these cells. Dynamic modeling brings the potential of exploring the most relevant mechanisms and designing optimal processes more systematically. In this work we explore mechanisms by means of a model selection, reduction and cross-validation pipeline which enables to dissect the most relevant fermentation features for the species under consideration, Saccharomyces cerevisiae T73 and Saccharomyces kudriavzevii CR85. The pipeline involved the comparison of a collection of models which incorporate several alternative mechanisms with emphasis on the inhibitory effects due to temperature and ethanol. We focused on defining a minimal model with the minimum number of parameters, to maximize the identifiability and the quality of cross-validation. The selected model was then used to highlight differences in behavior between species. The analysis of model parameters would indicate that the specific growth rate and the transport of hexoses at initial times are higher for S. cervisiae T73 while S. kudriavzevii CR85 diverts more flux for glycerol production and cellular maintenance. As a result, the fermentations with S. kudriavzevii CR85 are typically slower; produce less ethanol but higher glycerol. Finally, we also explored optimal initial inoculation and process temperature to find the best compromise between final product characteristics and fermentation duration. Results reveal that the production of glycerol is distinctive in S. kudriavzevii CR85, it was not possible to achieve the same production of glycerol with S. cervisiae T73 in any of the conditions tested. This result brings the idea that the optimal design of mixed cultures may have an enormous potential for the improvement of final wine quality.

Saccharomyces cerevisiae and S. kudriavzevii Synthetic Wine Fermentation Performance Dissected by Predictive Modeling

PubMed Central

Henriques, David; Alonso-del-Real, Javier; Querol, Amparo; Balsa-Canto, Eva

2018-01-01

Wineries face unprecedented challenges due to new market demands and climate change effects on wine quality. New yeast starters including non-conventional Saccharomyces species, such as S. kudriavzevii, may contribute to deal with some of these challenges. The design of new fermentations using non-conventional yeasts requires an improved understanding of the physiology and metabolism of these cells. Dynamic modeling brings the potential of exploring the most relevant mechanisms and designing optimal processes more systematically. In this work we explore mechanisms by means of a model selection, reduction and cross-validation pipeline which enables to dissect the most relevant fermentation features for the species under consideration, Saccharomyces cerevisiae T73 and Saccharomyces kudriavzevii CR85. The pipeline involved the comparison of a collection of models which incorporate several alternative mechanisms with emphasis on the inhibitory effects due to temperature and ethanol. We focused on defining a minimal model with the minimum number of parameters, to maximize the identifiability and the quality of cross-validation. The selected model was then used to highlight differences in behavior between species. The analysis of model parameters would indicate that the specific growth rate and the transport of hexoses at initial times are higher for S. cervisiae T73 while S. kudriavzevii CR85 diverts more flux for glycerol production and cellular maintenance. As a result, the fermentations with S. kudriavzevii CR85 are typically slower; produce less ethanol but higher glycerol. Finally, we also explored optimal initial inoculation and process temperature to find the best compromise between final product characteristics and fermentation duration. Results reveal that the production of glycerol is distinctive in S. kudriavzevii CR85, it was not possible to achieve the same production of glycerol with S. cervisiae T73 in any of the conditions tested. This result brings the idea that the optimal design of mixed cultures may have an enormous potential for the improvement of final wine quality. PMID:29456524

Optimization of lipase production by solid-state fermentation of olive pomace: from flask to laboratory-scale packed-bed bioreactor.

PubMed

Oliveira, Felisbela; Salgado, José Manuel; Abrunhosa, Luís; Pérez-Rodríguez, Noelia; Domínguez, José M; Venâncio, Armando; Belo, Isabel

2017-07-01

Lipases are versatile catalysts with many applications and can be produced by solid-state fermentation (SSF) using agro-industrial wastes. The aim of this work was to maximize the production of Aspergillus ibericus lipase under SSF of olive pomace (OP) and wheat bran (WB), evaluating the effect on lipase production of C/N ratio, lipids, phenols, content of sugars of substrates and nitrogen source addition. Moreover, the implementation of the SSF process in a packed-bed bioreactor and the improvement of lipase extraction conditions were assessed. Low C/N ratios and high content of lipids led to maximum lipase production. Optimum SSF conditions were achieved with a C/N mass ratio of 25.2 and 10.2% (w/w) lipids in substrate, by the mixture of OP:WB (1:1) and supplemented with 1.33% (w/w) (NH 4 ) 2 SO 4 . Studies in a packed-bed bioreactor showed that the lower aeration rates tested prevented substrate dehydration, improving lipase production. In this work, the important role of Triton X-100 on lipase extraction from the fermented solid substrate has been shown. A final lipase activity of 223 ± 5 U g -1 (dry basis) was obtained after 7 days of fermentation.

Metabolic and transcriptomic response of the wine yeast Saccharomyces cerevisiae strain EC1118 after an oxygen impulse under carbon-sufficient, nitrogen-limited fermentative conditions.

PubMed

Orellana, Marcelo; Aceituno, Felipe F; Slater, Alex W; Almonacid, Leonardo I; Melo, Francisco; Agosin, Eduardo

2014-05-01

During alcoholic fermentation, Saccharomyces cerevisiae is exposed to continuously changing environmental conditions, such as decreasing sugar and increasing ethanol concentrations. Oxygen, a critical nutrient to avoid stuck and sluggish fermentations, is only discretely available throughout the process after pump-over operation. In this work, we studied the physiological response of the wine yeast S. cerevisiae strain EC1118 to a sudden increase in dissolved oxygen, simulating pump-over operation. With this aim, an impulse of dissolved oxygen was added to carbon-sufficient, nitrogen-limited anaerobic continuous cultures. Results showed that genes related to mitochondrial respiration, ergosterol biosynthesis, and oxidative stress, among other metabolic pathways, were induced after the oxygen impulse. On the other hand, mannoprotein coding genes were repressed. The changes in the expression of these genes are coordinated responses that share common elements at the level of transcriptional regulation. Beneficial and detrimental effects of these physiological processes on wine quality highlight the dual role of oxygen in 'making or breaking wines'. These findings will facilitate the development of oxygen addition strategies to optimize yeast performance in industrial fermentations. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Combination of uniform design with artificial neural network coupling genetic algorithm: an effective way to obtain high yield of biomass and algicidal compound of a novel HABs control actinomycete

PubMed Central

2014-01-01

Controlling harmful algae blooms (HABs) using microbial algicides is cheap, efficient and environmental-friendly. However, obtaining high yield of algicidal microbes to meet the need of field test is still a big challenge since qualitative and quantitative analysis of algicidal compounds is difficult. In this study, we developed a protocol to increase the yield of both biomass and algicidal compound present in a novel algicidal actinomycete Streptomyces alboflavus RPS, which kills Phaeocystis globosa. To overcome the problem in algicidal compound quantification, we chose algicidal ratio as the index and used artificial neural network to fit the data, which was appropriate for this nonlinear situation. In this protocol, we firstly determined five main influencing factors through single factor experiments and generated the multifactorial experimental groups with a U15(155) uniform-design-table. Then, we used the traditional quadratic polynomial stepwise regression model and an accurate, fully optimized BP-neural network to simulate the fermentation. Optimized with genetic algorithm and verified using experiments, we successfully increased the algicidal ratio of the fermentation broth by 16.90% and the dry mycelial weight by 69.27%. These results suggested that this newly developed approach is a viable and easy way to optimize the fermentation conditions for algicidal microorganisms. PMID:24886410

Combination of uniform design with artificial neural network coupling genetic algorithm: an effective way to obtain high yield of biomass and algicidal compound of a novel HABs control actinomycete.

PubMed

Cai, Guanjing; Zheng, Wei; Yang, Xujun; Zhang, Bangzhou; Zheng, Tianling

2014-05-24

Controlling harmful algae blooms (HABs) using microbial algicides is cheap, efficient and environmental-friendly. However, obtaining high yield of algicidal microbes to meet the need of field test is still a big challenge since qualitative and quantitative analysis of algicidal compounds is difficult. In this study, we developed a protocol to increase the yield of both biomass and algicidal compound present in a novel algicidal actinomycete Streptomyces alboflavus RPS, which kills Phaeocystis globosa. To overcome the problem in algicidal compound quantification, we chose algicidal ratio as the index and used artificial neural network to fit the data, which was appropriate for this nonlinear situation. In this protocol, we firstly determined five main influencing factors through single factor experiments and generated the multifactorial experimental groups with a U15(155) uniform-design-table. Then, we used the traditional quadratic polynomial stepwise regression model and an accurate, fully optimized BP-neural network to simulate the fermentation. Optimized with genetic algorithm and verified using experiments, we successfully increased the algicidal ratio of the fermentation broth by 16.90% and the dry mycelial weight by 69.27%. These results suggested that this newly developed approach is a viable and easy way to optimize the fermentation conditions for algicidal microorganisms.

[Temperature-switched high-efficiency D-lactate production from glycerol].

PubMed

Tian, Kangming; Zhou, Li; Chen, Xianzhong; Shen, Wei; Shi, Guiyang; Singh, Suren; Lu, Fuping; Wang, Zhengxiang

2013-01-01

Glycerol from oil hydrolysis industry is being considered as one of the abundent raw materials for fermentation industry. In present study, the aerobic and anaerobic metabolism and growth properties on glycerol by Esherichia coli CICIM B0013-070, a D-lactate over-producing strain constructed previously, at different temperatures were investigated, followed by a novel fermentation process, named temperature-switched process, was established for D-lactate production from glycerol. Under the optimal condition, lactate yield was increased from 64.0% to 82.6%. Subsequently, the yield of D-lactate from glycerol was reached up to 88.9% while a thermo-inducible promoter was used to regulate D-lactate dehydrogenase transcription.

[Amperometric biosensor for ethanol analysis in wines and grape must during wine fermentation].

PubMed

Shkotova, L V; Slast'ia, E A; Zhyliakova, T A; Soldatkin, O P; Schuhmann, W; Dziadevych, S V

2005-01-01

The amperometric biosensor for ethanol determination based on alcohol oxidase immobilised by the method of electrochemical polymerization has been developed. The industrial screen-printed platinum electrodes were used as transducers for creation of amperometric alcohol biosensor. Optimal conditions for electrochemical deposition of an active membrane with alcohol oxidase has been determined. Biosensors are characterised by good reproducibility and operational stability with minimal detection limit of ethanol 8 x 10(-5) M. The good correlation of results for ethanol detection in wine and during wine fermentation by using the developed amperometric biosensor with the data obtained by the standard methods was shown (r = 0.995).

A new strategy of glucose supply in a microbial fermentation model

NASA Astrophysics Data System (ADS)

Kasbawati, Gunawan, A. Y.; Sidarto, K. A.; Hertadi, R.

2015-09-01

Strategy of glucose supply to achieve an optimal productivity of ethanol production of a yeast cell is one of the main features in a microbial fermentation process. Beside a known continuous glucose supply, in this study we consider a new supply strategy so called the on-off supply. An optimal control theory is applied to the fermentation system to find the optimal rate of glucose supply and time of supply. The optimization problem is solved numerically using Differential Evolutionary algorithm. We find two alternative solutions that we can choose to get the similar result: either long period process with low supply or short period process with high glucose supply.

Optimized batch fermentation of cheese whey. Supplemented feedlot waste filtrate to produce a nitrogen-rich feed supplement for ruminants

DOE Office of Scientific and Technical Information (OSTI.GOV)

Erdman, M.D.; Reddy, C.A.

1986-03-01

An optimized batch fermentation process for the conversion of cattle feedlot waste filtrate, supplemented with cheese whey, into a nitrogenous feed supplement for ruminants is described. Feedlot waste filtrate supplemented with cheese whey (5 g of whey per 100 ml) was fermented by the indigenous microbial flora in the feedlot waste filtrate. Ammonium hydroxide was added to the fermentation not only to maintain a constant pH but also to produce ammonium salts of organic acids, which have been shown to be valuable as nitrogenous feed supplements for ruminants. The utilization of substrate carbohydrate at pH 7.0 and 43 degrees Cmore » was greater than 94% within 8 h, and the crude protein (total N X 6.25) content of the product was 70 to 78% (dry weight basis). About 66 to 69% of the crude protein was in the form of ammonia nitrogen. Lactate and acetate were the predominant acids during the first 6 to 8 hours of fermentation, but after 24 hours, appreciable levels of propionate and butyrate were also present. The rate of fermentation and the crude protein content of the product were optimal at pH 7.0 and decreased at a lower pH. For example, fermentation did not go to completion even after 24 hours at pH 4.5. Fermentation proceeded optimally at 43 degrees C, less so at 37 degrees C, and considerably more slowly at 23 and 50 degrees C. Concentrations of up to 15 g of cheese whey per 100 ml of feedlot waste filtrate were fermented efficiently. Fermentation of feedlot waste filtrate obtained from animals fed low silage-high grain, high silage-low grain, or dairy rations resulted in similar products in terms of total nitrogen and organic acid composition.« less

Production, Purification and Characterisation of a Potential Fibrinolytic Protease from Endophytic Xylaria curta by Solid Substrate Fermentation.

PubMed

Meshram, Vineet; Saxena, Sanjai; Paul, Karan; Gupta, Mahiti; Kapoor, Neha

2017-04-01

The present investigation highlights the optimal conditions for production of a non-toxic, bi-functional fibrinolytic enzyme xylarinase produced by endophytic fungus Xylaria curta by solid substrate fermentation using rice chaff medium. The purified enzyme is a monomeric protein with a molecular mass of ∼33 kDa. The enzyme exhibits cleavage of Aα and Bβ chains of fibrin(ogen) and has no effect on γ chain. The optimal fibrinolytic activity of the enzyme was observed at 35 °C and pH 8. The fibrinolytic activity was enhanced in the presence of Ca 2+ , whereas it was completely inhibited in the presence of Fe 2+ and Zn 2+ ions and inhibitors like EDTA and EGTA suggesting it to be a metalloprotease. The K m and V max of the enzyme for azocasein were 326 μM and 0.13 μM min -1 . The N-terminal sequence of the enzyme (SNGPLPGGVVWAG) was same when compared to xylarinase isolated from culture broth of X. curta. Thus, xylarinase could be exploited as a potent clot busting enzyme which could be produced on large scale using solid substrate fermentation.

Production of L-asparaginase, an anticancer agent, from Aspergillus niger using agricultural waste in solid state fermentation.

PubMed

Mishra, Abha

2006-10-01

This article reports the production of high levels of L-asparaginase from a new isolate of Aspergillus niger in solid state fermentation (SSF) using agro-wastes from three leguminous crops (bran of Cajanus cajan, Phaseolus mungo, and Glycine max). When used as the sole source for growth in SSF, bran of G. max showed maximum enzyme production followed by that of P. mungo and C. cajan. A 96-h fermentation time under aerobic condition with moisture content of 70%, 30 min of cooking time and 1205-1405 micro range of particle size in SSF appeared optimal for enzyme production. Enzyme yield was maximum (40.9 +/- 3.35 U/g of dry substrate) at pH 6.5 and temperature 30 +/- 2 degrees C. The optimum temperature and pH for enzyme activity were 40 degrees C and 6.5, respectively. The study suggests that choosing an appropriate substrate when coupled with process level optimization improves enzyme production markedly. Developing an asparaginase production process based on bran of G. max as a substrate in SSF is economically attractive as it is a cheap and readily available raw material in agriculture-based countries.

Production of lipopeptides by Bacillus amyloliquefaciens XZ-173 in solid state fermentation using soybean flour and rice straw as the substrate.

PubMed

Zhu, Zhen; Zhang, Guoyi; Luo, Yi; Ran, Wei; Shen, Qirong

2012-05-01

This work was aimed to produce lipopeptides by Bacillus amyloliquefaciens XZ-173 in solid state fermentation using agro-industrial byproducts. A central composite design was used to get the highest lipopeptides production. Results revealed that the optimal conditions for maximum lipopeptides production were 1.79% starch and 1.91% yeast extract by employing 5.58 g soybean flour and 3.67 g rice straw as the solid substrate with initial pH 7.5, moisture content 55% and a 10% inoculum level at 30°C for 2 days. Under these conditions, the experimental yield of lipopeptides reached 50.01 mg/gds, which was very close to the predicted value (49.91 mg/gds). At high concentration, the lipopeptides extracted from fermented substrates showed strong antibiotic activity against Rhizoctonia solani and Ralstonia solanacearum and certain emulsification but good emulsion stability. This is the first report on lipopeptides production that uses rice straw as a major substrate. Copyright © 2012 Elsevier Ltd. All rights reserved.

[Biotechnological optimization of nutrient composition of fermented dairy drink].

PubMed

Donskaya, G A

2014-01-01

The receipt based on the results of carried out studies is substantiated and technology of the new fermented dairy drink containing whole milk and whey with inulin (Jerusalem artichoke extract) and optimizing initial mineral composition of raw material has been developed. The starters ascertaining optimal organoleptic properties of the drink have been selected. It has been established that Jerusalem artichoke and its derivatives in the form of syrups and extracts stimulate fermentative processes of technological microflora, with maximum activity observed with Jerusalem artichoke extract. Physical-chemical and microbiological characteristics of the drink have been defined during storage. The possibility to optimize the nutrient composition of fermented dairy product by means of introducing of Jerusalem artichoke extract into milk-protein base has been demonstrated. It has been calculated that consumption of 100 g of fermented dairy drink enriched with Jerusalem artichoke extract makes it possible to satisfy the physiological needs (recommended daily allowance--RDA) for babies from 0 to 3 months in vitamins B1, B2 and B6 by 25-35% and in minerals P, K, and Ca by 20, 68, 34, 26%. For adults receiving 250 g of fermented beverage meets RDA for vitamins B1, B2 and B6 by 10-19% and in the macronutrients P, K, Ca-by 25-35%. Designed fermented dairy drink supplemented with natural plant ingredient possesses increased antioxidant activity and may be recommended for mass consumption without any limitations.

Ethanol production from banana peels using statistically optimized simultaneous saccharification and fermentation process.

PubMed

Oberoi, Harinder Singh; Vadlani, Praveen V; Saida, Lavudi; Bansal, Sunil; Hughes, Joshua D

2011-07-01

Dried and ground banana peel biomass (BP) after hydrothermal sterilization pretreatment was used for ethanol production using simultaneous saccharification and fermentation (SSF). Central composite design (CCD) was used to optimize concentrations of cellulase and pectinase, temperature and time for ethanol production from BP using SSF. Analysis of variance showed a high coefficient of determination (R(2)) value of 0.92 for ethanol production. On the basis of model graphs and numerical optimization, the validation was done in a laboratory batch fermenter with cellulase, pectinase, temperature and time of nine cellulase filter paper unit/gram cellulose (FPU/g-cellulose), 72 international units/gram pectin (IU/g-pectin), 37 °C and 15 h, respectively. The experiment using optimized parameters in batch fermenter not only resulted in higher ethanol concentration than the one predicted by the model equation, but also saved fermentation time. This study demonstrated that both hydrothermal pretreatment and SSF could be successfully carried out in a single vessel, and use of optimized process parameters helped achieve significant ethanol productivity, indicating commercial potential for the process. To the best of our knowledge, ethanol concentration and ethanol productivity of 28.2 g/l and 2.3 g/l/h, respectively from banana peels have not been reported to date. Copyright © 2011 Elsevier Ltd. All rights reserved.

The preparation and characterization of chitin and chitosan under large-scale submerged fermentation level using shrimp by-products as substrate.

PubMed

Zhang, Hongcai; Yun, Sanyue; Song, Lingling; Zhang, Yiwen; Zhao, Yanyun

2017-03-01

The crustacean shells of crabs and shrimps produces quantities of by-products, leading to seriously environmental pollution and human health problems during industrial processing, yet they turned into high-value useful products, such as chitin and chitosan. To prepare them under large-scale submerged fermentation level, shrimp shell powders (SSPs) was fermented by successive three-step fermentation of Serratia marcescens B742, Lactobacillus plantarum ATCC 8014 and Rhizopus japonicus M193 to extract chitin and chitosan based on previously optimal conditions. Moreover, the key parameters was investigated to monitor the changes of resulted products during fermentation process. The results showed that the yield of prepared chitin and chitosan reached 21.35 and 13.11% with the recovery rate of 74.67 and 63.42%, respectively. The degree of deacetylation (DDA) and molecular mass (MM) of produced chitosan were 81.23% and 512.06kDa, respectively. The obtained chitin and chitosan was characterized using Fourier transform infrared spectrometer (FT-IR) and X-ray diffraction (XRD) analysis. The established microbial fermentation method can be applied for the industrial large-scale production of chitin and chitosan, while the use of chemical reagents was significantly reduced. Copyright © 2016 Elsevier B.V. All rights reserved.

Biogenic amines in table olives. Analysis by high-performance liquid chromatography.

PubMed

Hornero-Méndez, D; Garrido-Fernández, A

1994-09-01

Biogenic amines in fermented vegetables have scarcely been studied. Available data show that in table olives and fermented cucumbers their presence is rare and any determinations made have been restricted mainly to histamine. However, some microorganisms, especially those related to spoilage, found in the fermentation brines of such products may have amino acid decarboxylase activity and give rise to biogenic amines by unusual processes. A method for the simultaneous determination of eight biogenic amines (tryptamine, beta-phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine, and spermine) has been developed to study their occurrence in fermented vegetables in more detail. The method consists of extraction of the amines from olive paste with 5% m/v trichloracetic acid and successive transfers into water-saturated n-BuOH and 0.1 mol l-1 HCl. An aliquot of this mixture is dried and derivatized with dansyl chloride. The dansyl derivatives are then analysed by high-performance liquid chromatography. Special emphasis has been given to optimization of the n-BuOH and 0.1 mol l-1 HCl extractions and to the derivatization conditions. By applying this method to the analysis of spoilt olives, the presence of some biogenic amines has been demonstrated. Thus a new method for monitoring the presence of biogenic amines during the fermentation of olives and for detecting anomalous fermentations is envisaged.

Co-fermentation of cellobiose and xylose by mixed culture of recombinant Saccharomyces cerevisiae and kinetic modeling.

PubMed

Chen, Yingying; Wu, Ying; Zhu, Baotong; Zhang, Guanyu; Wei, Na

2018-01-01

Efficient conversion of cellulosic sugars in cellulosic hydrolysates is important for economically viable production of biofuels from lignocellulosic biomass, but the goal remains a critical challenge. The present study reports a new approach for simultaneous fermentation of cellobiose and xylose by using the co-culture consisting of recombinant Saccharomyces cerevisiae specialist strains. The co-culture system can provide competitive advantage of modularity compared to the single culture system and can be tuned to deal with fluctuations in feedstock composition to achieve robust and cost-effective biofuel production. This study characterized fermentation kinetics of the recombinant cellobiose-consuming S. cerevisiae strain EJ2, xylose-consuming S. cerevisiae strain SR8, and their co-culture. The motivation for kinetic modeling was to provide guidance and prediction of using the co-culture system for simultaneous fermentation of mixed sugars with adjustable biomass of each specialist strain under different substrate concentrations. The kinetic model for the co-culture system was developed based on the pure culture models and incorporated the effects of product inhibition, initial substrate concentration and inoculum size. The model simulations were validated by results from independent fermentation experiments under different substrate conditions, and good agreement was found between model predictions and experimental data from batch fermentation of cellobiose, xylose and their mixtures. Additionally, with the guidance of model prediction, simultaneous co-fermentation of 60 g/L cellobiose and 20 g/L xylose was achieved with the initial cell densities of 0.45 g dry cell weight /L for EJ2 and 0.9 g dry cell weight /L SR8. The results demonstrated that the kinetic modeling could be used to guide the design and optimization of yeast co-culture conditions for achieving simultaneous fermentation of cellobiose and xylose with improved ethanol productivity, which is critically important for robust and efficient renewable biofuel production from lignocellulosic biomass.

A new search for thermotolerant yeasts, its characterization and optimization using response surface methodology for ethanol production.

PubMed

Arora, Richa; Behera, Shuvashish; Sharma, Nilesh K; Kumar, Sachin

2015-01-01

The progressive rise in energy crisis followed by green house gas (GHG) emissions is serving as the driving force for bioethanol production from renewable resources. Current bioethanol research focuses on lignocellulosic feedstocks as these are abundantly available, renewable, sustainable and exhibit no competition between the crops for food and fuel. However, the technologies in use have some drawbacks including incapability of pentose fermentation, reduced tolerance to products formed, costly processes, etc. Therefore, the present study was carried out with the objective of isolating hexose and pentose fermenting thermophilic/thermotolerant ethanologens with acceptable product yield. Two thermotolerant isolates, NIRE-K1 and NIRE-K3 were screened for fermenting both glucose and xylose and identified as Kluyveromyces marxianus NIRE-K1 and K. marxianus NIRE-K3. After optimization using Face-centered Central Composite Design (FCCD), the growth parameters like temperature and pH were found to be 45.17°C and 5.49, respectively for K. marxianus NIRE-K1 and 45.41°C and 5.24, respectively for K. marxianus NIRE-K3. Further, batch fermentations were carried out under optimized conditions, where K. marxianus NIRE-K3 was found to be superior over K. marxianus NIRE-K1. Ethanol yield (Y x∕s ), sugar to ethanol conversion rate (%), microbial biomass concentration (X) and volumetric product productivity (Q p ) obtained by K. marxianus NIRE-K3 were found to be 9.3, 9.55, 14.63, and 31.94% higher than that of K. marxianus NIRE-K1, respectively. This study revealed the promising potential of both the screened thermotolerant isolates for bioethanol production.

Response surface optimization of the thermal acid pretreatment of sugar beet pulp for bioethanol production using Trichoderma viride and Saccharomyces cerevisiae.

PubMed

El-Gendy, Nour Sh; Madian, Hekmat R; Nassar, Hussein N; Abu Amr, Salem S

2015-01-01

Worldwide nowadays, relying on the second generation bioethanol from the lignocellulosic feedstock is a mandatory aim. However, one of the major drawbacks for high ethanol yield is the physical and chemical pretreatment of this kind of feedstock. As the pretreatment is a crucial process operation that modifies the lignocellulosic structure and enhances its accessibility for the high cost hydrolytic enzymes in an attempt to maximize the yield of the fermentable sugars. The objective of this work was to optimize and integrate a physicochemical pretreatment of one of the major agricultural wastes in Egypt; the sugar beet pulp (SBP) and the enzymatic saccharification of the pretreated SBP using a whole fungal cells with a separate bioethanol fermentation batch processes to maximize the bioethanol yield. The response surface methodology was employed in this study to statistically evaluate and optimize the conditions for a thermal acid pretreatment of SBP. The significance and the interaction effects of the concentrations of HCl and SBP and the reaction temperature and time were studied using a three-level central composite design of experiments. A quadratic model equation was obtained to maximize the production of the total reducing sugars. The validity of the predicted model was confirmed. The thermally acid pretreated SBP was further subjected to a solid state fermentation batch process using Trichoderma viride F94. The thermal acid pretreatment and fungal hydrolyzes were integrated with two parallel batch fermentation processes of the produced hydrolyzates using Saccharomyces cerevisiae Y39, that yielded a total of ≈ 48 g/L bioethanol, at a conversion rate of ≈ 0.32 g bioethanol/ g SBP. Applying the proposed integrated process, approximately 97.5 gallon of ethanol would be produced from a ton (dry weight) of SBP.

Response surface optimization of the thermal acid pretreatment of sugar beet pulp for bioethanol production using Trichoderma viride and Saccharomyces cerevisiae.

PubMed

El-Gendy, Nour Sh; Madian, Hekmat R; Nassar, Hussein N; Amr, Salem S Abu

2015-09-15

Worldwide nowadays, relying on the second generation bioethanol from the lignocellulosic feedstock is a mandatory aim. However, one of the major drawbacks for high ethanol yield is the physical and chemical pretreatment of this kind of feedstock. As the pretreatment is a crucial process operation that modifies the lignocellulosic structure and enhances its accessibility for the high cost hydrolytic enzymes in an attempt to maximize the yield of the fermentable sugars. The objective of this work was to optimize and integrate a physicochemical pretreatment of one of the major agricultural wastes in Egypt; the sugar beet pulp (SBP) and the enzymatic saccharification of the pretreated SBP using a whole fungal cells with a separate bioethanol fermentation batch processes to maximize the bioethanol yield. The response surface methodology was employed in this study to statistically evaluate and optimize the conditions for a thermal acid pretreatment of SBP. The significance and the interaction effects of the concentrations of HCl and SBP and the reaction temperature and time were studied using a three-level central composite design of experiments. A quadratic model equation was obtained to maximize the production of the total reducing sugars. The validity of the predicted model was confirmed. The thermally acid pretreated SBP was further subjected to a solid state fermentation batch process using Trichoderma viride F94. The thermal acid pretreatment and fungal hydrolyzes were integrated with two parallel batch fermentation processes of the produced hydrolyzates using Saccharomyces cerevisiae Y39, that yielded a total of ≈ 48 g/L bioethanol, at a conversion rate of ≈ 0.32 g bioethanol/ g SBP. Applying the proposed integrated process, approximately 97.5 gallon of ethanol would be produced from a ton (dry weight) of SBP.

A new search for thermotolerant yeasts, its characterization and optimization using response surface methodology for ethanol production

PubMed Central

Arora, Richa; Behera, Shuvashish; Sharma, Nilesh K.; Kumar, Sachin

2015-01-01

The progressive rise in energy crisis followed by green house gas (GHG) emissions is serving as the driving force for bioethanol production from renewable resources. Current bioethanol research focuses on lignocellulosic feedstocks as these are abundantly available, renewable, sustainable and exhibit no competition between the crops for food and fuel. However, the technologies in use have some drawbacks including incapability of pentose fermentation, reduced tolerance to products formed, costly processes, etc. Therefore, the present study was carried out with the objective of isolating hexose and pentose fermenting thermophilic/thermotolerant ethanologens with acceptable product yield. Two thermotolerant isolates, NIRE-K1 and NIRE-K3 were screened for fermenting both glucose and xylose and identified as Kluyveromyces marxianus NIRE-K1 and K. marxianus NIRE-K3. After optimization using Face-centered Central Composite Design (FCCD), the growth parameters like temperature and pH were found to be 45.17°C and 5.49, respectively for K. marxianus NIRE-K1 and 45.41°C and 5.24, respectively for K. marxianus NIRE-K3. Further, batch fermentations were carried out under optimized conditions, where K. marxianus NIRE-K3 was found to be superior over K. marxianus NIRE-K1. Ethanol yield (Yx∕s), sugar to ethanol conversion rate (%), microbial biomass concentration (X) and volumetric product productivity (Qp) obtained by K. marxianus NIRE-K3 were found to be 9.3, 9.55, 14.63, and 31.94% higher than that of K. marxianus NIRE-K1, respectively. This study revealed the promising potential of both the screened thermotolerant isolates for bioethanol production. PMID:26388844

Direct bioethanol production by amylolytic yeast Candida albicans.

PubMed

Aruna, A; Nagavalli, M; Girijashankar, V; Ponamgi, S P D; Swathisree, V; Rao, L Venkateswar

2015-03-01

An attempt was made to produce bioethanol using optimized fermentation parameters and mutationally improved strain of Candida albicans. The mutant strain OMC3E6 obtained by UV irradiation followed by ethidium bromide successive mutations showed 2.6 times more glucoamylase secretion and 1.5 times more bioethanol production via direct conversion of starch. Enhanced hydrolysis of insoluble starch (72%) and potato starch (70%) was achieved with glucoamylase enzyme preparation from mutant C. albicans. In fermentation medium, the use of maltose, corn steep liquor, NaH2 PO4 , NaCl + MgSO4 and Triton X-100 has increased the glucoamylase production by the microbe. Under optimized conditions, C. albicans eventually produced 437 g ethanol kg(-1) potatoes. Earlier reports mentioned the use of thrice the quantity of starch as reported by us followed by more fermentation period (3-4 days) and demanded pretreatment of starch sources with alpha-amylase as well. Here, we simplified these three steps and obtained 73% conversion of insoluble starch into ethanol via direct conversion method in a period of 2 days without the involvement of cell immobilizations or enzyme pretreatment steps. Due to fast depletion of fossil fuels in the modern world, bioethanol usage as an alternate energy source is the need of the hour. For the first time, we report bioethanol production by Candida albicans via direct conversion of starchy biomass into ethanol along with enhanced starch-hydrolysing capacity and ethanol conversion ratio. So far, C. albicans was dealt in the field of clinical pathology, but here we successfully employed this organism to produce bioethanol from starchy agri-substrates. Optimizing fermentation parameters and improving the microbial strains through successive mutagenesis can improve the end product yield. © 2014 The Society for Applied Microbiology.

The deep processing of seaweed industrial waste--Influence of several fermentation on seaweed waste of feed

NASA Astrophysics Data System (ADS)

Zhao, Shipeng; Zhang, Shuping

2018-02-01

This paper focuses on several factors on the effects of fermented seaweed feed, and obtains the optimal fermentation process through the analysis of nutrients. Through the experiment we can get, Seaweed waste fermented the best feed when adding 1% of microbial agents and 0.5% of corn powder, fermenting for 15 days.

PMKT2, a new killer toxin from Pichia membranifaciens, and its promising biotechnological properties for control of the spoilage yeast Brettanomyces bruxellensis.

PubMed

Santos, A; San Mauro, M; Bravo, E; Marquina, D

2009-02-01

Pichia membranifaciens CYC 1086 secretes a killer toxin (PMKT2) that is inhibitory to a variety of spoilage yeasts and fungi of agronomical interest. The killer toxin in the culture supernatant was concentrated by ultrafiltration and purified to homogeneity by two successive steps, including native electrophoresis and HPLC gel filtration. Biochemical characterization of the toxin showed it to be a protein with an apparent molecular mass of 30 kDa and an isoelectric point of 3.7. At pH 4.5, optimal killer activity was observed at temperatures up to 20 degrees C. Above approximately this pH, activity decreased sharply and was barely noticeable at pH 6. The toxin concentrations present in the supernatant during optimal production conditions exerted a fungicidal effect on a variety of fungal and yeast strains. The results obtained suggest that PMKT2 has different physico-chemical properties from PMKT as well as different potential uses in the biocontrol of spoilage yeasts. PMKT2 was able to inhibit Brettanomyces bruxellensis while Saccharomyces cerevisiae was fully resistant, indicating that PMKT2 could be used in wine fermentations to avoid the development of the spoilage yeast without deleterious effects on the fermentative strain. In small-scale fermentations, PMKT2, as well as P. membranifaciens CYC 1086, was able to inhibit B. bruxellensis, verifying the biocontrol activity of PMKT2 in simulated winemaking conditions.

High-theabrownins instant dark tea product by Aspergillus niger via submerged fermentation: α-glucosidase and pancreatic lipase inhibition and antioxidant activity.

PubMed

Wang, Yuwan; Zhang, Mingyue; Zhang, Zhengzhu; Lu, Hengqian; Gao, Xueling; Yue, Pengxiang

2017-12-01

Theabrownins (TB) are bioactive components that are usually extracted from Chinese dark tea, in which they are present at low concentrations. The present study aimed to produce an instant dark tea high in theabrownins via submerged fermentation by the fungus Aspergillus niger. Three fermentation parameters that affect theabrownins content (i.e. inoculum size, liquid-solid ratio and rotation speed) were optimized using response surface methodology. Optimum fermentation conditions were modeled to be an inoculum of 5.40% (v/v), a liquid-solid ratio of 27.45 mL g -1 and a rotation speed of 184 rpm and were predicted to yield 292.99 g kg -1 TB. Under these experimentally conditions, the TB content of the instant dark tea was 291.93 g kg -1 . The antioxidant capacity and α-glucosidase and pancreatic lipase inhibitory activities of the high-TB instant black tea were higher than four other typical instant dark tea products. The results of the present study show that careful management of culture conditions can produce a dark tea high in theabrownins. Furthermore, high-theabrownins instant dark tea could serve as a source of bioactive products and be used in functional foods as an ingredient imparting antioxidant properties and the ability to inhibit pancreatic lipase and α-glucosidase. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Process Optimization on Micro-Aeration Supply for High Production Yield of 2,3-Butanediol from Maltodextrin by Metabolically-Engineered Klebsiella oxytoca

PubMed Central

Jantama, Sirima Suvarnakuta; Kanchanatawee, Sunthorn

2016-01-01

An optimization process with a cheap and abundant substrate is considered one of the factors affecting the price of the production of economical 2,3-Butanediol (2,3-BD). A combination of the conventional method and response surface methodology (RSM) was applied in this study. The optimized levels of pH, aeration rate, agitation speed, and substrate concentration (maltodextrin) were investigated to determine the cost-effectiveness of fermentative 2,3-BD production by metabolically-engineered Klebsiella oxytoca KMS005. Results revealed that pH, aeration rate, agitation speed, and maltodextrin concentration at levels of 6.0, 0.8 vvm, 400 rpm, and 150 g/L respectively were the optimal conditions. RSM also indicated that the agitation speed was the most influential parameter when either agitation and aeration interaction or agitation and substrate concentration interaction played important roles for 2,3-BD production by the strain from maltodextrin. Under interim fed-batch fermentation, 2,3-BD concentration, yield, and productivity were obtained at 88.1±0.2 g/L, 0.412±0.001 g/g, and 1.13±0.01 g/L/h respectively within 78 h. PMID:27603922

Optimization of Nutrient Composition for Producing ACE Inhibitory Peptides from Goat Milk Fermented by Lactobacillus bulgaricus LB6.

PubMed

Shu, Guowei; Shi, Xiaoyu; Chen, He; Ji, Zhe; Meng, Jiangpeng

2018-03-23

Hypertension is a serious threat to human health and food-derived angiotensin converting enzyme (ACE; EC 3.4.15.1) inhibitory peptides can be used to regulate high blood pressure without side effects. The composition of the nutrient medium for the production of these peptides by fermenting goat milk with Lactobacillus bulgaricus LB6 was optimized to increase the ACE inhibitory activity by Box-Behnken design (BBD) of response surface methodology (RSM) in the present study. Soybean peptone, glucose, and casein had significant effects on both ACE inhibition rate and viable counts of L. bulgaricus LB6 during incubation. The results showed that the maximum values of ACE inhibition rate and viable counts for L. bulgaricus LB6 were reaching to 86.37 ± 0.53% and 8.06 × 10 7 under the optimal conditions, which were 0.35% (w/w) soybean peptone, 1.2% (w/w) glucose, and 0.15% (w/w) casein. The results were in close agreement with the model prediction. The optimal values of the medium component concentrations can be a good reference for obtaining ACE inhibitory peptides from goat milk.

Application of solid-acid catalyst and marine macro-algae Gracilaria verrucosa to production of fermentable sugars.

PubMed

Jeong, Gwi-Taek; Kim, Sung-Koo; Park, Don-Hee

2015-04-01

In this study, the hydrolysis of marine macro-algae Gracilaria verrucosa with a solid-acid catalyst was investigated. To optimize the hydrolysis, four reaction factors, including liquid-to-solid ratio, catalyst loading, reaction temperature, and reaction time, were investigated. In the results, the highest total reducing sugar (TRS) yield, 61 g/L (51.9%), was obtained under the following conditions: 1:7.5 solid-to-liquid ratio, 15% (w/v) catalyst loading, 140 °C reaction temperature, and 150 min reaction time. Under these conditions, 10.7 g/L of 5-HMF and 2.5 g/L of levulinic acid (LA) were generated. The application of solid-acid catalyst and marine macro-algae resources shows a very high potential for production of fermentable sugars. Copyright © 2015 Elsevier Ltd. All rights reserved.

Combinatorial pretreatment and fermentation optimization enabled a record yield on lignin bioconversion.

PubMed

Liu, Zhi-Hua; Xie, Shangxian; Lin, Furong; Jin, Mingjie; Yuan, Joshua S

2018-01-01

Lignin valorization has recently been considered to be an essential process for sustainable and cost-effective biorefineries. Lignin represents a potential new feedstock for value-added products. Oleaginous bacteria such as Rhodococcus opacus can produce intracellular lipids from biodegradation of aromatic substrates. These lipids can be used for biofuel production, which can potentially replace petroleum-derived chemicals. However, the low reactivity of lignin produced from pretreatment and the underdeveloped fermentation technology hindered lignin bioconversion to lipids. In this study, combinatorial pretreatment with an optimized fermentation strategy was evaluated to improve lignin valorization into lipids using R. opacus PD630. As opposed to single pretreatment, combinatorial pretreatment produced a 12.8-75.6% higher lipid concentration in fermentation using lignin as the carbon source. Gas chromatography-mass spectrometry analysis showed that combinatorial pretreatment released more aromatic monomers, which could be more readily utilized by lignin-degrading strains. Three detoxification strategies were used to remove potential inhibitors produced from pretreatment. After heating detoxification of the lignin stream, the lipid concentration further increased by 2.9-9.7%. Different fermentation strategies were evaluated in scale-up lipid fermentation using a 2.0-l fermenter. With laccase treatment of the lignin stream produced from combinatorial pretreatment, the highest cell dry weight and lipid concentration were 10.1 and 1.83 g/l, respectively, in fed-batch fermentation, with a total soluble substrate concentration of 40 g/l. The improvement of the lipid fermentation performance may have resulted from lignin depolymerization by the combinatorial pretreatment and laccase treatment, reduced inhibition effects by fed-batch fermentation, adequate oxygen supply, and an accurate pH control in the fermenter. Overall, these results demonstrate that combinatorial pretreatment, together with fermentation optimization, favorably improves lipid production using lignin as the carbon source. Combinatorial pretreatment integrated with fed-batch fermentation was an effective strategy to improve the bioconversion of lignin into lipids, thus facilitating lignin valorization in biorefineries.

Integrated process development-a robust, rapid method for inclusion body harvesting and processing at the microscale level.

PubMed

Walther, Cornelia; Kellner, Martin; Berkemeyer, Matthias; Brocard, Cécile; Dürauer, Astrid

2017-10-21

Escherichia coli stores large amounts of highly pure product within inclusion bodies (IBs). To take advantage of this beneficial feature, after cell disintegration, the first step to optimal product recovery is efficient IB preparation. This step is also important in evaluating upstream optimization and process development, due to the potential impact of bioprocessing conditions on product quality and on the nanoscale properties of IBs. Proper IB preparation is often neglected, due to laboratory-scale methods requiring large amounts of materials and labor. Miniaturization and parallelization can accelerate analyses of individual processing steps and provide a deeper understanding of up- and downstream processing interdependencies. Consequently, reproducible, predictive microscale methods are in demand. In the present study, we complemented a recently established high-throughput cell disruption method with a microscale method for preparing purified IBs. This preparation provided results comparable to laboratory-scale IB processing, regarding impurity depletion, and product loss. Furthermore, with this method, we performed a "design of experiments" study to demonstrate the influence of fermentation conditions on the performance of subsequent downstream steps and product quality. We showed that this approach provided a 300-fold reduction in material consumption for each fermentation condition and a 24-fold reduction in processing time for 24 samples.

Butanol production under microaerobic conditions with a symbiotic system of Clostridium acetobutylicum and Bacillus cereus.

PubMed

Wu, Pengfei; Wang, Genyu; Wang, Gehua; Børresen, Børre Tore; Liu, Hongjuan; Zhang, Jianan

2016-01-14

One major problem of ABE (acetone, butanol and ethanol) fermentation is high oxygen sensitivity of Clostridium acetobutylicum. Currently, no single strain has been isolated or genetically engineered to produce butanol effectively under aerobic conditions. In our previous work, a symbiotic system TSH06 has been developed successfully by our group, and two strains, C. acetobutylicum TSH1 and Bacillus cereus TSH2, were isolated from TSH06. Compared with single culture, TSH06 showed promotion on cell growth and solvent accumulation under microaerobic conditions. To simulate TSH06, a new symbiotic system was successfully re-constructed by adding living cells of B. cereus TSH2 into C. acetobutylicum TSH1 cultures. During the fermentation process, the function of B. cereus TSH2 was found to deplete oxygen and provide anaerobic environment for C. acetobutylicum TSH1. Furthermore, inoculation ratio of C. acetobutylicum TSH1 and B. cereus TSH2 affected butanol production. In a batch fermentation with optimized inoculation ratio of 5 % C. acetobutylicum TSH1 and 0.5 % B. cereus TSH2, 11.0 g/L butanol and 18.1 g/L ABE were produced under microaerobic static condition. In contrast to the single culture of C. acetobutylicum TSH1, the symbiotic system became more aerotolerant and was able to produce 11.2 g/L butanol in a 5 L bioreactor even with continuous 0.15 L/min air sparging. In addition, qPCR assay demonstrated that the abundance of B. cereus TSH2 increased quickly at first and then decreased sharply to lower than 1 %, whereas C. acetobutylicum TSH1 accounted for more than 99 % of the whole population in solventogenic phase. The characterization of a novel symbiotic system on butanol fermentation was studied. The new symbiotic system re-constructed by co-culture of C. acetobutylicum TSH1 and B. cereus TSH2 showed excellent performance on butanol production under microaerobic conditions. B. cereus TSH2 was a good partner for C. acetobutylicum TSH1 by providing an anaerobic environment. During fermentation process, the high ratio of Clostridium and low ratio of Bacillus composition indicated that this symbiotic system was an effective and easily controlled cultivation model for ABE fermentation under microaerobic conditions.

Simultaneously saccharification and fermentation approach as a tool for enhanced fossil fuels biodesulfurization.

PubMed

Paixão, Susana M; Arez, Bruno F; Roseiro, José C; Alves, Luís

2016-11-01

Biodesulfurization can be a complementary technology to the hydrodesulfurization, the commonly physical-chemical process used for sulfur removal from crude oil. The desulfurizing bacterium Gordonia alkanivorans strain 1B as a fructophilic microorganism requires fructose as C-source. In this context, the main goal of this work was the optimization of a simultaneous saccharification and fermentation (SSF) approach using the Zygosaccharomyces bailii strain Talf1 crude enzymes with invertase activity and sucrose as a cheaper fructose-rich commercial C-source (50% fructose) towards dibenzothiophene (DBT) desulfurization by strain 1B. The determination of optimal conditions, for both sucrose hydrolysis and DBT desulfurization was carried out through two sequential experimental uniform designs according to the Doehlert distribution for two factors: pH (5.5-7.5) and temperature (28-38 °C), with the enzyme load of 1.16 U/g/L; and enzyme load (0-4 U/g/L) and temperature (28-38 °C), with pH at 7.5. Based on 2-hydroxybiphenyl production, the analysis of the response surfaces obtained pointed out for pH 7.5, 32 °C and 1.8 U/g/L as optimal conditions. Further optimized SSF of sucrose during the DBT desulfurization process permitted to attain a 4-fold enhanced biodesulfurization. This study opens a new focus of research through the exploitation of sustainable low cost sucrose-rich feedstocks towards a more economical viable bioprocess scale-up. Copyright © 2016 Elsevier Ltd. All rights reserved.

Production of Self-Purifying Proteins in a Variety of Expression Hosts with Focus on Organophosphorus Hydrolase

DTIC Science & Technology

2012-08-17

cell-density fermentation at laboratory scale, and have provided evidence of their effectiveness. Our most recent work has been on the optimization...of the fermentation process itself, as well as a more biochemical optimization of the expression system. Overall, the ARO support on this project...large scale in high-density fermentation in microbial hosts, which is a critical gap in its appeal. The overall goals of our first renewal proposal

Xylose fermentation to ethanol by new Galactomyces geotrichum and Candida akabanensis strains.

PubMed

Valinhas, Raquel V; Pantoja, Lílian A; Maia, Ana Carolina F; Miguel, Maria Gabriela C P; Vanzela, Ana Paula F C; Nelson, David L; Santos, Alexandre S

2018-01-01

The conversion of pentoses into ethanol remains a challenge and could increase the supply of second-generation biofuels. This study sought to isolate naturally occurring yeasts from plant biomass and determine their capabilities for transforming xylose into ethanol. Three yeast strains with the ability to ferment xylose were isolated from pepper, tomato and sugarcane bagasse. The strains selected were characterized by morphological and auxanographic assays, and they were identified by homology analysis of 5.8 S and 26 S ribosomal RNA gene sequences. The identities of two lineages of microrganism were associated with Galactomyces geotrichum , and the other was associated with Candida akabanensis . Fermentative processes were conducted with liquid media containing only xylose as the carbon source. Y P/S values for the production of ethanol ranging between 0.29 and 0.35 g g -1 were observed under non-optimized conditions.

Improve the Recovery of Fermentable Sugar from Rice Straw by Sonication and Its Mathematical Modeling

NASA Astrophysics Data System (ADS)

Bhattacharyya, Saurav; Dutta, Somenath; Datta, Sidhartha; Bhattacharjee, Chiranjib

2012-08-01

Rice straw is waste renewable agricultural biomass, which contains sufficient amount of fermentable sugars like glucose, galactose fructose, xylose etc. These sugars can be treated with fermentation pathway to produce ethanol. Hydrolysis of pretreated rice straw in dilute sulfuric acid was investigated at different acid concentrations (0.25-0.75 % w/v), and sonication was carried out to improve the extent of sugar extraction. The current work examines the effect of sonication on extraction of total reducing sugar (TRS) and an empirical mathematical model has been established to predict it. Effects of various operating variables of sonication, including amplitude (60-100 %), cycle (0.6-1.0), treatment time (0-15 min) have been analyzed for each acid concentration. Observation shows that on optimization of the sonication conditions (100 % amplitude, 0.8 cycle and 10 min) around 90 % improvement of TRS extraction occurs at 0.5 % (w/v) acid concentration.

Conversion of lignocellulosic agave residues into liquid biofuels using an AFEX™-based biorefinery.

PubMed

Flores-Gómez, Carlos A; Escamilla Silva, Eleazar M; Zhong, Cheng; Dale, Bruce E; da Costa Sousa, Leonardo; Balan, Venkatesh

2018-01-01

Agave-based alcoholic beverage companies generate thousands of tons of solid residues per year in Mexico. These agave residues might be used for biofuel production due to their abundance and favorable sustainability characteristics. In this work, agave leaf and bagasse residues from species Agave tequilana and Agave salmiana were subjected to pretreatment using the ammonia fiber expansion (AFEX) process. The pretreatment conditions were optimized using a response surface design methodology. We also identified commercial enzyme mixtures that maximize sugar yields for AFEX-pretreated agave bagasse and leaf matter, at ~ 6% glucan (w/w) loading enzymatic hydrolysis. Finally, the pretreated agave hydrolysates (at a total solids loading of ~ 20%) were used for ethanol fermentation using the glucose- and xylose-consuming strain Saccharomyces cerevisiae 424A (LNH-ST), to determine ethanol yields at industrially relevant conditions. Low-severity AFEX pretreatment conditions are required (100-120 °C) to enable efficient enzymatic deconstruction of the agave cell wall. These studies showed that AFEX-pretreated A. tequilana bagasse, A. tequilana leaf fiber, and A. salmiana bagasse gave ~ 85% sugar conversion during enzyme hydrolysis and over 90% metabolic yields of ethanol during fermentation without any washing step or nutrient supplementation. On the other hand, although lignocellulosic A. salmiana leaf gave high sugar conversions, the hydrolysate could not be fermented at high solids loadings, apparently due to the presence of natural inhibitory compounds. These results show that AFEX-pretreated agave residues can be effectively hydrolyzed at high solids loading using an optimized commercial enzyme cocktail (at 25 mg protein/g glucan) producing > 85% sugar conversions and over 40 g/L bioethanol titers. These results show that AFEX technology has considerable potential to convert lignocellulosic agave residues to bio-based fuels and chemicals in a biorefinery.

Optimization of prehydrolysis time and substrate feeding to improve ethanol production by simultaneous saccharification and fermentation of furfural process residue.

PubMed

He, Jianlong; Zhang, Wenbo; Liu, Xiaoyan; Xu, Ning; Xiong, Peng

2016-11-01

Ethanol is a very important industrial chemical. In order to improve ethanol productivity using Saccharomyces cerevisiae in fermentation from furfural process residue, we developed a process of simultaneous saccharification and fermentation (SSF) of furfural process residue, optimizing prehydrolysis cellulase loading concentration, prehydrolysis time, and substrate feeding strategy. The ethanol concentration obtained from the optimized process was 19.3 g/L, corresponding 76.5% ethanol yield, achieved by running SSF for 48 h from 10% furfural process residue with prehydrolysis at 50°C for 4 h and cellulase loading of 15 FPU/g furfural process residue. For higher ethanol concentrations, fed-batch fermentation was performed. The optimized fed-batch process increased the ethanol concentration to 37.6 g/L, 74.5% yield, obtained from 10% furfural process residue with two additions of 5% substrate at 12 and 24 h. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Optimization of Aspergillus niger rock phosphate solubilization in solid-state fermentation and use of the resulting product as a P fertilizer

PubMed Central

Mendes, Gilberto de Oliveira; da Silva, Nina Morena Rêgo Muniz; Anastácio, Thalita Cardoso; Vassilev, Nikolay Bojkov; Ribeiro, José Ivo; da Silva, Ivo Ribeiro; Costa, Maurício Dutra

2015-01-01

A biotechnological strategy for the production of an alternative P fertilizer is described in this work. The fertilizer was produced through rock phosphate (RP) solubilization by Aspergillus niger in a solid-state fermentation (SSF) with sugarcane bagasse as substrate. SSF conditions were optimized by the surface response methodology after an initial screening of factors with significant effect on RP solubilization. The optimized levels of the factors were 865 mg of biochar, 250 mg of RP, 270 mg of sucrose and 6.2 ml of water per gram of bagasse. At this optimal setting, 8.6 mg of water-soluble P per gram of bagasse was achieved, representing an increase of 2.4 times over the non-optimized condition. The optimized SSF product was partially incinerated at 350°C (SB-350) and 500°C (SB-500) to reduce its volume and, consequently, increase P concentration. The post-processed formulations of the SSF product were evaluated in a soil–plant experiment. The formulations SB-350 and SB-500 increased the growth and P uptake of common bean plants (Phaseolus vulgaris L.) when compared with the non-treated RP. Furthermore, these two formulations had a yield relative to triple superphosphate of 60% (on a dry mass basis). Besides increasing P concentration, incineration improved the SSF product performance probably by decreasing microbial immobilization of nutrients during the decomposition of the remaining SSF substrate. The process proposed is a promising alternative for the management of P fertilization since it enables the utilization of low-solubility RPs and relies on the use of inexpensive materials. PMID:26112323

Optimization of Aspergillus niger rock phosphate solubilization in solid-state fermentation and use of the resulting product as a P fertilizer.

PubMed

Mendes, Gilberto de Oliveira; da Silva, Nina Morena Rêgo Muniz; Anastácio, Thalita Cardoso; Vassilev, Nikolay Bojkov; Ribeiro, José Ivo; da Silva, Ivo Ribeiro; Costa, Maurício Dutra

2015-11-01

A biotechnological strategy for the production of an alternative P fertilizer is described in this work. The fertilizer was produced through rock phosphate (RP) solubilization by Aspergillus niger in a solid-state fermentation (SSF) with sugarcane bagasse as substrate. SSF conditions were optimized by the surface response methodology after an initial screening of factors with significant effect on RP solubilization. The optimized levels of the factors were 865 mg of biochar, 250 mg of RP, 270 mg of sucrose and 6.2 ml of water per gram of bagasse. At this optimal setting, 8.6 mg of water-soluble P per gram of bagasse was achieved, representing an increase of 2.4 times over the non-optimized condition. The optimized SSF product was partially incinerated at 350°C (SB-350) and 500°C (SB-500) to reduce its volume and, consequently, increase P concentration. The post-processed formulations of the SSF product were evaluated in a soil-plant experiment. The formulations SB-350 and SB-500 increased the growth and P uptake of common bean plants (Phaseolus vulgaris L.) when compared with the non-treated RP. Furthermore, these two formulations had a yield relative to triple superphosphate of 60% (on a dry mass basis). Besides increasing P concentration, incineration improved the SSF product performance probably by decreasing microbial immobilization of nutrients during the decomposition of the remaining SSF substrate. The process proposed is a promising alternative for the management of P fertilization since it enables the utilization of low-solubility RPs and relies on the use of inexpensive materials. © 2015 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

Phenotypic selection of a wild Saccharomyces cerevisiae strain for simultaneous saccharification and co-fermentation of AFEX pretreated corn stover

Treesearch

Mingie Jin; Cory Sarks; Christa Gunawan; Benjamin D. Bice; Shane P. Simonett; Ragothaman Avanasi Narasimhan; Laura B. Willis; Bruce E. Dale; Venkatesh Balan; Trey K. Sato

2013-01-01

Simultaneous saccharification and co-fermentation (SSCF) process involves enzymatic hydrolysis of pretreated lignocellulosic biomass and fermentation of glucose and xylose in one bioreactor. The optimal temperatures for enzymatic hydrolysis are higher than the standard fermentation temperature of ethanologenic Saccharomyces cerevisiae. Moreover,...

[Optimization of succinic acid fermentation with Actinobacillus succinogenes by response surface methodology].

PubMed

Shen, Naikun; Qin, Yan; Wang, Qingyan; Xie, Nengzhong; Mi, Huizhi; Zhu, Qixia; Liao, Siming; Huang, Ribo

2013-10-01

Succinic acid is an important C4 platform chemical in the synthesis of many commodity and special chemicals. In the present work, different compounds were evaluated for succinic acid production by Actinobacillus succinogenes GXAS 137. Important parameters were screened by the single factor experiment and Plackeet-Burman design. Subsequently, the highest production of succinic acid was approached by the path of steepest ascent. Then, the optimum values of the parameters were obtained by Box-Behnken design. The results show that the important parameters were glucose, yeast extract and MgCO3 concentrations. The optimum condition was as follows (g/L): glucose 70.00, yeast extract 9.20 and MgCO3 58.10. Succinic acid yield reached 47.64 g/L at the optimal condition. Succinic acid increased by 29.14% than that before the optimization (36.89 g/L). Response surface methodology was proven to be a powerful tool to optimize succinic acid production.

Enhanced production of raw starch degrading enzyme using agro-industrial waste mixtures by thermotolerant Rhizopus microsporus for raw cassava chip saccharification in ethanol production.

PubMed

Trakarnpaiboon, Srisakul; Srisuk, Nantana; Piyachomkwan, Kuakoon; Sakai, Kenji; Kitpreechavanich, Vichien

2017-09-14

In the present study, solid-state fermentation for the production of raw starch degrading enzyme was investigated by thermotolerant Rhizopus microsporus TISTR 3531 using a combination of agro-industrial wastes as substrates. The obtained crude enzyme was applied for hydrolysis of raw cassava starch and chips at low temperature and subjected to nonsterile ethanol production using raw cassava chips. The agro-industrial waste ratio was optimized using a simplex axial mixture design. The results showed that the substrate mixture consisting of rice bran:corncob:cassava bagasse at 8 g:10 g:2 g yielded the highest enzyme production of 201.6 U/g dry solid. The optimized condition for solid-state fermentation was found as 65% initial moisture content, 35°C, initial pH of 6.0, and 5 × 10 6 spores/mL inoculum, which gave the highest enzyme activity of 389.5 U/g dry solid. The enzyme showed high efficiency on saccharification of raw cassava starch and chips with synergistic activities of commercial α-amylase at 50°C, which promotes low-temperature bioethanol production. A high ethanol concentration of 102.2 g/L with 78% fermentation efficiency was achieved from modified simultaneous saccharification and fermentation using cofermentation of the enzymatic hydrolysate of 300 g raw cassava chips/L with cane molasses.

Phytase production by solid-state fermentation of groundnut oil cake by Aspergillus niger: A bioprocess optimization study for animal feedstock applications.

PubMed

Buddhiwant, Priyanka; Bhavsar, Kavita; Kumar, V Ravi; Khire, Jayant M

2016-08-17

This investigation deals with the use of agro-industrial waste, namely groundnut oil cake (GOC), for phytase production by the fungi Aspergillus niger NCIM 563. Plackett-Burman design (PBD) was used to evaluate the effect of 11 process variables and studies here showed that phytase production was significantly influenced by glucose, dextrin, distilled water, and MgSO4 · 7H2O. The use of response surface methodology (RSM) by Box-Behnken design (BBD) of experiments further enhanced the production by a remarkable 36.67-fold from the original finding of 15 IU/gds (grams of dry substrate) to 550 IU/gds. This is the highest solid-state fermentation (SSF) phytase production reported when compared to other microorganisms and in fact betters the best known by a factor of 2. Experiments carried out using dried fermented koji for phosphorus and mineral release and also thermal stability have shown the phytase to be as efficient as the liquid enzyme extract. Also, the enzyme, while exhibiting optimal activity under acidic conditions, was found to have significant activity in a broad range of pH values (1.5-6.5). The studies suggest the suitability of the koji supplemented with phytase produced in an SSF process by the "generally regarded as safe" (GRAS) microorganism A. niger as a cost-effective value-added livestock feed when compared to that obtained by submerged fermentation (SmF).

Recovery of nitrogen and phosphorus from alkaline fermentation liquid of waste activated sludge and application of the fermentation liquid to promote biological municipal wastewater treatment.

PubMed

Tong, Juan; Chen, Yinguang

2009-07-01

In previous publications we reported that by controlling the pH at 10.0 the accumulation of short-chain fatty acids (SCFA) during waste activated sludge (WAS) fermentation was remarkably improved [Yuan, H., Chen, Y., Zhang, H., Jiang, S., Zhou, Q., Gu, G., 2006. Improved bioproduction of short-chain fatty acids (SCFAs) from excess sludge under alkaline conditions. Environ. Sci. Technol. 40, 2025-2029], but significant ammonium nitrogen (NH(4)-N) and soluble ortho-phosphorus (SOP) were released [Chen, Y., Jiang, S., Yuan, H., Zhou, Q., Gu, G., 2007. Hydrolysis and acidification of waste activated sludge at different pHs. Water Res. 41, 683-689]. This paper investigated the simultaneous recovery of NH(4)-N and SOP from WAS alkaline fermentation liquid and the application of the fermentation liquid as an additional carbon source for municipal wastewater biological nitrogen and phosphorus removal. The central composite design (CCD) of the response surface methodology (RSM) was employed to optimize and model the simultaneous NH(4)-N and SOP recovery from WAS alkaline fermentation liquid. Under the optimum conditions, the predicted and experimental recovery efficiency was respectively 73.4 and 75.7% with NH(4)-N, and 82.0 and 83.2% with SOP, which suggested that the developed models described the experiments well. After NH(4)-N and SOP recovery, the alkaline fermentation liquid was added to municipal wastewater, and the influence of volume ratio of fermentation liquid to municipal wastewater (FL/MW) on biological nitrogen and phosphorus removal was investigated. The addition of fermentation liquid didn't significantly affect nitrification. Both SOP and total nitrogen (TN) removal were increased with fermentation liquid, but there was no significant increase at FL/MW greater than 1/35. Compared to the blank test, the removal efficiency of SOP and TN at FL/MW=1/35 was improved from 44.0 to 92.9%, and 63.3 to 83.2%, respectively. The enhancement of phosphorus and nitrogen removal was mainly attributed to the increase of influent SCFA, or rather, the increase of intracellular polyhydroxyalkanoates (PHA) which served as the carbon and energy sources for denitrification and phosphorus uptake. The addition of alkaline fermentation liquid to municipal wastewater, however, increased the effluent COD, which was caused mainly by the increase of influent humic acid, not protein or carbohydrate.

Production of high optical purity l-lactic acid from waste activated sludge by supplementing carbohydrate: effect of temperature and pretreatment time.

PubMed

Jian, Qiwei; Li, Xiang; Chen, Yinguang; Liu, Yanan; Pan, Yin

2016-10-01

It has been widely accepted that the most environmentally beneficial way to treat waste activated sludge (WAS), the byproduct of municipal wastewater treatment plant, is to recover the valuable organic acid. However, the bio-conversion of lactic acid, one of the high added-value chemical, is seldom reported from WAS fermentation. In this paper, l-lactic acid was observed dominant in the WAS fermentation liquid with carbohydrate addition at ambient temperature. Furthermore, the effect of temperature on l-lactic acid and d-lactic acid production was fully discussed: two isomers were rapidly produced and consumed up in one day at mesophilic condition; and almost optically pure l-lactic acid was generated at thermophilic condition, yet time-consuming with yield of l-lactic acid enhancing by 52.9% compared to that at ambient temperature. The study mechanism showed that mesophilic condition was optimal for both production and consumption of l-lactic acid and d-lactic acid, while consumption of l-lactic acid and production of d-lactic acid were severely inhibited at thermophilic condition. Therefore, by maintaining thermophilic for 4 h in advance and subsequently fermenting mesophilic for 34 h, the concentration of l-lactic acid with optical activity of 98.3% was improved to 16.6 ± 0.5 g COD/L at a high specific efficiency of 0.6097/d.

Biotransformation of 5-hydroxymethylfurfural (HMF) by Scheffersomyces stipitis during ethanol fermentation of hydrolysate of the seaweed Gelidium amansii.

PubMed

Ra, Chae Hun; Jeong, Gwi-Taek; Shin, Myung Kyo; Kim, Sung-Koo

2013-07-01

The seaweed, Gelidium amansii, was fermented to produce bioethanol. Optimal pretreatment condition was determined as 94 mM H2SO4 and 10% (w/v) seaweed slurry at 121°C for 60 min. The mono sugars of 43.5 g/L with 57.4% of conversion from total carbohydrate of 75.8 g/L with G. amansii slurry 100g dcw/L were obtained by thermal acid hydrolysis pretreatment and enzymatic saccharification. G. amansii hydrolysate was used as the substrate for ethanol production by separate hydrolysis and fermentation (SHF). The ethanol concentration of 20.5 g/L was produced by Scheffersomyces stipitis KCTC 7228. The effect of HMF on ethanol production by S. stipitis KCTC 7228 was evaluated and 5-hydroxymethylfurfural (HMF) was converted to 2,5-bis-hydroxymethylfuran. The accumulated 2,5-bis-hydroxymethylfuran in the medium did not affect galactose and glucose uptakes and ethanol production. Biotransformation of HMF to less inhibitory compounds by S. stipitis KCTC 7228 could enhance overall fermentation yields of seaweed hydrolysates to ethanol. Copyright © 2013 Elsevier Ltd. All rights reserved.

Open fermentative production of L-lactic acid with high optical purity by thermophilic Bacillus coagulans using excess sludge as nutrient.

PubMed

Ma, Kedong; Maeda, Toshinari; You, Huiyan; Shirai, Yoshihito

2014-01-01

The development of a low-cost polymer-grade L-lactic acid production process was achieved in this study. Excess sludge hydrolyzate (ESH) was chosen as nutrient source for the objective of reducing nutrient cost in lactic acid production. 1% of ESH had high performance in lactic acid production relative to 2g/l yeast extract (YE) while the production cost of ESH was much lower than that of YE, indicating ESH was a promising substitute of YE. By employing a thermophilic strain of Bacillus coagulans (NBRC 12583), non-sterilized batch and repeated batch L-lactic acid fermentation was successfully performed, and the optical purity of L-lactic acid accumulated was more than 99%. Moreover, the factors associated with cell growth and lactic acid fermentation was investigated through a two-stage lactic acid production strategy. Oxygen played an important role in cell growth, and the optimal condition for cell growth and fermentation was pH 7.0 and 50°C. Copyright © 2013 Elsevier Ltd. All rights reserved.

Computer Simulation Elucidates Yeast Flocculation and Sedimentation for Efficient Industrial Fermentation.

PubMed

Liu, Chen-Guang; Li, Zhi-Yang; Hao, Yue; Xia, Juan; Bai, Feng-Wu; Mehmood, Muhammad Aamer

2018-05-01

Flocculation plays an important role in the immobilized fermentation of biofuels and biochemicals. It is essential to understand the flocculation phenomenon at physical and molecular scale; however, flocs cannot be studied directly due to fragile nature. Hence, the present study is focused on the morphological specificities of yeast flocs formation and sedimentation via the computer simulation by a single floc growth model, based on Diffusion-Limited Aggregation (DLA) model. The impact of shear force, adsorption, and cell propagation on porosity and floc size is systematically illustrated. Strong shear force and weak adsorption reduced floc size but have little impact on porosity. Besides, cell propagation concreted the compactness of flocs enabling them to gain a larger size. Later, a multiple flocs growth model is developed to explain sedimentation at various initial floc sizes. Both models exhibited qualitative agreements with available experimental data. By regulating the operation constraints during fermentation, the present study will lead to finding optimal conditions to control the floc size distribution for efficient fermentation and harvesting. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

The effect of pH on solubilization of organic matter and microbial community structures in sludge fermentation.

PubMed

Maspolim, Yogananda; Zhou, Yan; Guo, Chenghong; Xiao, Keke; Ng, Wun Jern

2015-08-01

Sludge fermentation between pH 4 and 11 was investigated to generate volatile fatty acids (VFA). Despite the highest sludge solubilization of 25.9% at pH 11, VFA accumulation was optimized at pH 8 (12.5% out of 13.1% sludge solubilization). 454 pyrosequencing identified wide diversity of acidogens in bioreactors operated at the various pHs, with Tissierella, Petrimonas, Proteiniphilum, Levilinea, Proteiniborus and Sedimentibacter enriched and contributing to the enhanced fermentation at pH 8. Hydrolytic enzymatic assays determined abiotic effect to be the leading cause for improved solubilization under high alkaline condition but the environmental stress at pH 9 and above might lead to disrupt biological activities and eventually VFA production. Furthermore, molecular weight (MW) characterization of the soluble fractions found large MW aromatic substances at pH 9 and above, that is normally associated with poor biodegradability, making them disadvantageous for subsequent bioprocesses. The findings provided information to better understand and control sludge fermentation. Copyright © 2015 Elsevier Ltd. All rights reserved.

Optimization of Fermentation Conditions and Rheological Properties of Exopolysaccharide Produced by Deep-Sea Bacterium Zunongwangia profunda SM-A87

PubMed Central

Liu, Sheng-Bo; Qiao, Li-Ping; He, Hai-Lun; Zhang, Qian; Chen, Xiu-Lan; Zhou, Wei-Zhi; Zhou, Bai-Cheng; Zhang, Yu-Zhong

2011-01-01

Zunongwangia profunda SM-A87 isolated from deep-sea sediment can secrete large quantity of exopolysaccharide (EPS). Response surface methodology was applied to optimize the culture conditions for EPS production. Single-factor experiment showed that lactose was the best carbon source. Based on the Plackett–Burman design, lactose, peptone and temperature were selected as significant variables, which were further optimized by the steepest ascent (descent) method and central composite design. The optimal culture conditions for EPS production and broth viscosity were determined as 32.21 g/L lactose, 8.87 g/L peptone and an incubation temperature of 9.8°C. Under these conditions, the maximum EPS yield and broth viscosity were 8.90 g/L and 6551 mPa•s, respectively, which is the first report of such high yield of EPS from a marine bacterium. The aqueous solution of the EPS displayed high viscosity, interesting shearing thinning property and great tolerance to high temperature, a wide range of pH, and high salinity. PMID:22096500

Development of a model system for the study of spoilage associated secondary cucumber fermentation during long-term storage.

PubMed

Franco, Wendy; Pérez-Díaz, Ilenys M

2012-10-01

Calcium chloride fermentations represent an alternative to reduce chloride concentrations in the wastewaters generated from commercial cucumber fermentations, currently performed in cover brine solutions containing 6% to 12% sodium chloride. However, preliminary attempts to commercially ferment the cucumbers in the presence of oxygen led to the development of a secondary cucumber fermentation or spoilage. The development of cucumber secondary fermentation has also been occasionally reported by processors using cover brine solutions containing sodium chloride. This study focused on the development of a model system to characterize CaCl(2) and NaCl secondary cucumber fermentations under conditions similar to those present on the commercial scale. Cucumber fruits mixed with cover brine solutions, containing 100 mM CaCl(2) or 1.03 M NaCl, and 25 mM acetic acid, were fermented in 2 L fermentation vessels subjected to air-purging at a rate of 5 mL/min. Microorganisms and selected biochemical changes detected in the experimental cucumber fermentations had been previously observed in commercial spoilage samples, suggesting the successful reproduction of the secondary fermentation in the laboratory. Experimental secondary fermentations were characterized by the rapid oxidation of the lactic acid produced during the primary fermentation, which, in turn, increased pH. Lactic acid disappearance seemed to be the result of yeast metabolism that also led to the chemical reduction of the environment to levels at which other bacteria could become established and produce butyric, propionic, and acetic acids. This model system will be applied for the identification of strategies to prevent the initiation of the cucumber secondary fermentation and reduce economic losses in the pickling industry. The study of secondary cucumber fermentation has represented a challenge for many years. The successful development of a model system for the study of this phenomenon in the laboratory is instrumental in furthering the study of the event and in optimizing the sodium-chloride-free fermentation at the commercial scale. Journal of Food Science © 2012 Institute of Food Technologists® No claim to original US government works.

Statistical optimization of medium composition for bacterial cellulose production by Gluconacetobacter hansenii UAC09 using coffee cherry husk extract--an agro-industry waste.

PubMed

Rani, Mahadevaswamy Usha; Rastogi, Navin K; Appaiah, K A Anu

2011-07-01

During the production of grape wine, the formation of thick leathery pellicle/bacterial cellulose (BC) at the airliquid interface was due to the bacterium, which was isolated and identified as Gluconacetobacter hansenii UAC09. Cultural conditions for bacterial cellulose production from G. hansenii UAC09 were optimized by central composite rotatable experimental design. To economize the BC production, coffee cherry husk (CCH) extract and corn steep liquor (CSL) were used as less expensive sources of carbon and nitrogen, respectively. CCH and CSL are byproducts from the coffee processing and starch processing industry, respectively. The interactions between pH (4.5- 8.5), CSL (2-10%), alcohol (0.5-2%), acetic acid (0.5- 2%), and water dilution rate to CCH ratio (1:1 to 1:5) were studied using response surface methodology. The optimum conditions for maximum BC production were pH (6.64), CSL (10%), alcohol (0.5%), acetic acid (1.13%), and water to CCH ratio (1:1). After 2 weeks of fermentation, the amount of BC produced was 6.24 g/l. This yield was comparable to the predicted value of 6.09 g/l. This is the first report on the optimization of the fermentation medium by RSM using CCH extract as the carbon source for BC production by G. hansenii UAC09.

Novel Bacillus subtilis IND19 cell factory for the simultaneous production of carboxy methyl cellulase and protease using cow dung substrate in solid-substrate fermentation.

PubMed

Vijayaraghavan, Ponnuswamy; Arun, Arumugaperumal; Al-Dhabi, Naif Abdullah; Vincent, Samuel Gnana Prakash; Arasu, Mariadhas Valan; Choi, Ki Choon

2016-01-01

Hydrolytic enzymes, such as cellulases and proteases, have various applications, including bioethanol production, extraction of fruit and vegetable juice, detergent formulation, and leather processing. Solid-substrate fermentation has been an emerging method to utilize low-cost agricultural residues for the production of these enzymes. Although the production of carboxy methyl cellulase (CMCase) and protease in solid state fermentation (SSF) have been studied extensively, research investigating multienzyme production in a single fermentation process is limited. The production of multienzymes from a single fermentation system could reduce the overall production cost of enzymes. In order to achieve enhanced production of enzymes, the response surface methodology (RSM) was applied. Bacillus subtilis IND19 utilized cow dung substrates for the production of CMCase and protease. A central composite design and a RSM were used to determine the optimal concentrations of peptone, NaH2PO4, and medium pH. Maximum productions of CMCase and protease were observed at 0.9 % peptone, 0.78 % NaH2PO4, and medium pH of 8.41, and 1 % peptone, 0.72 % NaH2PO4, and medium pH of 8.11, respectively. Under the optimized conditions, the experimental yield of CMCase and protease reached 473.01 and 4643 U/g, which were notably close to the predicted response (485.05 and 4710 U/g). These findings corresponded to an overall increase of 2.1- and 2.5-fold in CMCase and protease productions, respectively. Utilization of cow dung for the production of enzymes is critical to producing multienzymes in a single fermentation step. Cow dung is available in large quantity throughout the year. This report is the first to describe simultaneous production of CMCase and protease using cow dung. This substrate could be directly used as the culture medium without any pretreatment for the production of these enzymes at an industrial scale.

Maximizing power generation from dark fermentation effluents in microbial fuel cell by selective enrichment of exoelectrogens and optimization of anodic operational parameters.

PubMed

Varanasi, Jhansi L; Sinha, Pallavi; Das, Debabrata

2017-05-01

To selectively enrich an electrogenic mixed consortium capable of utilizing dark fermentative effluents as substrates in microbial fuel cells and to further enhance the power outputs by optimization of influential anodic operational parameters. A maximum power density of 1.4 W/m 3 was obtained by an enriched mixed electrogenic consortium in microbial fuel cells using acetate as substrate. This was further increased to 5.43 W/m 3 by optimization of influential anodic parameters. By utilizing dark fermentative effluents as substrates, the maximum power densities ranged from 5.2 to 6.2 W/m 3 with an average COD removal efficiency of 75% and a columbic efficiency of 10.6%. A simple strategy is provided for selective enrichment of electrogenic bacteria that can be used in microbial fuel cells for generating power from various dark fermentative effluents.

Development and Optimization of a Fluorescent Differential Display PCR System for Analyzing the Stress Response in Lactobacillus sakei Strains

PubMed Central

Bonomo, Maria Grazia; Sico, Maria Anna; Grieco, Simona; Salzano, Giovanni

2009-01-01

Lactobacillus sakei is widely used as starter in the production process of Italian fermented sausages and its growth and survival are affected by various factors. We studied the differential expression of genome in response to different stresses by the fluorescent differential display (FDD) technique. This study resulted in the development and optimization of an innovative technique, with a high level of reproducibility and quality, which allows the identification of gene expression changes associated with different microbial behaviours under different growth conditions. PMID:22253979

Performance study of sugar-yeast-ethanol bio-hybrid fuel cells

NASA Astrophysics Data System (ADS)

Jahnke, Justin P.; Mackie, David M.; Benyamin, Marcus; Ganguli, Rahul; Sumner, James J.

2015-05-01

Renewable alternatives to fossil hydrocarbons for energy generation are of general interest for a variety of political, economic, environmental, and practical reasons. In particular, energy from biomass has many advantages, including safety, sustainability, and the ability to be scavenged from native ecosystems or from waste streams. Microbial fuel cells (MFCs) can take advantage of microorganism metabolism to efficiently use sugar and other biomolecules as fuel, but are limited by low power densities. In contrast, direct alcohol fuel cells (DAFCs) take advantage of proton exchange membranes (PEMs) to generate electricity from alcohols at much higher power densities. Here, we investigate a novel bio-hybrid fuel cell design prepared using commercial off-the-shelf DAFCs. In the bio-hybrid fuel cells, biomass such as sugar is fermented by yeast to ethanol, which can be used to fuel a DAFC. A separation membrane between the fermentation and the DAFC is used to purify the fermentate while avoiding any parasitic power losses. However, shifting the DAFCs from pure alcohol-water solutions to filtered fermented media introduces complications related to how the starting materials, fermentation byproducts, and DAFC waste products affect both the fermentation and the long-term DAFC performance. This study examines the impact of separation membrane pore size, fermentation/fuel cell byproducts, alcohol and salt concentrations, and load resistance on fuel cell performance. Under optimized conditions, the performance obtained is comparable to that of a similar DAFC run with a pure alcohol-water mixture. Additionally, the modified DAFC can provide useable amounts of power for weeks.

Redesigning Escherichia coli Metabolism for Anaerobic Production of Isobutanol▿†

PubMed Central

Trinh, Cong T.; Li, Johnny; Blanch, Harvey W.; Clark, Douglas S.

2011-01-01

Fermentation enables the production of reduced metabolites, such as the biofuels ethanol and butanol, from fermentable sugars. This work demonstrates a general approach for designing and constructing a production host that uses a heterologous pathway as an obligately fermentative pathway to produce reduced metabolites, specifically, the biofuel isobutanol. Elementary mode analysis was applied to design an Escherichia coli strain optimized for isobutanol production under strictly anaerobic conditions. The central metabolism of E. coli was decomposed into 38,219 functional, unique, and elementary modes (EMs). The model predictions revealed that during anaerobic growth E. coli cannot produce isobutanol as the sole fermentative product. By deleting 7 chromosomal genes, the total 38,219 EMs were constrained to 12 EMs, 6 of which can produce high yields of isobutanol in a range from 0.29 to 0.41 g isobutanol/g glucose under anaerobic conditions. The remaining 6 EMs rely primarily on the pyruvate dehydrogenase enzyme complex (PDHC) and are typically inhibited under anaerobic conditions. The redesigned E. coli strain was constrained to employ the anaerobic isobutanol pathways through deletion of 7 chromosomal genes, addition of 2 heterologous genes, and overexpression of 5 genes. Here we present the design, construction, and characterization of an isobutanol-producing E. coli strain to illustrate the approach. The model predictions are evaluated in relation to experimental data and strategies proposed to improve anaerobic isobutanol production. We also show that the endogenous alcohol/aldehyde dehydrogenase AdhE is the key enzyme responsible for the production of isobutanol and ethanol under anaerobic conditions. The glycolytic flux can be controlled to regulate the ratio of isobutanol to ethanol production. PMID:21642415

High-loading-substrate enzymatic hydrolysis of palm plantation waste followed by unsterilized-mixed-culture fermentation for bio-ethanol production

NASA Astrophysics Data System (ADS)

Bardant, Teuku Beuna; Winarni, Ina; Sukmana, Hadid

2017-01-01

It was desired to obtain a general formula for producing bio-ethanol from any part of lignocelluloses wastes that came from palm oil industries due to its abundance. Optimum condition that obtained by using RSM for conducting high-loading-substrate enzymatic hydrolysis of palm oil empty fruit bunch was applied to palm oil trunks and then followed by unsterilized fermentation for producing bio-ethanol. From several optimized conditions investigated, the resulted ethanol concentration could reach 7.92 %v by using 36.5 %w of palm oil trunks but the results were averagely 2.46 %v lower than palm oil empty fruit bunch. The results was statistically compared and showed best correlative coefficient at 0.808 (in scale 0-1) which support the conclusion that the optimum condition for empty fruit bunch and trunks are similar. Utilization of mixed-culture yeast was investigated to produce ethanol from unsterilized hydrolysis product but the improvement wasn't significant compares to single culture yeast.

Identification of newly isolated Talaromyces pinophilus and statistical optimization of β-glucosidase production under solid-state fermentation.

PubMed

El-Naggar, Noura El-Ahmady; Haroun, S A; Oweis, Eman A; Sherief, A A

2015-01-01

Fungi able to degrade agriculture wastes were isolated from different soil samples, rice straw, and compost; these isolates were screened for their ability to produce β-glucosidase. The most active fungal isolate was identified as Talaromyces pinophilus strain EMOO 13-3. The Plackett-Burman design is used for identifying the significant variables that influence β-glucosidase production under solid-state fermentation. Fifteen variables were examined for their significances on the production of β-glucosidase in 20 experimental runs. Among the variables screened, moisture content, Tween 80, and (NH4)2SO4 had significant effects on β-glucosidase production with confidence levels above 90% (p < 0.1). The optimal levels of these variables were further optimized using Box-Behnken statical design. As a result, the maximal β-glucosidase activity is 3648.519 U g(-1), which is achieved at the following fermentation conditions: substrate amount 0.5 (g/250 mL flask), NaNO3 0.5 (%), KH2PO4 0.3 (%), KCl 0.02 (%), MgSO4 · 7H2O 0.01 (%), CaCl2 0.01 (%), yeast extract 0.07 (%), FeSO4 · 7H2O 0.0002 (%), Tween 80 0.02 (%), (NH4)2SO4 0.3 (%), pH 6.5, temperature 25°C, moisture content 1 (mL/g dry substrate), inoculum size 0.5 (mL/g dry substrate), and incubation period 5 days.

An obligately aerobic soil bacterium activates fermentative hydrogen production to survive reductive stress during hypoxia.

PubMed

Berney, Michael; Greening, Chris; Conrad, Ralf; Jacobs, William R; Cook, Gregory M

2014-08-05

Oxygen availability is a major factor and evolutionary force determining the metabolic strategy of bacteria colonizing an environmental niche. In the soil, conditions can switch rapidly between oxia and anoxia, forcing soil bacteria to remodel their energy metabolism accordingly. Mycobacterium is a dominant genus in the soil, and all its species are obligate aerobes. Here we show that an obligate aerobe, the soil actinomycete Mycobacterium smegmatis, adopts an anaerobe-type strategy by activating fermentative hydrogen production to adapt to hypoxia. This process is controlled by the two-component system DosR-DosS/DosT, an oxygen and redox sensor that is well conserved in mycobacteria. We show that DosR tightly regulates the two [NiFe]-hydrogenases: Hyd3 (MSMEG_3931-3928) and Hyd2 (MSMEG_2719-2718). Using genetic manipulation and high-sensitivity GC, we demonstrate that Hyd3 facilitates the evolution of H2 when oxygen is depleted. Combined activity of Hyd2 and Hyd3 was necessary to maintain an optimal NAD(+)/NADH ratio and enhanced adaptation to and survival of hypoxia. We demonstrate that fermentatively-produced hydrogen can be recycled when fumarate or oxygen become available, suggesting Mycobacterium smegmatis can switch between fermentation, anaerobic respiration, and aerobic respiration. Hydrogen metabolism enables this obligate aerobe to rapidly meet its energetic needs when switching between microoxic and anoxic conditions and provides a competitive advantage in low oxygen environments.

Benzoic Acid Production with Respect to Starter Culture and Incubation Temperature during Yogurt Fermentation using Response Surface Methodology.

PubMed

Yu, Hyung-Seok; Lee, Na-Kyoung; Jeon, Hye-Lin; Eom, Su Jin; Yoo, Mi-Young; Lim, Sang-Dong; Paik, Hyun-Dong

2016-01-01

Benzoic acid is occasionally used as a raw material supplement in food products and is sometimes generated during the fermentation process. In this study, the production of naturally occurring yogurt preservatives was investigated for various starter cultures and incubation temperatures, and considered food regulations. Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus reuteri, Lactobacillus plantarum, Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium bifidum, Bifidobacterium infantis, and Bifidobacterium breve were used as yogurt starter cultures in commercial starters. Among these strains, L. rhamnosus and L. paracasei showed the highest production of benzoic acid. Therefore, the use of L. rhamnosus, L. paracasei, S. thermophilus, and different incubation temperatures were examined to optimize benzoic acid production. Response surface methodology (RSM) based on a central composite design was performed for various incubation temperatures (35-44℃) and starter culture inoculum ratios (0-0.04%) in a commercial range of dairy fermentation processes. The optimum conditions were 0.04% L. rhamnosus, 0.01% L. paracasei, 0.02% S. thermophilus, and 38.12℃, and the predicted and estimated concentrations of benzoic acid were 13.31 and 13.94 mg/kg, respectively. These conditions maximized naturally occurring benzoic acid production during the yogurt fermentation process, and the observed production levels satisfied regulatory guidelines for benzoic acid in dairy products.

Benzoic Acid Production with Respect to Starter Culture and Incubation Temperature during Yogurt Fermentation using Response Surface Methodology

PubMed Central

Yoo, Mi-Young; Lim, Sang-Dong

2016-01-01

Benzoic acid is occasionally used as a raw material supplement in food products and is sometimes generated during the fermentation process. In this study, the production of naturally occurring yogurt preservatives was investigated for various starter cultures and incubation temperatures, and considered food regulations. Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus reuteri, Lactobacillus plantarum, Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium bifidum, Bifidobacterium infantis, and Bifidobacterium breve were used as yogurt starter cultures in commercial starters. Among these strains, L. rhamnosus and L. paracasei showed the highest production of benzoic acid. Therefore, the use of L. rhamnosus, L. paracasei, S. thermophilus, and different incubation temperatures were examined to optimize benzoic acid production. Response surface methodology (RSM) based on a central composite design was performed for various incubation temperatures (35-44℃) and starter culture inoculum ratios (0-0.04%) in a commercial range of dairy fermentation processes. The optimum conditions were 0.04% L. rhamnosus, 0.01% L. paracasei, 0.02% S. thermophilus, and 38.12℃, and the predicted and estimated concentrations of benzoic acid were 13.31 and 13.94 mg/kg, respectively. These conditions maximized naturally occurring benzoic acid production during the yogurt fermentation process, and the observed production levels satisfied regulatory guidelines for benzoic acid in dairy products. PMID:27433115

Preparation of hydrolytic liquid from dried distiller's grains with solubles and fumaric acid fermentation by Rhizopus arrhizus RH 7-13.

PubMed

Liu, Huan; Yue, Xuemin; Jin, Yuhan; Wang, Meng; Deng, Li; Wang, Fang; Tan, Tianwei

2017-10-01

Fumaric acid production from lignocellulosic materials is an alternative chemicals production system. This work investigated the suitable conditions for hydrolysis of dried distiller's grains with solubles (DDGS). The hydrolytic liquid was subsequently used for the production of fumaric acid. After optimizing the hydrolysis conditions, the most suitable concentration of H 2 SO 4 (2%), hydrolysis temperature (120 °C), hydrolysis time (100min) and solid/liquid ratio (1:10) were obtained. The yield of monosaccharides reached 258 mg/g DDGS and 15.88 g/L glucose, 7.53 g/L xylose and 2.35 g/L arabinose were obtained in unprocessed hydrolytic liquid. The furfural inhibitor in the hydrolytic liquid was also detected and the yield of it was reducing progressively in the pretreatment process. The ferment ability of the hydrolytic liquid from DDGS was tested through the process of fumaric acid production by Rhizopus arrhizus RH 7-13. The unprocessed hydrolytic liquid was not appropriate for the fermentation process. The yield of fumaric acid from the concentrated processed hydrolytic liquid reached 18.93 g/L, which was close to the yield of fermenting 80 g/L glucose. This result indicated that the commonly used carbon resource glucose could to some extent be replaced by processed hydrolytic liquid. Copyright © 2017 Elsevier Ltd. All rights reserved.

Production of laccase by Coriolus versicolor and its application in decolorization of dyestuffs: (I). Production of laccase by batch and repeated-batch processes.

PubMed

Lin, Jian-Ping; Wei, Lian; Xia, Li-Ming; Cen, Pei-Lin

2003-01-01

The production of laccase by Coriolus versicolor was studied. The effect of cultivation conditions on laccase production by Coriolus versicolor was examined to obtain optimal medium and cultivation conditions. Both batch and repeated-batch processes were performed for laccase production. In repeated-batch fermentation with self-immobilized mycelia, total of 14 cycles were performed with laccase activity in the range between 3.4 and 14.8 U/ml.

Bioflavoring by non-conventional yeasts in sequential beer fermentations.

PubMed

Holt, Sylvester; Mukherjee, Vaskar; Lievens, Bart; Verstrepen, Kevin J; Thevelein, Johan M

2018-06-01

Non-conventional yeast species have great capacity for producing diverse flavor profiles in production of alcoholic beverages, but their potential for beer brewing, in particular in consecutive fermentations with Saccharomyces cerevisiae, has only poorly been explored. We have screened 17 non-conventional yeast species for production of an appealing profile of flavor esters and phenolics in the first phase of alcoholic fermentation, followed by inoculation with S. cerevisiae to complete the fermentation. For measurement of phenolic compounds and their precursors we developed an improved and highly sensitive methodology. The results show that non-conventional yeast species possess promising potential for enhancement of desirable flavors in beer production. Notable examples are increasing isoamyl acetate (fruity, banana flavor) by application of P. kluyverii, augmenting ethyl phenolic compounds (spicy notes) with Brettanomyces species and enhancing 4-vinyl guaiacol (clove-like aroma) with T. delbrueckii. All Pichia strains also produced high levels of ethyl acetate (solvent-like flavor). This might be selectively counteracted by selection of an appropriate S. cerevisiae strain for the second fermentation phase, which lowers total ester profile. Hence, optimization of the process conditions and/or proper strain selection in sequentially inoculated fermentations are required to unlock the full potential for aroma improvement by the non-conventional yeast species. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

Detoxification of Aflatoxin B₁ by Zygosaccharomyces rouxii with Solid State Fermentation in Peanut Meal.

PubMed

Zhou, Guanghui; Chen, Yujie; Kong, Qing; Ma, Yunxiao; Liu, Yang

2017-01-20

Aflatoxins are highly carcinogenic, teratogenetic, and morbigenous secondary metabolites of Aspergillus flavus and A. parasiticus that can contaminate multiple staple foods, such as peanut, maize, and tree nuts. In this study, Zygosaccharomyces rouxii was screened out and identified from fermented soy paste-one kind of traditional Chinese food-to detoxify aflatoxin B₁ (AFB₁) by aerobic solid state fermentation in peanut meal. The optimal degradation condition was chosen from single factor experiment, and the most effective detoxification rate was about 97%. As for liquid fermentation, we tested the binding ability of Z. rouxii , and the highest binding rate reached was 74.3% (nonviable cells of Z. rouxii ) in phosphate-buffered saline (PBS). Moreover, the biotransformation of AFB₁ through fermentation of Z. rouxii in peanut meal was further verified by liquid chromatography/mass spectrometry (LC/MS). According to TIC scan, after fermentation by Z. rouxii, the AFB₁ in peanut meal was prominently degraded to the lowering peaks of AFB₁. Additionally, m / s statistics demonstrated that AFB₁ may be degraded to some new products whose structural properties may be different from AFB₁, or the degradation products may be dissolved in the aqueous phase rather than the organic phase. As far as we know, this is the first report indicating that the safe strain of Z. rouxii has the ability to detoxify AFB₁.

Detoxification of Aflatoxin B1 by Zygosaccharomyces rouxii with Solid State Fermentation in Peanut Meal

PubMed Central

Zhou, Guanghui; Chen, Yujie; Kong, Qing; Ma, Yunxiao; Liu, Yang

2017-01-01

Aflatoxins are highly carcinogenic, teratogenetic, and morbigenous secondary metabolites of Aspergillus flavus and A. parasiticus that can contaminate multiple staple foods, such as peanut, maize, and tree nuts. In this study, Zygosaccharomyces rouxii was screened out and identified from fermented soy paste—one kind of traditional Chinese food—to detoxify aflatoxin B1 (AFB1) by aerobic solid state fermentation in peanut meal. The optimal degradation condition was chosen from single factor experiment, and the most effective detoxification rate was about 97%. As for liquid fermentation, we tested the binding ability of Z. rouxii, and the highest binding rate reached was 74.3% (nonviable cells of Z. rouxii) in phosphate-buffered saline (PBS). Moreover, the biotransformation of AFB1 through fermentation of Z. rouxii in peanut meal was further verified by liquid chromatography/mass spectrometry (LC/MS). According to TIC scan, after fermentation by Z. rouxii, the AFB1 in peanut meal was prominently degraded to the lowering peaks of AFB1. Additionally, m/s statistics demonstrated that AFB1 may be degraded to some new products whose structural properties may be different from AFB1, or the degradation products may be dissolved in the aqueous phase rather than the organic phase. As far as we know, this is the first report indicating that the safe strain of Z. rouxii has the ability to detoxify AFB1. PMID:28117705

Fermentation of sugarcane bagasse and chicken manure to calcium carboxylates under thermophilic conditions.

PubMed

Fu, Zhihong; Holtzapple, Mark T

2010-09-01

Sugarcane bagasse and chicken manure were anaerobically fermented to carboxylic acids using a mixed culture of marine microorganisms at 55 degrees C. Using the MixAlco process--an example of consolidated bioprocessing--the resulting carboxylate salts can be converted to mixed alcohol fuels or gasoline. To enhance digestibility, sugarcane bagasse was lime pretreated with 0.1 g Ca(OH)(2)/g dry biomass at 100 degrees C for 2 h. Four-stage countercurrent fermentation of 80% sugarcane bagasse/20% chicken manure was performed at various volatile solids (VS) loading rates and liquid residence times. Calcium carbonate was used as a buffer during fermentation. The highest acid productivity of 0.79 g/(L day) occurred at a total acid concentration of 21.5 g/L. The highest conversion (0.59 g VS digested/g VS fed) and yield (0.18 g total acids/g VS fed) occurred at a total acid concentration of 15.5 g/L. The continuum particle distribution model (CPDM) predicted the experimental total acid concentrations and conversions at an average error of 10.14% and 12.68%, respectively. CPDM optimizations show that high conversion (>80%) and total acid concentration of 21.3 g/L are possible with 300 g substrate/(L liquid), 30 days liquid residence time, and 3 g/(L day) solid loading rate. Thermophilic fermentation has a higher acetate content (approximately 63 wt%) than mesophilic fermentation (approximately 39 wt%).

Identification of candidate genes for yeast engineering to improve bioethanol production in very high gravity and lignocellulosic biomass industrial fermentations.

PubMed

Pereira, Francisco B; Guimarães, Pedro Mr; Gomes, Daniel G; Mira, Nuno P; Teixeira, Miguel C; Sá-Correia, Isabel; Domingues, Lucília

2011-12-09

The optimization of industrial bioethanol production will depend on the rational design and manipulation of industrial strains to improve their robustness against the many stress factors affecting their performance during very high gravity (VHG) or lignocellulosic fermentations. In this study, a set of Saccharomyces cerevisiae genes found, through genome-wide screenings, to confer resistance to the simultaneous presence of different relevant stresses were identified as required for maximal fermentation performance under industrial conditions. Chemogenomics data were used to identify eight genes whose expression confers simultaneous resistance to high concentrations of glucose, acetic acid and ethanol, chemical stresses relevant for VHG fermentations; and eleven genes conferring simultaneous resistance to stresses relevant during lignocellulosic fermentations. These eleven genes were identified based on two different sets: one with five genes granting simultaneous resistance to ethanol, acetic acid and furfural, and the other with six genes providing simultaneous resistance to ethanol, acetic acid and vanillin. The expression of Bud31 and Hpr1 was found to lead to the increase of both ethanol yield and fermentation rate, while Pho85, Vrp1 and Ygl024w expression is required for maximal ethanol production in VHG fermentations. Five genes, Erg2, Prs3, Rav1, Rpb4 and Vma8, were found to contribute to the maintenance of cell viability in wheat straw hydrolysate and/or the maximal fermentation rate of this substrate. The identified genes stand as preferential targets for genetic engineering manipulation in order to generate more robust industrial strains, able to cope with the most significant fermentation stresses and, thus, to increase ethanol production rate and final ethanol titers.

Statistical optimization of bioprocess parameters for enhanced gallic acid production from coffee pulp tannins by Penicillium verrucosum.

PubMed

Bhoite, Roopali N; Navya, P N; Murthy, Pushpa S

2013-01-01

Gallic acid (3,4,5-trihydroxybenzoic acid) was produced by microbial biotransformation of coffee pulp tannins by Penicillium verrucosum. Gallic acid production was optimized using response surface methodology (RSM) based on central composite rotatable design. Process parameters such as pH, moisture, and fermentation period were considered for optimization. Among the various fungi isolated from coffee by-products, Penicillium verrucosum produced 35.23 µg/g of gallic acid on coffee pulp as sole carbon source in solid-state fermentation. The optimum values of the parameters obtained from the RSM were pH 3.32, moisture 58.40%, and fermentation period of 96 hr. Gallic acid production with an increase of 4.6-fold was achieved upon optimization of the process parameters. The results optimized could be translated to 1-kg tray fermentation. High-performance liquid chromatography (HPLC) analysis and spectral studies such as mass spectroscopy (MS) and (1)H-nuclear magnetic resonance (NMR) confirmed that the bioactive compound isolated was gallic acid. Thus, coffee pulp, which is available in enormous quantity, could be used for the production of value-added products that can find avenues in food, pharmaceutical, and chemical industries.

Effect of oxygen supply on Monascus pigments and citrinin production in submerged fermentation.

PubMed

Yang, Jian; Chen, Qi; Wang, Weiping; Hu, Jiajun; Hu, Chuan

2015-05-01

The influence of oxygen supply on Monascus pigments and citrinin production by Monascus ruber HS.4000 in submerged fermentation was studied. For Monascus cultivation with high pigments and low citrinin production, the initial growth phase, mid-stage phase, and later-stage production phase were separated by shifting oxygen supply. The optimal condition for the fermentation process in shake-flask fermentation was a three-stage rotating rate controlled strategy (0-48 h at 150 rpm, 48-108 h at 250 rpm, 108-120 h at 200 rpm) with medium volume of 100 mL added to 250 mL Erlenmeyer flasks at 30°C for 120 h cultivation. Compared to constant one-stage cultivation (medium volume of 100 mL, rotating rate of 250 rpm), the pigments were reduced by 40.4%, but citrinin was reduced by 64.2%. The most appropriate condition for the fermentation process in a 10 L fermentor is also a three-stage aeration process (0-48 h at 300 L/h, 48-96 h at 500 L/h, 96-120 h at 200 L/h) with agitation of 300 rpm at 30°C for 120 h cultivation, and 237.3 ± 5.7 U/mL pigments were produced in 120 h with 6.05 ± 0.19 mg/L citrinin in a 10 L fermentor. Compared to aeration-constant (500 L/h) cultivation, pigment production was increased by 29.6% and citrinin concentration was reduced by 79.5%. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Treatment of antibiotic fermentation wastewater by combined polyferric sulfate coagulation, Fenton and sedimentation process.

PubMed

Xing, Zi-Peng; Sun, De-Zhi

2009-09-15

Attempts were made in this study to examine the combined polyferric sulfate (PFS) coagulation, Fenton and sedimentation process for treatment of non-degradable antibiotic fermentation wastewater. The experimental results indicated that 66.6% of color and 72.4% of chemical oxygen demand (COD) were removed under the optimum conditions of PFS dosage 200mg/L and pH 4.0. In addition, optimal parameters of Fenton process were determined to be 150 mg/L of H(2)O(2) dosage, 120 mg/L of FeSO(4) and 1h of reaction time. When Fenton treated effluent was controlled at pH 7.0, the pollutants could be further removed by sedimentation process. The overall color, COD and suspended solids (SS) removal reached 97.3%, 96.9% and 86.7% under selected conditions, respectively. Thus this study might offer an effective way for wastewater treatment of antibiotics manufacturer and pharmaceutical industry.

Application of response surface methodology in optimization of lactic acid fermentation of radish: effect of addition of salt, additives and growth stimulators.

PubMed

Joshi, V K; Chauhan, Arjun; Devi, Sarita; Kumar, Vikas

2015-08-01

Lactic acid fermentation of radish was conducted using various additive and growth stimulators such as salt (2 %-3 %), lactose, MgSO4 + MnSO4 and Mustard (1 %, 1.5 % and 2 %) to optimize the process. Response surface methodology (Design expert, Trial version 8.0.5.2) was applied to the experimental data for the optimization of process variables in lactic acid fermentation of radish. Out of various treatments studied, only the treatments having ground mustard had an appreciable effect on lactic acid fermentation. Both linear and quadratic terms of the variables studied had a significant effect on the responses studied. The interactions between the variables were found to contribute to the response at a significant level. The best results were obtained in the treatment with 2.5 % salt, 1.5 % lactose, 1.5 % (MgSO4 + MnSO4) and 1.5 % mustard. These optimized concentrations increased titrable acidity and LAB count, but lowered pH. The second-order polynomial regression model determined that the highest titrable acidity (1.69), lowest pH (2.49) and maximum LAB count (10 × 10(8) cfu/ml) would be obtained at these concentrations of additives. Among 30 runs conducted, run 2 has got the optimum concentration of salt- 2.5 %, lactose- 1.5 %, MgSO4 + MnSO4- 1.5 % and mustard- 1.5 % for lactic acid fermentation of radish. The values for different additives and growth stimulators optimized in this study could successfully be employed for the lactic acid fermentation of radish as a postharvest reduction tool and for product development.

Media optimization for laccase production by Trichoderma harzianum ZF-2 using response surface methodology.

PubMed

Gao, Huiju; Chu, Xiang; Wang, Yanwen; Zhou, Fei; Zhao, Kai; Mu, Zhimei; Liu, Qingxin

2013-12-01

Trichoderma harzianum ZF-2 producing laccase was isolated from decaying samples from Shandong, China, and showed dye decolorization activities. The objective of this study was to optimize its culture conditions using a statistical analysis of its laccase production. The interactions between different fermentation parameters for laccase production were characterized using a Plackett-Burman design and the response surface methodology. The different media components were initially optimized using the conventional one-factor-at-a-time method and an orthogonal test design, and a Plackett-Burman experiment was then performed to evaluate the effects on laccase production. Wheat straw powder, soybean meal, and CuSO4 were all found to have a significant influence on laccase production, and the optimal concentrations of these three factors were then sequentially investigated using the response surface methodology with a central composite design. The resulting optimal medium components for laccase production were determined as follows: wheat straw powder 7.63 g/l, soybean meal 23.07 g/l, (NH4)2SO4 1 g/l, CuSO4 0.51 g/l, Tween-20 1 g/l, MgSO4 1 g/l, and KH2PO4 0.6 g/l. Using this optimized fermentation method, the yield of laccase was increased 59.68 times to 67.258 U/ml compared with the laccase production with an unoptimized medium. This is the first report on the statistical optimization of laccase production by Trichoderma harzianum ZF-2.

Heterologous expression of a recombinant lactobacillal β-galactosidase in Lactobacillus plantarum: effect of different parameters on the sakacin P-based expression system.

PubMed

Nguyen, Tien-Thanh; Nguyen, Hoang-Minh; Geiger, Barbara; Mathiesen, Geir; Eijsink, Vincent G H; Peterbauer, Clemens K; Haltrich, Dietmar; Nguyen, Thu-Ha

2015-03-07

Two overlapping genes lacL and lacM (lacLM) encoding for heterodimeric β-galactosidase from Lactobacillus reuteri were previously cloned and over-expressed in the food-grade host strain Lactobacillus plantarum WCFS1, using the inducible lactobacillal pSIP expression system. In this study, we analyzed different factors that affect the production of recombinant L. reuteri β-galactosidase. Various factors related to the cultivation, i.e. culture pH, growth temperature, glucose concentration, as well as the induction conditions, including cell concentration at induction point and inducer concentration, were tested. Under optimal fermentation conditions, the maximum β-galactosidase levels obtained were 130 U/mg protein and 35-40 U/ml of fermentation broth corresponding to the formation of approximately 200 mg of recombinant protein per litre of fermentation medium. As calculated from the specific activity of the purified enzyme (190 U/mg), β-galactosidase yield amounted to roughly 70% of the total soluble intracellular protein of the host organism. It was observed that pH and substrate (glucose) concentration are the most prominent factors affecting the production of recombinant β-galactosidase. The over-expression of recombinant L. reuteri β-galactosidase in a food-grade host strain was optimized, which is of interest for applications of this enzyme in the food industry. The results provide more detailed insight into these lactobacillal expression systems and confirm the potential of the pSIP system for efficient, tightly controlled expression of enzymes and proteins in lactobacilli.

A simple and cost-saving approach to optimize the production of subtilisin NAT by submerged cultivation of Bacillus subtilis natto.

PubMed

Ku, Ting-Wei; Tsai, Ruei-Lan; Pan, Tzu-Ming

2009-01-14

Subtilisin NAT, formerly designated nattokinase or subtilisin BSP, is a potent cardiovascular drug because of its strong fibrinolytic activity and safety. In this study, one Bacillus subtilis natto strain with high fibrinolytic activity was isolated. We further studied the optimal conditions for subtilisin NAT production by submerged cultivation and three variables/three levels of response surface methodology (RSM) using various inoculum densities, glucose concentrations, and defatted soybean concentrations as the three variables. According to the RSM analysis, while culturing by 2.93% defatted soybean, 1.75% glucose, and 4.00% inoculum density, we obtained an activity of 13.78 SU/mL. Processing the batch fermentation with this optimal condition, the activity reached 13.69 SU/mL, which is equal to 99.3% of the predicted value.

Development of a simple intensified fermentation strategy for growth of Magnetospirillum gryphiswaldense MSR-1: Physiological responses to changing environmental conditions.

PubMed

Fernández-Castané, Alfred; Li, Hong; Thomas, Owen R T; Overton, Tim W

2018-06-01

The development of a simple pH-stat fed-batch fermentation strategy for the production of Magnetospirillum gryphiswaldense MSR-1 and magnetosomes (nanoscale magnetic organelles with biotechnological applications) is described. Flow cytometry was exploited as a powerful analytical tool for process development, enabling rapid monitoring of cell morphology, physiology and polyhydroxyalkanoate production. The pH-stat fed-batch growth strategy was developed by varying the concentrations of the carbon source (lactic acid) and the alternative electron acceptor (sodium nitrate) in the feed. Growth conditions were optimized on the basis of biomass concentration, cellular magnetism (indicative of magnetosome production), and intracellular iron concentration. The highest biomass concentration and cellular iron content achieved were an optical density at 565 nm of 15.5 (equivalent to 4.2 g DCW·L -1 ) and 33.1 mg iron·g -1 DCW, respectively. This study demonstrates the importance of analyzing bacterial physiology during fermentation development and will potentially aid the industrial production of magnetosomes, which can be used in a wide range of biotechnology and healthcare applications. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Enhanced enzymatic hydrolysis and ethanol production from cashew apple bagasse pretreated with alkaline hydrogen peroxide.

PubMed

da Costa, Jessyca Aline; Marques, José Edvan; Gonçalves, Luciana Rocha Barros; Rocha, Maria Valderez Ponte

2015-03-01

The effect of combinations and ratios between different enzymes has been investigated in order to assess the optimal conditions for hydrolysis of cashew apple bagasse pretreated with alkaline hydrogen peroxide (the solids named CAB-AHP). The separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) processes were evaluated in the ethanol production. The enzymatic hydrolysis conducted with cellulase complex and β-glucosidase in a ratio of 0.61:0.39, enzyme loading of 30FPU/g(CAB-AHP) and 66CBU/g(CAB-AHP), respectively, using 4% cellulose from CAB-AHP, turned out to be the most effective conditions, with glucose and xylose yields of 511.68 mg/g(CAB-AHP) and 237.8 mg/g(CAB-AHP), respectively. Fermentation of the pure hydrolysate by Kluyveromyces marxianus ATCC 36907 led to an ethanol yield of 61.8kg/ton(CAB), corresponding to 15 g/L ethanol and productivity of 3.75 g/( Lh). The ethanol production obtained for SSF process using K. marxianus ATCC 36907 was 18 g/L corresponding to 80% yield and 74.2kg/ton(CAB). Copyright © 2014 Elsevier Ltd. All rights reserved.

Improving cellulase productivity of Penicillium oxalicum RE-10 by repeated fed-batch fermentation strategy.

PubMed

Han, Xiaolong; Song, Wenxia; Liu, Guodong; Li, Zhonghai; Yang, Piao; Qu, Yinbo

2017-03-01

Medium optimization and repeated fed-batch fermentation were performed to improve the cellulase productivity by P. oxalicum RE-10 in submerged fermentation. First, Plackett-Burman design (PBD) and central composite design (CCD) were used to optimize the medium for cellulase production. PBD demonstrated wheat bran and NaNO 3 had significant influences on cellulase production. The CCD results showed the maximum filter paper activity (FPA) production of 8.61U/mL could be achieved in Erlenmeyer flasks. The maximal FPA reached 12.69U/mL by submerged batch fermentation in a 7.5-L stirred tank, 1.76-fold higher than that on the original medium. Then, the repeated fed-batch fermentation strategy was performed successfully for increasing the cellulase productivity from 105.75U/L/h in batch fermentation to 158.38U/L/h. The cellulase activity and the glucan conversion of delignined corn cob residue hydrolysis had no significant difference between the enzymes sampled from different cycles of the repeated fed-batch fermentation and that from batch culture. Copyright © 2016 Elsevier Ltd. All rights reserved.

Changes in selected physical property and enzyme activity of rice and barley koji during fermentation and storage.

PubMed

Bechman, Allison; Phillips, Robert D; Chen, Jinru

2012-06-01

Koji are solid-state fermentation products made by inoculating steamed grains with the spores of fungi, particularly Aspergillus spp. This research was undertaken to identify the fermentation and storage conditions optimal for the production and maintenance of selected hydrolytic enzymes, such as α-amlyase and protease, in koji. Steamed rice and barley were inoculated with 2 × 10 ¹¹ Aspergillus oryzae spores per kilogram of grains and fermented for 118 h in a growth chamber at 28 to 32 °C with controlled relative humidities. Samples were drawn periodically during fermentation and storage at -20, 4, or 32 °C, and α-amylase and protease activity, mold counts, a(w), moisture contents, and pH of collected samples were determined. It was observed that the a(w), moisture contents, and pH of the koji were influenced by the duration of fermentation and temperature of storage. The α-amylase activity of both koji increased as the populations of A. oryzae increased during the exponential growth phase. The enzyme activity of barley koji was significantly higher than that of rice koji, reaching a peak activity of 211.87 or 116.57 U at 46 and 58 h, respectively, into the fermentation process. The enzyme activity in both products started to decrease once the mold culture entered the stationary growth phase. The protease activities of both koji were low and remained relatively stable during fermentation and storage. These results suggest that rice and barley koji can be used as sources of α-amylase and desired enzyme activity can be achieved by controlling the fermentation and storage conditions. Amylases and proteases are 2 important hydrolytic enzymes. In the food industry, these enzymes are used to break down starches and proteins while reducing the viscosity of foods. Although amylases and proteases are found in plants and animals, commercial enzymes are often produced using bacteria or molds through solid state fermentation, which is designed to use natural microbial process to produce enzymes in a controlled environment. A properly produced and maintained koji with a high hydrolytic enzyme activity can serve as an important source of the enzymes for the food industry. © 2012 Institute of Food Technologists®

A two-step optimization strategy for 2nd generation ethanol production using softwood hemicellulosic hydrolysate as fermentation substrate.

PubMed

Boboescu, Iulian-Zoltan; Gélinas, Malorie; Beigbeder, Jean-Baptiste; Lavoie, Jean-Michel

2017-11-01

Ethanol production using waste biomass represents a very attractive approach. However, there are considerable challenges preventing a wide distribution of these novel technologies. Thus, a fractional-factorial screening of process variables and Saccharomyces cerevisiae yeast inoculum conditions was performed using a synthetic fermentation media. Subsequently, a response-surface methodology was developed for maximizing ethanol yields using a hemicellulosic solution generated through the chemical hydrolysis of steam treatment broth obtained from residual softwood biomass. In addition, nutrient supplementation using starch-based ethanol production by-products was investigated. An ethanol yield of 74.27% of the theoretical maximum was observed for an initial concentration of 65.17g/L total monomeric sugars. The two-step experimental strategy used in this work represents the first successful attempt to developed and use a model to make predictions regarding the optimal ethanol production using both softwood feedstock residues as well as 1st generation ethanol production by-products. Copyright © 2017 Elsevier Ltd. All rights reserved.

Optimization of a natural medium for cellulase by a marine Aspergillus niger using response surface methodology.

PubMed

Xue, Dong-Sheng; Chen, Hui-Yin; Lin, Dong-Qiang; Guan, Yi-Xin; Yao, Shan-Jing

2012-08-01

The components of a natural medium were optimized to produce cellulase from a marine Aspergillus niger under solid state fermentation conditions by response surface methodology. Eichhornia crassipes and natural seawater were used as a major substrate and a source of mineral salts, respectively. Mineral salts of natural seawater could increase cellulase production. Raw corn cob and raw rice straw showed a significant positive effect on cellulase production. The optimum natural medium consisted of 76.9 % E. crassipes (w/w), 8.9 % raw corn cob (w/w), 3.5 % raw rice straw (w/w), 10.7 % raw wheat bran (w/w), and natural seawater (2.33 times the weight of the dry substrates). Incubation for 96 h in the natural medium increased the biomass to the maximum. The cellulase production was 17.80 U/g the dry weight of substrates after incubation for 144 h. The natural medium avoided supplying chemicals and pretreating substrates. It is promising for future practical fermentation of environment-friendly producing cellulase.

Adding Value to Goat Meat: Biochemical and Technological Characterization of Autochthonous Lactic Acid Bacteria to Achieve High-Quality Fermented Sausages

PubMed Central

Nediani, Miriam T.; García, Luis; Saavedra, Lucila; Martínez, Sandra; López Alzogaray, Soledad; Fadda, Silvina

2017-01-01

Quality and safety are important challenges in traditional fermented sausage technology. Consequently, the development of a tailored starter culture based on indigenous microbiota constitutes an interesting alternative. In the present study, spontaneously fermented goat meat sausages were created and analyzed using a physicochemical and microbiological approach. Thereafter 170 lactic acid bacteria (LAB) strains were isolated and preliminary characterized by phenotypic assays. The hygienic and technological properties, and growth and fermentative potential of isolates using a goat-meat-based culture medium were evaluated. All strains proved to have bioprotective features due to their acidogenic metabolism. Almost all grew optimally in meat environments. LAB isolates presented proteolytic activity against meat proteins and enriched amino acid contents of the goat-meat-based model. The most efficient strains were four different Lactobacillus sakei isolates, as identified by genotyping and RAPD analysis. L. sakei strains are proposed as optimal candidates to improve the production of fermented goat meat sausages, creating a new added-value fermented product. PMID:28513575

Adding Value to Goat Meat: Biochemical and Technological Characterization of Autochthonous Lactic Acid Bacteria to Achieve High-Quality Fermented Sausages.

PubMed

Nediani, Miriam T; García, Luis; Saavedra, Lucila; Martínez, Sandra; López Alzogaray, Soledad; Fadda, Silvina

2017-05-17

Quality and safety are important challenges in traditional fermented sausage technology. Consequently, the development of a tailored starter culture based on indigenous microbiota constitutes an interesting alternative. In the present study, spontaneously fermented goat meat sausages were created and analyzed using a physicochemical and microbiological approach. Thereafter 170 lactic acid bacteria (LAB) strains were isolated and preliminary characterized by phenotypic assays. The hygienic and technological properties, and growth and fermentative potential of isolates using a goat-meat-based culture medium were evaluated. All strains proved to have bioprotective features due to their acidogenic metabolism. Almost all grew optimally in meat environments. LAB isolates presented proteolytic activity against meat proteins and enriched amino acid contents of the goat-meat-based model. The most efficient strains were four different Lactobacillus sakei isolates, as identified by genotyping and RAPD analysis. L. sakei strains are proposed as optimal candidates to improve the production of fermented goat meat sausages, creating a new added-value fermented product.

Enhanced viability of Lactobacillus reuteri for probiotics production in mixed solid-state fermentation in the presence of Bacillus subtilis.

PubMed

Zhang, Yi-Ran; Xiong, Hai-Rong; Guo, Xiao-Hua

2014-01-01

In order to develop a multi-microbe probiotic preparation of Lactobacillus reuteri G8-5 and Bacillus subtilis MA139 in solid-state fermentation, a series of parameters were optimized sequentially in shake flask culture. The effect of supplementation of B. subtilis MA139 as starters on the viability of L. reuteri G8-5 was also explored. The results showed that the optimized process was as follows: water content, 50 %; initial pH of diluted molasses, 6.5; inocula volume, 2 %; flask dry contents, 30∼35 g/250 g without sterilization; and fermentation time, 2 days. The multi-microbial preparations finally provided the maximum concentration of Lactobacillus of about 9.01 ± 0.15 log CFU/g and spores of Bacillus of about 10.30 ± 0.08 log CFU/g. Compared with pure fermentation of L. reuteri G8-5, significantly high viable cells, low value of pH, and reducing sugar in solid substrates were achieved in mixed fermentation in the presence of B. subtilis MA139 (P < 0.05). Meanwhile, the mixed fermentation showed the significantly higher antimicrobial activity against E. coli K88 (P < 0.05). Based on the overall results, the optimized process enhanced the production of multi-microbe probiotics in solid-state fermentation with low cost. Moreover, the viability of L. reuteri G8-5 could be significantly enhanced in the presence of B. subtilis MA139 in solid-state fermentation, which favored the production of probiotics for animal use.

Medium Optimization and Fermentation Kinetics for κ-Carrageenase Production by Thalassospira sp. Fjfst-332.

PubMed

Guo, Juanjuan; Zhang, Longtao; Lu, Xu; Zeng, Shaoxiao; Zhang, Yi; Xu, Hui; Zheng, Baodong

2016-11-05

Effective degradation of κ-carrageenan by isolated Thalassospira sp. fjfst-332 is reported for the first time in this paper. It was identified by 16S rDNA sequencing and morphological observation using Transmission Electron Microscopy (TEM). Based on a Plackett-Burman design for significant variables, Box-Behnken experimental design and response surface methodology were used to optimize the culture conditions. Through statistical optimization, the optimum medium components were determined as follows: 2.0 g/L κ-carrageenan, 1.0 g/L yeast extract, 1.0 g/L FOS, 20.0 g/L NaCl, 2.0 g/L NaNO₃, 0.5 g/L MgSO₄·7H₂O, 0.1 g/L K₂HPO₄, and 0.1 g/L CaCl₂. The highest activity exhibited by Thalassospira sp. fjfst-332 was 267 U/mL, which makes it the most vigorous wild bacterium for κ-carrageenan production. In order to guide scaled-up production, two empirical models-the logistic equation and Luedeking-Piretequation-were proposed to predict the strain growth and enzyme production, respectively. Furthermore, we report the fermentation kinetics and every empirical equation of the coefficients (α, β, X ₀, X m and μ m ) for the two models, which could be used to design and optimize industrial processes.

Lignin blockers and uses thereof

DOEpatents

Yang, Bin; Wyman, Charles E

2013-11-12

Disclosed is a method for converting cellulose in a lignocellulosic biomass. The method provides for a lignin-blocking polypeptide and/or protein treatment of high lignin solids. The treatment enhances cellulase availability in cellulose conversion and allows for the determination of optimized pretreatment conditions. Additionally, ethanol yields from a Simultaneous Saccharification and Fermentation process are improved 5-25% by treatment with a lignin-blocking polypeptide and/or protein.

[Screening endophytic bacteria against plant-parasitic nematodes].

PubMed

Peng, Shuang; Yan, Shuzhen; Chen, Shuanglin

2011-03-01

Plant-parasite nematode is one of the most important pathogens in plant. Our objective is to screen endophytic bacteria against plant-parasitic nematodes from plant. Endophytic bacteria were isolated and screened by testing their metabolite against Bursaphelenchus xylophilus in vitro. Those strains inhibiting B. xylophilus were selected to culture in liquid medium and fermentation conditions were optimized by orthogonal test. The stability of the antinematode substances was evaluated by various. In addition, four strains were identified by 16SrDNA sequence analysis. In total 13 strains of endophytic bacteria secreting antinematode metabolite were isolated from 6 species of plant. The supernatant of the fermentation broth of these endophytic bacteria gave 100% mortality of nematodes after treated as the follows: 1 ml each was mixed with 0.2 ml of the suspension of nematodes (2000 nematodes/ml) then incubated at 250C for 24 h, some of which could led to leakage or dissolution of nematodes. Among them, four strains, BCM2, SZ5, CCM7 and DP1, showed stronger activity than others. The supernatants diluted three times also gave not less than 95% mortality after 24 h treatment, and those from DP1 and SZ5 even gave 100% mortality. The fermentation conditions of the four strains were optimized and the antinematode activity grew up four times after optimization. The antinematode substances of these strains were found stable when treated with protease or heating or stored at 4 degrees C after 100 days, while instable when treated with acid or alkali. DP1 and CCM7 were identified to be Bacillus subtilis, while SZ5 and BCM2 to be Bacillus cereus. Endophytic bacteria secreting antinematode metabolite were found in economic crops. The metabolite of some strains showed strong and stable antinematode activity. Our results indicate the real potential of biocontrol by endophytic bacteria.

Change of pH during excess sludge fermentation under alkaline, acidic and neutral conditions.

PubMed

Yuan, Yue; Peng, Yongzhen; Liu, Ye; Jin, Baodan; Wang, Bo; Wang, Shuying

2014-12-01

The change in pH during excess sludge (ES) fermentation of varying sludge concentrations was investigated in a series of reactors at alkaline, acidic, and neutral pHs. The results showed that the changes were significantly affected by fermentative conditions. Under different conditions, pH exhibited changing profiles. When ES was fermented under alkaline conditions, pH decreased in a range of (10±1). At the beginning of alkaline fermentation, pH dropped significantly, at intervals of 4h, 4h, and 5h with sludge concentrations of 8665.6mg/L, 6498.8mg/L, and 4332.5mg/L, then it would become moderate. However, under acidic conditions, pH increased from 4 to 5. Finally, under neutral conditions pH exhibited a decrease then an increase throughout entire fermentation process. Further study showed short-chain fatty acids (SCFAs), ammonia nitrogen and cations contributed to pH change under various fermentation conditions. This study presents a novel strategy based on pH change to predict whether SCFAs reach their stable stage. Copyright © 2014 Elsevier Ltd. All rights reserved.

Enzymatic hydrolysis optimization of microwave alkali pretreated wheat straw and ethanol production by yeast.

PubMed

Singh, Anita; Bishnoi, Narsi R

2012-03-01

Microwave alkali pretreated wheat straw was used for in-house enzyme production by Aspergillusflavus and Trichodermareesei. Produced enzymes were concentrated, pooled and assessed for the hydrolysis of pretreated wheat straw. Factors affecting hydrolysis were screened out by Placket-Burman design (PBD) and most significant factors were further optimized by Box-Behnken design (BBD). Under optimum conditions, 82% efficiency in hydrolysis yield was observed. After the optimization by response surface methodology (RSM), a model was proposed to predict the optimum value confirmed by the experimental results. The concentrated enzymatic hydrolyzate was fermented for ethanol production by Saccharomyces cerevisiae, Pichia stipitis and co-culture of both. The yield of ethanol was found to be 0.48 g(p)/g(s), 0.43 g(p)/g(s) and 0.40 g(p)/g(s) by S. cerevisiae, P. stipitis and by co-culture, respectively, using concentrated enzymatic hydrolyzate. During anaerobic fermentation 42.31 μmol/mL, 36.69 μmol/mL, 43.35 μmol/mL CO(2) was released by S. cerevisiae, P. stipitis and by co-culture, respectively. Copyright © 2011 Elsevier Ltd. All rights reserved.

Production of antioxidant compounds of grape seed skin by fermentation and its optimization using response surface method

NASA Astrophysics Data System (ADS)

Andayani, D. G. S.; Risdian, C.; Saraswati, V.; Primadona, I.; Mawarda, P. C.

2017-03-01

Skins and seeds of grape are waste generated from food industry. These wastes contain nutrients of which able to be utilized as an important source for antioxidant metabolite production. Through an environmentally friendly process, natural antioxidant material was produced. This study aimed to generate antioxidant compounds by liquid fermentation. Optimization was carried out by using Schizosaccharomyces cerevisiae in Katu leaf substrate. Optimization variables through response surface methodology (RSM) were of sucrose concentration, skins and seeds of grape concentration, and pH. Results showed that the optimum conditions for antioxidant production were of 5 g/L sucrose, 5 g/L skins and seed at pH 5, respectively. The resulted antioxidant activity was of 1.62 mg/mL. Mathematical model of variance analysis using a second order polynomial corresponding to the resulted data for the antioxidant was of 20.70124 - 3.86997 A - 0.65996 B - 1.88367 C + 0.19634 A2 - 0.016638 B2 + 0.28848 C2 + 0.26980 AB - 0.068333 AC - 0.12367 BC. From the gained equation, the optimum yield from all variables was significant. Chemical analysis of the antioxidant was carried out using 2,2-Diphenyl-1-picrylhydrazyl (DPPH).

Waste-Activated Sludge Fermentation for Polyacrylamide Biodegradation Improved by Anaerobic Hydrolysis and Key Microorganisms Involved in Biological Polyacrylamide Removal.

PubMed

Dai, Xiaohu; Luo, Fan; Zhang, Dong; Dai, Lingling; Chen, Yinguang; Dong, Bin

2015-07-06

During the anaerobic digestion of dewatered sludge, polyacrylamide (PAM), a chemical conditioner, can usually be consumed as a carbon and nitrogen source along with other organic matter (e.g., proteins and carbohydrates in the sludge). However, a significant accumulation of acrylamide monomers (AMs) was observed during the PAM biodegradation process. To improve the anaerobic hydrolysis of PAM, especially the amide hydrolysis process, and to avoid the generation of the intermediate product AM, a new strategy is reported herein that uses an initial pH of 9, 200 mg COD/L of PAM and a fermentation time of 17 d. First, response surface methodology (RSM) was applied to optimize PAM removal in the anaerobic digestion of the sludge. The biological hydrolysis of PAM reached 86.64% under the optimal conditions obtained from the RSM. Then, the mechanisms for the optimized parameters that significantly improved the biological hydrolysis of PAM were investigated by the synergistic effect of the main organic compounds in the sludge, the floc size distribution, and the enzymatic activities. Finally, semi-continuous-flow experiments for a microbial community study were investigated based on the determination of key microorganisms involved in the biological hydrolysis of PAM.

Chitinolytic and chitosanolytic activities from crude cellulase extract produced by A. niger grown on apple pomace through Koji fermentation.

PubMed

Dhillon, Gurpreet Singh; Brar, Satinder Kaur; Kaur, Surinder; Valero, Jose R; Verma, Mausam

2011-12-01

Enzyme extracts of cellulase [filter paper cellulase (FPase) and carboxymethyl cellulase (CMCase)], chitinase, and chitosanase produced by Aspergillus niger NRRL-567 were evaluated. The interactive effects of initial moisture and different inducers for FP cellulase and CMCase production were optimized using response surface methodology. Higher enzyme activities [FPase 79.24+/- 4.22 IU/gram fermented substrate (gfs) and CMCase 124.04+/-7.78 IU/gfs] were achieved after 48 h fermentation in solid-state medium containing apple pomace supplemented with rice husk [1% (w/w)] under optimized conditions [pH 4.5, moisture 55% (v/w), and inducers veratryl alcohol (2 mM/kg), copper sulfate (1.5 mM/kg), and lactose 2% (w/w)] (p<0.05). Koji fermentation in trays was carried out and higher enzyme activities (FPase 96.67+/-4.18 IU/gfs and CMCase 146.50+/-11.92 IU/gfs) were achieved. The nonspecific chitinase and chitosanase activities of cellulase enzyme extract were analyzed using chitin and chitosan substrates with different physicochemical characteristics, such as degree of deacetylation, molecular weight, and viscosity. Higher chitinase and chitosanase activities of 70.28+/-3.34 IU/gfs and 60.18+/-3.82 to 64.20+/-4.12 IU/gfs, respectively, were achieved. Moreover, the enzyme was stable and retained 92-94% activity even after one month. Cellulase enzyme extract obtained from A. niger with chitinolytic and chitosanolytic activities could be potentially used for making low-molecular-weight chitin and chitosan oligomers, having promising applications in biomedicine, pharmaceuticals, food, and agricultural industries, and in biocontrol formulations.

Modeling and experimental studies on intermittent starch feeding and citrate addition in simultaneous saccharification and fermentation of starch to flavor compounds.

PubMed

Chavan, Abhijit R; Raghunathan, Anuradha; Venkatesh, K V

2009-04-01

Simultaneous saccharification and fermentation (SSF) is a combined process of saccharification of a renewable bioresource and fermentation process to produce products, such as lactic acid and ethanol. Recently, SSF has been extensively used to convert various sources of cellulose and starch into fermentative products. Here, we present a study on production of buttery flavors, namely diacetyl and acetoin, by growing Lactobacillus rhamnosus on a starch medium containing the enzyme glucoamylase. We further develop a structured kinetics for the SSF process, which includes enzyme and growth kinetics. The model was used to simulate the effect of pH and temperature on the SSF process so as to obtain optimum operating conditions. The model was experimentally verified by conducting SSF using an initial starch concentration of 100 g/L. The study demonstrated that the developed kinetic was able to suggest strategies for improved productivities. The developed model was able to accurately predict the enhanced productivity of flavors in a three stage process with intermittent addition of starch. Experimental and simulations demonstrated that citrate addition can also lead to enhanced productivity of flavors. The developed optimal model for SSF was able to capture the dynamics of SSF in batch mode as well as in a three stage process. The structured kinetics was also able to quantify the effect of multiple substrates present in the medium. The study demonstrated that structured kinetic models can be used in the future for design and optimization of SSF as a batch or a fed-batch process.

Analysis of antimycin A by reversed-phase liquid chromatography/nuclear magnetic-resonance spectrometry

USGS Publications Warehouse

Ha, Steven T.K.; Wilkins, Charles L.; Abidi, Sharon L.

1989-01-01

A mixture of closely related streptomyces fermentation products, antimycin A, Is separated, and the components are identified by using reversed-phase high-performance liquid chromatography with directly linked 400-MHz proton nuclear magnetic resonance detection. Analyses of mixtures of three amino acids, alanine, glycine, and valine, are used to determine optimal measurement conditions. Sensitivity increases of as much as a factor of 3 are achieved, at the expense of some loss in chromatographic resolution, by use of an 80-μL NMR cell, Instead of a smaller 14-μL cell. Analysis of the antimycin A mixture, using the optimal analytical high performance liquid chromatography/nuclear magnetic resonance conditions, reveals it to consist of at least 10 closely related components.

Nutrient assessment of olive leaf residues processed by solid-state fermentation as an innovative feedstuff additive.

PubMed

Xie, P-J; Huang, L-X; Zhang, C-H; Zhang, Y-L

2016-07-01

Olive leaf residue feedstuff additives were prepared by solid-state fermentation (SSF), and its feeding effects on broiler chickens were examined. The fermentation's nutrient value, that is, protein enrichment, cellulase activity, tannic acid degradation and amino acid enhancement, was determined. The effect of different strains, including molds (Aspergillus niger, Aspergillus oryzae and Trichoderma viride) and yeasts (Candida utilis, Candida tropicalis and Geotrichum candidum), and the fermentation time on the nutrient values of the feedstuff additives was investigated. The experimental results showed that the optimal parameters for best performance were A. niger and C. utilis in a 1 : 1 ratio (v/v) in co-culture fermentation for 5 days. Under these conditions, the total content of amino acids in the fermented olive leaf residues increased by 22·0% in comparison with that in the raw leaf residues. Both Glutamic acid and Aspartic acid contents were increased by more than 25·4%. Broiler chickens fed with different amounts of feedstuff additives were assessed. The results demonstrated that the chicken weight gains increased by 120%, and normal serum biochemical parameters were improved significantly after 10% of the feedstuff additives were supplemented to the daily chicken feed for 28 days. The co-culture combination of A. niger and C. utilis with SSF for olive leaf residue had the best nutrient values. The addition of 10% fermented olive leaf residue facilitated the chicken growth and development. This study reveals that olive leaf residues fermented by SSF exhibited considerable potential as feed additives for feeding poultry. © 2016 The Society for Applied Microbiology.

Systematic optimization of fed-batch simultaneous saccharification and fermentation at high-solid loading based on enzymatic hydrolysis and dynamic metabolic modeling of Saccharomyces cerevisiae.

PubMed

Unrean, Pornkamol; Khajeeram, Sutamat; Laoteng, Kobkul

2016-03-01

An integrative simultaneous saccharification and fermentation (SSF) modeling is a useful guiding tool for rapid process optimization to meet the techno-economic requirement of industrial-scale lignocellulosic ethanol production. In this work, we have developed the SSF model composing of a metabolic network of a Saccharomyces cerevisiae cell associated with fermentation kinetics and enzyme hydrolysis model to quantitatively capture dynamic responses of yeast cell growth and fermentation during SSF. By using model-based design of feeding profiles for substrate and yeast cell in the fed-batch SSF process, an efficient ethanol production with high titer of up to 65 g/L and high yield of 85 % of theoretical yield was accomplished. The ethanol titer and productivity was increased by 47 and 41 %, correspondingly, in optimized fed-batch SSF as compared to batch process. The developed integrative SSF model is, therefore, considered as a promising approach for systematic design of economical and sustainable SSF bioprocessing of lignocellulose.

[Determination of process variable pH in solid-state fermentation by FT-NIR spectroscopy and extreme learning machine (ELM)].

PubMed

Liu, Guo-hai; Jiang, Hui; Xiao, Xia-hong; Zhang, Dong-juan; Mei, Cong-li; Ding, Yu-han

2012-04-01

Fourier transform near-infrared (FT-NIR) spectroscopy was attempted to determine pH, which is one of the key process parameters in solid-state fermentation of crop straws. First, near infrared spectra of 140 solid-state fermented product samples were obtained by near infrared spectroscopy system in the wavelength range of 10 000-4 000 cm(-1), and then the reference measurement results of pH were achieved by pH meter. Thereafter, the extreme learning machine (ELM) was employed to calibrate model. In the calibration model, the optimal number of PCs and the optimal number of hidden-layer nodes of ELM network were determined by the cross-validation. Experimental results showed that the optimal ELM model was achieved with 1040-1 topology construction as follows: R(p) = 0.961 8 and RMSEP = 0.104 4 in the prediction set. The research achievement could provide technological basis for the on-line measurement of the process parameters in solid-state fermentation.

Study of the rheological properties of a fermentation broth of the fungus Beauveria bassiana in a bioreactor under different hydrodynamic conditions.

PubMed

Núñez-Ramírez, Diola Marina; Medina-Torres, Luis; Valencia-López, José Javier; Calderas, Fausto; López Miranda, Javier; Medrano-Roldán, Hiram; Solís-Soto, Aquiles

2012-11-01

Fermentation with filamentous fungi in a bioreactor is a complex dynamic process that is affected by flow conditions and the evolution of the rheological properties of the medium. These properties are mainly affected by the biomass concentration and the morphology of the fungus. In this work, the rheological properties of a fermentation with the fungus Beauveria bassiana under different hydrodynamic conditions were studied and the rheological behavior of this broth was simulated through a mixture of carboxymethyl cellulose sodium and cellulose fibers (CMCNa-SF). The bioreactor was a 10 L CSTR tank operated at different stir velocities. Rheological results were similar at 100 and 300 rpm for both systems. However, there was a significant increase in the viscosity accompanied by a change in the consistence index, calculated according to the power law model, for both systems at 800 rpm. The systems exhibited shear-thinning behavior at all stir velocities, which was determined with the power law model. The mixing time was observed to increase as the cellulose content in the system increased and, consequently, the efficiency of mixing diminished. These results are thought to be due to the rheological and morphological similarities of the two fungal systems. These results will help in the optimization of scale-up production of these fungi.

A neural network strategy for end-point optimization of batch processes.

PubMed

Krothapally, M; Palanki, S

1999-01-01

The traditional way of operating batch processes has been to utilize an open-loop "golden recipe". However, there can be substantial batch to batch variation in process conditions and this open-loop strategy can lead to non-optimal operation. In this paper, a new approach is presented for end-point optimization of batch processes by utilizing neural networks. This strategy involves the training of two neural networks; one to predict switching times and the other to predict the input profile in the singular region. This approach alleviates the computational problems associated with the classical Pontryagin's approach and the nonlinear programming approach. The efficacy of this scheme is illustrated via simulation of a fed-batch fermentation.

Formation of ethyl carbamate and changes during fermentation and storage of yellow rice wine.

PubMed

Wu, Pinggu; Cai, Chenggang; Shen, Xianghong; Wang, Liyuan; Zhang, Jing; Tan, Ying; Jiang, Wei; Pan, Xiaodong

2014-01-01

Ethyl carbamate (EC) was analyzed during yellow rice wine production and storage. EC increased slowly during fermentation and rapidly after frying and sterilization. Less amount of EC was formed when cooled rapidly to 30 °C than when cooled naturally. High temperature and long storage time increased EC formation. After 400 days storage, EC increased from 74.0 to 84.2, 131.8 and 509.4 μg/kg at 4 °C, room temperature and 37 °C, respectively, and there was significantly difference between the fried wine and the wine on sale from 2011 (p<0.01). Urea increased during yellow rice wine fermentation and was above 20 mg/kg after the wine was fried; urea contributed to EC formation when the fried wine was cooled slowly. These results indicate that it is necessary for industry to optimize the wine frying conditions, such as temperature, time and cooling process in order to decrease EC formation. Copyright © 2013 Elsevier Ltd. All rights reserved.

Waste molasses alone displaces glucose-based medium for microalgal fermentation towards cost-saving biodiesel production.

PubMed

Yan, Dong; Lu, Yue; Chen, Yi-Feng; Wu, Qingyu

2011-06-01

The by-product of sugar refinery-waste molasses was explored as alternative to glucose-based medium of Chlorella protothecoides in this study. Enzymatic hydrolysis is required for waste molasses suitable for algal growth. Waste molasses hydrolysate was confirmed as a sole source of full nutrients to totally replace glucose-based medium in support of rapid growth and high oil yield from algae. Under optimized conditions, the maximum algal cell density, oil content, and oil yield were respectively 70.9 g/L, 57.6%, and 40.8 g/L. The scalability of the waste molasses-fed algal system was confirmed from 0.5L flasks to 5L fermenters. The quality of biodiesel from waste molasses-fed algae was probably comparable to that from glucose-fed ones. Economic analysis indicated the cost of oil production from waste molasses-fed algae reduced by 50%. Significant cost reduction of algal biodiesel production through fermentation engineering based on the approach is expected. Copyright © 2011 Elsevier Ltd. All rights reserved.

Extractive fermentation of xylanase from Aspergillus tamarii URM 4634 in a bioreactor.

PubMed

da Silva, Anna Carolina; Soares de França Queiroz, Alana Emília; Evaristo dos Santos Nascimento, Talita Camila; Rodrigues, Cristine; Gomes, José Erick Galindo; Souza-Motta, Cristina Maria; Porto de Souza Vandenberghe, Luciana; Valente de Medeiros, Erika; Moreira, Keila Aparecida; Herculano, Polyanna Nunes

2014-08-01

Of the many reported applications for xylanase, its use as a food supplement has played an important role for monogastric animals, because it can improve the utilisation of nutrients. The aim of this work was to produce xylanase by extractive fermentation in an aqueous two-phase system using Aspergillus tamarii URM 4634, increasing the scale of production in a bioreactor, partially characterising the xylanase and evaluating its influence on monogastric digestion in vitro. Through extractive fermentation in a bioreactor, xylanase was obtained with an activity of 331.4 U mL(-1) and 72% yield. The xylanase was stable under variable pH and temperature conditions, and it was optimally active at pH 3.6 and 90 °C. Xylanase activity potentiated the simulation of complete monogastric digestion by 6%, and only Mg2+ inhibited its activity. This process provides a system for efficient xylanase production by A. tamarii URM 4634 that has great potential for industrial use.

Effect of production conditions on the stability of a human bifidobacterial species Bifidobacterium longum in yogurt.

PubMed

Abe, F; Tomita, S; Yaeshima, T; Iwatsuki, K

2009-12-01

Human bifidobacteria are more sensitive to external environmental factors than animal bifidobacteria, and it is difficult to ensure their stable survival in yogurt. The purpose of this investigation was to observe the survival of human bifidobacteria in yogurts produced under various production conditions. Frozen or lyophilized bifidobacteria starters containing Bifidobacterium longum BB536 originally isolated from an infant, and commercial lyophilized yogurt starters were used for yogurt preparation. After producing yogurts under various conditions, the survival of bifidobacteria in these yogurts over various storage periods was observed. Although there were some differences in bifidobacterial survival in yogurt between various production conditions, more than 1.0 x 10(7) CFU g(-1) of Bif. longum survived in yogurt after 35 days' storage at 5 degrees C. Lower fermentation temperature (37 degrees C) and inclusion of Lactococcus lactis in the starter significantly (P < 0.05) improved survival of Bif. longum in the yogurt. In this investigation, the human bifidobacterial strain Bif. longum survived adequately in yogurt, although the fermentation temperature and starter composition affect bifidobacterial survival. This investigation indicates that stable probiotic yogurt using human bifidobacteria can be produced by choosing optimal production conditions.

Enhancement of L(+)-Lactic Acid Production of Immobilized Rhizopus Oryzae Implanted by Ion Beams

NASA Astrophysics Data System (ADS)

Fan, Yonghong; Yang, Yingge; Zheng, Zhiming; Li, Wen; Wang, Peng; Yao, Liming; Yu, Zengliang

2008-02-01

Immobilized Rhizopus oryzae culturing may be a solution to the inhibited production of L(+)-lactic acid in submerged fermentation, which is caused by aggregated mycelia floc. In the present study, a R. oryzae mutant (RL6041) with a 90% conversion rate of glucose into L-lactic acid was obtained by N+ implantation under the optimized conditions of a beam energy of 15 keV and a dose of 2.6 × 1015 ions/cm2. Using polyurethane foam as the immobilization matrix, the optimal L-lactic acid production conditions were determined as 4 mm polyurethane foam, 150 r/min, 50 g/L ~ 80 g/L of initial glucose, 38°C and pH 6.0. 15-cycle repeated productions of L-lactic acid by immobilized RL6041 were performed under the optimized culturing conditions and over 80% of the glucose was converted into L-lactic acid in 30 hours on average. The results show that immobilized RL6041 is a promising candidate for continuous L-lactic acid production.

Coproduction of xylose, lignosulfonate and ethanol from wheat straw.

PubMed

Zhu, Shengdong; Huang, Wangxiang; Huang, Wenjing; Wang, Ke; Chen, Qiming; Wu, Yuanxin

2015-06-01

A novel integrated process to coproduce xylose, lignosulfonate and ethanol from wheat straw was investigated. Firstly, wheat straw was treated by dilute sulfuric acid and xylose was recovered from its hydrolyzate. Its optimal conditions were 1.0wt% sulfuric acid, 10% (w/v) wheat straw loading, 100°C, and 2h. Then the acid treated wheat straw was treated by sulfomethylation reagent and its hydrolyzate containing lignosulfonate was directly recovered. Its optimal conditions were 150°C, 15% (w/v) acid treated wheat straw loading, and 5h. Finally, the two-step treated wheat straw was converted to ethanol through enzymatic hydrolysis and microbial fermentation. Under optimal conditions, 1kg wheat straw could produce 0.225kg xylose with 95% purity, 4.16kg hydrolyzate of sulfomethylation treatment containing 5.5% lignosulfonate, 0.183kg ethanol and 0.05kg lignin residue. Compared to present technology, this process is a potential economically profitable wheat straw biorefinery. Copyright © 2015 Elsevier Ltd. All rights reserved.

Ethanol and Methanol Can Improve Huperzine A Production from Endophytic Colletotrichum gloeosporioides ES026

PubMed Central

Zhao, Xin-Mei; Wang, Zhang-Qian; Shu, Shao-Hua; Wang, Wen-Juan; Xu, Hai-Jie; Ahn, Young-Joon; Wang, Mo; Hu, Xuebo

2013-01-01

Huperzine A (HupA) is a plant alkaloid that is of great interest as a therapeutic candidate for the treatment of Alzheimer's disease. However, the current production of HupA from plants in large quantity is unsustainable because the plant resource is scarce and the content of HupA in plants is extremely low. Surprisingly, this compound was recently found to be produced by various endophytic fungi, which are much more controllable than the plants due to simpler genetics and ease of manipulation. However, it might be due to the innate properties of endophytic symbiosis, that production of this chemical in large quantity from endophytes has not yet been put into practice. Endophytic Colletotrichum gloeosporioides ES026 was previously isolated from a HupA producing plant and the fungi also proved to produce HupA. In this study, various fermentation conditions were tried to optimize the production of HupA from C. gloeosporioides ES026. Optimization of these parameters resulted in a 25.58% increase in HupA yield. Potato extracts supplemented with glucose or sucrose but not maltose facilitated HupA producing from the fungi. A final concentration of 0.5–2% ethanol stimulated the growth of fungi while methanol with the same treatment slightly inhibited the growth. However, both methanol and ethanol greatly increased the HupA production with the highest yield of HupA (51.89% increment) coming from ethanol treatment. Further analysis showed that both ethanol and methanol were strong inducers of HupA production, while ethanol was partially used as a carbon source during fermentation. It was noticed that the color of that ethanol treated mycelia gradually became dark while methanol treated ones stayed grey during fermentation. The present study sheds light on the importance of optimizing the fermentation process, which, combined with effective inducers, maximizes production of chemicals of important economic interest from endophytic fungi. PMID:23613930

Ethanol and methanol can improve huperzine A production from endophytic Colletotrichum gloeosporioides ES026.

PubMed

Zhao, Xin-Mei; Wang, Zhang-Qian; Shu, Shao-Hua; Wang, Wen-Juan; Xu, Hai-Jie; Ahn, Young-Joon; Wang, Mo; Hu, Xuebo

2013-01-01

Huperzine A (HupA) is a plant alkaloid that is of great interest as a therapeutic candidate for the treatment of Alzheimer's disease. However, the current production of HupA from plants in large quantity is unsustainable because the plant resource is scarce and the content of HupA in plants is extremely low. Surprisingly, this compound was recently found to be produced by various endophytic fungi, which are much more controllable than the plants due to simpler genetics and ease of manipulation. However, it might be due to the innate properties of endophytic symbiosis, that production of this chemical in large quantity from endophytes has not yet been put into practice. Endophytic Colletotrichum gloeosporioides ES026 was previously isolated from a HupA producing plant and the fungi also proved to produce HupA. In this study, various fermentation conditions were tried to optimize the production of HupA from C. gloeosporioides ES026. Optimization of these parameters resulted in a 25.58% increase in HupA yield. Potato extracts supplemented with glucose or sucrose but not maltose facilitated HupA producing from the fungi. A final concentration of 0.5-2% ethanol stimulated the growth of fungi while methanol with the same treatment slightly inhibited the growth. However, both methanol and ethanol greatly increased the HupA production with the highest yield of HupA (51.89% increment) coming from ethanol treatment. Further analysis showed that both ethanol and methanol were strong inducers of HupA production, while ethanol was partially used as a carbon source during fermentation. It was noticed that the color of that ethanol treated mycelia gradually became dark while methanol treated ones stayed grey during fermentation. The present study sheds light on the importance of optimizing the fermentation process, which, combined with effective inducers, maximizes production of chemicals of important economic interest from endophytic fungi.

Enhancement of volatile fatty acid production by co-fermentation of food waste and excess sludge without pH control: The mechanism and microbial community analyses.

PubMed

Wu, Qing-Lian; Guo, Wan-Qian; Zheng, He-Shan; Luo, Hai-Chao; Feng, Xiao-Chi; Yin, Ren-Li; Ren, Nan-Qi

2016-09-01

The study provided a cost-effective and high-efficiency volatile fatty acid (VFA) production strategy by co-fermentation of food waste (FW) and excess sludge (ES) without artificial pH control. VFA production of 867.42mg COD/g-VS was obtained under the optimized condition: FW/ES 5, solid retention time 7d, organic loading rate 9g VS/L-d and temperature 40°C. Mechanism exploration revealed that the holistic biodegradability of substrate was greatly enhanced, and proper pH range (5.2-6.4) was formed by the high buffering capacity of the co-fermentation system itself, which effectively enhanced hydrolysis yield (63.04%) and acidification yield (83.46%) and inhibited methanogenesis. Moreover, microbial community analysis manifested that co-fermentation raised the relative abundances of hydrolytic and acidogenic bacteria including Clostridium, Sporanaerobacter, Tissierella and Bacillus, but suppressed the methanogen Anaerolineae, which also facilitated high VFA production. These results were of great guiding significance aiming for VFA recovery from FW and ES in large-scale. Copyright © 2016 Elsevier Ltd. All rights reserved.

Hyper-thermal acid hydrolysis and adsorption treatment of red seaweed, Gelidium amansii for butyric acid production with pH control.

PubMed

Ra, Chae Hun; Jeong, Gwi-Taek; Kim, Sung-Koo

2017-03-01

Optimal hyper-thermal (HT) acid hydrolysis conditions for Gelidium amansii were determined to be 12% (w/v) seaweed slurry content and 144 mM H 2 SO 4 at 150 °C for 10 min. HT acid hydrolysis-treated G. amansii hydrolysates produced low concentrations of inhibitory compounds and adsorption treatment using 3% activated carbon. An adsorption time of 5 min was subsequently used to remove the inhibitory 5-hydroxymethylfurfural from the medium. A final maximum monosaccharide concentration of 44.6 g/L and 79.1% conversion from 56.4 g/L total fermentable monosaccharides with 120 g dw/L G. amansii slurry was obtained from HT acid hydrolysis, enzymatic saccharification, and adsorption treatment. This study demonstrates the potential for butyric acid production from G. amansii hydrolysates under non-pH-controlled as well as pH-controlled fermentation using Clostridium acetobutylicum KCTC 1790. The activated carbon treatment and pH-controlled fermentation showed synergistic effects and produced butyric acid at a concentration of 11.2 g/L after 9 days of fermentation.

A novel one-stage cultivation/fermentation strategy for improved biogas production with microalgal biomass.

PubMed

Klassen, Viktor; Blifernez-Klassen, Olga; Hoekzema, Yoep; Mussgnug, Jan H; Kruse, Olaf

2015-12-10

The use of alga biomass for biogas generation has been studied for over fifty years but until today, several distinct features, like inefficient degradation and low C/N ratios, limit the applicability of algal biomass for biogas production in larger scale. In this work we investigated a novel, one-stage combined cultivation/fermentation strategy including inherently progressing nitrogen starvation conditions to generate improved microalgal biomass substrates. For this strategy, comparable low amounts of nitrogen fertilizers were applied during cultivation and no additional enzymatic, chemical or physical pretreatments had to be performed. The results of this study demonstrate that progressing nitrogen limitation leads to continuously increasing C/N ratios of the biomass up to levels of 24-26 for all three tested alga strains (Chlamydomonas reinhardtii, Parachlorella kessleri and Scenedesmus obliquus). Importantly, the degradation efficiency of the algal cells increased with progressing starvation, leading to strain-specific cell disintegration efficiencies of 35%-100% during the fermentation process. Nitrogen limitation treatment resulted in a 65% increase of biogas yields for C. reinhardtii biomass (max. 698±23mL biogas g(-1) VS) when compared to replete conditions. For P. kessleri and S. obliquus, yields increased by 94% and 106% (max. 706±39mL and 586±36mL biogas g(-1) VS, respectively). From these results we conclude that this novel one-stage cultivation strategy with inherent nitrogen limitation can be used as a pretreatment for microalgal biomass generation, in order to produce accessible substrates with optimized C/N ratios for the subsequent anaerobic fermentation process, thus increasing methane production and avoiding the risk of ammonia inhibition effects within the fermenter. Copyright © 2015 Elsevier B.V. All rights reserved.

An obligately aerobic soil bacterium activates fermentative hydrogen production to survive reductive stress during hypoxia

PubMed Central

Berney, Michael; Greening, Chris; Conrad, Ralf; Jacobs, William R.; Cook, Gregory M.

2014-01-01

Oxygen availability is a major factor and evolutionary force determining the metabolic strategy of bacteria colonizing an environmental niche. In the soil, conditions can switch rapidly between oxia and anoxia, forcing soil bacteria to remodel their energy metabolism accordingly. Mycobacterium is a dominant genus in the soil, and all its species are obligate aerobes. Here we show that an obligate aerobe, the soil actinomycete Mycobacterium smegmatis, adopts an anaerobe-type strategy by activating fermentative hydrogen production to adapt to hypoxia. This process is controlled by the two-component system DosR-DosS/DosT, an oxygen and redox sensor that is well conserved in mycobacteria. We show that DosR tightly regulates the two [NiFe]-hydrogenases: Hyd3 (MSMEG_3931-3928) and Hyd2 (MSMEG_2719-2718). Using genetic manipulation and high-sensitivity GC, we demonstrate that Hyd3 facilitates the evolution of H2 when oxygen is depleted. Combined activity of Hyd2 and Hyd3 was necessary to maintain an optimal NAD+/NADH ratio and enhanced adaptation to and survival of hypoxia. We demonstrate that fermentatively-produced hydrogen can be recycled when fumarate or oxygen become available, suggesting Mycobacterium smegmatis can switch between fermentation, anaerobic respiration, and aerobic respiration. Hydrogen metabolism enables this obligate aerobe to rapidly meet its energetic needs when switching between microoxic and anoxic conditions and provides a competitive advantage in low oxygen environments. PMID:25049411

Optimization of pretreatment and fermentation conditions for production of extracellular cellulase complex using sugarcane bagasse.

PubMed

Ashfaque, Mohammad; Solomon, Sushil; Pathak, Neelam

2014-01-01

Sugarcane bagasse (SCB), a lignocellulosic byproduct of juice extraction from sugarcane, is rich in cellulose (40-42%). This could be used as a substrate for the production of cellulase complex. Fermentation conditions were optimized for production of cellulase complex (CMCase, Cellulobiase and FPase) by wild type Trichoderma sp. using sugarcane bagasse as sole carbon source. Alkaline treatment (2% NaOH) of bagasse (AlSCB) was found suitable for the production of reducing sugar over the acidic pretreatment method. After 5 days of incubation period, 5% substrate concentration at pH 5.0 and 400C resulted in maximum production of CMCase (0.622 U), while maximum (3.388 U) production of cellulobiase was obtained at 300C. The CMCase was precipitated and purified to the extent of 59.06 fold by affinity chromatography with 49.09% recovery. On 12% SDS-PAGE, a single band corresponding to 33 kDa was observed. The Km and Vmax for CMCase from Trichoderma was found 507.04 mg/ml and 65.32 mM/min, respectively. The enzyme exhibited maximum activity at 300C at pH-5.0 (0.363 U) and was stable over range of 20-60°C and pH 5.0-7.5.

Experimental design data for the biosynthesis of citric acid using Central Composite Design method.

PubMed

Kola, Anand Kishore; Mekala, Mallaiah; Goli, Venkat Reddy

2017-06-01

In the present investigation, we report that statistical design and optimization of significant variables for the microbial production of citric acid from sucrose in presence of filamentous fungi A. niger NCIM 705. Various combinations of experiments were designed with Central Composite Design (CCD) of Response Surface Methodology (RSM) for the production of citric acid as a function of six variables. The variables are; initial sucrose concentration, initial pH of medium, fermentation temperature, incubation time, stirrer rotational speed, and oxygen flow rate. From experimental data, a statistical model for this process has been developed. The optimum conditions reported in the present article are initial concentration of sucrose of 163.6 g/L, initial pH of medium 5.26, stirrer rotational speed of 247.78 rpm, incubation time of 8.18 days, fermentation temperature of 30.06 °C and flow rate of oxygen of 1.35 lpm. Under optimum conditions the predicted maximum citric acid is 86.42 g/L. The experimental validation carried out under the optimal values and reported citric acid to be 82.0 g/L. The model is able to represent the experimental data and the agreement between the model and experimental data is good.

The effect of fermented cocoa pod (Theobroma cacao) husk supplemented with mineral on in vitro digestibility, rumen bacteria population and rumen liquid characteristics

NASA Astrophysics Data System (ADS)

Nurhaita; Definiati, N.; Santoso, U.; Akbar, S. A.; Henuk, Y. L.

2018-02-01

This study aimed to determine the effect of mineral supplementation, such as S, P and Zn on the nutrients digestibility of fermented cocoa pod husk, the population of rumen bacteria and rumen liquid characteristics in vitro. The study used a randomized block design with 5 treatments and 4 replicates. The treatments tested were: T0 = without minerals; T1 = 0.2% S mineral; T2 = 0.27% P mineral; T3 = S and P; and T4 = S, P and Zn at 50 ppm. Parameters measured were: (1) digestibility of dry matter and organic matter; (2) rumen bacterial and cellulolytic bacterial populations; (3) characteristics of rumen liquid in vitro. The results of the study showed that mineral supplementation significantly (P <0.05) improved dry matter and organic matter digestibility. Mineral supplementation had no effect on the total population of rumen bacteria and cellulolytic rumen bacterial populations. The characteristics of rumen liquid such pH, VFA and NH3 were in optimal condition. In conclusion supplementation of S, P and Zn simultaneously gave the best results to improve the digestibility of dry matter and organic matter and to maintain rumen liquid characteristics under optimal conditions for growth and microbial activity

A novel method to prepare L-Arabinose from xylose mother liquor by yeast-mediated biopurification

PubMed Central

2011-01-01

Background L-arabinose is an important intermediate for anti-virus drug synthesis and has also been used in food additives for diets-controlling in recent years. Commercial production of L-arabinose is a complex progress consisting of acid hydrolysis of gum arabic, followed by multiple procedures of purification, thus making high production cost. Therefore, there is a biotechnological and commercial interest in the development of new cost-effective and high-performance methods for obtaining high purity grade L-arabinose. Results An alternative, economical method for purifying L-arabinose from xylose mother liquor was developed in this study. After screening 306 yeast strains, a strain of Pichia anomala Y161 was selected as it could effectively metabolize other sugars but not L-arabinose. Fermentation in a medium containing xylose mother liquor permitted enrichment of L-arabinose by a significant depletion of other sugars. Biochemical analysis of this yeast strain confirmed that its poor capacity for utilizing L-arabinose was due to low activities of the enzymes required for the metabolism of this sugar. Response surface methodology was employed for optimization the fermentation conditions in shake flask cultures. The optimum conditions were: 75 h fermentation time, at 32.5°C, in a medium containing 21% (v/v) xylose mother liquor. Under these conditions, the highest purity of L-arabinose reached was 86.1% of total sugar, facilitating recovery of white crystalline L-arabinose from the fermentation medium by simple methods. Conclusion Yeast-mediated biopurification provides a dynamic method to prepare high purity of L-arabinose from the feedstock solution xylose mother liqour, with cost-effective and high-performance properties. PMID:21649890

Omega-3 production by fermentation of Yarrowia lipolytica: From fed-batch to continuous.

PubMed

Xie, Dongming; Miller, Edward; Sharpe, Pamela; Jackson, Ethel; Zhu, Quinn

2017-04-01

The omega-3 fatty acid, cis-5,8,11,14,17-eicosapentaenoic acid (C20:5; EPA) has wide-ranging benefits in improving heart health, immune function, and mental health. A sustainable source of EPA production through fermentation of metabolically engineered Yarrowia lipolytica has been developed. In this paper, key fed-batch fermentation conditions were identified to achieve 25% EPA in the yeast biomass, which is so far the highest EPA titer reported in the literature. Dynamic models of the EPA fermentation process were established for analyzing, optimizing, and scaling up the fermentation process. In addition, model simulations were used to develop a two-stage continuous process and compare to single-stage continuous and fed- batch processes. The two stage continuous process, which is equipped with a smaller growth fermentor (Stage 1) and a larger production fermentor (Stage 2), was found to be a superior process to achieve high titer, rate, and yield of EPA. A two-stage continuous fermentation experiment with Y. lipolytica strain Z7334 was designed using the model simulation and then tested in a 2 L and 5 L fermentation system for 1,008 h. Compared with the standard 2 L fed-batch process, the two-stage continuous fermentation process improved the overall EPA productivity by 80% and EPA concentration in the fermenter by 40% while achieving comparable EPA titer in biomass and similar conversion yield from glucose. During the long-term experiment it was also found that the Y. lipolytica strain evolved to reduce byproduct and increase lipid production. This is one of the few continuous fermentation examples that demonstrated improved productivity and concentration of a final product with similar conversion yield compared with a fed-batch process. This paper suggests the two-stage continuous fermentation could be an effective process to achieve improved production of omega-3 and other fermentation products where non-growth or partially growth associated kinetics characterize the process. Biotechnol. Bioeng. 2017;114: 798-812. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Engineering Escherichia coli for high-yield geraniol production with biotransformation of geranyl acetate to geraniol under fed-batch culture.

PubMed

Liu, Wei; Xu, Xin; Zhang, Rubing; Cheng, Tao; Cao, Yujin; Li, Xiaoxiao; Guo, Jiantao; Liu, Huizhou; Xian, Mo

2016-01-01

Geraniol is an acyclic monoterpene alcohol, which exhibits good prospect as a gasoline alternative. Geraniol is naturally encountered in plants at low concentrations and an attractive target for microbial engineering. Geraniol has been heterologously produced in Escherichia coli, but the low titer hinders its industrial applications. Moreover, bioconversion of geraniol by E. coli remains largely unknown. Recombinant overexpression of Ocimum basilicum geraniol synthase, Abies grandis geranyl diphosphate synthase, and a heterotic mevalonate pathway in E. coli BL21 (DE3) enabled the production of up to 68.6 ± 3 mg/L geraniol in shake flasks. Initial fed-batch fermentation only increased geraniol production to 78.8 mg/L. To further improve the production yield, the fermentation conditions were optimized. Firstly, 81.4 % of volatile geraniol was lost during the first 5 h of fermentation in a solvent-free system. Hence, isopropyl myristate was added to the culture medium to form an aqueous-organic two-phase culture system, which effectively prevented volatilization of geraniol. Secondly, most of geraniol was eventually biotransformed into geranyl acetate by E. coli, thus decreasing geraniol production. For the first time, we revealed the role of acetylesterase (Aes, EC 3.1.1.6) from E. coli in hydrolyzing geranyl acetate to geraniol, and production of geraniol was successfully increased to 2.0 g/L under controlled fermentation conditions. An efficient geraniol production platform was established by overexpressing several key pathway proteins in engineered E. coli strain combined with a controlled fermentation system. About 2.0 g/L geraniol was obtained using our controllable aqueous-organic two-phase fermentation system, which is the highest yield to date. In addition, the interconversion between geraniol and geranyl acetate by E. coli was first elucidated. This study provided a new and promising strategy for geraniol biosynthesis, which laid a basis for large-scale industrial application.

Genetic programming assisted stochastic optimization strategies for optimization of glucose to gluconic acid fermentation.

PubMed

Cheema, Jitender Jit Singh; Sankpal, Narendra V; Tambe, Sanjeev S; Kulkarni, Bhaskar D

2002-01-01

This article presents two hybrid strategies for the modeling and optimization of the glucose to gluconic acid batch bioprocess. In the hybrid approaches, first a novel artificial intelligence formalism, namely, genetic programming (GP), is used to develop a process model solely from the historic process input-output data. In the next step, the input space of the GP-based model, representing process operating conditions, is optimized using two stochastic optimization (SO) formalisms, viz., genetic algorithms (GAs) and simultaneous perturbation stochastic approximation (SPSA). These SO formalisms possess certain unique advantages over the commonly used gradient-based optimization techniques. The principal advantage of the GP-GA and GP-SPSA hybrid techniques is that process modeling and optimization can be performed exclusively from the process input-output data without invoking the detailed knowledge of the process phenomenology. The GP-GA and GP-SPSA techniques have been employed for modeling and optimization of the glucose to gluconic acid bioprocess, and the optimized process operating conditions obtained thereby have been compared with those obtained using two other hybrid modeling-optimization paradigms integrating artificial neural networks (ANNs) and GA/SPSA formalisms. Finally, the overall optimized operating conditions given by the GP-GA method, when verified experimentally resulted in a significant improvement in the gluconic acid yield. The hybrid strategies presented here are generic in nature and can be employed for modeling and optimization of a wide variety of batch and continuous bioprocesses.

Effect of pH on ethanol-type acidogenic fermentation of fruit and vegetable waste.

PubMed

Wu, Yuanyuan; Wang, Cuiping; Zheng, Mingyue; Zuo, Jiane; Wu, Jing; Wang, Kaijun; Yang, Boqiong

2017-02-01

The aim of this study was to investigate the possibility and optimal controlling strategy for ethanol-type acidogenic fermentation of fruit and vegetable waste by mixed microbial cultures. Four continuous stirred tank reactors (CSTR) were operated at various pHs (4.0, 5.0, 5.5, and 6.0) with an organic loading rate of 13gVS/(Ld) and hydraulic retention time of 3d. Butyrate-type fermentation was observed at pH 5.0, 5.5, and 6.0. Conversely, at pH 4.0, ethanol-type fermentation was observed with a high mass concentration and proportion (of total fermentative products) of ethanol, which were 6.7g/L and 88.8%, respectively. However, the total concentration of ethanol-type fermentative products substantially decreased from days 22-25. The optimal pH of ethanol-type fermentative microorganisms was investigated by using batch experiments with pH controlled at 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, and 7.0 and results showed that the maximum ethanol concentration and relatively highest acidogenic rate were found at pH of 5.5. The pH in the long term CSTR was changed from 4.0 to 5.5 to improve ethanol-type fermentation and results showed that ethanol-type fermentation was improved temporarily, however, was followed by the reappearance of butyrate-type fermentation. In addition, ethanol-type fermentation recovered once more when pH was reverted to 4.0. Therefore, the results of this study suggest that a process of dynamic, sequenced pH control with the order pH 4.0, 5.5 and 4.0 might be a feasible controlling strategy for continuous and stable ethanol-type fermentation. Copyright © 2016 Elsevier Ltd. All rights reserved.

Use of Response Surface Methodology to Optimize Culture Conditions for Hydrogen Production by an Anaerobic Bacterial Strain from Soluble Starch

NASA Astrophysics Data System (ADS)

Kieu, Hoa Thi Quynh; Nguyen, Yen Thi; Dang, Yen Thi; Nguyen, Binh Thanh

2016-05-01

Biohydrogen is a clean source of energy that produces no harmful byproducts during combustion, being a potential sustainable energy carrier for the future. Therefore, biohydrogen produced by anaerobic bacteria via dark fermentation has attracted attention worldwide as a renewable energy source. However, the hydrogen production capability of these bacteria depends on major factors such as substrate, iron-containing hydrogenase, reduction agent, pH, and temperature. In this study, the response surface methodology (RSM) with central composite design (CCD) was employed to improve the hydrogen production by an anaerobic bacterial strain isolated from animal waste in Phu Linh, Soc Son, Vietnam (PL strain). The hydrogen production process was investigated as a function of three critical factors: soluble starch concentration (8 g L-1 to 12 g L-1), ferrous iron concentration (100 mg L-1 to 200 mg L-1), and l-cysteine concentration (300 mg L-1 to 500 mg L-1). RSM analysis showed that all three factors significantly influenced hydrogen production. Among them, the ferrous iron concentration presented the greatest influence. The optimum hydrogen concentration of 1030 mL L-1 medium was obtained with 10 g L-1 soluble starch, 150 mg L-1 ferrous iron, and 400 mg L-1 l-cysteine after 48 h of anaerobic fermentation. The hydrogen concentration produced by the PL strain was doubled after using RSM. The obtained results indicate that RSM with CCD can be used as a technique to optimize culture conditions for enhancement of hydrogen production by the selected anaerobic bacterial strain. Hydrogen production from low-cost organic substrates such as soluble starch using anaerobic fermentation methods may be one of the most promising approaches.

Optimization of phytase production by Penicillium purpurogenum GE1 under solid state fermentation by using Box-Behnken design.

PubMed

Awad, Ghada E A; Helal, Mohamed M I; Danial, Enas N; Esawy, Mona A

2014-01-01

Phytase production by Penicillium purpurogenum GE1 isolated from soil around bean root nodules was investigated by solid state fermentation (SSF) using mixed substrates consisted of corn cob and corn bran. The SSF conditions were optimized by using one-variable-at-a-time strategy. The optimum conditions for phytase production were at 27 °C, pH 8 and 66% moisture content. The study of different carbon and nitrogen sources revealed that glucose and peptone registered the highest enzyme productivity (92 ± 5.6 U/g ds, 125 ± 4.9 U/g ds). Among different surfactants, maximum phytase productivity was observed with Tween 80 at 0.001 concentrations (170 ± 4.2 U/g ds). A Box-Behnken design was employed to investigate the optimization of the most significant variables affecting the enzyme production. Maximal phytase production was detected after the addition of (g/5 g ds): 0.75 glucose, 0.375 peptone and 0, 01 tween 80. This result represented an improvement in phytase production of 2.6 folds when compared to that previously obtained using the basal medium under the same cultivation conditions. The generated model was found to be very adequate for phytase production (90% accuracy) as the experimental value was 444 ± 3.5 U/g ds compared to 401 U/g ds for the predicted value. In brief, the production of phytase using corn cob and corn bran is a novel and cheap way for the production of this important enzyme and opens a new way for researchers to discover and explore this arena.

[Optimization of fermentation conditions for cold-adapted amylase production by Micrococcus antarcticus and its enzymatic properties].

PubMed

Fan, Hong-xi; Liu, Ying; Liu, Zhi-pei

2009-08-15

By single factor experiments, the fermentation conditions for cold-adapted amylase production from Micrococcus antarcticus were determined as follows(medium g/L): Na2 HPO4 2.0, KH2PO4 1.0, MgSO4 x 7H2O 0.1, NaCl 5.0, (NH4)2SO4 2.5, maltose 5.0, trace element solution 5.0 mL, pH 8.0, 100 mL/Erlenmeyer flask (500 mL); cultivation was in a rotating shaker at 12 degrees C and 160 r/min for 64 h.Under those conditions,the highest total enzyme activity (2.6 U/mL) was obtained and increased by 10.8 fold compared with the original value of 0.24 U/mL before optimization. This amylase was purified by concentration with ultrafiltration membrane module, Hitrap Q anion exchange chromatography and Superdex 200 gel filtration chromatography. The optimal temperature and pH for the purified amylase were 30 degrees C and 6.0, respectively.It still showed high activity at low temperature 10-15 degrees C. It was sensitive to high temperature but was stable at pH 6.0-10.0 with at least 70% activity remained. These results indicated that it was a typical cold-adapted enzyme. The enzyme activity was stimulated by Ca2+, Mn2+, Co2+ and Mg2+; but inhibited by Zn2+, Ba2+, Ag+, Cu2+, Al3+, Fe2, Fe3+, Hg2+, EDTA and citrate. This cold-adapted amylase showed resistance to inactivation of 0.1% nonionic surfactants such as Tween 80, TrintonX-100, etc. Its Km was 0.90 mg/mL.

Optimization of a low-cost defined medium for alcoholic fermentation--a case study for potential application in bioethanol production from industrial wastewaters.

PubMed

Comelli, Raúl N; Seluy, Lisandro G; Isla, Miguel A

2016-01-25

In bioethanol production processes, the media composition has an impact on product concentration, yields and the overall process economics. The main purpose of this research was to develop a low-cost mineral-based supplement for successful alcoholic fermentation in an attempt to provide an economically feasible alternative to produce bioethanol from novel sources, for example, sugary industrial wastewaters. Statistical experimental designs were used to select essential nutrients for yeast fermentation, and its optimal concentrations were estimated by Response Surface Methodology. Fermentations were performed on synthetic media inoculated with 2.0 g L(-1) of yeast, and the evolution of biomass, sugar, ethanol, CO2 and glycerol were monitored over time. A mix of salts [10.6 g L(-1) (NH4)2HPO4; 6.4 g L(-1) MgSO4·7H2O and 7.5 mg L(-1) ZnSO4·7H2O] was found to be optimal. It led to the complete fermentation of the sugars in less than 12h with an average ethanol yield of 0.42 g ethanol/g sugar. A general C-balance indicated that no carbonaceous compounds different from biomass, ethanol, CO2 or glycerol were produced in significant amounts in the fermentation process. Similar results were obtained when soft drink wastewaters were tested to evaluate the potential industrial application of this supplement. The ethanol yields were very close to those obtained when yeast extract was used as the supplement, but the optimized mineral-based medium is six times cheaper, which favorably impacts the process economics and makes this supplement more attractive from an industrial viewpoint. Copyright © 2015 Elsevier B.V. All rights reserved.

Microbial bioinformatics for food safety and production

PubMed Central

Alkema, Wynand; Boekhorst, Jos; Wels, Michiel

2016-01-01

In the production of fermented foods, microbes play an important role. Optimization of fermentation processes or starter culture production traditionally was a trial-and-error approach inspired by expert knowledge of the fermentation process. Current developments in high-throughput ‘omics’ technologies allow developing more rational approaches to improve fermentation processes both from the food functionality as well as from the food safety perspective. Here, the authors thematically review typical bioinformatics techniques and approaches to improve various aspects of the microbial production of fermented food products and food safety. PMID:26082168

Cellulolytic enzymes production by utilizing agricultural wastes under solid state fermentation and its application for biohydrogen production.

PubMed

Saratale, Ganesh D; Kshirsagar, Siddheshwar D; Sampange, Vilas T; Saratale, Rijuta G; Oh, Sang-Eun; Govindwar, Sanjay P; Oh, Min-Kyu

2014-12-01

Phanerochaete chrysosporium was evaluated for cellulase and hemicellulase production using various agricultural wastes under solid state fermentation. Optimization of various environmental factors, type of substrate, and medium composition was systematically investigated to maximize the production of enzyme complex. Using grass powder as a carbon substrate, maximum activities of endoglucanase (188.66 U/gds), exoglucanase (24.22 U/gds), cellobiase (244.60 U/gds), filter paperase (FPU) (30.22 U/gds), glucoamylase (505.0 U/gds), and xylanase (427.0 U/gds) were produced under optimized conditions. The produced crude enzyme complex was employed for hydrolysis of untreated and mild acid pretreated rice husk. The maximum amount of reducing sugar released from enzyme treated rice husk was 485 mg/g of the substrate. Finally, the hydrolysates of rice husk were used for hydrogen production by Clostridium beijerinckii. The maximum cumulative H2 production and H2 yield were 237.97 mL and 2.93 mmoL H2/g of reducing sugar, (or 2.63 mmoL H2/g of cellulose), respectively. Biohydrogen production performance obtained from this work is better than most of the reported results from relevant studies. The present study revealed the cost-effective process combining cellulolytic enzymes production under solid state fermentation (SSF) and the conversion of agro-industrial residues into renewable energy resources.

Optimizing bio-physical conditions and pre-treatment options for breaking lignin barrier of maize stover feed using white rot fungi.

PubMed

Atuhaire, Andrew M; Kabi, Fred; Okello, Samuel; Mugerwa, Swidiq; Ebong, Cyprian

2016-12-01

The greatest limitation to utilization of maize stover by ruminants as a feed is the high concentration of lignin, which limits fibre digestibility. However, ruminants can effectively utilize maize stover if its nutritive value is improved using white rot fungal species. This study was designed to determine optimal bio-physical conditions for mycelial growth and select the most ideal fungal species and pre-treatment options for improving nutritive value of maize stover. Four popular edible Pleurotus fungal species (viz. Pleurotus florida, Pleurotus ostreatus, Pleurotus sajor caju and Pleurotus pulmonarius ) were subjected to varying temperatures, pH levels, hydrogen peroxide (H 2 O 2 ) concentration and illumination to establish the extent of mycelial growth rate. Inclusion of H 2 O 2 was used to determine optimal levels for preservation and prevention of contamination from other indigenous microbiota. Effects of pre-treatment options on chemical composition and nutritive value of maize stover were also examined. Mycelial growth rate of Pleurotus species on potato dextrose agar (PDA) varied ( P  < 0.05) with temperature, pH level and H 2 O 2 concentration following a quadratic trend. Optimal temperature, pH and H 2 O 2 concentration for mycelial growth on PDA were 25 °C, 5 and 0.01 mL/L, respectively. Under the different bio-physical conditions, P. sajor caju had the highest mycelia density and growth rate. Chemical composition of solid-state fermented maize stover differed ( P  < 0.05) among the Pleurotus species. Maize stover fermented with P. sajor caju had the highest crude protein (CP) of 86.6 g/kg DM, in-vitro dry matter digestibility (IVDMD) of 731 g/kg DM, in-vitro organic matter digestibility (IVOMD) of 670.4 g/kg DM and metabolizable energy (ME) of 10.0 MJ/kg DM but with the lowest lignin (sa) of 50 g/kg DM. At 25 °C, P. sajor caju had the highest mycelial growth rate on PDA and highest lignin (sa) breakdown in the maize stover substrate. It was, therefore, selected as the most ideal fungal species for improving nutritive value of maize stover. Pre-treatment of maize stover with Lactobacillus plantarum and molasses under anaerobic condition for 7 days before inoculation with P. sajor caju resulted into a substrate with the highest ( P  < 0.05) CP (96.6 g/kg DM), IVDMD (752.3 g/kg DM), IVOMD (687.2 g/kg DM) and ME (10.2 MJ/kg DM). However, neutral detergent fiber exclusive of residual ash (NDFom) and lignin (sa) fractions decreased ( P  < 0.05) as a result of subjecting maize stover to pre-treatment with L. plantarum and molasses prior to fermentation with P. sajor caju. Therefore, pre-treatment of maize stover with L. plantarum and molasses for 7 days prior to fermentation with P. sajor caju for 14 days in darkness at 25 °C offered the greatest potential for breaking the lignin barrier.

Impact of initial lipid content and oxygen supply on alcoholic fermentation in champagne-like musts.

PubMed

Ochando, Thomas; Mouret, Jean-Roch; Humbert-Goffard, Anne; Sablayrolles, Jean-Marie; Farines, Vincent

2017-08-01

Available nitrogen, lipids, or oxygen are nutrients with major impact on the kinetics of winemaking fermentation. Assimilable nitrogen is usually the growth-limiting nutrient which availability determines the fermentation rate and therefore the fermentation duration. In some particular cases, as in Champagne, grape musts have high available nitrogen content and low turbidity, i.e., below 50 Nephelometric Turbidity Unit (NTU). In the case of low turbidity, the availability of lipids, particularly phytosterols, becomes limiting. In this situation, control of oxygenation, which is necessary for lipid synthesis by yeast, is particularly crucial during fermentation. To mimic and understand these situations, a synthetic medium simulating the average composition of a Champagne must was used. This medium contained phytosterol (mainly β-sitosterol) concentrations ranging from 0 to 8mg/L corresponding to turbidity between 10 and 90 NTU. Population reached during the stationary phase and the maximum fermentation rate are conditioned by the initial phytosterol concentration determining the amount of nitrogen consumption. An early loss of viability was observed when the lipid concentrations were very low. For example, the viability continuously decreased during the stationary phase to a final value of 50% for an initial phytosterol concentration of 1mg/L. In some fermentations, 10mg/L oxygen were added at the end of the growth phase to combine the effects of initial content of phytosterols in the musts and the de novo synthesis of ergosterol and unsaturated fatty acids induced by oxygen addition. Effect of oxygen supply on the fermentation kinetics was particularly significant for media with low phytosterol contents. For example, the maximum fermentation rate was increased by 1.4-fold and the fermentation time was 70h shorter with oxygen addition in the medium containing 2mg/L of phytosterols. As a consequence of the oxygen supply, for the media containing 3, 5 and 8mg/L of phytosterols, the assimilable nitrogen was completely exhausted and the fermentation kinetics, as well as the final populations and viabilities (greater than 90%), were identical for the 3 conditions. The impacts of the lipid content and additional oxygen on acetate, glycerol and succinate synthesis were also studied. The phytosterols decreased the acetate and increased the succinate synthesis, and oxygenation resulted in a decrease in succinate formation. This work highlights the similarities and differences between the effects of lipids and oxygen on fermentation kinetics and yeast metabolism. This research highlights the need for an optimal combined management of lipid content in the must via turbidity and oxygenation, particularly in nitrogen-rich musts. Copyright © 2016 Elsevier Ltd. All rights reserved.

Optimization of biotransformation from phytosterol to androstenedione by a mutant Mycobacterium neoaurum ZJUVN-08.

PubMed

Zhang, Xiao-yan; Peng, Yong; Su, Zhong-rui; Chen, Qi-he; Ruan, Hui; He, Guo-qing

2013-02-01

Biotransformation of phytosterol (PS) by a newly isolated mutant Mycobacterium neoaurum ZJUVN-08 to produce androstenedione has been investigated in this paper. The parameters of the biotransformation process were optimized using fractional factorial design and response surface methodology. Androstenedione was the sole product in the fermentation broth catalyzed by the mutant M. neoaurum ZJUVN-08 strain. Results showed that molar ratio of hydroxypropyl-β-cyclodextrin (HP-β-CD) to PS and substrate concentrations were the two most significant factors affecting androstenedione production. By analyzing the statistical model of three-dimensional surface plot, the optimal process conditions were observed at 0.1 g/L inducer, pH 7.0, molar ratio of HP-β-CD to PS 1.92:1, 8.98 g/L PS, and at 120 h of incubation time. Under these conditions, the maximum androstenedione yield was 5.96 g/L and nearly the same with the non-optimized (5.99 g/L), while the maximum PS conversion rate was 94.69% which increased by 10.66% compared with the non-optimized (84.03%). The predicted optimum conditions from the mathematical model were in agreement with the verification experimental results. It is considered that response surface methodology was a powerful and efficient method to optimize the parameters of PS biotransformation process.

Optimization of biotransformation from phytosterol to androstenedione by a mutant Mycobacterium neoaurum ZJUVN-08*

PubMed Central

Zhang, Xiao-yan; Peng, Yong; Su, Zhong-rui; Chen, Qi-he; Ruan, Hui; He, Guo-qing

2013-01-01

Biotransformation of phytosterol (PS) by a newly isolated mutant Mycobacterium neoaurum ZJUVN-08 to produce androstenedione has been investigated in this paper. The parameters of the biotransformation process were optimized using fractional factorial design and response surface methodology. Androstenedione was the sole product in the fermentation broth catalyzed by the mutant M. neoaurum ZJUVN-08 strain. Results showed that molar ratio of hydroxypropyl-β-cyclodextrin (HP-β-CD) to PS and substrate concentrations were the two most significant factors affecting androstenedione production. By analyzing the statistical model of three-dimensional surface plot, the optimal process conditions were observed at 0.1 g/L inducer, pH 7.0, molar ratio of HP-β-CD to PS 1.92:1, 8.98 g/L PS, and at 120 h of incubation time. Under these conditions, the maximum androstenedione yield was 5.96 g/L and nearly the same with the non-optimized (5.99 g/L), while the maximum PS conversion rate was 94.69% which increased by 10.66% compared with the non-optimized (84.03%). The predicted optimum conditions from the mathematical model were in agreement with the verification experimental results. It is considered that response surface methodology was a powerful and efficient method to optimize the parameters of PS biotransformation process. PMID:23365012

Statistical Optimization of Laccase Production and Delignification of Sugarcane Bagasse by Pleurotus ostreatus in Solid-State Fermentation

PubMed Central

Karp, Susan Grace; Faraco, Vincenza; Amore, Antonella; Letti, Luiz Alberto Junior; Thomaz Soccol, Vanete; Soccol, Carlos Ricardo

2015-01-01

Laccases are oxidative enzymes related to the degradation of phenolic compounds, including lignin units, with concomitant reduction of oxygen to water. Delignification is a necessary pretreatment step in the process of converting plant biomass into fermentable sugars. The objective of this work was to optimize the production of laccases and to evaluate the delignification of sugarcane bagasse by Pleurotus ostreatus in solid-state fermentation. Among eight variables (pH, water activity, temperature, and concentrations of CuSO4, (NH4)2SO4, KH2PO4, asparagine, and yeast extract), copper sulfate and ammonium sulfate concentrations were demonstrated to significantly influence laccase production. The replacement of ammonium sulfate by yeast extract and the addition of ferulic acid as inducer provided increases of 5.7- and 2.0-fold, respectively, in laccase activity. Optimization of laccase production as a function of yeast extract, copper sulfate, and ferulic acid concentrations was performed by response surface methodology and optimal concentrations were 6.4 g/L, 172.6 μM, and 1.86 mM, respectively. Experimentally, the maximum laccase activity of 151.6 U/g was produced at the 5th day of solid-state fermentation. Lignin content in sugarcane bagasse was reduced from 31.89% to 26.36% after 5 days and to 20.79% after 15 days by the biological treatment of solid-state fermentation. PMID:26180784

High-throughput fermentation screening for the yeast Yarrowia lipolytica with real-time monitoring of biomass and lipid production.

PubMed

Back, Alexandre; Rossignol, Tristan; Krier, François; Nicaud, Jean-Marc; Dhulster, Pascal

2016-08-23

Because the model yeast Yarrowia lipolytica can synthesize and store lipids in quantities up to 20 % of its dry weight, it is a promising microorganism for oil production at an industrial scale. Typically, optimization of the lipid production process is performed in the laboratory and later scaled up for industrial production. However, the scale-up process can be complicated by genetic modifications that are optimized for one set of growing conditions can confer a less-than-optimal phenotype in a different environment. To address this issue, small cultivation systems have been developed that mimic the conditions in benchtop bioreactors. In this work, we used one such microbioreactor system, the BioLector, to develop high-throughput fermentation procedures that optimize growth and lipid accumulation in Y. lipolytica. Using this system, we were able to monitor lipid and biomass production in real time throughout the culture duration. The BioLector can monitor the growth of Y. lipolytica in real time by evaluating scattered light; this produced accurate measurements until cultures reached an equivalent of OD600nm = 115 and a cell dry weight of 100 g L(-1). In addition, a lipid-specific fluorescent probe was applied which reliably monitored lipid production up to a concentration of 12 g L(-1). Through screening various growing conditions, we determined that a carbon/nitrogen ratio of 35 was the most efficient for lipid production. Further screening showed that ammonium chloride and glycerol were the most valuable nitrogen and carbon sources, respectively, for growth and lipid production. Moreover, a carbon concentration above 1 M appeared to impair growth and lipid accumulation. Finally, we used these optimized conditions to screen engineered strains of Y. lipolytica with high lipid-accumulation capability. The growth and lipid content of the strains cultivated in the BioLector were compared to those grown in benchtop bioreactors. To our knowledge, this is the first time that the BioLector has been used to track lipid production in real time and to monitor the growth of Y. lipolytica. The present study also showed the efficacy of the BioLector in screening growing conditions and engineered strains prior to scale-up. The method described here could be applied to other oleaginous microorganisms.

Sequence and transcriptional analysis of the genes responsible for curdlan biosynthesis in Agrobacterium sp. ATCC 31749 under simulated dissolved oxygen gradients conditions.

PubMed

Zhang, Hong-Tao; Zhan, Xiao-Bei; Zheng, Zhi-Yong; Wu, Jian-Rong; Yu, Xiao-Bin; Jiang, Yun; Lin, Chi-Chung

2011-07-01

Expression at the mRNA level of ten selected genes in Agrobacterium sp. ATCC 31749 under various dissolved oxygen (DO) levels during curdlan fermentation related to electron transfer chain (ETC), tricarboxylic acid (TCA) cycle, peptidoglycan/lipopolysaccharide biosynthesis, and uridine diphosphate (UDP)-glucose biosynthesis were determined by qRT-PCR. Experiments were performed at DO levels of 30%, 50%, and 75%, as well as under low-oxygen conditions. The effect of high cell density on transcriptional response of the above genes under low oxygen was also studied. Besides cytochrome d (cyd A), the transcription levels of all the other genes were increased at higher DO and reached maximum at 50% DO. Under 75% DO, the transcriptional levels of all the genes were repressed. In addition, transcription levels of icd, sdh, cyo A, and fix N genes did not exhibit significant fluctuation with high cell density culture under low oxygen. These results suggested a mechanism for DO regulation of curdlan synthesis through regulation of transcriptional levels of ETCs, TCA, and UDP-glucose synthesis genes during curdlan fermentation. To our knowledge, this is the first report that DO concentration apparently regulates curdlan biosynthesis in Agrobacterium sp. ATCC 31749 providing essential lead for the optimization of the fermentation at the industrial scale.

Improving production of volatile fatty acids from food waste fermentation by hydrothermal pretreatment.

PubMed

Yin, Jun; Wang, Kun; Yang, Yuqiang; Shen, Dongsheng; Wang, Meizhen; Mo, Han

2014-11-01

Food waste (FW) was pretreated by a hydrothermal method and then fermented for volatile fatty acid (VFAs) production. The soluble substance in FW increased after hydrothermal pretreatment (⩽200 °C). Higher hydrothermal temperature would lead to mineralization of the organic compounds. The optimal temperature for organic dissolution was 180 °C, at which FW dissolved 42.5% more soluble chemical oxygen demand than the control. VFA production from pretreated FW fermentation was significantly enhanced compared with the control. The optimal hydrothermal temperature was 160 °C with a VFA yield of 0.908 g/g VSremoval. Butyrate and acetate were the prevalent VFAs followed by propionate and valerate. FW fermentation was inhibited after 200 °C pretreatment. The VFAs were extracted from the fermentation broth by liquid-liquid extraction. The VFA recovery was 50-70%. Thus, 0.294-0.411 g VFAs could be obtained per gram of hydrothermally pretreated FW (in dry weight) by this method. Copyright © 2014 Elsevier Ltd. All rights reserved.

Comparison of solid-state and submerged-state fermentation for the bioprocessing of switchgrass to ethanol and acetate by Clostridium phytofermentans.

PubMed

Jain, Abhiney; Morlok, Charles K; Henson, J Michael

2013-01-01

The conversion of sustainable energy crops using microbiological fermentation to biofuels and bioproducts typically uses submerged-state processes. Alternatively, solid-state fermentation processes have several advantages when compared to the typical submerged-state processes. This study compares the use of solid-state versus submerged-state fermentation using the mesophilic anaerobic bacterium Clostridium phytofermentans in the conversion of switchgrass to the end products of ethanol, acetate, and hydrogen. A shift in the ratio of metabolic products towards more acetate and hydrogen production than ethanol production was observed when C. phytofermentans was grown under solid-state conditions as compared to submerged-state conditions. Results indicated that the end product concentrations (in millimolar) obtained using solid-state fermentation were higher than using submerged-state fermentation. In contrast, the total fermentation products (in weight of product per weight of carbohydrates consumed) and switchgrass conversion were higher for submerged-state fermentation. The conversion of xylan was greater than glucan conversion under both fermentation conditions. An initial pH of 7 and moisture content of 80 % resulted in maximum end products formation. Scanning electron microscopy study showed the presence of biofilm formed by C. phytofermentans growing on switchgrass under submerged-state fermentation whereas bacterial cells attached to surface and no apparent biofilm was observed when grown under solid-state fermentation. To our knowledge, this is the first study reporting consolidated bioprocessing of a lignocellulosic substrate by a mesophilic anaerobic bacterium under solid-state fermentation conditions.

Modelling and optimization of environmental conditions for kefiran production by Lactobacillus kefiranofaciens.

PubMed

Cheirsilp, B; Shimizu, H; Shioya, S

2001-12-01

A mathematical model for kefiran production by Lactobacillus kefiranofaciens was established, in which the effects of pH, substrate and product on cell growth, exopolysaccharide formation and substrate assimilation were considered. The model gave a good representation both of the formation of exopolysaccharides (which are not only attached to cells but also released into the medium) and of the time courses of the production of galactose and glucose in the medium (which are produced and consumed by the cells). Since pH and both lactose and lactic acid concentrations differently affected production and growth activity, the model included the effects of pH and the concentrations of lactose and lactic acid. Based on the mathematical model, an optimal pH profile for the maximum production of kefiran in batch culture was obtained. In this study, a simplified optimization method was developed, in which the optimal pH profile was determined at a particular final fermentation time. This was based on the principle that, at a certain time, switching from the maximum specific growth rate to the critical one (which yields the maximum specific production rate) results in maximum production. Maximum kefiran production was obtained, which was 20% higher than that obtained in the constant-pH control fermentation. A genetic algorithm (GA) was also applied to obtain the optimal pH profile; and it was found that practically the same solution was obtained using the GA.

Modeling and investigation of submerged fermentation process to produce extracellular polysaccharide using Lactobacillus confusus.

PubMed

Thirugnanasambandham, K; Sivakumar, V; Prakash Maran, J

2014-12-19

The main objective of the present study is to investigate and optimize the Submerged fermentation (SMF) process parameters such as addition of coconut water, NaCl dose, incubation time and temperature on the production of extracellular polysaccharide (EPS) and biomass production using Lactobacillus confuses. Response surface methodology (RSM) coupled with four factors three level Box-Behnken design (BBD) was employed to model the SMF process. RSM analysis indicated good correspondence between experimental and predicted values. Three dimentional (3D) response surface plots were used to study the interactive effects of process variables on SMF process. The optimum process conditions for the maximum production of EPS and biomass were found to be as follows; addition of coconut water of 40%, NaCl dose of 15%, incubation time of 24h and temperature of 35°C. Under these conditions, 10.57 g/L of EPS and 3.9 g/L of biomass were produced. Copyright © 2014 Elsevier Ltd. All rights reserved.

Effect of ultrasonic extraction conditions on antioxidative and immunomodulatory activities of a Ganoderma lucidum polysaccharide originated from fermented soybean curd residue.

PubMed

Shi, Min; Yang, Yingnan; Hu, Xuansheng; Zhang, Zhenya

2014-07-15

A crude Ganoderma lucidum polysaccharide (GLPL) was extracted from fermented soybean curd residue by ultrasonic assisted extraction. The optimal extraction conditions were 30 min at 80 °C with 80 W and water to solid ratio of 10, and with this method 115.47 ± 2.95 mg/g of GLPL yield was obtained. Additionally, the antioxidant and immunomodulatory activities of GLPL were investigated. The results showed that GLPL exhibited strong antioxidant effects, which included scavenging activities against DPPH radicals, hydrogen oxide and ABTS radicals with IC50 values of 0.23, 0.48 and 0.69 mg/mL, respectively. For immunomodulatory activities, GLPL was shown to strongly stimulate the proliferation of macrophages (158.02 ± 13.12%), the production of nitric oxide and phagocytosis (21.16 ± 1.65 μM), and, at 40.00 μg/mL, protected macrophage from Doxorubicin (DOX) (0.16 ± 0.003). Copyright © 2014 Elsevier Ltd. All rights reserved.

Influence of temperature and hydraulic retention on the production of volatile fatty acids during anaerobic fermentation of cow manure and maize silage.

PubMed

Cavinato, Cristina; Da Ros, Cinzia; Pavan, Paolo; Bolzonella, David

2017-01-01

The aim of this study was to verify the efficiency of a separate hydrolysis step by testing different working temperatures (37-55°C) and hydraulic retention times (two, four and six days) and by evaluating readily biodegradable carbon production. The fermentation products included primarily acetic, propionic and butyric acids. These acids can be easily converted into biogas or can be recovered in a biorefinery approach, for example, to produce polyhydroxyalkanoates. The optimal condition was found by applying an organic loading rate of 17.9gTVSm -3 with a four-day retention time at 37°C for an acidification yield of 183.2gCOD VFA kgVS fed -1 . Copyright © 2016 Elsevier Ltd. All rights reserved.

Kinetic Analysis of Strains of Lactic Acid Bacteria and Acetic Acid Bacteria in Cocoa Pulp Simulation Media toward Development of a Starter Culture for Cocoa Bean Fermentation ▿

PubMed Central

Lefeber, Timothy; Janssens, Maarten; Camu, Nicholas; De Vuyst, Luc

2010-01-01

The composition of cocoa pulp simulation media (PSM) was optimized with species-specific strains of lactic acid bacteria (PSM-LAB) and acetic acid bacteria (PSM-AAB). Also, laboratory fermentations were carried out in PSM to investigate growth and metabolite production of strains of Lactobacillus plantarum and Lactobacillus fermentum and of Acetobacter pasteurianus isolated from Ghanaian cocoa bean heap fermentations, in view of the development of a defined starter culture. In a first step, a selection of strains was made out of a pool of strains of these LAB and AAB species, obtained from previous studies, based on their fermentation kinetics in PSM. Also, various concentrations of citric acid in the presence of glucose and/or fructose (PSM-LAB) and of lactic acid in the presence of ethanol (PSM-AAB) were tested. These data could explain the competitiveness of particular cocoa-specific strains, namely, L. plantarum 80 (homolactic and acid tolerant), L. fermentum 222 (heterolactic, citric acid fermenting, mannitol producing, and less acid tolerant), and A. pasteurianus 386B (ethanol and lactic acid oxidizing, acetic acid overoxidizing, acid tolerant, and moderately heat tolerant), during the natural cocoa bean fermentation process. For instance, it turned out that the capacity to use citric acid, which was exhibited by L. fermentum 222, is of the utmost importance. Also, the formation of mannitol was dependent not only on the LAB strain but also on environmental conditions. A mixture of L. plantarum 80, L. fermentum 222, and A. pasteurianus 386B can now be considered a mixed-strain starter culture for better controlled and more reliable cocoa bean fermentation processes. PMID:20889778

Kinetic analysis of strains of lactic acid bacteria and acetic acid bacteria in cocoa pulp simulation media toward development of a starter culture for cocoa bean fermentation.

PubMed

Lefeber, Timothy; Janssens, Maarten; Camu, Nicholas; De Vuyst, Luc

2010-12-01

The composition of cocoa pulp simulation media (PSM) was optimized with species-specific strains of lactic acid bacteria (PSM-LAB) and acetic acid bacteria (PSM-AAB). Also, laboratory fermentations were carried out in PSM to investigate growth and metabolite production of strains of Lactobacillus plantarum and Lactobacillus fermentum and of Acetobacter pasteurianus isolated from Ghanaian cocoa bean heap fermentations, in view of the development of a defined starter culture. In a first step, a selection of strains was made out of a pool of strains of these LAB and AAB species, obtained from previous studies, based on their fermentation kinetics in PSM. Also, various concentrations of citric acid in the presence of glucose and/or fructose (PSM-LAB) and of lactic acid in the presence of ethanol (PSM-AAB) were tested. These data could explain the competitiveness of particular cocoa-specific strains, namely, L. plantarum 80 (homolactic and acid tolerant), L. fermentum 222 (heterolactic, citric acid fermenting, mannitol producing, and less acid tolerant), and A. pasteurianus 386B (ethanol and lactic acid oxidizing, acetic acid overoxidizing, acid tolerant, and moderately heat tolerant), during the natural cocoa bean fermentation process. For instance, it turned out that the capacity to use citric acid, which was exhibited by L. fermentum 222, is of the utmost importance. Also, the formation of mannitol was dependent not only on the LAB strain but also on environmental conditions. A mixture of L. plantarum 80, L. fermentum 222, and A. pasteurianus 386B can now be considered a mixed-strain starter culture for better controlled and more reliable cocoa bean fermentation processes.

Extracellular production of human parathyroid hormone as a thioredoxin fusion form in Escherichia coli by chemical permeabilization combined with heat treatment.

PubMed

Fu, Xiang-Yang; Tong, Wang-Yu; Wei, Dong-Zhi

2005-01-01

A pET system encoding the fusion protein gene of thioredoxin (Trx) and human parathyroid hormone (hPTH) was introduced into Escherichia coli BL21 (DE3). Recombinant Trx-hPTH fusion protein was expressed in soluble form in the cytoplasm of the E. coli transformant. To recover Trx-hPTH from the E. coli culture efficiently, a novel tactic was developed by adding Triton X-100 into the fermentation culture at the exponential growth phase of E. coli and by heat treatment of the culture at the end of the fermentation. A concentration of 1% (v/v) Triton X-100 was added into the culture at the same time as IPTG addition after optimization. Under these conditions, addition of Triton X-100 had little effect on the cell growth, but more than 75% of the total recombinant Trx-hPTH was released into the fermentation broth. Also, a much higher volumetric yield of recombinant Trx-hPTH could be obtained with protein release compared to yield without protein release. Simultaneously, owing to the highly thermal stability of Trx-hPTH fusion protein, heat treatment of the fermentation broth at 80 degrees C for 15 min at the end of fermentation was employed for primary purification. Results demonstrated that heat treatment not only boosted further release of the recombinant Trx-hPTH fusion protein into the fermentation broth but also precipitated/denatured most of the nontarget proteins released in the broth. The tactics described herein integrated the fermentation process with subsequent recovery steps and thus provided a valuable and economical method for the production of Trx-hPTH and maybe some other Trx fusions in E. coli.

Investigation of nutrient feeding strategies in a countercurrent mixed-acid multi-staged fermentation: experimental data.

PubMed

Smith, Aaron Douglas; Lockman, Nur Ain; Holtzapple, Mark T

2011-06-01

Nutrients are essential for microbial growth and metabolism in mixed-culture acid fermentations. Understanding the influence of nutrient feeding strategies on fermentation performance is necessary for optimization. For a four-bottle fermentation train, five nutrient contacting patterns (single-point nutrient addition to fermentors F1, F2, F3, and F4 and multi-point parallel addition) were investigated. Compared to the traditional nutrient contacting method (all nutrients fed to F1), the near-optimal feeding strategies improved exit yield, culture yield, process yield, exit acetate-equivalent yield, conversion, and total acid productivity by approximately 31%, 39%, 46%, 31%, 100%, and 19%, respectively. There was no statistical improvement in total acid concentration. The traditional nutrient feeding strategy had the highest selectivity and acetate-equivalent selectivity. Total acid productivity depends on carbon-nitrogen ratio.

Effect of in situ exopolysaccharide production on physicochemical, rheological, sensory, and microstructural properties of the yogurt drink ayran: an optimization study based on fermentation kinetics.

PubMed

Yilmaz, M T; Dertli, E; Toker, O S; Tatlisu, N B; Sagdic, O; Arici, M

2015-03-01

Exopolysaccharide (EPS)-producing starter cultures are preferred for the manufacture of fermented milk products to improve rheological and technological properties. However, no clear correlation exists between EPS production and the rheological and technological properties of fermented milk products such as the yogurt drink ayran. In this study, 4 different strain conditions (EPS- and EPS+ Streptococcus thermophilus strains) were tested as a function of incubation temperature (32, 37, or 42°C) and time (2, 3, or 4 h) to determine the effect of culture type and in situ EPS production on physicochemical, rheological, sensory, and microstructural properties of ayran. Furthermore, we assessed the effect of fermentation conditions on amounts of EPS production by different EPS-producing strains during ayran production. A multifactorial design of response surface methodology was used to model linear, interaction, and quadratic effects of these variables on steady shear rheological properties of ayran samples and in situ EPS production levels. The physicochemical and microbiological characteristics of ayran samples altered depending on incubation conditions and strain selection. Steady shear tests showed that ayran samples inoculated with EPS+ strains exhibited pseudoplastic flow behavior. Production of ayran with EPS- strain (control sample) resulted in the lowest apparent viscosity values (η50), whereas those produced with the combination of 2 EPS+ strains yielded ayran with notably increased η50 values. We concluded that incubation time was the variable with the greatest effect on η50, consistency coefficient (K), and flow behavior index (n) values. In situ EPS production was also affected by these conditions during ayran fermentation in which strain-specific metabolism conditions were found to be the most important factor for EPS production. In addition, these findings correlated the amount of in situ EPS produced with the rheological properties of ayran. Scanning electron microscopy images of the samples showed differences in structural features, revealing a prominent network strand structure in the ayran samples inoculated with the admixture of 2 EPS-producing strains incubated at 37°C for 3 h. These results provide useful information for large-scale production of ayran by the dairy industry. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Solid-substrate bioprocessing of cow dung for the production of carboxymethyl cellulase by Bacillus halodurans IND18.

PubMed

Vijayaraghavan, P; Prakash Vincent, S G; Dhillon, G S

2016-02-01

The production of carboxymethyl cellulase (CMCase) by Bacillus halodurans IND18 under solid substrate fermentation (SSF) using cow dung was optimized through two level full factorial design and second order response surface methodology (RSM). The central composite design (CCD) was employed to optimize the vital fermentation parameters, such as pH of the substrate, concentration of nitrogen source (peptone) and ion (sodium dihydrogen phosphate) sources in medium for achieving higher enzyme production. The optimum medium composition was found to be 1.46% (w/w) peptone, 0.095% (w/w) sodium dihydrogen phosphate and pH 8.0. The model prediction of 4210IU/g enzyme activity at optimum conditions was verified experimentally as 4140IU/g. The enzyme was active over a broad temperature range (40-60±1°C) and pH (7.0-9.0) with maximal activity at 60±1°C and pH 8.0. This study demonstrated the potential of cow dung as novel substrate for CMCase production. Copyright © 2015 Elsevier Ltd. All rights reserved.

Xylanase production by Burkholderia sp. DMAX strain under solid state fermentation using distillery spent wash.

PubMed

Mohana, Sarayu; Shah, Amita; Divecha, Jyoti; Madamwar, Datta

2008-11-01

Xylanase production by a newly isolated strain of Burkholderia sp. was studied under solid state fermentation using anaerobically treated distillery spent wash. Response surface methodology (RSM) involving Box-Behnken design was employed for optimizing xylanase production. The interactions between distillery effluent concentration, initial pH, moisture ratio and inoculum size were investigated and modeled. Under optimized conditions, xylanase production was found to be in the range of 5200-5600 U/g. The partially purified enzyme recovered after ammonium sulphate fractionation showed maximum activity at 50 degrees C and pH 8.6. Kinetic parameters like Km and Vmax for xylan were found to be 12.75 mg/ml and 165 micromol/mg/min. In the presence of metal ions such as Ca2+, Co2+, Mn2+, Ba2+, Mg2+ and protein disulphide reducing agents such as beta-mercaptoethanol and dithiotheritol (DTT) the activity of enzyme increased, where as strong inhibition of enzyme activity was observed in the presence of Cu2+, Ag+, Fe2+ and SDS. The crude enzyme hydrolysed lignocellulosic substrate, wheat bran as well as industrial pulp.

An economical approach for d-lactic acid production utilizing unpolished rice from aging paddy as major nutrient source.

PubMed

Lu, Zhengdong; Lu, Mingbo; He, Feng; Yu, Longjiang

2009-03-01

In order to reduce the raw material cost of d-lactic acid fermentation, the unpolished rice from aging paddy was used as major nutrient source in this study. The unpolished rice saccharificate, wheat bran powder and yeast extract were employed as carbon source, nitrogen source and growth factors, respectively. Response surface methodology (RSM) was applied to optimize the dosages of medium compositions. As a result, when the fermentation was carried out under the optimal conditions for wheat bran powder (29.10g/l) and yeast extract (2.50g/l), the d-lactic acid yield reached 731.50g/kg unpolished rice with a volumetric production rate of 1.50g/(lh). In comparison with fresh corn and polished rice, the d-lactic acid yield increased by 5.79% and 8.71%, and the raw material cost decreased by 65% and 52%, respectively, when the unpolished rice was used as a major nutrient source. These results might provide a reference for the industrial production of d-lactic acid.

Green and sustainable succinic acid production from crude glycerol by engineered Yarrowia lipolytica via agricultural residue based in situ fibrous bed bioreactor.

PubMed

Li, Chong; Gao, Shi; Yang, Xiaofeng; Lin, Carol Sze Ki

2018-02-01

In situ fibrous bed bioreactor (isFBB) for efficient succinic acid (SA) production by Yarrowia lipolytica was firstly developed in our former study. In this study, agricultural residues including wheat straw, corn stalk and sugarcane bagasse were investigated for the improvement of isFBB, and sugarcane bagasse was demonstrated to be the best immobilization material. With crude glycerol as the sole carbon source, optimization for isFBB batch fermentation was carried out. Under the optimal conditions of 20g sugarcane bagasse as immobilization material, 120gL -1 crude glycerol as carbon source and 4Lmin -1 of aeration rate, the resultant SA concentration was 53.6gL -1 with an average productivity of 1.45gL -1 h -1 and a SA yield of 0.45gg -1 . By feeding crude glycerol, SA titer up to 209.7gL -1 was obtained from fed batch fermentation, which was the highest value that ever reported. Copyright © 2017 Elsevier Ltd. All rights reserved.

Modelling and optimisation of enzymatic extrusion pretreatment of broken rice for rice wine manufacture.

PubMed

Li, Hongyan; Wei, Benxi; Wu, Chunsen; Zhang, Bao; Xu, Xueming; Jin, Zhengyu; Tian, Yaoqi

2014-05-01

The manufacture of Chinese rice wine involves an uneconomical, time-consuming, and environmentally unfriendly pretreatment process. In this study, the enzymatic extrusion of broken rice was applied to the brewing of rice wine. The response surface methodology was used to study the effects of the barrel temperature (BT), moisture content (MC), and amylase concentration (AC) on the alcohol yield. A second-order polynomial model had a good fit to the experimental data and the coefficient of determination (R(2)) was 0.9879. According to the model, the optimal parameters required to obtain the highest alcoholic degree of 17.94% were: BT=100.14°C, MC=43%, and AC=1.45‰. Under these optimal conditions, the alcoholic degree actually reached 18.3%, which was close to the value predicted by the model. Enzymatic extrusion improved the yeast growth and alcohol yield during the fermentation process. The fermentation recovery and efficiency of processed rice wine were 38.07% and 94.66%, respectively. Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.

High-level expression of recombinant thermostable β-glucosidase in Escherichia coli by regulating acetic acid.

PubMed

Shi, Xuejia; Xie, Jingcong; Liao, Shiyong; Wu, Tao; Zhao, Lin-Guo; Ding, Gang; Wang, Zhenzhong; Xiao, Wei

2017-10-01

In the fermentation progress, fermentation parameters including the feed rate, induction temperature, and induction pH evidently regulate the accumulation of acetic acid generated by recombinant E. coli in the medium. The production of thermostable β-glucosidase (Tpebgl3) was increased by optimizing the parameters mentioned step by step. The optimal conditions were obtained with the highest enzyme expression (560.4U/mL) and the maximum DCW (65g/L) at the pre-induction specific growth rate of 0.2h -1 followed by a post-induction specific growth rate (0.18h -1 ); induction temperature is 39°C; the pH is 7.2; the concentration of acetic acid was maintained all along below 0.9g/L. Results show it is necessary for the synthesis of Tpebgl3 to regulate the accumulation of acetic acid at the premise of feeding to meet the normal growth of E. coli. The production of Tpebgl3 by recombinant E. coli is the highest reported to date. Copyright © 2017 Elsevier Ltd. All rights reserved.

Optimization of succinic acid fermentation with Actinobacillus succinogenes by response surface methodology (RSM)*

PubMed Central

Zhang, Yun-jian; Li, Qiang; Zhang, Yu-xiu; Wang, Dan; Xing, Jian-min

2012-01-01

Succinic acid is considered as an important platform chemical. Succinic acid fermentation with Actinobacillus succinogenes strain BE-1 was optimized by central composite design (CCD) using a response surface methodology (RSM). The optimized production of succinic acid was predicted and the interactive effects between glucose, yeast extract, and magnesium carbonate were investigated. As a result, a model for predicting the concentration of succinic acid production was developed. The accuracy of the model was confirmed by the analysis of variance (ANOVA), and the validity was further proved by verification experiments showing that percentage errors between actual and predicted values varied from 3.02% to 6.38%. In addition, it was observed that the interactive effect between yeast extract and magnesium carbonate was statistically significant. In conclusion, RSM is an effective and useful method for optimizing the medium components and investigating the interactive effects, and can provide valuable information for succinic acid scale-up fermentation using A. succinogenes strain BE-1. PMID:22302423

Realm of Thermoalkaline Lipases in Bioprocess Commodities.

PubMed

Lajis, Ahmad Firdaus B

2018-01-01

For decades, microbial lipases are notably used as biocatalysts and efficiently catalyze various processes in many important industries. Biocatalysts are less corrosive to industrial equipment and due to their substrate specificity and regioselectivity they produced less harmful waste which promotes environmental sustainability. At present, thermostable and alkaline tolerant lipases have gained enormous interest as biocatalyst due to their stability and robustness under high temperature and alkaline environment operation. Several characteristics of the thermostable and alkaline tolerant lipases are discussed. Their molecular weight and resistance towards a range of temperature, pH, metal, and surfactants are compared. Their industrial applications in biodiesel, biodetergents, biodegreasing, and other types of bioconversions are also described. This review also discusses the advance of fermentation process for thermostable and alkaline tolerant lipases production focusing on the process development in microorganism selection and strain improvement, culture medium optimization via several optimization techniques (i.e., one-factor-at-a-time, surface response methodology, and artificial neural network), and other fermentation parameters (i.e., inoculums size, temperature, pH, agitation rate, dissolved oxygen tension (DOT), and aeration rate). Two common fermentation techniques for thermostable and alkaline tolerant lipases production which are solid-state and submerged fermentation methods are compared and discussed. Recent optimization approaches using evolutionary algorithms (i.e., Genetic Algorithm, Differential Evolution, and Particle Swarm Optimization) are also highlighted in this article.

Optimization strategies based on sequential quadratic programming applied for a fermentation process for butanol production.

PubMed

Pinto Mariano, Adriano; Bastos Borba Costa, Caliane; de Franceschi de Angelis, Dejanira; Maugeri Filho, Francisco; Pires Atala, Daniel Ibraim; Wolf Maciel, Maria Regina; Maciel Filho, Rubens

2009-11-01

In this work, the mathematical optimization of a continuous flash fermentation process for the production of biobutanol was studied. The process consists of three interconnected units, as follows: fermentor, cell-retention system (tangential microfiltration), and vacuum flash vessel (responsible for the continuous recovery of butanol from the broth). The objective of the optimization was to maximize butanol productivity for a desired substrate conversion. Two strategies were compared for the optimization of the process. In one of them, the process was represented by a deterministic model with kinetic parameters determined experimentally and, in the other, by a statistical model obtained using the factorial design technique combined with simulation. For both strategies, the problem was written as a nonlinear programming problem and was solved with the sequential quadratic programming technique. The results showed that despite the very similar solutions obtained with both strategies, the problems found with the strategy using the deterministic model, such as lack of convergence and high computational time, make the use of the optimization strategy with the statistical model, which showed to be robust and fast, more suitable for the flash fermentation process, being recommended for real-time applications coupling optimization and control.

Bacteriophages BCP1-1 and BCP8-2 require divalent cations for efficient control of Bacillus cereus in fermented foods.

PubMed

Bandara, Nadeeka; Jo, Junhee; Ryu, Sangryeol; Kim, Kwang-Pyo

2012-08-01

Bacillus cereus is a foodborne bacterial pathogen that causes diarrhea and vomiting. In this study, the usefulness of bacteriophages to eradicate B. cereus from fermented foods was investigated. A total of 13 phages were isolated from Korean fermented food products, and 2 (BCP1-1 and BCP8-2) were further characterized. Transmission electron microscopy (TEM), restriction enzyme digestion pattern analysis, and SDS-PAGE of the structural proteins suggest that both phages belong to the family Myoviridae, containing approximately 150 kbp-long genomes. The host ranges of both phages were limited to B. cereus group species (12/13), as they were not able to lyse other Gram-positive or negative strains including Bacillus subtilis. Purified phages were used to inhibit B. cereus growth in a model fermented food system, cheonggukjang, a fast-fermented soybean paste product. BCP1-1 and BCP8-2 were able to effectively eradicate B. cereus from the food only if divalent cations (Ca²⁺, Mg²⁺, or Mn²⁺) were added to the medium. Further studies reveal that divalent cations are essential for phage adsorption, while a monovalent cation (Na⁺) is required for the post-adsorption phase of phage infection. Taken together, our findings imply that a phage could be an ideal anti-bacterial agent for use in fermented food products that require the presence of beneficial microflora and, during phage application, optimization of phage reaction conditions is critical for the successful utilization of phage biocontrol. Copyright © 2012 Elsevier Ltd. All rights reserved.

Influence of Culturing Conditions on Bioprospecting and the Antimicrobial Potential of Endophytic Fungi from Schinus terebinthifolius.

PubMed

Tonial, Fabiana; Maia, Beatriz H L N S; Gomes-Figueiredo, Josiane A; Sobottka, Andrea M; Bertol, Charise D; Nepel, Angelita; Savi, Daiani C; Vicente, Vânia A; Gomes, Renata R; Glienke, Chirlei

2016-02-01

In this study, we analyzed the antimicrobial activity of extracts harvested from 17 endophytic fungi isolated from the medicinal plant Schinus terebinthifolius. Morphological and molecular analyses indicated that these fungal species belonged to the genera Alternaria, Bjerkandera, Colletotrichum, Diaporthe, Penicillium, and Xylaria. Of the endophytes analyzed, 64.7 % produced antimicrobial compounds under at least one of the fermentation conditions tested. Nine isolates produced compounds that inhibited growth of Staphylococcus aureus, four produced compounds that inhibited Candida albicans, and two that inhibited Pseudomonas aeruginosa. The fermentation conditions of the following endophytes were optimized: Alternaria sp. Sect. Alternata-LGMF626, Xylaria sp.-LGMF673, and Bjerkandera sp.-LGMF713. Specifically, the carbon and nitrogen sources, initial pH, temperature, and length of incubation were varied. In general, production of antimicrobial compounds was greatest when galactose was used as a carbon source, and acidification of the growth medium enhanced the production of compounds that inhibited C. albicans. Upon large-scale fermentation, Alternaria sp. Sect. Alternata-LGMF626 produced an extract containing two fractions that were active against methicillin-resistant S. aureus. One of the extracts exhibited high activity (minimum inhibitory concentration of 18.52 µg/mL), and the other exhibited moderate activity (minimum inhibitory concentration of 55.55 µg/mL). The compounds E-2-hexyl-cinnamaldehyde and two compounds of the pyrrolopyrazine alkaloids class were identified in the active fractions by gas chromatography-mass spectrometry.

An Energy Balance Model to Predict Chemical Partitioning in a Photosynthetic Microbial Mat

NASA Technical Reports Server (NTRS)

Hoehler, Tori M.; Albert, Daniel B.; DesMarais, David J.

2006-01-01

Studies of biosignature formation in photosynthetic microbial mat communities offer potentially useful insights with regards to both solar and extrasolar astrobiology. Biosignature formation in such systems results from the chemical transformation of photosynthetically fixed carbon by accessory microorganisms. This fixed carbon represents a source not only of reducing power, but also energy, to these organisms, so that chemical and energy budgets should be coupled. We tested this hypothesis by applying an energy balance model to predict the fate of photosynthetic productivity under dark, anoxic conditions. Fermentation of photosynthetically fixed carbon is taken to be the only source of energy available to cyanobacteria in the absence of light and oxygen, and nitrogen fixation is the principal energy demand. The alternate fate for fixed carbon is to build cyanobacterial biomass with Redfield C:N ratio. The model predicts that, under completely nitrogen-limited conditions, growth is optimized when 78% of fixed carbon stores are directed into fermentative energy generation, with the remainder allocated to growth. These predictions were compared to measurements made on microbial mats that are known to be both nitrogen-limited and populated by actively nitrogen-fixing cyanobacteria. In these mats, under dark, anoxic conditions, 82% of fixed carbon stores were diverted into fermentation. The close agreement between these independent approaches suggests that energy balance models may provide a quantitative means of predicting chemical partitioning within such systems - an important step towards understanding how biological productivity is ultimately partitioned into biosignature compounds.

Screening of Ganoderma strains with high polysaccharides and ganoderic acid contents and optimization of the fermentation medium by statistical methods.

PubMed

Wei, Zhen-hua; Duan, Ying-yi; Qian, Yong-qing; Guo, Xiao-feng; Li, Yan-jun; Jin, Shi-he; Zhou, Zhong-Xin; Shan, Sheng-yan; Wang, Chun-ru; Chen, Xue-Jiao; Zheng, Yuguo; Zhong, Jian-Jiang

2014-09-01

Polysaccharides and ganoderic acids (GAs) are the major bioactive constituents of Ganoderma species. However, the commercialization of their production was limited by low yield in the submerged culture of Ganoderma despite improvement made in recent years. In this work, twelve Ganoderma strains were screened to efficiently produce polysaccharides and GAs, and Ganoderma lucidum 5.26 (GL 5.26) that had been never reported in fermentation process was found to be most efficient among the tested stains. Then, the fermentation medium was optimized for GL 5.26 by statistical method. Firstly, glucose and yeast extract were found to be the optimum carbon source and nitrogen source according to the single-factor tests. Ferric sulfate was found to have significant effect on GL 5.26 biomass production according to the results of Plackett-Burman design. The concentrations of glucose, yeast extract and ferric sulfate were further optimized by response surface methodology. The optimum medium composition was 55 g/L of glucose, 14 g/L of yeast extract, 0.3 g/L of ferric acid, with other medium components unchanged. The optimized medium was testified in the 10-L bioreactor, and the production of biomass, IPS, total GAs and GA-T enhanced by 85, 27, 49 and 93 %, respectively, compared to the initial medium. The fermentation process was scaled up to 300-L bioreactor; it showed good IPS (3.6 g/L) and GAs (670 mg/L) production. The biomass was 23.9 g/L in 300-L bioreactor, which was the highest biomass production in pilot scale. According to this study, the strain GL 5.26 showed good fermentation property by optimizing the medium. It might be a candidate industrial strain by further process optimization and scale-up study.

Improved fed-batch production of high-purity PHB (poly-3 hydroxy butyrate) by Cupriavidus necator (MTCC 1472) from sucrose-based cheap substrates under response surface-optimized conditions.

PubMed

Dey, Pinaki; Rangarajan, Vivek

2017-10-01

Experimental investigations were carried out for Cupriavidus necator (MTCC 1472)-based improved production of poly-3 hydroxy butyrate (PHB) through induced nitrogen limiting fed-batch cultivation strategies. Initially Plackett-Burman design and response surface methodology were implemented to optimize most influencing process parameters. With optimized process parameter values, continuous feeding strategies ware applied in a 5-l fermenter with table sugar concentration of 100 g/l, nitrogen concentration of 0.12 g/l for fed-batch fermentation with varying dilution rates of 0.02 and 0.046 1/h. To get enriched production of PHB, concentration of the sugar was further increased to 150 and 200 g/l in feeding. Maximum concentrations of PHB achieved were 22.35 and 23.07 g/l at those dilution rates when sugar concentration maintains at 200 g/l in feeding. At maximum concentration of PHB (23.07 g/l), productivity of 0.58 g/l h was achieved with maximum PHB accumulation efficiency up to 64% of the dry weight of biomass. High purity of PHB, close to medical grade was achieved after surfactant hypochlorite extraction method, and it was further confirmed by SEM, EDX, and XRD studies.

Waste-Activated Sludge Fermentation for Polyacrylamide Biodegradation Improved by Anaerobic Hydrolysis and Key Microorganisms Involved in Biological Polyacrylamide Removal

PubMed Central

Dai, Xiaohu; Luo, Fan; Zhang, Dong; Dai, Lingling; Chen, Yinguang; Dong, Bin

2015-01-01

During the anaerobic digestion of dewatered sludge, polyacrylamide (PAM), a chemical conditioner, can usually be consumed as a carbon and nitrogen source along with other organic matter (e.g., proteins and carbohydrates in the sludge). However, a significant accumulation of acrylamide monomers (AMs) was observed during the PAM biodegradation process. To improve the anaerobic hydrolysis of PAM, especially the amide hydrolysis process, and to avoid the generation of the intermediate product AM, a new strategy is reported herein that uses an initial pH of 9, 200 mg COD/L of PAM and a fermentation time of 17 d. First, response surface methodology (RSM) was applied to optimize PAM removal in the anaerobic digestion of the sludge. The biological hydrolysis of PAM reached 86.64% under the optimal conditions obtained from the RSM. Then, the mechanisms for the optimized parameters that significantly improved the biological hydrolysis of PAM were investigated by the synergistic effect of the main organic compounds in the sludge, the floc size distribution, and the enzymatic activities. Finally, semi-continuous-flow experiments for a microbial community study were investigated based on the determination of key microorganisms involved in the biological hydrolysis of PAM. PMID:26144551

Improvement of Vitamin K2 Production by Escherichia sp. with Nitrogen Ion Beam Implantation Induction

NASA Astrophysics Data System (ADS)

Liu, Yan; Wang, Li; Zheng, Zhiming; Wang, Peng; Zhao, Genhai; Liu, Hui; Gong, Guohong; Wu, Hefang; Liu, Hongxia; Tan, Mu; Li, Zhemin

2015-02-01

Low-energy ion implantation as a novel mutagen has been increasingly applied in the microbial mutagenesis for its higher mutation frequency and wider mutation spectra. In this work, N+ ion beam implantation was used to enhance Escherichia sp. in vitamin K2 yield. Optimization of process parameters under submerged fermentation was carried out to improve the vitamin K2 yield of mutant FM5-632. The results indicate that an excellent mutant FM5-632 with a yield of 123.2±1.6 μg/L, that is four times that of the original strain, was achieved by eight successive implantations under the conditions of 15 keV and 60×2.6×1013 ions/cm2. A further optimization increased the yield of the mutant by 39.7%, i.e. 172.1±1.2 μg/L which occurred in the mutant cultivated in the optimal fermentation culture medium composed of (per liter): 15.31 g glycerol, 10 g peptone, 2.89 g yeast extract, 5 g K2HPO4, 1 g NaCl, 0.5 g MgSO4·7H2O and 0.04 g cedar wood oil, incubated at 33 °C, pH 7.0 and 180 rpm for 120 h.

Performance comparison of ethanol and butanol production in a continuous and closed-circulating fermentation system with membrane bioreactor.

PubMed

Chen, Chunyan; Long, Sihua; Li, Airong; Xiao, Guoqing; Wang, Linyuan; Xiao, Zeyi

2017-03-16

Since both ethanol and butanol fermentations are urgently developed processes with the biofuel-demand increasing, performance comparison of aerobic ethanol fermentation and anerobic butanol fermentation in a continuous and closed-circulating fermentation (CCCF) system was necessary to achieve their fermentation characteristics and further optimize the fermentation process. Fermentation and pervaporation parameters including the average cell concentration, glucose consumption rate, cumulated production concentration, product flux, and separation factor of ethanol fermentation were 11.45 g/L, 3.70 g/L/h, 655.83 g/L, 378.5 g/m 2 /h, and 4.83, respectively, the corresponding parameters of butanol fermentation were 2.19 g/L, 0.61 g/L/h, 28.03 g/L, 58.56 g/m 2 /h, and 10.62, respectively. Profiles of fermentation and pervaporation parameters indicated that the intensity and efficiency of ethanol fermentation was higher than butanol fermentation, but the stability of butanol fermentation was superior to ethanol fermentation. Although the two fermentation processes had different features, the performance indicated the application prospect of both ethanol and butanol production by the CCCF system.

Optimization of L-asparaginase production from novel Enterobacter sp., by submerged fermentation using response surface methodology.

PubMed

Erva, Rajeswara Reddy; Goswami, Ajgebi Nath; Suman, Priyanka; Vedanabhatla, Ravali; Rajulapati, Satish Babu

2017-03-16

The culture conditions and nutritional rations influencing the production of extra cellular antileukemic enzyme by novel Enterobacter aerogenes KCTC2190/MTCC111 were optimized in shake-flask culture. Process variables like pH, temperature, incubation time, carbon and nitrogen sources, inducer concentration, and inoculum size were taken into account. In the present study, finest enzyme activity achieved by traditional one variable at a time method was 7.6 IU/mL which was a 2.6-fold increase compared to the initial value. Further, the L-asparaginase production was optimized using response surface methodology, and validated experimental result at optimized process variables gave 18.35 IU/mL of L-asparaginase activity, which is 2.4-times higher than the traditional optimization approach. The study explored the E. aerogenes MTCC111 as a potent and potential bacterial source for high yield of antileukemic drug.

Isolation and Characterization of Brewer's Yeast Variants with Improved Fermentation Performance under High-Gravity Conditions▿

PubMed Central

Blieck, Lies; Toye, Geert; Dumortier, Françoise; Verstrepen, Kevin J.; Delvaux, Freddy R.; Thevelein, Johan M.; Van Dijck, Patrick

2007-01-01

To save energy, space, and time, today's breweries make use of high-gravity brewing in which concentrated medium (wort) is fermented, resulting in a product with higher ethanol content. After fermentation, the product is diluted to obtain beer with the desired alcohol content. While economically desirable, the use of wort with an even higher sugar concentration is limited by the inability of brewer's yeast (Saccharomyces pastorianus) to efficiently ferment such concentrated medium. Here, we describe a successful strategy to obtain yeast variants with significantly improved fermentation capacity under high-gravity conditions. We isolated better-performing variants of the industrial lager strain CMBS33 by subjecting a pool of UV-induced variants to consecutive rounds of fermentation in very-high-gravity wort (>22° Plato). Two variants (GT336 and GT344) showing faster fermentation rates and/or more-complete attenuation as well as improved viability under high ethanol conditions were identified. The variants displayed the same advantages in a pilot-scale stirred fermenter under high-gravity conditions at 11°C. Microarray analysis identified several genes whose altered expression may be responsible for the superior performance of the variants. The role of some of these candidate genes was confirmed by genetic transformation. Our study shows that proper selection conditions allow the isolation of variants of commercial brewer's yeast with superior fermentation characteristics. Moreover, it is the first study to identify genes that affect fermentation performance under high-gravity conditions. The results are of interest to the beer and bioethanol industries, where the use of more-concentrated medium is economically advantageous. PMID:17158628

Kinetics of growth and sugar consumption in yeasts.

PubMed

van Dijken, J P; Weusthuis, R A; Pronk, J T

1993-01-01

An overview is presented of the steady- and transient state kinetics of growth and formation of metabolic byproducts in yeasts. Saccharomyces cerevisiae is strongly inclined to perform alcoholic fermentation. Even under fully aerobic conditions, ethanol is produced by this yeast when sugars are present in excess. This so-called 'Crabtree effect' probably results from a multiplicity of factors, including the mode of sugar transport and the regulation of enzyme activities involved in respiration and alcoholic fermentation. The Crabtree effect in S. cerevisiae is not caused by an intrinsic inability to adjust its respiratory activity to high glycolytic fluxes. Under certain cultivation conditions, for example during growth in the presence of weak organic acids, very high respiration rates can be achieved by this yeast. S. cerevisiae is an exceptional yeast since, in contrast to most other species that are able to perform alcoholic fermentation, it can grow under strictly anaerobic conditions. 'Non-Saccharomyces' yeasts require a growth-limiting supply of oxygen (i.e. oxygen-limited growth conditions) to trigger alcoholic fermentation. However, complete absence of oxygen results in cessation of growth and therefore, ultimately, of alcoholic fermentation. Since it is very difficult to reproducibly achieve the right oxygen dosage in large-scale fermentations, non-Saccharomyces yeasts are therefore not suitable for large-scale alcoholic fermentation of sugar-containing waste streams. In these yeasts, alcoholic fermentation is also dependent on the type of sugar. For example, the facultatively fermentative yeast Candida utilis does not ferment maltose, not even under oxygen-limited growth conditions, although this disaccharide supports rapid oxidative growth.

β-(1→6)-D-glucan secreted during the optimised production of exopolysaccharides by Paecilomyces variotii has immunostimulatory activity.

PubMed

Osaku, Erica F; Menolli, Rafael A; de M Costa, Claudia R L; Tessaro, Fernando Henrique G; de Melo, Renan H; do Amaral, Alex E; Duarte, Péricles A D; de Santana Filho, Arquimedes Paixão; Ruthes, Andrea Caroline; da C Silva, José Luis; Mello, Rosiane G

2018-06-01

Paecilomyces variotii is a filamentous fungus that occurs worldwide in soil and decaying vegetation. Optimization of the fermentation process for exopolysaccharide (EPS) production from the fungus P. variotii, structure determination and immuno-stimulating activity of EPS were performed. Response surface methodology (RSM) coupled with central composite design (CCD) was used to optimize the physical and chemical factors required to produce EPS in submerged fermentation. Preliminary investigations to choose the three factors for the present work were made using a factorial experimental design. Glucose, ammonium nitrate (NH 4 NO 3 ) and pH were used as variables for which, with constant temperature of 28 °C and agitation of 90 rpm, the optimal process parameters were determined as glucose values of 0.96%, NH 4 NO 3 0.26% and pH 8.0. The three parameters presented significant effects. In this condition of culture, the main composition of the isolated EPS was a linear β-(1 → 6)-linked-D-glucan, as determined by Nuclear Magnetic Resonance (NMR) and methylation analysis. This polysaccharide is a very unusual as an EPS from fungi, especially a filamentous fungus such as P. variotii. Murine peritoneal macrophages cultivated with β-glucan for 6 and 48 h showed an increase in TNF-α, IL-6 and nitric oxide release with increased polysaccharide concentrations. Therefore, we conclude that the β-(1 → 6)-linked-D-glucan produced in optimised conditions of P. variotii cultivation has an immune-stimulatory activity on murine macrophages.

Optimized production of Serratia marcescens B742 mutants for preparing chitin from shrimp shells powders.

PubMed

Zhang, Hongcai; Fang, Jiyang; Deng, Yun; Zhao, Yanyun

2014-08-01

To improve the deproteinization (DP) efficacy of shrimp shell powders (SSP) for preparing chitin, Serratia marcescens B742 mutants were prepared using 2% diethyl sulfate (DES), UV-irradiation, and/or microwave heating treatments. Both single-stage and multi-stage mutations were investigated for optimizing S. marcescens B742 mutation conditions. Under the optimized mutation conditions (2% DES treatment for 30min plus successive 20min UV-irradiation), the protease and chitosanase activity produced by mutant S. marcescens B742 was 240.15 and 170.6mU/mL, respectively, as compared with 212.58±1.51 and 83.75±6.51mU/mL, respectively, by wild S. marcescens B742. DP efficacy of SSP by mutant S. marcescens B742 reached 91.4±4.6% after 3d of submerged fermentation instead of 83.4±4.7% from the wild S. marcescens B742 after 4d of submerged fermentation. Molecular mass of chitosanase and protease was 41.20 and 47.10kDa, respectively, and both enzymes were verified by mass spectrometry analysis. The chitosanase from both wild and mutant S. marcescens B742 was activated by sodium dodecyl sulfate (SDS), Tween 20, Tween 40, and Triton-100, and the protease and chitosanase were strongly inhibited by ethylenediaminetetraacetic acid (EDTA). These results suggested that S. marcescens B742 mutants can be used in the biological production of chitin through deproteinization of SSP. Copyright © 2014 Elsevier B.V. All rights reserved.

Fed batch fermentation and purification strategy for high yield production of Brucella melitensis recombinant Omp 28 kDa protein and its application in disease diagnosis.

PubMed

Karothia, B S; Athmaram, T N; D, Thavaselvam; Ashu, Kumar; Tiwari, Sapna; Singh, Anil K; Sathyaseelan, K; Gopalan, N

2013-07-01

Brucellosis is a disease caused by bacteria belonging to the genus Brucella. It affects cattle, goat, sheep, dog and humans. The serodiagnosis of brucellosis involves detection of antibodies generated against the LPS or whole cell bacterial extracts, however these tests lack sensitivity and specificity. The present study was performed to optimize the culture condition for the production of recombinant Brucella melitensis outer membrane protein 28 kDa protein in E.coli via fed batch fermentation. Expression was induced with 1.5mM isopropyl β thiogalactoside and the expressed recombinant protein was purified using Ni-NTA affinity chromatography. After fed-batch fermentation the dry cell weight of 17.81 g/L and a purified protein yield of 210.10 mg/L was obtained. The purified Brucella melitensis recombinant Omp 28 kDa protein was analyzed through SDS- poly acrylamide gel electrophoresis and western blotting. The obtained recombinant protein was evaluated for its diagnostic application through Indirect ELISA using brucellosis suspected human sera samples. Our results clearly indicate that recombinant Omp28 produced via fed batch fermentation has immense potential as a diagnostic reagent that could be employed in sero monitoring of brucellosis.

Acid hydrolysis of Jerusalem artichoke for ethanol fermentation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kim, K.; Hamdy, M.K.

1986-01-01

An excellent substrate for ethanol production is the Jerusalem artichoke (JA) tuber (Helianthus tuberosus). This crop contains a high level of inulin that can be hydrolyzed mainly to D-fructose and has several distinct advantages as an energy source compared to others. The potential ethanol yield of ca. 4678 L/ha on good agricultural land is equivalent to that obtained from sugar beets and twice that of corn. When JA is to be used for ethanol fermentation by conventional yeast, it is first converted to fermentable sugars by enzymes or acids although various strains of yeast were used for the direct fermentationmore » of JA extracts. Fleming and GrootWassink compared various acids (hydrochloric, sulfuric, citric, and phosphoric) and strong cation exchange resin for their effectiveness on inulin hydrolysis and reported that no differences were noted among the acids or resin in their influence on inulin hydrolysis. Undesirable side reactions were noted during acid hydrolysis leading to the formation of HMF and 2-(2-hydroxy acetyl) furan. The HMF at a level of 0.1% is known to inhibit growth and ethanol fermentation by yeast. In this study the authors established optimal conditions for complete acid-hydrolysis of JA with minimum side reactions and maximum sugar-ethanol production. A material balance for the ethanol production was also determined.« less

Production of gaba (γ – Aminobutyric acid) by microorganisms: a review

PubMed Central

Dhakal, Radhika; Bajpai, Vivek K.; Baek, Kwang-Hyun

2012-01-01

GABA (γ-aminobutyric acid) is a four carbon non-protein amino acid that is widely distributed in plants, animals and microorganisms. As a metabolic product of plants and microorganisms produced by the decarboxylation of glutamic acid, GABA functions as an inhibitory neurotransmitter in the brain that directly affects the personality and the stress management. A wide range of traditional foods produced by microbial fermentation contain GABA, in which GABA is safe and eco-friendly, and also has the possibility of providing new health-benefited products enriched with GABA. Synthesis of GABA is catalyzed by glutamate decarboxylase, therefore, the optimal fermentation condition is mainly based on the biochemical properties of the enzyme. Major GABA producing microorganisms are lactic acid bacteria (LAB), which make food spoilage pathogens unable to grow and act as probiotics in the gastrointestinal tract. The major factors affecting the production of GABA by microbial fermentation are temperature, pH, fermentation time and different media additives, therefore, these factors are summarized to provide the most up-dated information for effective GABA synthesis. There has been a huge accumulation of knowledge on GABA application for human health accompanying with a demand on natural GABA supply. Only the GABA production by microorganisms can fulfill the demand with GABA-enriched health beneficial foods. PMID:24031948

Production of gaba (γ - Aminobutyric acid) by microorganisms: a review.

PubMed

Dhakal, Radhika; Bajpai, Vivek K; Baek, Kwang-Hyun

2012-10-01

GABA (γ-aminobutyric acid) is a four carbon non-protein amino acid that is widely distributed in plants, animals and microorganisms. As a metabolic product of plants and microorganisms produced by the decarboxylation of glutamic acid, GABA functions as an inhibitory neurotransmitter in the brain that directly affects the personality and the stress management. A wide range of traditional foods produced by microbial fermentation contain GABA, in which GABA is safe and eco-friendly, and also has the possibility of providing new health-benefited products enriched with GABA. Synthesis of GABA is catalyzed by glutamate decarboxylase, therefore, the optimal fermentation condition is mainly based on the biochemical properties of the enzyme. Major GABA producing microorganisms are lactic acid bacteria (LAB), which make food spoilage pathogens unable to grow and act as probiotics in the gastrointestinal tract. The major factors affecting the production of GABA by microbial fermentation are temperature, pH, fermentation time and different media additives, therefore, these factors are summarized to provide the most up-dated information for effective GABA synthesis. There has been a huge accumulation of knowledge on GABA application for human health accompanying with a demand on natural GABA supply. Only the GABA production by microorganisms can fulfill the demand with GABA-enriched health beneficial foods.

Influence of aeration during propagation of pitching yeast on fermentation and beer flavor.

PubMed

Cheong, Chul; Wackerbauer, Karl; Kang, Soon Ah

2007-02-01

The effect of yeast propagated at different aeration conditions on yeast physiology, fermentation ability, and beer quality was investigated using three strains of Saccharomyces cerevisiae. It was shown that yeast cells grown under continuous aeration conditions during propagation were almost two times higher as compared with discontinuous aeration conditions. The maximum of cell growth of all samples reached between 36 h and 48 h. The concentration of trehalose was increased under continuous aerated yeasts, whereas glycogen was decreased. It was also observed that the concentration of glycogen and trehalose in yeast cells had no direct effect on subsequent fermentation ability. The effect of yeast propagated under different aeration conditions on subsequent fermentation ability was different from yeast strains, in which the influence will be most pronounced at the first fermentation. Later, the yeasts might regain its original characteristics in the following fermentations. Generally, continuously propagated yeast had a positive effect on beer quality in subsequent fermentation. Hence, the concentration of aroma compounds obtained with yeast propagated under 6 1/h for 48 h aeration was lower than those grown under other aeration conditions in the bottom yeasts; in particular, the amounts of phenylethyl alcohol, ester, and fatty acids were decreased.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Miller, J.E.; Weathers, P.J.; McConville, F.X.

Apple pomace (the pulp residue from pressing apple juice) is an abundant waste product and presents an expensive disposal problem. A typical (50,000 gal. juice/day) apple juice company in central Massachusetts produces 100 tons of pomace per day. Some of it is used as pig feed, but it is poor quality feed because of its low protein content. Most of the pomace is hauled away (at a cost of $4/ton) and landfilled (at a cost of $10/ton). If 5% (w/w) conversion of pomace to ethanol could be achieved, the need for this company to purchase No. 6 fuel oil (1000more » gal/day) for cooking during processing would be eliminated. Our approach was to saccharify the pomace enzymatically, and then to carry out a yeast fermentation on the hydrolysate. We chose to use enzymatic hydrolysis instead of dilute acid hydrolysis in order to minimize pH control problems both in the fermentation phase and in the residue. The only chemical studies have concerned small subfractions of apple material: for example, cell walls have been analyzed but they constitute only 1 to 2% of the fresh weight of the apple (about 15 to 30% of the pomace fraction). Therefore, our major problems were: (1) to optimize hydrolysis by enzyme mixtures, using weight loss and ultimate ethanol production as optimization criteria; (2) to optimize ethanol production from the hydrolysate by judicious choice of yeast strains and fermentation conditions; and (3) achieve these optimizations consistent with minimum processing cost and energy input. We have obtained up to 5.1% (w/w) of ethanol without saccharification. We show here that hydrolysis with high levels of enzyme can enhance ethanol yield by up to 27%, to a maximum level of 6% (w/w); however, enzyme treament may be cost-effective only a low levels, for improvement of residue compaction. 3 figures, 4 tables.« less

Solid-state fermentation of Jatropha seed cake for optimization of lipase, protease and detoxification of anti-nutrients in Jatropha seed cake using Aspergillus versicolor CJS-98.

PubMed

Veerabhadrappa, Mohankumar Bavimane; Shivakumar, Sharath Belame; Devappa, Somashekar

2014-02-01

This study focused on the solid-state fermentation of Jatropha seed cake (JSC), a byproduct generated after biodiesel production. Presence of anti-nutritional compounds and toxins restricts its application in livestock feed. The disposal of the JSC is a major environmental problem in the future, due to the generation of huge quantity of JSC after biodiesel extraction. Hence the JSC was assessed for its suitability as substrate for production and optimization of lipase and protease from Aspergillus versicolor CJS-98 by solid-state fermentation (SSF). The present study was also focused on the biodetoxification of anti-nutrients and toxins in JSC. The SSF parameters were optimized for maximum production of lipase and protease. Under the optimized conditions, the JSC supplemented with maltose and peptone (2%), adjusted to pH 7.0, moisture content 40%, inoculated with 1 × 10(7) spores per 5 g cake and incubated at 25°C, produced maximum lipase, 1288 U/g and protease, 3366 U/g at 96 h. The anti-nutrients like phytic acid (6.08%), tannins (0.37%), trypsin inhibitors (697.5 TIU/g), cyanogenic glucosides (692.5 μg/100 g), and lectins (0.309 mg/ml), were reduced to 1.70%, 0.23%, 12.5 TIU/g, 560.6 μg/100 g and 0.034 mg/ml respectively. The main toxic compound phorbol esters content in the JSC was reduced from 0.083% to 0.015% after SSF. Our results indicate that viability of SSF to utilize the huge amount of seed cake generated after extraction of biodiesel, for production of industrial enzymes and biodetoxification of anti-nutrients, toxins. Copyright © 2013 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

First report of the production of a potent biosurfactant with α,β-trehalose by Fusarium fujikuroi under optimized conditions of submerged fermentation.

PubMed

Reis, Cristiane Bianchi Loureiro Dos; Morandini, Liziane Maria Barassuol; Bevilacqua, Caroline Borges; Bublitz, Fabricio; Ugalde, Gustavo; Mazutti, Marcio Antonio; Jacques, Rodrigo Josemar Seminoti

2018-04-24

Biosurfactants have many advantages over synthetic surfactants but have higher production costs. Identifying microorganisms with high production capacities for these molecules and optimizing their growth conditions can reduce cost. The present work aimed to isolate and identify a fungus with high biosurfactant production capacity, optimize its growth conditions in a low cost culture medium, and characterize the chemical structure of the biosurfactant molecule. The fungal strain UFSM-BAS-01 was isolated from soil contaminated with hydrocarbons and identified as Fusarium fujikuroi. To optimize biosurfactant production, a Plackett-Burman design and a central composite rotational design were used. The variables evaluated were pH, incubation period, temperature, agitation and amount of inoculum in a liquid medium containing glucose. The partial structure of the biosurfactant molecule was identified by nuclear magnetic resonance spectrometry. F. fujikuroi reduced surface tension from 72 to 20mNm -1 under the optimized conditions of pH 5.0, 37°C and 7 days of incubation with 190rpm agitation. The partial identification of the structure of the biosurfactant demonstrated the presence of an α,β-trehalose. The present study is the first report of the biosynthesis of this compound by F. fujikuroi, suggesting that the biosurfactant produced belongs to the class of trehalolipids. Copyright © 2018 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Optimization and shelf life of a low-lactose yogurt with Lactobacillus rhamnosus HN001.

PubMed

Ibarra, A; Acha, R; Calleja, M-T; Chiralt-Boix, A; Wittig, E

2012-07-01

Lactose intolerance results in gastrointestinal discomfort and the malabsorption of certain nutrients, such as calcium. The replacement of milk with low-lactose and probiotic-enriched dairy products is an effective strategy of mitigating the symptoms of lactose intolerance. Lactobacillus rhamnosus HN001 (HN001) is a safe, immunity-stimulating probiotic. We have developed a process to increase the hydrolysis of lactose and HN001 growth in yogurt versus β-galactosidase (βG) concentration and enzymatic hydrolysis time (EHT) before bacterial fermentation. The objective of this study was to optimize the conditions by which yogurt is processed as a function of βG and EHT using a multifactorial design, with lactose content, HN001 growth, process time, and sensory quality as dependent variables. Further, the shelf life of the optimized yogurt was evaluated. In the optimization study, polynomials explained the dependent variables. Based on Pearson correlation coefficients, HN001 growth correlated positively with the hydrolysis of lactose. However, low lactose content and high HN001 count increased the fermentation time and lowered the sensory quality. The optimized conditions-using polynomials to obtain yogurt with >1 × 10(7) cfu of HN001/mL, <10 g of lactose/L, and a minimum overall sensory quality of 7 on the Karlsruhe scale-yielded a theoretical value of 910 neutral lactose units/kg for βG and 2.3h for EHT, which were validated in an industrial-scale assay. Based on a shelf-life study at 3 temperatures, the hydrolysis of lactose and the growth of HN001 continue during storage. Arrhenius equations were developed for the variables in the shelf-life study. Our results demonstrate that it is feasible to develop a low-lactose yogurt to which HN001 has been added for lactose-intolerant persons who wish to strengthen their immune system. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Bioconversion of renewable resources into lactic acid: an industrial view.

PubMed

Yadav, A K; Chaudhari, A B; Kothari, R M

2011-03-01

Lactic acid, an anaerobic product of glycolysis, can be theoretically produced by synthetic route; however, it is commercially produced by homo-fermentative batch mode of operations. Factors affecting its production and strategies improving it are considered while devising an optimized protocol. Although a hetero-fermentative mode of production exists, it is rarely used for commercial production. Attempts to use Rhizopus sp. for lactic acid production through either hetero-fermentative or thermophilic conditions were not economical. Since almost 70% of the cost of its production is accounted by raw materials, R & D efforts are still focused to find economically attractive agri-products to serve as sources of carbon and complex nitrogen inputs to meet fastidious nutrient needs for microbial growth and lactic acid production. Therefore, need exists for using multi-pronged strategies for higher productivity. Its present production and consumption scenario is examined. Its optically active isomers and chemical structure permit its use for the production of several industrially important chemicals, health products (probiotics), food preservatives, and bio-plastics. In addition, its salts and esters appear to have a variety of applications.

Ion-exclusion chromatography determination of organic acid in uridine 5'-monophosphate fermentation broth.

PubMed

Niu, Huanqing; Chen, Yong; Xie, Jingjing; Chen, Xiaochun; Bai, Jianxin; Wu, Jinglan; Liu, Dong; Ying, Hanjie

2012-09-01

Simultaneous determination of organic acids using ion-exclusion liquid chromatography and ultraviolet detection is described. The chromatographic conditions are optimized when an Aminex HPX-87H column (300 × 7.8 mm) is employed, with a solution of 3 mmol/L sulfuric acid as eluent, a flow rate of 0.4 mL/min and a column temperature of 60°C. Eight organic acids (including orotic acid, α-ketoglutaric acid, citric acid, pyruvic acid, malic acid, succinic acid, lactic acid and acetic acid) and one nucleotide are successfully quantified. The calibration curves for these analytes are linear, with correlation coefficients exceeding 0.999. The average recovery of organic acids is in the range of 97.6% ∼ 103.1%, and the relative standard deviation is in the range of 0.037% ∼ 0.38%. The method is subsequently applied to obtain organic acid profiles of uridine 5'-monophosphate culture broth fermented from orotic acid by Saccharomyces cerevisiae. These data demonstrate the quantitative accuracy for nucleotide fermentation mixtures, and suggest that the method may also be applicable to other biological samples.

Co-production of bio-ethanol, xylonic acid and slow-release nitrogen fertilizer from low-cost straw pulping solid residue.

PubMed

Huang, Chen; Ragauskas, Arthur J; Wu, Xinxing; Huang, Yang; Zhou, Xuelian; He, Juan; Huang, Caoxing; Lai, Chenhuan; Li, Xin; Yong, Qiang

2018-02-01

A novel bio-refinery sequence yielding varieties of co-products was developed using straw pulping solid residue. This process utilizes neutral sulfite pretreatment which under optimal conditions (160 °C and 3% (w/v) sulfite charge) provides 64.3% delignification while retaining 90% of cellulose and 67.3% of xylan. The pretreated solids exhibited excellent enzymatic digestibility, with saccharification yields of 86.9% and 81.1% for cellulose and xylan, respectively. After pretreatment, the process of semi-simultaneous saccharification and fermentation (S-SSF) and bio-catalysis was investigated. The results revealed that decreased ethanol yields were achieved when solid loading increased from 5% to 30%. An acceptable ethanol yield of 76.8% was obtained at 20% solid loading. After fermentation, bio-catalysis of xylose remaining in fermentation broth resulted in near 100% xylonic acid (XA) yield at varied solid loadings. To complete the co-product portfolio, oxidation ammoniation of the dissolved lignin successfully transformed it into biodegradable slow-release nitrogen fertilizer with excellent agricultural properties. Copyright © 2017 Elsevier Ltd. All rights reserved.

Direct microbial conversion of wheat straw into lipid by a cellulolytic fungus of Aspergillus oryzae A-4 in solid-state fermentation.

PubMed

Lin, Hui; Cheng, Wan; Ding, Hai-tao; Chen, Xue-jiao; Zhou, Qi-fa; Zhao, Yu-hua

2010-10-01

Direct microbial conversion of wheat straw into lipid by a cellulolytic fungus of Aspergillus oryzae A-4 in solid-state fermentation (SSF) was investigated. In submerged fermentation, A. oryzae A-4 accumulated lipid to 15-18.15% of biomass when pure cellulose was utilized as the sole substrate. In SSF of the wheat straw and bran mixture, A. oryzae A-4 yielded lipid of 36.6mg/g dry substrate (gds), and a cellulase activity of 1.82 FPU/gds with 25.25% of holocellulose utilization in the substrates were detected on the 6th day. The lipid yield reached 62.87 mg/gds in SSF on the 6th day under the optimized conditions from Plackett-Burman design (PBD). Cellulase secretion of A. oryzae A-4 was found to influence the lipid yield. Dilute acid pretreatment of the straw and addition of some agro-industrial wastes to the straw could enhance lipid production of A. oryzae A-4. Copyright 2010 Elsevier Ltd. All rights reserved.

D-Lactic acid production by Sporolactobacillus inulinus YBS1-5 with simultaneous utilization of cottonseed meal and corncob residue.

PubMed

Bai, Zhongzhong; Gao, Zhen; Sun, Junfei; Wu, Bin; He, Bingfang

2016-05-01

d-Lactic acid, is an important organic acid produced from agro-industrial wastes by Sporolactobacillus inulinus YBS1-5 was investigated to reduce the raw material cost of fermentation. The YBS1-5 strain could produce d-lactic acid by using cottonseed meal as the sole nitrogen source. For efficient utilization, the cottonseed meal was enzymatically hydrolyzed and simultaneously utilized during d-lactic acid fermentation. Corncob residues are rich in cellulose and can be enzymatically hydrolyzed without pretreatment. The hydrolysate of this lignocellulosic waste could be utilized by strain YBS1-5 as a carbon source for d-lactic acid production. Under optimal conditions, a high d-lactic acid concentration (107.2g/L) was obtained in 7-L fed-batch fermenter, with an average productivity of 1.19g/L/h and a yield of 0.85g/g glucose. The optical purity of d-lactic acid in the broth was 99.2%. This study presented a new approach for low-cost production of d-lactic acid for an industrial application. Copyright © 2016 Elsevier Ltd. All rights reserved.

Biohydrogen from thermophilic co-fermentation of swine manure with fruit and vegetable waste: maximizing stable production without pH control.

PubMed

Tenca, A; Schievano, A; Perazzolo, F; Adani, F; Oberti, R

2011-09-01

Hydrogen production by dark fermentation may suffer of inhibition or instability due to pH deviations from optimality. The co-fermentation of promptly degradable feedstock with alkali-rich materials, such as livestock wastes, may represent a feasible and easy to implement approach to avoid external adjustments of pH. Experiments were designed to investigate the effect of the mixing ratio of fruit-vegetable waste with swine manure with the aim of maximizing biohydrogen production while obtaining process stability through the endogenous alkalinity of manure. Fruit-vegetable/swine manure ratio of 35/65 and HRT of 2d resulted to give the highest production rate of 3.27 ± 0.51 L(H2)L(-1)d(-1), with a corresponding hydrogen yield of 126 ± 22 mL(H2)g(-1)(VS-added) and H2 content in the biogas of 42 ± 5%. At these operating conditions the process exhibited also one of the highest measured stability, with daily productions deviating for less than 14% from the average. Copyright © 2011 Elsevier Ltd. All rights reserved.

High-level pullulan production by Aureobasidium pullulans var. melanogenium P16 isolated from mangrove system.

PubMed

Ma, Zai-Chao; Fu, Wen-Juan; Liu, Guang-Lei; Wang, Zhi-Peng; Chi, Zhen-Ming

2014-06-01

After over 100 strains of Aureobasidium spp. isolated from mangrove system were screened for their ability to produce exopolysaccharide (EPS), it was found that Aureobasidium pullulans var. melanogenium P16 strain among them could produce high level of EPS. Under the optimal conditions, 65.3 g/L EPS was produced by the P16 strain within 120 h at flask level. During 10-L batch fermentation, when the medium contained 120.0 g/L sucrose, 67.4 g/L of EPS and 23.1 g/L of cell dry weight in the culture were obtained within 120 h, leaving 0.78 g/L of reducing sugar and 11.4 g/L of total sugar in the fermented medium. It should be stressed that during the fermentation, no melanin was observed. After purification, the purified EPS was confirmed to be pullulan. This is the first time to report that A. pullulans var. melanogenium P16 strain isolated from the mangrove system can produce high level of pullulan.

Simultaneous enrichment of cereals with polyunsaturated fatty acids and pigments by fungal solid state fermentations.

PubMed

Čertík, Milan; Adamechová, Zuzana; Guothová, Lucia

2013-10-20

Four Mucor strains were tested for their ability to grow on four cereal substrates and enriched them with gamma-linolenic acid (GLA) and β-carotene. M. circinelloides CCF-2617 as the best producer accumulated of both GLA and β-carotene in high amounts during utilization of rye bran/spent malt grains (3:1). The first growth phase was characterized by rapid GLA biosynthesis, while distinct β-carotene formation was found in the stationary fungal growth. Therefore various cultivation conditions were tested in order to optimize the yield of either GLA or β-carotene. The fungus grown on cereal substrate supplemented with glucose produced maximal 8.5 mg β-carotene and 12.1 g GLA in 1 kg fermented substrate, respectively. On the other hand, the highest amount of GLA in the fermented substrate (24.2 g/kg) was achieved when 30% of sunflower oil was employed to the substrate. Interestingly, β-carotene biosynthesis was completely inhibited when either whey or linseed oil were added to the substrate. Copyright © 2013 Elsevier B.V. All rights reserved.

Evaluation and simultaneous optimization of bio-hydrogen production using 3 2 factorial design and the desirability function

NASA Astrophysics Data System (ADS)

Cuetos, M. J.; Gómez, X.; Escapa, A.; Morán, A.

Various mixtures incorporating a simulated organic fraction of municipal solid wastes and blood from a poultry slaughterhouse were used as substrate in a dark fermentation process for the production of hydrogen. The individual and interactive effects of hydraulic retention time (HRT), solid content in the feed (%TS) and proportion of residues (%Blood) on bio-hydrogen production were studied in this work. A central composite design and response surface methodology were employed to determine the optimum conditions for the hydrogen production process. Experimental results were approximated to a second-order model with the principal effects of the three factors considered being statistically significant (P < 0.05). The production of hydrogen obtained from the experimental point at conditions close to best operability was 0.97 L Lr -1 day -1. Moreover, a desirability function was employed in order to optimize the process when a second, methanogenic, phase is coupled with it. In this last case, the optimum conditions lead to a reduction in the production of hydrogen when the optimization process involves the maximization of intermediary products.

Influence of nitrogen sources on growth and fermentation performance of different wine yeast species during alcoholic fermentation.

PubMed

Kemsawasd, Varongsiri; Viana, Tiago; Ardö, Ylva; Arneborg, Nils

2015-12-01

In this study, the influence of twenty different single (i.e. 19 amino acids and ammonium sulphate) and two multiple nitrogen sources (N-sources) on growth and fermentation (i.e. glucose consumption and ethanol production) performance of Saccharomyces cerevisiae and of four wine-related non-Saccharomyces yeast species (Lachancea thermotolerans, Metschnikowia pulcherrima, Hanseniaspora uvarum and Torulaspora delbrueckii) was investigated during alcoholic fermentation. Briefly, the N-sources with beneficial effects on all performance parameters (or for the majority of them) for each yeast species were alanine, arginine, asparagine, aspartic acid, glutamine, isoleucine, ammonium sulphate, serine, valine and mixtures of 19 amino acids and of 19 amino acids plus ammonium sulphate (for S. cerevisiae), serine (for L. thermotolerans), alanine (for H. uvarum), alanine and asparagine (for M. pulcherrima), arginine, asparagine, glutamine, isoleucine and mixture of 19 amino acids (for T. delbrueckii). Furthermore, our results showed a clear positive effect of complex mixtures of N-sources on S. cerevisiae and on T. delbrueckii (although to a lesser extent) as to all performance parameters studied, whereas for L. thermotolerans, H. uvarum and M. pulcherrima, single amino acids affected growth and fermentation performance to the same extent as the mixtures. Moreover, we found groups of N-sources with similar effects on the growth and/or fermentation performance of two or more yeast species. Finally, the influences of N-sources observed for T. delbrueckii and H. uvarum resembled those of S. cerevisiae the most and the least, respectively. Overall, this work contributes to an improved understanding of how different N-sources affect growth, glucose consumption and ethanol production of wine-related yeast species under oxygen-limited conditions, which, in turn, may be used to, e.g. optimize growth and fermentation performance of the given yeast upon N-source supplementation during wine fermentations.

Novel optimization strategy for tannase production through a modified solid-state fermentation system.

PubMed

Wu, Changzheng; Zhang, Feng; Li, Lijun; Jiang, Zhedong; Ni, Hui; Xiao, Anfeng

2018-01-01

High amounts of insoluble substrates exist in the traditional solid-state fermentation (SSF) system. The presence of these substrates complicates the determination of microbial biomass. Thus, enzyme activity is used as the sole index for the optimization of the traditional SSF system, and the relationship between microbial growth and enzyme synthesis is always ignored. This study was conducted to address this deficiency. All soluble nutrients from tea stalk were extracted using water. The aqueous extract was then mixed with polyurethane sponge to establish a modified SSF system, which was then used to conduct tannase production. With this system, biomass, enzyme activity, and enzyme productivity could be measured rationally and accurately. Thus, the association between biomass and enzyme activity could be easily identified, and the shortcomings of traditional SSF could be addressed. Different carbon and nitrogen sources exerted different effects on microbial growth and enzyme production. Single-factor experiments showed that glucose and yeast extract greatly improved microbial biomass accumulation and that tannin and (NH 4 ) 2 SO 4 efficiently promoted enzyme productivity. Then, these four factors were optimized through response surface methodology. Tannase activity reached 19.22 U/gds when the added amounts of tannin, glucose, (NH 4 ) 2 SO 4 , and yeast extract were 7.49, 8.11, 9.26, and 2.25%, respectively. Tannase activity under the optimized process conditions was 6.36 times higher than that under the initial process conditions. The optimized parameters were directly applied to the traditional tea stalk SSF system. Tannase activity reached 245 U/gds, which is 2.9 times higher than our previously reported value. In this study, a modified SSF system was established to address the shortcomings of the traditional SSF system. Analysis revealed that enzymatic activity and microbial biomass are closely related, and different carbon and nitrogen sources have different effects on microbial growth and enzyme production. The maximal tannase activity was obtained under the optimal combination of nutrient sources that enhances cell growth and tannase accumulation. Moreover, tannase production through the traditional tea stalk SSF was markedly improved when the optimized parameters were applied. This work provides an innovative approach to bioproduction research through SSF.

Characteristics and optimized fermentation of a novel magnetotactic bacterium, Magnetospirillum sp. ME-1.

PubMed

Ke, Linfeng; Chen, Yajun; Liu, Pengming; Liu, Shan; Wu, Dandan; Yuan, Yihui; Wu, Yan; Gao, Meiying

2018-03-04

Magnetotactic bacteria (MTB) can biosynthesize magnetosomes, which have great potential for applications. A new MTB strain, Magnetospirillum sp. ME-1, was isolated and cultivated from freshwater sediments of East Lake (Wuhan, China) using the limiting dilution method. ME-1 had a chain of 17 ± 4 magnetosomes in the form of cubooctahedral crystals with a shape factor of 0.89. ME-1 was closest to Magnetospirillum sp. XM-1 according to 16S rRNA gene sequence similarity. Compared with XM-1, ME-1 possessed additional copy of mamPA and a larger mamO in magnetosome-specific genes. ME-1 had an intact citric acid cycle, and complete pathway models of ammonium assimilation and dissimilatory nitrate reduction. Potential carbon and nitrogen sources in these pathways were confirmed to be used in ME-1. Adipate was determined to be used in the fermentation medium as a new kind of dicarboxylic acid. The optimized fermentation medium was determined by orthogonal tests. The large-scale production of magnetosomes was achieved and the magnetosome yield (wet weight) reached 120 mg/L by fed-batch cultivation of ME-1 at 49 h in a 10-L fermenter with the optimized fermentation medium. This study may provide insights into the isolation and cultivation of other new MTB strains and the production of magnetosomes.

East Europe Report. Scientific Affairs, No. 785.

DTIC Science & Technology

1983-08-11

in close cooperation with the Institute of Microbiology, is developing an ASU for fermentation processes. Contracts have already been entered into...and such sys- tems are being devised for the plants in Peshtera and in Razgrad. Associated with fermentation processes is the technology for...experimental fermentation system is already in operation in Vidin and methane is being produced. But a number of problems in optimizing these processes with

Role of a membrane-bound aldehyde dehydrogenase complex AldFGH in acetic acid fermentation with Acetobacter pasteurianus SKU1108.

PubMed

Yakushi, Toshiharu; Fukunari, Seiya; Kodama, Tomohiro; Matsutani, Minenosuke; Nina, Shun; Kataoka, Naoya; Theeragool, Gunjana; Matsushita, Kazunobu

2018-05-01

Acetic acid fermentation is widely considered a consequence of ethanol oxidation by two membrane-bound enzymes-alcohol dehydrogenase and aldehyde dehydrogenase (ALDH)-of acetic acid bacteria. Here, we used a markerless gene disruption method to construct a mutant of the Acetobacter pasteurianus strain SKU1108 with a deletion in the aldH gene, which encodes the large catalytic subunit of a heterotrimeric ALDH complex (AldFGH), to examine the role of AldFGH in acetic acid fermentation. The ΔaldH strain grew less on ethanol-containing medium, i.e., acetic acid fermentation conditions, than the wild-type strain and significantly accumulated acetaldehyde in the culture medium. Unexpectedly, acetaldehyde oxidase activity levels of the intact ΔaldH cells and the ΔaldH cell membranes were similar to those of the wild-type strain, which might be attributed to an additional ALDH isozyme (AldSLC). The apparent K M values of the wild-type and ΔaldH membranes for acetaldehyde were similar to each other, when the cells were cultured in nonfermentation conditions, where ΔaldH cells grow as well as the wild-type cells. However, the membranes of the wild-type cells grown under fermentation conditions showed a 10-fold lower apparent K M value than those of the cells grown under nonfermentation conditions. Under fermentation conditions, transcriptional levels of a gene for AldSLC were 10-fold lower than those under nonfermentation conditions, whereas aldH transcript levels were not dramatically changed under the two conditions. We suggest that A. pasteurianus SKU1108 has two ALDHs, and the AldFGH complex is indispensable for acetic acid fermentation and is the major enzyme under fermentation conditions.

Preparation of stable isotope-labeled peripheral cannabinoid receptor CB2 by bacterial fermentation

PubMed Central

Berger, Christian; Ho, Jenny T.C.; Kimura, Tomohiro; Hess, Sonja; Gawrisch, Klaus; Yeliseev, Alexei

2010-01-01

We developed a bacterial fermentation protocol for production of a stable isotope-labeled cannabinoid receptor CB2 for subsequent structural studies of this protein by nuclear magnetic resonance spectroscopy. The human peripheral cannabinoid receptor was expressed in Escherichia coli as a fusion with maltose binding protein and two affinity tags. The fermentation was performed in defined media comprised of mineral salts, glucose and 15N2-L-tryptophan to afford incorporation of the labeled amino acid into the protein. Medium, growth and expression conditions were optimized so that the fermentation process produced about 2 mg of purified, labeled CB2 per liter of culture medium. By performing a mass spectroscopic characterization of the purified CB2, we determined that one of the two 15N atoms in tryptophan was incorporated into the recombinant protein. NMR analysis of 15N chemical shifts strongly suggests that the 15N atoms are located in Trp-indole rings. Importantly, analysis of the peptides derived from the CNBr cleavage of the purified protein confirmed a minimum of 95% incorporation of the labeled tryptophan into the CB2 sequence. The labeled CB2, purified and reconstituted into liposomes at a protein-to-lipid molar ratio of 1:500, was functional as confirmed by activation of cognate G proteins in an in vitro coupled assay. To our knowledge, this is the first reported production of a biologically active, stable isotope-labeled G protein-coupled receptor by bacterial fermentation. PMID:20044006

Reconstruction of a metabolic regulatory network in Escherichia coli for purposeful switching from cell growth mode to production mode in direct GABA fermentation from glucose.

PubMed

Soma, Yuki; Fujiwara, Yuri; Nakagawa, Takuya; Tsuruno, Keigo; Hanai, Taizo

2017-09-01

γ-aminobutyric acid (GABA) is a drug and functional food additive and is used as a monomer for producing the biodegradable plastic, polyamide 4. Recently, direct GABA fermentation from glucose has been developed as an alternative to glutamate-based whole cell bioconversion. Although total productivity in fermentation is determined by the specific productivity and cell amount responsible for GABA production, the optimal metabolic state for GABA production conflicts with that for bacterial cell growth. Herein, we demonstrated metabolic state switching from the cell growth mode based on the metabolic pathways of the wild type strain to a GABA production mode based on a synthetic metabolic pathway in Escherichia coli through rewriting of the metabolic regulatory network and pathway engineering. The GABA production mode was achieved by multiple strategies such as conditional interruption of the TCA and glyoxylate cycles, engineering of GABA production pathway including a bypass for precursor metabolite supply, and upregulation of GABA transporter. As a result, we achieved 3-fold improvement in total GABA production titer and yield (4.8g/L, 49.2% (mol/mol glucose)) in batch fermentation compared to the case without metabolic state switching (1.6g/L, 16.4% (mol/mol glucose)). This study reports the highest GABA production performance among previous reports on GABA fermentation from glucose using engineered E. coli. Copyright © 2017 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

Evaluation of model parameters for growth, tannic acid utilization and tannase production in Bacillus gottheilii M2S2 using polyurethane foam blocks as support.

PubMed

Selvaraj, Subbalaxmi; Vytla, Ramachandra Murty

2017-10-01

Production of tannase from B. gottheilii M2S2 was studied under solid-state fermentation with an optimized medium consisting of polyurethane foam matrix of dimension 40 × 40 × 5 mm, impregnated with a liquid medium comprising (w/v): 4% tannic acid; 2% NH 4 NO 3 ; 0.1% KH 2 PO 4 ; 0.2% MgSO 4 ; 0.1% NaCl and 0.05% CaCl 2 ·2H 2 O in distilled water, having a pH of 4.7. Maximum tannase production of 56.87 U/L was obtained after 32 h of fermentation at 32 °C in static condition. This study deals with the evaluation of unstructured kinetic models to understand the behavior of biomass, tannase production and tannic acid degradation, with the fermentation time. The growth rate of B. gottheilii M2S2 was 0.0703 h -1 at 32 h of fermentation. Product ( Y x/s ) and biomass yield ( Y p/s ) coefficients were estimated as 1.77 U/g of tannic acid and 0.276 g of biomass/g of tannic acid. All the kinetic constants µ , α , β , m and n were evaluated using MATLAB 2015Rb program. The experimental and model-generated data showed a good correlation, which indicated that these models will describe tannase production and fermentation process.

Transcriptomes Reveal Genetic Signatures Underlying Physiological Variations Imposed by Different Fermentation Conditions in Lactobacillus plantarum

PubMed Central

Bongers, Roger S.; van Bokhorst-van de Veen, Hermien; Wiersma, Anne; Overmars, Lex; Marco, Maria L.; Kleerebezem, Michiel

2012-01-01

Lactic acid bacteria (LAB) are utilized widely for the fermentation of foods. In the current post-genomic era, tools have been developed that explore genetic diversity among LAB strains aiming to link these variations to differential phenotypes observed in the strains investigated. However, these genotype-phenotype matching approaches fail to assess the role of conserved genes in the determination of physiological characteristics of cultures by environmental conditions. This manuscript describes a complementary approach in which Lactobacillus plantarum WCFS1 was fermented under a variety of conditions that differ in temperature, pH, as well as NaCl, amino acid, and O2 levels. Samples derived from these fermentations were analyzed by full-genome transcriptomics, paralleled by the assessment of physiological characteristics, e.g., maximum growth rate, yield, and organic acid profiles. A data-storage and -mining suite designated FermDB was constructed and exploited to identify correlations between fermentation conditions and industrially relevant physiological characteristics of L. plantarum, as well as the associated transcriptome signatures. Finally, integration of the specific fermentation variables with the transcriptomes enabled the reconstruction of the gene-regulatory networks involved. The fermentation-genomics platform presented here is a valuable complementary approach to earlier described genotype-phenotype matching strategies which allows the identification of transcriptome signatures underlying physiological variations imposed by different fermentation conditions. PMID:22802930

Production of volatile fatty acids from sewage organic matter by combined bioflocculation and alkaline fermentation.

PubMed

Khiewwijit, Rungnapha; Temmink, Hardy; Labanda, Alvaro; Rijnaarts, Huub; Keesman, Karel J

2015-12-01

This study explored the potential of volatile fatty acids (VFA) production from sewage by a combined high-loaded membrane bioreactor and sequencing batch fermenter. VFA production was optimized with respect to SRT and alkaline pH (pH 8-10). Application of pH shock to a value of 9 at the start of a sequencing batch cycle, followed by a pH uncontrolled phase for 7days, gave the highest VFA yield of 440mgVFA-COD/g VSS. This yield was much higher than at fermentation without pH control or at a constant pH between 8 and 10. The high yield in the pH 9 shocked system could be explained by (1) a reduction of methanogenic activity, or (2) a high degree of solids degradation or (3) an enhanced protein hydrolysis and fermentation. VFA production can be further optimized by fine-tuning pH level and longer operation, possibly allowing enrichment of alkalophilic and alkali-tolerant fermenting microorganisms. Copyright © 2015 Elsevier Ltd. All rights reserved.

Fermentation of Saccharomyces cerevisiae - Combining kinetic modeling and optimization techniques points out avenues to effective process design.

PubMed

Scheiblauer, Johannes; Scheiner, Stefan; Joksch, Martin; Kavsek, Barbara

2018-09-14

A combined experimental/theoretical approach is presented, for improving the predictability of Saccharomyces cerevisiae fermentations. In particular, a mathematical model was developed explicitly taking into account the main mechanisms of the fermentation process, allowing for continuous computation of key process variables, including the biomass concentration and the respiratory quotient (RQ). For model calibration and experimental validation, batch and fed-batch fermentations were carried out. Comparison of the model-predicted biomass concentrations and RQ developments with the corresponding experimentally recorded values shows a remarkably good agreement for both batch and fed-batch processes, confirming the adequacy of the model. Furthermore, sensitivity studies were performed, in order to identify model parameters whose variations have significant effects on the model predictions: our model responds with significant sensitivity to the variations of only six parameters. These studies provide a valuable basis for model reduction, as also demonstrated in this paper. Finally, optimization-based parametric studies demonstrate how our model can be utilized for improving the efficiency of Saccharomyces cerevisiae fermentations. Copyright © 2018 Elsevier Ltd. All rights reserved.

Enhanced candicidal compound production by a new soil isolate Penicillium verruculosum MKH7 under submerged fermentation.

PubMed

Talukdar, Shruti; Talukdar, Madhumita; Buragohain, Manorama; Yadav, Archana; Yadav, R N S; Bora, T C

2016-12-09

Microorganisms are a rich source of structurally diverse secondary metabolites that exert a major impact on the control of infectious diseases and other medical conditions. The biosynthesis of these metabolites can be improved by manipulating the nutritional or environmental factors. This work evaluates the effects of fermentation parameters on the production of a lactone compound effective against Candida albicans by Penicillium verruculosum MKH7 under submerged fermentation. Design-Expert version8.0 software was used for construction of the experimental design and statistical analysis of the experimental data. The important factors influencing antibiotic production selected in accordance with the Plackett-Burman design were found to be initial pH, temperature, peptone, MgSO 4 .7H 2 O. Orthogonal central composite design and response surface methodology were adopted to further investigate the mutual interaction between the variables and identify the optimum values that catalyse maximum metabolite production. The determination coefficient (R 2 ) of the fitted second order model was 0.9852. The validation experiments using optimized conditions of initial pH 7.4, temperature 27 °C, peptone 9.2 g/l and MgSO 4 .7H 2 O 0.39 g/l resulted in a significant increase (almost 7 fold from 30 to 205.5 mg/l) in the metabolite production which was in agreement with the prediction (211.24 mg/l). Stability of the compound was also assessed on the basis of its response to physical and chemical stresses. So far as our knowledge goes, till date there are no reports available on the production of antibiotics by Penicillium verruculosum through media optimization using RSM. Optimization not only led to a 7 fold increase in metabolite yield but the same was achieved at much lesser time (8-10 days compared to the earlier 12-15 days). The enhanced yield of the antibiotic strongly suggests that the fungus P. verruculosum MKH7 can be efficiently used for antibiotic production on a large scale.

Engineering of a Highly Efficient Escherichia coli Strain for Mevalonate Fermentation through Chromosomal Integration

PubMed Central

Wang, Jilong; Niyompanich, Suthamat; Tai, Yi-Shu; Wang, Jingyu; Bai, Wenqin; Mahida, Prithviraj; Gao, Tuo

2016-01-01

ABSTRACT Chromosomal integration of heterologous metabolic pathways is optimal for industrially relevant fermentation, as plasmid-based fermentation causes extra metabolic burden and genetic instabilities. In this work, chromosomal integration was adapted for the production of mevalonate, which can be readily converted into β-methyl-δ-valerolactone, a monomer for the production of mechanically tunable polyesters. The mevalonate pathway, driven by a constitutive promoter, was integrated into the chromosome of Escherichia coli to replace the native fermentation gene adhE or ldhA. The engineered strains (CMEV-1 and CMEV-2) did not require inducer or antibiotic and showed slightly higher maximal productivities (0.38 to ∼0.43 g/liter/h) and yields (67.8 to ∼71.4% of the maximum theoretical yield) than those of the plasmid-based fermentation. Since the glycolysis pathway is the first module for mevalonate synthesis, atpFH deletion was employed to improve the glycolytic rate and the production rate of mevalonate. Shake flask fermentation results showed that the deletion of atpFH in CMEV-1 resulted in a 2.1-fold increase in the maximum productivity. Furthermore, enhancement of the downstream pathway by integrating two copies of the mevalonate pathway genes into the chromosome further improved the mevalonate yield. Finally, our fed-batch fermentation showed that, with deletion of the atpFH and sucA genes and integration of two copies of the mevalonate pathway genes into the chromosome, the engineered strain CMEV-7 exhibited both high maximal productivity (∼1.01 g/liter/h) and high yield (86.1% of the maximum theoretical yield, 30 g/liter mevalonate from 61 g/liter glucose after 48 h in a shake flask). IMPORTANCE Metabolic engineering has succeeded in producing various chemicals. However, few of these chemicals are commercially competitive with the conventional petroleum-derived materials. In this work, chromosomal integration of the heterologous pathway and subsequent optimization strategies ensure stable and efficient (i.e., high-titer, high-yield, and high-productivity) production of mevalonate, which demonstrates the potential for scale-up fermentation. Among the optimization strategies, we demonstrated that enhancement of the glycolytic flux significantly improved the productivity. This result provides an example of how to tune the carbon flux for the optimal production of exogenous chemicals. PMID:27736790

[Optimization on the production of analgesic peptide from Buthus martensii Karsch in Pichia pastoris].

PubMed

Yang, Jin-ling; He, Hui-xia; Zhu, Hui-xin; Cheng, Ke-di; Zhu, Ping

2009-01-01

The technology of liquid fermentation for producing the recombinant analgesic peptide BmK AngM1 from Buthus martensii Karsch in Pichia pastoris was studied by single-factor and orthogonal test. The results showed that the optimal culture conditions were as follows: 1.2% methanol, 0.6% casamino acids, initial pH 6.0, and three times of basal inoculation volume. Under the above culture conditions, the expression level of recombinant BmK AngM1 in Pichia pastoris was above 500 mg x L(-1), which was more than three times of the control. The study has laid a foundation for the large-scale preparation of BmK AngM1 to meet the needs of theoretical research of BmK AngM1 and development of new medicines.

Acidic pH shock induced overproduction of ε-poly-L-lysine in fed-batch fermentation by Streptomyces sp. M-Z18 from agro-industrial by-products.

PubMed

Ren, Xi-Dong; Chen, Xu-Sheng; Zeng, Xin; Wang, Liang; Tang, Lei; Mao, Zhong-Gui

2015-06-01

ε-Poly-L-lysine (ε-PL) is produced by Streptomyces as a secondary metabolite with wide industrial applications, but its production still needs to be further enhanced. Environmental stress is an important approach for the promotion of secondary metabolites production by Streptomyces. In this study, the effect of acidic pH shock on enhancing ε-PL production by Streptomyces sp. M-Z18 was investigated in a 5-L fermenter. Based on the evaluation of acidic pH shock on mycelia metabolic activity and shock parameters optimization, an integrated pH-shock strategy was developed as follows: pre-acid-shock adaption at pH 5.0 to alleviate the damage caused by the followed pH shock, and then acidic pH shock at 3.0 for 12 h (including pH decline from 4.0 to 3.0) to positively regulate mycelia metabolic activity, finally restoring pH to 4.0 to provide optimal condition for ε-PL production. After 192 h of fed-batch fermentation, the maximum ε-PL production and productivity reached 54.70 g/L and 6.84 g/L/day, respectively, which were 52.50 % higher than those of control without pH shock. These results demonstrated that acidic pH shock is an efficient approach for improving ε-PL production. The information obtained should be useful for ε-PL production by other Streptomyces.

Analysis of microbial diversity in Shenqu with different fermentation times by PCR-DGGE.

PubMed

Liu, Tengfei; Jia, Tianzhu; Chen, Jiangning; Liu, Xiaoyu; Zhao, Minjie; Liu, Pengpeng

Shenqu is a fermented product that is widely used in traditional Chinese medicine (TCM) to treat indigestion; however, the microbial strains in the fermentation process are still unknown. The aim of this study was to investigate microbial diversity in Shenqu using different fermentation time periods. DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) profiles indicated that a strain of Pediococcus acidilactici (band 9) is the predominant bacteria during fermentation and that the predominant fungi were uncultured Rhizopus, Aspergillus oryzae, and Rhizopus oryzae. In addition, pathogenic bacteria, such as Enterobacter cloacae, Klebsiella oxytoca, Erwinia billingiae, and Pantoea vagan were detected in Shenqu. DGGE analysis showed that bacterial and fungal diversity declined over the course of fermentation. This determination of the predominant bacterial and fungal strains responsible for fermentation may contribute to further Shenqu research, such as optimization of the fermentation process. Copyright © 2017. Published by Elsevier Editora Ltda.

Optimization of Enzymatic Saccharification of Alkali Pretreated Parthenium sp. Using Response Surface Methodology

PubMed Central

Pandiyan, K.; Tiwari, Rameshwar; Singh, Surender; Nain, Pawan K. S.; Rana, Sarika; Arora, Anju; Singh, Shashi B.; Nain, Lata

2014-01-01

Parthenium sp. is a noxious weed which threatens the environment and biodiversity due to its rapid invasion. This lignocellulosic weed was investigated for its potential in biofuel production by subjecting it to mild alkali pretreatment followed by enzymatic saccharification which resulted in significant amount of fermentable sugar yield (76.6%). Optimization of enzymatic hydrolysis variables such as temperature, pH, enzyme, and substrate loading was carried out using central composite design (CCD) in response to surface methodology (RSM) to achieve the maximum saccharification yield. Data obtained from RSM was validated using ANOVA. After the optimization process, a model was proposed with predicted value of 80.08% saccharification yield under optimum conditions which was confirmed by the experimental value of 85.80%. This illustrated a good agreement between predicted and experimental response (saccharification yield). The saccharification yield was enhanced by enzyme loading and reduced by temperature and substrate loading. This study reveals that under optimized condition, sugar yield was significantly increased which was higher than earlier reports and promises the use of Parthenium sp. biomass as a feedstock for bioethanol production. PMID:24900917

Batch fermentation options for high titer bioethanol production from a SPORL pretreated Douglas-Fir forest residue without detoxification

Treesearch

Mingyan Yang; Hairui Ji; Junyong Zhu

2016-01-01

This study evaluated batch fermentation modes, namely, separate hydrolysis and fermentation (SHF), quasi-simultaneous saccharification and fermentation (Q-SSF), and simultaneous saccharification and fermentation (SSF), and fermentation conditions, i.e., enzyme and yeast loadings, nutrient supplementation and sterilization, on high titer bioethanol production from SPORL...

Application of response surface methodology for optimization of polygalacturonase production by Aspergillus niger.

PubMed

Yadav, Kaushlesh K; Garg, Neelima; Kumar, Devendra; Kumar, Sanjay; Singh, Achal; Muthukumar, M

2015-01-01

Polygalacturonase (PG) degrades pectin into D-galacturonic acid monomers and is used widely in food industry especially for juice clarification. In the present study,. fermentation conditions for polygalacturonase production by Asgergillus niger NAIMCCF-02958, using mango peel as substrate, were optimized using the 2(3) factorial design with central composite rotatable experimental design (CCRD) of response surface methodology (RSM). The maximum PG activity 723.66 U g(-1) was achieved under pH 4.0, temperature 30 degrees C and 2% inoculum by response surface curve. The experimental value of PG activity wkas higher 607.65 U g(-1) than the predicted value 511.75 U g(-1). Under the proposed optimized conditions, the determination coefficient (R2) was equal to 0.66 indicating that the model could explain 66% of the total variation as well as establish the relationship between the variables and the responses. ANOVA analysis and the three dimensional plots also confirmed interactions among the parameters.

Statistical optimization for improved production of cyclosporin a in solid-state fermentation.

PubMed

Survase, Shrikant A; Annapure, Uday S; Singhal, Rekha S

2009-11-01

This work evaluates the effect of different amino acids on production of CyA production in solid-state fermentation that was previously optimized for different fermentation parameters by one-factor-at-a-time for the maximum production of CyA by Tolypocladium inflatum MTCC 557. Based on the Plackett-Burman design, glycerol, ammonium sulfate, FeCl3, and inoculum size were selected for further optimization by response surface methodology (RSM). After identifying effective nutrients, RSM was used to develop mathematical model equations, study responses, and establish the optimum concentrations of the key nutrients for higher CyA production. It was observed that supplementation of medium containing (% w/w) glycerol, 1.53; ammonium sulfate, 0.95; FeCl3, 0.18; and inoculum size 6.4 ml/5g yielded a maximum of 7,106 mg/kg as compared with 6,480 mg CyA/kg substrate using one factor at a time. In the second step, the effect of amino acids on the production of CyA was studied. Addition of L-valine and L-leucine in combination after 20 h of fermentation resulted in maximum production of 8,166 mg/kg.

High level production of β-galactosidase exhibiting excellent milk-lactose degradation ability from Aspergillus oryzae by codon and fermentation optimization.

PubMed

Zhao, Qianqian; Liu, Fei; Hou, Zhongwen; Yuan, Chao; Zhu, Xiqiang

2014-03-01

A β-galactosidase gene from Aspergillus oryzae was engineered utilizing codon usage optimization to be constitutively and highly expressed in the Pichia pastoris SMD1168H strain in a high-cell-density fermentation. After fermentation for 96 h in a 50-L fermentor using glucose and glycerol as combined carbon sources, the recombinant enzyme in the culture supernatant had an activity of 4,239.07 U mL(-1) with o-nitrophenyl-β-D-galactopyranoside as the substrate, and produced a total of extracellular protein content of 7.267 g L(-1) in which the target protein (6.24 g L(-1)) occupied approximately 86 %. The recombinant β-galactosidase exhibited an excellent lactose hydrolysis ability. With 1,000 U of the enzyme in 100 mL milk, 92.44 % lactose was degraded within 24 h at 60 °C, and the enzyme could also accomplish the hydrolysis at low temperatures of 37, 25, and 10 °C. Thus, this engineered strain had significantly higher fermentation level of A. oryzae lactase than that before optimization and the β-galactosidase may have a good application potential in whey and milk industries.

Process engineering and scale-up of autotrophic Clostridium strain P11 syngas fermentation

NASA Astrophysics Data System (ADS)

Kundiyana, Dimple Kumar Aiyanna

Scope and Method of Study. Biomass gasification followed by fermentation of syngas to ethanol is a potential process to produce bioenergy. The process is currently being researched under laboratory- and pilot-scale in an effort to optimize the process conditions and make the process feasible for commercial production of ethanol and other biofuels such as butanol and propanol. The broad research objectives for the research were to improve ethanol yields during syngas fermentation and to design a economical fermentation process. The research included four statistically designed experimental studies in serum bottles, bench-scale and pilot-scale fermentors to screen alternate fermentation media components, to determine the effect of process parameters such as pH, temperature and buffer on syngas fermentation, to determine the effect of key limiting nutrients of the acetyl-CoA pathway in a continuous series reactor design, and to scale-up the syngas fermentation in a 100-L pilot scale fermentor. Findings and Conclusions. The first experimental study identified cotton seed extract (CSE) as a feasible medium for Clostridium strain P11 fermentation. The study showed that CSE at 0.5 g L-1 can potentially replace all the standard Clostridium strain P11 fermentation media components while using a media buffer did not significantly improve the ethanol production when used in fermentation with CSE. Scale-up of the CSE fermentation in 2-L and 5-L stirred tank fermentors showed 25% increase in ethanol yield. The second experimental study showed that syngas fermentation at 32°C without buffer was associated with higher ethanol concentration and reduced lag time in switching to solventogenesis. Conducting fermentation at 40°C or by lowering incubation pH to 5.0 resulted in reduced cell growth and no production of ethanol or acetic acid. The third experiment studied the effect of three limiting nutrients, calcium pantothenate, vitamin B12 and CoCl2 on syngas fermentation. Results indicated that it is possible to modulate the product formation by limiting key nutrients of acetyl-CoA pathway and using a continuous fermentation in two-stage fermentor design to improve ethanol yields. The last experimental study was conducted to commission a pilot scale fermentor, and subsequently scale-up the Clostridium strain P11 fermentation from a bench-scale to a pilot scale 100-L fermentor. Results indicated a six-fold improvement in ethanol concentration (25.3 g L-1 at the end of 59 d) compared to previous Clostridium strain P11 and Clostridium carboxidivorans fermentations plus the formation of other compounds such as isopropyl alcohol, acetic acid and butanol, which are of commercial importance.

Promoter engineering of the Saccharomyces cerevisiae RIM15 gene for improvement of alcoholic fermentation rates under stress conditions.

PubMed

Watanabe, Daisuke; Kaneko, Akie; Sugimoto, Yukiko; Ohnuki, Shinsuke; Takagi, Hiroshi; Ohya, Yoshikazu

2017-02-01

A loss-of-function mutation in the RIM15 gene, which encodes a Greatwall-like protein kinase, is one of the major causes of the high alcoholic fermentation rates in Saccharomyces cerevisiae sake strains closely related to Kyokai no. 7 (K7). However, impairment of Rim15p may not be beneficial under more severe fermentation conditions, such as in the late fermentation stage, as it negatively affects stress responses. To balance stress tolerance and fermentation performance, we inserted the promoter of a gluconeogenic gene, PCK1, into the 5'-untranslated region (5'-UTR) of the RIM15 gene in a laboratory strain to achieve repression of RIM15 gene expression in the glucose-rich early stage with its induction in the stressful late stage of alcoholic fermentation. The promoter-engineered strain exhibited a fermentation rate comparable to that of the RIM15-deleted strain with no decrease in cell viability. The engineered strain achieved better alcoholic fermentation performance than the RIM15-deleted strain under repetitive and high-glucose fermentation conditions. These data demonstrated the validity of promoter engineering of the RIM15 gene that governs inhibitory control of alcoholic fermentation. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Realm of Thermoalkaline Lipases in Bioprocess Commodities

PubMed Central

2018-01-01

For decades, microbial lipases are notably used as biocatalysts and efficiently catalyze various processes in many important industries. Biocatalysts are less corrosive to industrial equipment and due to their substrate specificity and regioselectivity they produced less harmful waste which promotes environmental sustainability. At present, thermostable and alkaline tolerant lipases have gained enormous interest as biocatalyst due to their stability and robustness under high temperature and alkaline environment operation. Several characteristics of the thermostable and alkaline tolerant lipases are discussed. Their molecular weight and resistance towards a range of temperature, pH, metal, and surfactants are compared. Their industrial applications in biodiesel, biodetergents, biodegreasing, and other types of bioconversions are also described. This review also discusses the advance of fermentation process for thermostable and alkaline tolerant lipases production focusing on the process development in microorganism selection and strain improvement, culture medium optimization via several optimization techniques (i.e., one-factor-at-a-time, surface response methodology, and artificial neural network), and other fermentation parameters (i.e., inoculums size, temperature, pH, agitation rate, dissolved oxygen tension (DOT), and aeration rate). Two common fermentation techniques for thermostable and alkaline tolerant lipases production which are solid-state and submerged fermentation methods are compared and discussed. Recent optimization approaches using evolutionary algorithms (i.e., Genetic Algorithm, Differential Evolution, and Particle Swarm Optimization) are also highlighted in this article. PMID:29666707

Evaluation of Ethanol Production Activity by Engineered Saccharomyces cerevisiae Fermenting Cellobiose through the Phosphorolytic Pathway in Simultaneous Saccharification and Fermentation of Cellulose.

PubMed

Lee, Won-Heong; Jin, Yong-Su

2017-09-28

In simultaneous saccharification and fermentation (SSF) for production of cellulosic biofuels, engineered Saccharomyces cerevisiae capable of fermenting cellobiose has provided several benefits, such as lower enzyme costs and faster fermentation rate compared with wild-type S. cerevisiae fermenting glucose. In this study, the effects of an alternative intracellular cellobiose utilization pathway-a phosphorolytic pathway based on a mutant cellodextrin transporter (CDT-1 (F213L)) and cellobiose phosphorylase (SdCBP)-was investigated by comparing with a hydrolytic pathway based on the same transporter and an intracellular β-glucosidase (GH1-1) for their SSF performances under various conditions. Whereas the phosphorolytic and hydrolytic cellobiose-fermenting S. cerevisiae strains performed similarly under the anoxic SSF conditions, the hydrolytic S. cerevisiae performed slightly better than the phosphorolytic S. cerevisiae under the microaerobic SSF conditions. Nonetheless, the phosphorolytic S. cerevisiae expressing the mutant CDT-1 showed better ethanol production than the glucose-fermenting S. cerevisiae with an extracellular β-glucosidase, regardless of SSF conditions. These results clearly prove that introduction of the intracellular cellobiose metabolic pathway into yeast can be effective on cellulosic ethanol production in SSF. They also demonstrate that enhancement of cellobiose transport activity in engineered yeast is the most important factor affecting the efficiency of SSF of cellulose.

Phenotypic evaluation and characterization of 21 industrial Saccharomyces cerevisiae yeast strains.

PubMed

Kong, In Iok; Turner, Timothy Lee; Kim, Heejin; Kim, Soo Rin; Jin, Yong-Su

2018-02-01

Microorganisms have been studied and used extensively to produce value-added fuels and chemicals. Yeasts, specifically Saccharomyces cerevisiae, receive industrial attention because of their well-known ability to ferment glucose and produce ethanol. Thousands of natural or genetically modified S. cerevisiae have been found in industrial environments for various purposes. These industrial strains are isolated from industrial fermentation sites, and they are considered as potential host strains for superior fermentation processes. In many cases, industrial yeast strains have higher thermotolerance, increased resistances towards fermentation inhibitors and increased glucose fermentation rates under anaerobic conditions when compared with laboratory yeast strains. Despite the advantages of industrial strains, they are often not well characterized. Through screening and phenotypic characterization of commercially available industrial yeast strains, industrial fermentation processes requiring specific environmental conditions may be able to select an ideal starting yeast strain to be further engineered. Here, we have characterized and compared 21 industrial S. cerevisiae strains under multiple conditions, including their tolerance to varying pH conditions, resistance to fermentation inhibitors, sporulation efficiency and ability to ferment lignocellulosic sugars. These data may be useful for the selection of a parental strain for specific biotechnological applications of engineered yeast. © FEMS 2018. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

[Production and partial characterization of beta-galactosidase from Kluyveromyces lactis grown in deproteinized whey].

PubMed

Ramírez Matheus, Alejandra O; Rivas, Nilo

2003-06-01

The purpose of this work was to optimize the beta-galactosidase production by Kluyveromyces lactis, applying the Surface Response Methodology (SRM) and using deproteinized whey as fermentation medium. An Orthogonal Central Compound Design (OCCD) was used without repetition, with four factors: temperature, pH, agitation speed and fermentation time. Then, enzyme activity (U/ml) as response variable was used. Thirty trials in twenty-five treatments, with six repetitions at the central point, were carried out, in a New Brunswick Bioflo 2000 fermentor with a volume of 2 liters. The deproteinized whey obtained by thermocoagulation was chemically analyzed. The results were: moisture 93.83%, total solids 6.17%, protein 0.44%, lactose 4.85%, acidity 0.43% and pH 4.58. The best conditions in the enzyme production were: temperature 30.3 degrees C, pH 4.68, agitation speed 191 r.p.m. and fermentation time 18.5 h. with an enzyme production of 8.3 U/ml. The degree of purification obtained was 7.4 times and the yield was 50.8%. The purified enzyme had an optimum temperature of 60 degrees C and a pH of 6.2. This work shows that the yeast Kluyveromyces lactis grown in deproteinized whey is able to produce the enzyme beta-galactosidase and SRM can be used in the fermentology processes, specifically in determining the best suitable operation conditions.

Lovastatin Production by Aspergillus terreus Using Agro-Biomass as Substrate in Solid State Fermentation

PubMed Central

Faseleh Jahromi, Mohammad; Liang, Juan Boo; Ho, Yin Wan; Mohamad, Rosfarizan; Goh, Yong Meng; Shokryazdan, Parisa

2012-01-01

Ability of two strains of Aspergillus terreus (ATCC 74135 and ATCC 20542) for production of lovastatin in solid state fermentation (SSF) using rice straw (RS) and oil palm frond (OPF) was investigated. Results showed that RS is a better substrate for production of lovastatin in SSF. Maximum production of lovastatin has been obtained using A. terreus ATCC 74135 and RS as substrate without additional nitrogen source (157.07 mg/kg dry matter (DM)). Although additional nitrogen source has no benefit effect on enhancing the lovastatin production using RS substrate, it improved the lovastatin production using OPF with maximum production of 70.17 and 63.76 mg/kg DM for A. terreus ATCC 20542 and A. terreus ATCC 74135, respectively (soybean meal as nitrogen source). Incubation temperature, moisture content, and particle size had shown significant effect on lovastatin production (P < 0.01) and inoculums size and pH had no significant effect on lovastatin production (P > 0.05). Results also have shown that pH 6, 25°C incubation temperature, 1.4 to 2 mm particle size, 50% initial moisture content, and 8 days fermentation time are the best conditions for lovastatin production in SSF. Maximum production of lovastatin using optimized condition was 175.85 and 260.85 mg/kg DM for A. terreus ATCC 20542 and ATCC 74135, respectively, using RS as substrate. PMID:23118499

Dynamic metabolic modeling of heterotrophic and mixotrophic microalgal growth on fermentative wastes

PubMed Central

Baroukh, Caroline; Turon, Violette; Bernard, Olivier

2017-01-01

Microalgae are promising microorganisms for the production of numerous molecules of interest, such as pigments, proteins or triglycerides that can be turned into biofuels. Heterotrophic or mixotrophic growth on fermentative wastes represents an interesting approach to achieving higher biomass concentrations, while reducing cost and improving the environmental footprint. Fermentative wastes generally consist of a blend of diverse molecules and it is thus crucial to understand microalgal metabolism in such conditions, where switching between substrates might occur. Metabolic modeling has proven to be an efficient tool for understanding metabolism and guiding the optimization of biomass or target molecule production. Here, we focused on the metabolism of Chlorella sorokiniana growing heterotrophically and mixotrophically on acetate and butyrate. The metabolism was represented by 172 metabolic reactions. The DRUM modeling framework with a mildly relaxed quasi-steady-state assumption was used to account for the switching between substrates and the presence of light. Nine experiments were used to calibrate the model and nine experiments for the validation. The model efficiently predicted the experimental data, including the transient behavior during heterotrophic, autotrophic, mixotrophic and diauxic growth. It shows that an accurate model of metabolism can now be constructed, even in dynamic conditions, with the presence of several carbon substrates. It also opens new perspectives for the heterotrophic and mixotrophic use of microalgae, especially for biofuel production from wastes. PMID:28582469

Enhanced acetic acid production from manalagi apple (Malus sylvestris mill) by mixed cultures of Saccharomyces cerevisiae and Acetobacter aceti in submerged fermentation

NASA Astrophysics Data System (ADS)

Rosada, K. K.

2018-05-01

The production of acetic acid from Manalagi apple was studied using a mixed culture of S. cerevisiae and A. aceti by submerged fermentation technique. Determination of the best conditions for producing acetic acid was performed by stratified optimization with variations that were made on the concentration of the initial sugar addition to the medium (0%, 10%, 20% w/v), the ratio of the number of inocula S. cerevisiae and A. aceti (7:3, 1:1, 3:7), and agitation rate (80 and 160 rpm). All experiments were done by using the initial pH medium of 4.5 and incubated at room temperature (28±2oC) for 14 days. The concentration of reducing sugar, alcohol, acetic acid, and the pH were measured every 48 hours. The efficiency of sugar conversion to acetic acid with the addition of initial sugar 0%, 10%, and20%were 233%, 46.6%, and 6.4% respectively after ten days of incubation. Overall, the result showed that the highest acetic acid was produced from Manalagi apple juice when no sugar was added, using seven parts of S. cerevisiae to three parts of A. aceti and agitation rate of 160 rpm on the tenth day of fermentation. Under these conditions, glucose conversion efficiency to acetic acid increased to 362%.

Evaluation of nanoparticle-immobilized cellulase for improved ethanol yield in simultaneous saccharification and fermentation reactions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lupoi, Jason; Smith, Emily

2011-12-01

Ethanol yields were 2.1 (P = 0.06) to 2.3 (P = 0.01) times higher in simultaneous saccharification and fermentation (SSF) reactions of microcrystalline cellulose when cellulase was physisorbed on silica nanoparticles compared to enzyme in solution. In SSF reactions, cellulose is hydrolyzed to glucose by cellulase while yeast simultaneously ferments glucose to ethanol. The 35 C temperature and the presence of ethanol in SSF reactions are not optimal conditions for cellulase. Immobilization onto solid supports can stabilize the enzyme and promote activity at non-optimum reaction conditions. Mock SSF reactions that did not contain yeast were used to measure saccharification productsmore » and identify the mechanism for the improved ethanol yield using immobilized cellulase. Cellulase adsorbed to 40 nm silica nanoparticles produced 1.6 times (P = 0.01) more glucose than cellulase in solution in 96 h at pH 4.8 and 35 C. There was no significant accumulation (<250 {mu}g) of soluble cellooligomers in either the solution or immobilized enzyme reactions. This suggests that the mechanism for the immobilized enzyme's improved glucose yield compared to solution enzyme is the increased conversion of insoluble cellulose hydrolysis products to soluble cellooligomers at 35 C and in the presence of ethanol. The results show that silica-immobilized cellulase can be used to produce increased ethanol yields in the conversion of lignocellulosic materials by SSF.« less

Statistical optimization of medium composition and culture condition for the production of recombinant anti-lipopolysaccharide factor of Eriocheir sinensis in Escherichia coli

NASA Astrophysics Data System (ADS)

Jiang, Shan; Liu, Mei; Wang, Baojie; Jiang, Keyong; Wang, Lei

2011-11-01

Anti-lipopolysaccharide factors (ALFs) are important antimicrobial peptides that are isolated from some aquatic species. In a previous study, we isolated ALF genes from Chinese mitten crab, Eriocheir sinensis. In this study, we optimized the production of a recombinant ALF by expressing E. sinensis ALF genes in Escherichia coli maintained in shake-flasks. In particular, we focused on optimization of both the medium composition and the culture condition. Various medium components were analyzed by the Plackett-Burman design, and two significant screened factors, (NH4)2SO4 and KH2PO4, were further optimized via the central composite design (CCD). Based on the CCD analysis, we investigated the induction start-up time, the isopropylthio-D-galactoside (IPTG) concentration, the post-induction time, and the temperature by response surface methodology. We found that the highest level of ALF fusion protein was achieved in the medium containing 1.89 g/L (NH4)2SO4 and 3.18 g/L KH2PO4, with a cell optical density of 0.8 at 600 nm before induction, an IPTG concentration of 0.5 mmol/L, a post-induction temperature of 32.7°C, and a post-induction time of 4 h. Applying the whole optimization strategy using all optimal factors improved the target protein content from 6.1% (without optimization) to 13.2%. We further applied the optimized medium and conditions in high cell density cultivation, and determined that the soluble target protein constituted 10.5% of the total protein. Our identification of the economic medium composition, optimal culture conditions, and details of the fermentation process should facilitate the potential application of ALF for further research.

Use of chemostat cultures mimicking different phases of wine fermentations as a tool for quantitative physiological analysis

PubMed Central

2014-01-01

Background Saccharomyces cerevisiae is the most relevant yeast species conducting the alcoholic fermentation that takes place during winemaking. Although the physiology of this model organism has been extensively studied, systematic quantitative physiology studies of this yeast under winemaking conditions are still scarce, thus limiting the understanding of fermentative metabolism of wine yeast strains and the systematic description, modelling and prediction of fermentation processes. In this study, we implemented and validated the use of chemostat cultures as a tool to simulate different stages of a standard wine fermentation, thereby allowing to implement metabolic flux analyses describing the sequence of metabolic states of S. cerevisae along the wine fermentation. Results Chemostat cultures mimicking the different stages of standard wine fermentations of S. cerevisiae EC1118 were performed using a synthetic must and strict anaerobic conditions. The simulated stages corresponded to the onset of the exponential growth phase, late exponential growth phase and cells just entering stationary phase, at dilution rates of 0.27, 0.04, 0.007 h−1, respectively. Notably, measured substrate uptake and product formation rates at each steady state condition were generally within the range of corresponding conversion rates estimated during the different batch fermentation stages. Moreover, chemostat data were further used for metabolic flux analysis, where biomass composition data for each condition was considered in the stoichiometric model. Metabolic flux distributions were coherent with previous analyses based on batch cultivations data and the pseudo-steady state assumption. Conclusions Steady state conditions obtained in chemostat cultures reflect the environmental conditions and physiological states of S. cerevisiae corresponding to the different growth stages of a typical batch wine fermentation, thereby showing the potential of this experimental approach to systematically study the effect of environmental relevant factors such as temperature, sugar concentration, C/N ratio or (micro) oxygenation on the fermentative metabolism of wine yeast strains. PMID:24928139

Production of ethanol and arabitol by Debaryomyces nepalensis: influence of process parameters

PubMed Central

2013-01-01

Debaryomyces nepalensis, osmotolerant yeast isolated from rotten apple, is known to utilize both hexoses and pentoses and produce industrially important metabolites like ethanol, xylitol and arabitol. In the present study, the effect of different growth substrates, trace elements, nitrogen concentration and initial pH on growth and formation of ethanol and arabitol were examined. Optimum conditions for maximizing the product yields were established: glucose as carbon source, an initial pH of 6.0, 6 g/L of ammonium sulphate and addition of micronutrients. Under these best suited conditions, a concentration of 11g/L of arabitol and 19 g/L of ethanol was obtained in shake flask fermentations. The fermentation was scaled up to 2.5 L bioreactor and the influence of aeration, agitation and initial substrate concentration was also determined. Under optimal conditions (150 g/L glucose, 400 rpm and 0.5 vvm) ethanol concentration reached 52 g/L, which corresponds to a yield of 0.34 g/g and volumetric productivity of 0.28 g/L/h, whereas arabitol production reached a maximum of 14 g/L with a yield and volumetric productivity of 0.1 g/g and 0.07 g/L/h respectively. PMID:23659479

Optimization of Banana Juice Fermentation for the Production of Microbial Oil †

PubMed Central

Vega, Esther Z.; Glatz, Bonita A.; Hammond, Earl G.

1988-01-01

Apiotrichum curvatum ATCC 20509 (formerly Candida curvata D), a lipid-accumulating yeast, was grown in banana juice. The optimum conditions for biomass production in shake flasks were 30°C growth temperature, efficient aeration, a juice concentration of 25%, and preliminary heat treatment at less than sterilization conditions. Under controlled conditions in a fermentor, 20% banana juice was optimum. High concentrations of yeast extract (0.3%) increased biomass production by 40% but decreased oil production by 30%. A lower yeast extract concentration (0.05%) increased biomass production by 2% and oil production by 25%. The best growth and oil production were observed when asparagine (1.4 g/liter) and mineral salts were added to the banana juice. The addition of minerals seemed to improve the utilization of carbon. Growth inhibition was observed when the fermentor was aerated with pure oxygen, even when additional nutrients were present. A fed-batch process permitted the juice concentration to be increased from 15 to 82%; biomass accumulation was three times higher than in batch fermentations. However, the cellular lipid content was only 30% of dry weight, and chemical oxygen demand reduction was slow and inefficient. PMID:16347584

Heat Pre-Treatment of Beverages Wastewater on Hydrogen Production

NASA Astrophysics Data System (ADS)

Uyub, S. Z.; Mohd, N. S.; Ibrahim, S.

2017-06-01

At present, a large variety of alternative fuels have been investigated and hydrogen gas is considered as the possible solution for the future due to its unique characteristics. Through dark fermentation process, several factors were found to have significant impact on the hydrogen production either through process enhancement or inhibition and degradation rates or influencing parameters. This work was initiated to investigate the optimum conditions for heat pre-treatment and initial pH for the dark fermentative process under mesophilic condition using a central composite design and response surface methodology (RSM). Different heat treatment conditions and pH were performed on the seed sludge collected from the anaerobic digester of beverage wastewater treatment plant. Heat treatment of inoculum was optimized at different exposure times (30, 90, 120 min), temperatures (80, 90 and 100°C) and pH (4.5, 5.5, 6.5) in order to maximize the biohydrogen production and methanogens activity inhibition. It was found that the optimum heat pre-treatment condition and pH occurred at 100°C for 50 min and the pH of 6.00. At this optimum condition the hydrogen yield was 63.0476 ml H2/mol glucose (H2 Yield) and the COD removal efficiency was 90.87%. In conclusion, it can be hypothesized that different heat treatment conditions led to differences in the initial microbial communities (hydrogen producing bacteria) which resulted in the different hydrogen yields.

Optimization of fermentation medium for the production of atrazine degrading strain Acinetobacter sp. DNS(32) by statistical analysis system.

PubMed

Zhang, Ying; Wang, Yang; Wang, Zhi-Gang; Wang, Xi; Guo, Huo-Sheng; Meng, Dong-Fang; Wong, Po-Keung

2012-01-01

Statistical experimental designs provided by statistical analysis system (SAS) software were applied to optimize the fermentation medium composition for the production of atrazine-degrading Acinetobacter sp. DNS(32) in shake-flask cultures. A "Plackett-Burman Design" was employed to evaluate the effects of different components in the medium. The concentrations of corn flour, soybean flour, and K(2)HPO(4) were found to significantly influence Acinetobacter sp. DNS(32) production. The steepest ascent method was employed to determine the optimal regions of these three significant factors. Then, these three factors were optimized using central composite design of "response surface methodology." The optimized fermentation medium composition was composed as follows (g/L): corn flour 39.49, soybean flour 25.64, CaCO(3) 3, K(2)HPO(4) 3.27, MgSO(4)·7H(2)O 0.2, and NaCl 0.2. The predicted and verifiable values in the medium with optimized concentration of components in shake flasks experiments were 7.079 × 10(8) CFU/mL and 7.194 × 10(8) CFU/mL, respectively. The validated model can precisely predict the growth of atrazine-degraing bacterium, Acinetobacter sp. DNS(32).

Enzymatic saccharification and lactic acid production from banana pseudo-stem through optimized pretreatment at lowest catalyst concentration

PubMed Central

Idrees, Muhammad; Adnan, Ahmad; Malik, Farnaz; Qureshi, Fahim Ashraf

2013-01-01

This work estimates the potential of banana pseudo-stem with high cellulosic content 42.2-63 %, for the production of fermentable sugars for lactic acid production through statistically optimized pretreatment method. To evaluate the catalyzed pretreatment efficiency of banana pseudo stem based on the enzymatic digestibility, Response Surface Methodology (RSM) was employed for the optimization of pretreatment temperature and time using lowest concentrations of H2SO4, NaOH, NaOH catalyzed Na2S and Na2SO3 that seemed to be significant variables with P<0.05. High F and R2 values and low p-value for hydrolysis yield indicated the model predictability. The optimized condition for NaOH was determined to be conc. 1 %, temperature 130 oC for 2.6 hr; Na2S; conc. 1 %, temperature 130 oC for 2.29 hr; Na2SO3; conc. 1 %, temperature 130 oC for 2.41 hr and H2SO4; conc. 1 %, temperature 129.45 oC for 2.18 hr, produced 84.91 %, 85.23 %, 81.2 % and 76.02 % hydrolysis yield, respectively. Sulphuric acid provided 33+1 gL-1 reducing sugars in pretreatment step along with 38+0.5 gL-1 during enzymatic hydrolysis. Separate hydrolysis and fermentation of resulting sugars showed that the conversion of glucans into lactic acid reached 92 % of the theoretical yield of glucose. PMID:26966423

Effect of mixing during fermentation in yogurt manufacturing.

PubMed

Aguirre-Ezkauriatza, E J; Galarza-González, M G; Uribe-Bujanda, A I; Ríos-Licea, M; López-Pacheco, F; Hernández-Brenes, C M; Alvarez, M M

2008-12-01

In traditional yogurt manufacturing, the yogurt is not agitated during fermentation. However, stirring could be beneficial, particularly for improving heat and mass transport across the fermentation tank. In this contribution, we studied the effect of low-speed agitation during fermentation on process time, acidity profile, and microbial dynamics during yogurt fermentation in 2 laboratory-scale fermenters (3 and 5 L) with different heat-transfer characteristics. Lactobacillus bulgaricus and Streptococcus thermophilus were used as fermenting bacteria. Curves of pH, lactic acid concentration, lactose concentration, and bacterial population profiles during fermentation are presented for static and low-agitation conditions during fermentation. At low-inoculum conditions, agitation reduced the processing time by shortening the lag phase. However, mixing did not modify the duration or the shape of the pH profiles during the exponential phase. In fermentors with poor heat-transfer characteristics, important differences in microbial dynamics were observed between the agitated and nonagitated fermentation experiments; that is, agitation significantly increased the observable specific growth rate and the final microbial count of L. bulgaricus.

The production of coenzyme Q10 in microorganisms.

PubMed

Cluis, Corinne P; Pinel, Dominic; Martin, Vincent J

2012-01-01

Coenzyme Q10 has emerged as a valuable molecule for pharmaceutical and cosmetic applications. Therefore, research into producing and optimizing coenzyme Q10 via microbial fermentation is ongoing. There are two major paths being explored for maximizing production of this molecule to commercially advantageous levels. The first entails using microbes that naturally produce coenzyme Q10 as fermentation biocatalysts and optimizing the fermentation parameters in order to reach industrial levels of production. However, the natural coenzyme Q10-producing microbes tend to be intractable for industrial fermentation settings. The second path to coenzyme Q10 production being explored is to engineer Escherichia coli with the ability to biosynthesize this molecule in order to take advantage of its more favourable fermentation characteristics and the well-understood array of genetic tools available for this bacteria. Although many studies have attempted to over-produce coenzyme Q10 in E. coli through genetic engineering, production titres still remain below those of the natural coenzyme Q10-producing microorganisms. Current research is providing the knowledge needed to alleviate the bottlenecks involved in producing coenzyme Q10 from an E. coli strain platform and the fermentation parameters that could dramatically increase production titres from natural microbial producers. Synthesizing the lessons learned from both approaches may be the key towards a more cost-effective coenzyme Q10 industry.

Noteworthy Facts about a Methane-Producing Microbial Community Processing Acidic Effluent from Sugar Beet Molasses Fermentation.

PubMed

Chojnacka, Aleksandra; Szczęsny, Paweł; Błaszczyk, Mieczysław K; Zielenkiewicz, Urszula; Detman, Anna; Salamon, Agnieszka; Sikora, Anna

2015-01-01

Anaerobic digestion is a complex process involving hydrolysis, acidogenesis, acetogenesis and methanogenesis. The separation of the hydrogen-yielding (dark fermentation) and methane-yielding steps under controlled conditions permits the production of hydrogen and methane from biomass. The characterization of microbial communities developed in bioreactors is crucial for the understanding and optimization of fermentation processes. Previously we developed an effective system for hydrogen production based on long-term continuous microbial cultures grown on sugar beet molasses. Here, the acidic effluent from molasses fermentation was used as the substrate for methanogenesis in an upflow anaerobic sludge blanket bioreactor. This study focused on the molecular analysis of the methane-yielding community processing the non-gaseous products of molasses fermentation. The substrate for methanogenesis produces conditions that favor the hydrogenotrophic pathway of methane synthesis. Methane production results from syntrophic metabolism whose key process is hydrogen transfer between bacteria and methanogenic Archaea. High-throughput 454 pyrosequencing of total DNA isolated from the methanogenic microbial community and bioinformatic sequence analysis revealed that the domain Bacteria was dominated by Firmicutes (mainly Clostridia), Bacteroidetes, δ- and γ-Proteobacteria, Cloacimonetes and Spirochaetes. In the domain Archaea, the order Methanomicrobiales was predominant, with Methanoculleus as the most abundant genus. The second and third most abundant members of the Archaeal community were representatives of the Methanomassiliicoccales and the Methanosarcinales. Analysis of the methanogenic sludge by scanning electron microscopy with Energy Dispersive X-ray Spectroscopy and X-ray diffraction showed that it was composed of small highly heterogeneous mineral-rich granules. Mineral components of methanogenic granules probably modulate syntrophic metabolism and methanogenic pathways. A rough functional analysis from shotgun data of the metagenome demonstrated that our knowledge of methanogenesis is poor and/or the enzymes responsible for methane production are highly effective, since despite reasonably good sequencing coverage, the details of the functional potential of the microbial community appeared to be incomplete.

Noteworthy Facts about a Methane-Producing Microbial Community Processing Acidic Effluent from Sugar Beet Molasses Fermentation

PubMed Central

Chojnacka, Aleksandra; Szczęsny, Paweł; Błaszczyk, Mieczysław K.; Zielenkiewicz, Urszula; Detman, Anna; Salamon, Agnieszka; Sikora, Anna

2015-01-01

Anaerobic digestion is a complex process involving hydrolysis, acidogenesis, acetogenesis and methanogenesis. The separation of the hydrogen-yielding (dark fermentation) and methane-yielding steps under controlled conditions permits the production of hydrogen and methane from biomass. The characterization of microbial communities developed in bioreactors is crucial for the understanding and optimization of fermentation processes. Previously we developed an effective system for hydrogen production based on long-term continuous microbial cultures grown on sugar beet molasses. Here, the acidic effluent from molasses fermentation was used as the substrate for methanogenesis in an upflow anaerobic sludge blanket bioreactor. This study focused on the molecular analysis of the methane-yielding community processing the non-gaseous products of molasses fermentation. The substrate for methanogenesis produces conditions that favor the hydrogenotrophic pathway of methane synthesis. Methane production results from syntrophic metabolism whose key process is hydrogen transfer between bacteria and methanogenic Archaea. High-throughput 454 pyrosequencing of total DNA isolated from the methanogenic microbial community and bioinformatic sequence analysis revealed that the domain Bacteria was dominated by Firmicutes (mainly Clostridia), Bacteroidetes, δ- and γ-Proteobacteria, Cloacimonetes and Spirochaetes. In the domain Archaea, the order Methanomicrobiales was predominant, with Methanoculleus as the most abundant genus. The second and third most abundant members of the Archaeal community were representatives of the Methanomassiliicoccales and the Methanosarcinales. Analysis of the methanogenic sludge by scanning electron microscopy with Energy Dispersive X-ray Spectroscopy and X-ray diffraction showed that it was composed of small highly heterogeneous mineral-rich granules. Mineral components of methanogenic granules probably modulate syntrophic metabolism and methanogenic pathways. A rough functional analysis from shotgun data of the metagenome demonstrated that our knowledge of methanogenesis is poor and/or the enzymes responsible for methane production are highly effective, since despite reasonably good sequencing coverage, the details of the functional potential of the microbial community appeared to be incomplete. PMID:26000448

Multi-objective optimization of an industrial penicillin V bioreactor train using non-dominated sorting genetic algorithm.

PubMed

Lee, Fook Choon; Rangaiah, Gade Pandu; Ray, Ajay Kumar

2007-10-15

Bulk of the penicillin produced is used as raw material for semi-synthetic penicillin (such as amoxicillin and ampicillin) and semi-synthetic cephalosporins (such as cephalexin and cefadroxil). In the present paper, an industrial penicillin V bioreactor train is optimized for multiple objectives simultaneously. An industrial train, comprising a bank of identical bioreactors, is run semi-continuously in a synchronous fashion. The fermentation taking place in a bioreactor is modeled using a morphologically structured mechanism. For multi-objective optimization for two and three objectives, the elitist non-dominated sorting genetic algorithm (NSGA-II) is chosen. Instead of a single optimum as in the traditional optimization, a wide range of optimal design and operating conditions depicting trade-offs of key performance indicators such as batch cycle time, yield, profit and penicillin concentration, is successfully obtained. The effects of design and operating variables on the optimal solutions are discussed in detail. Copyright 2007 Wiley Periodicals, Inc.

Glycerol Production by Fermenting Yeast Cells Is Essential for Optimal Bread Dough Fermentation

PubMed Central

Aslankoohi, Elham; Rezaei, Mohammad Naser; Vervoort, Yannick; Courtin, Christophe M.; Verstrepen, Kevin J.

2015-01-01

Glycerol is the main compatible solute in yeast Saccharomyces cerevisiae. When faced with osmotic stress, for example during semi-solid state bread dough fermentation, yeast cells produce and accumulate glycerol in order to prevent dehydration by balancing the intracellular osmolarity with that of the environment. However, increased glycerol production also results in decreased CO2 production, which may reduce dough leavening. We investigated the effect of yeast glycerol production level on bread dough fermentation capacity of a commercial bakery strain and a laboratory strain. We find that Δgpd1 mutants that show decreased glycerol production show impaired dough fermentation. In contrast, overexpression of GPD1 in the laboratory strain results in increased fermentation rates in high-sugar dough and improved gas retention in the fermenting bread dough. Together, our results reveal the crucial role of glycerol production level by fermenting yeast cells in dough fermentation efficiency as well as gas retention in dough, thereby opening up new routes for the selection of improved commercial bakery yeasts. PMID:25764309

Glycerol production by fermenting yeast cells is essential for optimal bread dough fermentation.

PubMed

Aslankoohi, Elham; Rezaei, Mohammad Naser; Vervoort, Yannick; Courtin, Christophe M; Verstrepen, Kevin J

2015-01-01

Glycerol is the main compatible solute in yeast Saccharomyces cerevisiae. When faced with osmotic stress, for example during semi-solid state bread dough fermentation, yeast cells produce and accumulate glycerol in order to prevent dehydration by balancing the intracellular osmolarity with that of the environment. However, increased glycerol production also results in decreased CO2 production, which may reduce dough leavening. We investigated the effect of yeast glycerol production level on bread dough fermentation capacity of a commercial bakery strain and a laboratory strain. We find that Δgpd1 mutants that show decreased glycerol production show impaired dough fermentation. In contrast, overexpression of GPD1 in the laboratory strain results in increased fermentation rates in high-sugar dough and improved gas retention in the fermenting bread dough. Together, our results reveal the crucial role of glycerol production level by fermenting yeast cells in dough fermentation efficiency as well as gas retention in dough, thereby opening up new routes for the selection of improved commercial bakery yeasts.

Production and Characterization of Lipases by Two New Isolates of Aspergillus through Solid-State and Submerged Fermentation

PubMed Central

Colla, Luciane Maria; Ficanha, Aline M. M.; Rizzardi, Juliana; Bertolin, Telma Elita; Reinehr, Christian Oliveira; Costa, Jorge Alberto Vieira

2015-01-01

Due to the numerous applications of lipases in industry, there is a need to study their characteristics, because lipases obtained from different sources may present different properties. The aim of this work was to accomplish the partial characterization of lipases obtained through submerged fermentation and solid-state fermentation by two species of Aspergillus. Fungal strains were isolated from a diesel-contaminated soil and selected as good lipases producers. Lipases obtained through submerged fermentation presented optimal activities at 37°C and pH 7.2 and those obtained through solid-state fermentation at 35°C and pH 6.0. The enzymes produced by submerged fermentation were more temperature-stable than those obtained by solid-state fermentation, presenting 72% of residual activity after one hour of exposition at 90°C. Lipases obtained through submerged fermentation had 80% of stability in acidic pH and those obtained through solid-state fermentation had stability greater than 60% in alkaline pH. PMID:26180809

Fermentation of lactose to ethanol in cheese whey permeate and concentrated permeate by engineered Escherichia coli.

PubMed

Pasotti, Lorenzo; Zucca, Susanna; Casanova, Michela; Micoli, Giuseppina; Cusella De Angelis, Maria Gabriella; Magni, Paolo

2017-06-02

Whey permeate is a lactose-rich effluent remaining after protein extraction from milk-resulting cheese whey, an abundant dairy waste. The lactose to ethanol fermentation can complete whey valorization chain by decreasing dairy waste polluting potential, due to its nutritional load, and producing a biofuel from renewable source at the same time. Wild type and engineered microorganisms have been proposed as fermentation biocatalysts. However, they present different drawbacks (e.g., nutritional supplements requirement, high transcriptional demand of recombinant genes, precise oxygen level, and substrate inhibition) which limit the industrial attractiveness of such conversion process. In this work, we aim to engineer a new bacterial biocatalyst, specific for dairy waste fermentation. We metabolically engineered eight Escherichia coli strains via a new expression plasmid with the pyruvate-to-ethanol conversion genes, and we carried out the selection of the best strain among the candidates, in terms of growth in permeate, lactose consumption and ethanol formation. We finally showed that the selected engineered microbe (W strain) is able to efficiently ferment permeate and concentrated permeate, without nutritional supplements, in pH-controlled bioreactor. In the conditions tested in this work, the selected biocatalyst could complete the fermentation of permeate and concentrated permeate in about 50 and 85 h on average, producing up to 17 and 40 g/l of ethanol, respectively. To our knowledge, this is the first report showing efficient ethanol production from the lactose contained in whey permeate with engineered E. coli. The selected strain is amenable to further metabolic optimization and represents an advance towards efficient biofuel production from industrial waste stream.

Physiological and fermentation properties of Bacillus coagulans and a mutant lacking fermentative lactate dehydrogenase activity.

PubMed

Su, Yue; Rhee, Mun Su; Ingram, Lonnie O; Shanmugam, K T

2011-03-01

Bacillus coagulans, a sporogenic lactic acid bacterium, grows optimally at 50-55 °C and produces lactic acid as the primary fermentation product from both hexoses and pentoses. The amount of fungal cellulases required for simultaneous saccharification and fermentation (SSF) at 55 °C was previously reported to be three to four times lower than for SSF at the optimum growth temperature for Saccharomyces cerevisiae of 35 °C. An ethanologenic B. coagulans is expected to lower the cellulase loading and production cost of cellulosic ethanol due to SSF at 55 °C. As a first step towards developing B. coagulans as an ethanologenic microbial biocatalyst, activity of the primary fermentation enzyme L-lactate dehydrogenase was removed by mutation (strain Suy27). Strain Suy27 produced ethanol as the main fermentation product from glucose during growth at pH 7.0 (0.33 g ethanol per g glucose fermented). Pyruvate dehydrogenase (PDH) and alcohol dehydrogenase (ADH) acting in series contributed to about 55% of the ethanol produced by this mutant while pyruvate formate lyase and ADH were responsible for the remainder. Due to the absence of PDH activity in B. coagulans during fermentative growth at pH 5.0, the l-ldh mutant failed to grow anaerobically at pH 5.0. Strain Suy27-13, a derivative of the l-ldh mutant strain Suy27, that produced PDH activity during anaerobic growth at pH 5.0 grew at this pH and also produced ethanol as the fermentation product (0.39 g per g glucose). These results show that construction of an ethanologenic B. coagulans requires optimal expression of PDH activity in addition to the removal of the LDH activity to support growth and ethanol production.

Limitation of thiamine pyrophosphate supply to growing Escherichia coli switches metabolism to efficient D-lactate formation.

PubMed

Tian, Kangming; Niu, Dandan; Liu, Xiaoguang; Prior, Bernard A; Zhou, Li; Lu, Fuping; Singh, Suren; Wang, Zhengxiang

2016-01-01

Efficient production of D-lactate by engineered Escherichia coli entails balancing cell growth and product synthesis. To develop a metabolic switch to implement a desirable transition from cell growth to product fermentation, a thiamine auxotroph B0013-080A was constructed in a highly efficient D-lactate producer E. coli strain B0013-070. This was achieved by inactivation of thiE, a gene encoding a thiamine phosphate synthase for biosynthesis of thiamine monophosphate. The resultant mutant B0013-080A failed to grow on the medium in the absence of thiamine yet growth was restored when exogenous thiamine was provided. A linear relationship between cell mass formation and amount of thiamine supplemented was mathematically determined in a shake flask experiment and confirmed in a 7-L bioreactor system. This calculation revealed that ∼ 95-96 thiamine molecules per cell were required to satisfy cell growth. This relationship was employed to develop a novel fermentation process for D-lactate production by using thiamine as a limiting condition. A D-lactate productivity of 4.11 g · L(-1) · h(-1) from glycerol under microaerobic condition and 3.66 g · L(-1) · h(-1) from glucose under anaerobic condition was achieved which is 19.1% and 10.2% higher respectively than the parental strain. These results revealed a convenient and reliable method to control cell growth and improve D-lactate fermentation. This control strategy could be applied to other biotechnological processes that require optimal allocation of carbon between cell growth and product formation. © 2015 Wiley Periodicals, Inc.

γ-Aminobutyric Acid (GABA) Production and Angiotensin-I Converting Enzyme (ACE) Inhibitory Activity of Fermented Soybean Containing Sea Tangle by the Co-Culture of Lactobacillus brevis with Aspergillus oryzae.

PubMed

Jang, Eun Kyeong; Kim, Nam Yeun; Ahn, Hyung Jin; Ji, Geun Eog

2015-08-01

To enhance the γ-aminobutyric acid (GABA) content, the optimized fermentation of soybean with added sea tangle extract was evaluated at 30°C and pH 5.0. The medium was first inoculated with Aspergillus oryzae strain FMB S46471 and fermented for 3 days, followed by the subsequent inoculation with Lactobacillus brevis GABA 100. After fermentation for 7 days, the fermented soybean showed approximately 1.9 g/kg GABA and exhibited higher ACE inhibitory activity than the traditional soybean product. Furthermore, several peptides in the fraction containing the highest ACE inhibitory activity were identified. The novel fermented soybean enriched with GABA and ACE inhibitory components has great pharmaceutical and functional food values.

Microbiota Dynamics Associated with Environmental Conditions and Potential Roles of Cellulolytic Communities in Traditional Chinese Cereal Starter Solid-State Fermentation

PubMed Central

Li, Pan; Liang, Hebin; Lin, Wei-Tie; Feng, Feng

2015-01-01

Traditional Chinese solid-state fermented cereal starters contain highly complex microbial communities and enzymes. Very little is known, however, about the microbial dynamics related to environmental conditions, and cellulolytic communities have never been proposed to exist during cereal starter fermentation. In this study, we performed Illumina MiSeq sequencing combined with PCR-denaturing gradient gel electrophoresis to investigate microbiota, coupled with clone library construction to trace cellulolytic communities in both fermentation stages. A succession of microbial assemblages was observed during the fermentation of starters. Lactobacillales and Saccharomycetales dominated the initial stages, with a continuous decline in relative abundance. However, thermotolerant and drought-resistant Bacillales, Eurotiales, and Mucorales were considerably accelerated during the heating stages, and these organisms dominated until the end of fermentation. Enterobacteriales were consistently ubiquitous throughout the process. For the cellulolytic communities, only the genera Sanguibacter, Beutenbergia, Agrobacterium, and Erwinia dominated the initial fermentation stages. In contrast, stages at high incubation temperature induced the appearance and dominance of Bacillus, Aspergillus, and Mucor. The enzymatic dynamics of amylase and glucoamylase also showed a similar trend, with the activities clearly increased in the first 7 days and subsequently decreased until the end of fermentation. Furthermore, β-glucosidase activity continuously and significantly increased during the fermentation process. Evidently, cellulolytic potential can adapt to environmental conditions by changes in the community structure during the fermentation of starters. PMID:26002897

U.S. Army Medical Research Institute of Infectious Disease Annual Progress Report, Fiscal Year 1985

DTIC Science & Technology

1985-10-01

response. 25. (U) 8410-8509-Tangential flow filtration was developed as a safer method of harvesting the 50-L anthrax fermenter cultures...yearly boosters. Efforts over several years have sought to improve this chemical vaccine through optimization of fermenter and recovery techniques so...A collaborative effort with WLAIR led to production of candidate human hybridomas to PA. Progrses: The 50-liter fermenter continues to be used

Bagasse hydrolyzates from Agave tequilana as substrates for succinic acid production by Actinobacillus succinogenes in batch and repeated batch reactor.

PubMed

Corona-González, Rosa Isela; Varela-Almanza, Karla María; Arriola-Guevara, Enrique; Martínez-Gómez, Álvaro de Jesús; Pelayo-Ortiz, Carlos; Toriz, Guillermo

2016-04-01

The aim of this work was to obtain fermentable sugars by enzymatic or acid hydrolyses of Agave tequilana Weber bagasse in order to produce succinic acid with Actinobacillus succinogenes. Hydrolyses were carried out with mineral acids (sulfuric and hydrochloric acids) or a commercial cellulolytic enzyme, and were optimized statistically by a response surface methodology, having as factors the concentration of acid/enzyme and time of hydrolysis. The concentration of sugars obtained at optimal conditions for each hydrolysis were 21.7, 22.4y 19.8g/L for H2SO4, HCl and the enzymatic preparation respectively. Concerning succinic acid production, the enzymatic hydrolyzates resulted in the highest yield (0.446g/g) and productivity (0.57g/Lh) using A. succinogenes in a batch reactor system. Repeated batch fermentation with immobilized A. succinogenes in agar and with the enzymatic hydrolyzates resulted in a maximum concentration of succinic acid of 33.6g/L from 87.2g/L monosaccharides after 5 cycles in 40h, obtaining a productivity of 1.32g/Lh. Copyright © 2016. Published by Elsevier Ltd.

Production of a cellulase-free alkaline xylanase from Bacillus pumilus MTCC 5015 by submerged fermentation and its application in biobleaching.

PubMed

Thomas, Leya; Sindhu, Raveendran; Binod, Parameswaran; Pandey, Ashok

2015-06-01

Here, we described the production of a cellulase-free alkaline xylanase from Bacillus pumilus MTCC 5015 by submerged fermentation and its application in biobleaching. Various process parameters affecting xylanase production by B. pumilus were optimized by adopting a Plackett-Burman design (PBD) as well as Response surface methodology (RSM). These statistical methods aid in improving the enzyme yield by analysing the individual crucial components of the medium. Maximum production was obtained with 4% yeast extract, 0.08% magnesium sulphate, 30 h of inoculum age, incubation temperature of 33.5 degrees C and pH 9.0. Under optimized conditions, the xylanase activity was 372 IU/ml. Media engineering improved a 5-fold increase in the enzyme production. Scanning electron microscopy (SEM) showed significant changes on the surface of xylanase treated pulps as a result of xylan hydrolysis. Increased roughness of paper carton fibres was apparent in scanning electron micrograph due to opening of the micro fibrils present on the surface by xylanase action. The untreated pulp did not show any such change. These results demonstrated that the B. pumilus MTCC 5015 xylanase was effective in bio-bleaching of paper carton.

Characterization of Lignocellulolytic Activities from a Moderate Halophile Strain of Aspergillus caesiellus Isolated from a Sugarcane Bagasse Fermentation

PubMed Central

Miranda-Miranda, Estefan; Sánchez-Reyes, Ayixón; Cuervo-Soto, Laura; Aceves-Zamudio, Denise; Atriztán-Hernández, Karina; Morales-Herrera, Catalina; Rodríguez-Hernández, Rocío; Folch-Mallol, Jorge

2014-01-01

A moderate halophile and thermotolerant fungal strain was isolated from a sugarcane bagasse fermentation in the presence of 2 M NaCl that was set in the laboratory. This strain was identified by polyphasic criteria as Aspergillus caesiellus. The fungus showed an optimal growth rate in media containing 1 M NaCl at 28°C and could grow in media added with up to 2 M NaCl. This strain was able to grow at 37 and 42°C, with or without NaCl. A. caesiellus H1 produced cellulases, xylanases, manganese peroxidase (MnP) and esterases. No laccase activity was detected in the conditions we tested. The cellulase activity was thermostable, halostable, and no differential expression of cellulases was observed in media with different salt concentrations. However, differential band patterns for cellulase and xylanase activities were detected in zymograms when the fungus was grown in different lignocellulosic substrates such as wheat straw, maize stover, agave fibres, sugarcane bagasse and sawdust. Optimal temperature and pH were similar to other cellulases previously described. These results support the potential of this fungus to degrade lignocellulosic materials and its possible use in biotechnological applications. PMID:25162614

Production, characteristics and applications of phytase from a rhizosphere isolated Enterobacter sp. ACSS.

PubMed

Chanderman, Ashira; Puri, Adarsh Kumar; Permaul, Kugen; Singh, Suren

2016-10-01

Optimization of process parameters for phytase production by Enterobacter sp. ACSS led to a 4.6-fold improvement in submerged fermentation, which was enhanced further in fed-batch fermentation. The purified 62 kDa monomeric phytase was optimally active at pH 2.5 and 60 °C and retained activity over a wide range of temperature (40-80 °C) and pH (2.0-6.0) with a half-life of 11.3 min at 80 °C. The kinetic parameters K m, V max, K cat, and K cat/K m of the pure phytase were 0.21 mM, 131.58 nmol mg(-1) s(-1), 1.64 × 10(3) s(-1), and 7.81 × 10(6) M(-1) s(-1), respectively. The enzyme was fairly stable in the presence of pepsin under physiological conditions. It was stimulated by Ca(+2), Mg(+2) and Mn(+2), but inhibited by Zn(+2), Cu(+2), Fe(+2), Pb(+2), Ba(+2) and surfactants. The enzyme can be applied in dephytinizing animal feeds, and the baking industry.

Integrated use of residues from olive mill and winery for lipase production by solid state fermentation with Aspergillus sp.

PubMed

Salgado, José Manuel; Abrunhosa, Luís; Venâncio, Armando; Domínguez, José Manuel; Belo, Isabel

2014-02-01

Two-phase olive mill waste (TPOMW) is presently the major waste produced by the olive mill industry. This waste has potential to be used as substrate for solid state fermentation (SSF) despite of its high concentration of phenolic compounds and low nitrogen content. In this work, it is demonstrated that mixtures of TPOMW with winery wastes support the production of lipase by Aspergillus spp. By agar plate screening, Aspergillus niger MUM 03.58, Aspergillus ibericus MUM 03.49, and Aspergillus uvarum MUM 08.01 were chosen for lipase production by SSF. Plackett-Burman experimental design was employed to evaluate the effect of substrate composition and time on lipase production. The highest amounts of lipase were produced by A. ibericus on a mixture of TPOMW, urea, and exhausted grape mark (EGM). Urea was found to be the most influent factor for the lipase production. Further optimization of lipase production by A. ibericus using a full factorial design (3(2)) conducted to optimal conditions of substrate composition (0.073 g urea/g and 25 % of EGM) achieve 18.67 U/g of lipolytic activity.

Transcription profile of brewery yeast under fermentation conditions.

PubMed

James, T C; Campbell, S; Donnelly, D; Bond, U

2003-01-01

Yeast strains, used in the brewing industry, experience distinctive physiological conditions. During a brewing fermentation, yeast are exposed to anaerobic conditions, high pressure, high specific gravity and low temperatures. The purpose of this study was to examine the global gene expression profile of yeast subjected to brewing stress. We have carried out a microarray analysis of a typical brewer's yeast during the course of an 8-day fermentation in 15 degrees P wort. We used the probes derived from Saccharomyces cerevisiae genomic DNA on the chip and RNA isolated from three stages of brewing. This analysis shows a high level of expression of genes involved in fatty acid and ergosterol biosynthesis early in fermentation. Furthermore, genes involved in respiration and mitochondrial protein synthesis also show higher levels of expression. Surprisingly, we observed a complete repression of many stress response genes and genes involved in protein synthesis throughout the 8-day period compared with that at the start of fermentation. This microarray data set provides an analysis of gene expression under brewing fermentation conditions. The data provide an insight into the various metabolic processes altered or activated by brewing conditions of growth. This study leads to future experiments whereby selective alterations in brewing conditions could be introduced to take advantage of the changing transcript profile to improve the quality of the brew.

Impact of an energy-conserving strategy on succinate production under weak acidic and anaerobic conditions in Enterobacter aerogenes.

PubMed

Tajima, Yoshinori; Yamamoto, Yoko; Fukui, Keita; Nishio, Yousuke; Hashiguchi, Kenichi; Usuda, Yoshihiro; Sode, Koji

2015-06-11

Succinate is an important C4 building block chemical, and its production via fermentative processes in bacteria has many practical applications in the biotechnology field. One of the major goals of optimizing the bacterium-based succinate production process is to lower the culture pH from the current neutral conditions, as this would reduce total production costs. In our previous studies, we selected Enterobacter aerogenes, a rapid glucose assimilator at pH 5.0, in order to construct a metabolically engineered strain that could produce succinate under weakly acidic conditions. This engineered strain produced succinate from glucose with a 72.7% (g/g) yield at pH 5.7, with a volumetric productivity of 0.23 g/L/h. Although this demonstrates proof-of-concept that bacterium-based succinate fermentation can be improved under weakly acidic conditions, several parameters still required further optimization. In this study, we genetically modified an E. aerogenes strain previously developed in our laboratory in order to increase the production of ATP during succinate synthesis, as we inferred that this would positively impact succinate biosynthesis. This led to the development of the ES08ΔptsG strain, which contains the following modifications: chromosomally expressed Actinobacillus succinogenes phosphoenolpyruvate carboxykinase, enhanced fumarate reductase, inactivated pyruvate formate lyase, pyruvate oxidase, and glucose-phosphotransferase permease (enzyme IIBC(Glc)). This strain produced 55.4 g/L succinate from glucose, with 1.8 g/L acetate as the major byproduct at pH 5.7 and anaerobic conditions. The succinate yield and volumetric productivity of this strain were 86.8% and 0.92 g/L/h, respectively. Focusing on increasing net ATP production during succinate synthesis leads to increased succinate yield and volumetric productivity in E. aerogenes. We propose that the metabolically engineered E. aerogenes ES08ΔptsG strain, which effectively produces succinate under weakly acidic and anaerobic conditions, has potential utility for economical succinate production.

Volatile fatty acids productions by mesophilic and thermophilic sludge fermentation: Biological responses to fermentation temperature.

PubMed

Hao, Jiuxiao; Wang, Hui

2015-01-01

The volatile fatty acids (VFAs) productions, as well as hydrolases activities, microbial communities, and homoacetogens, of mesophilic and thermophilic sludge anaerobic fermentation were investigated to reveal the microbial responses to different fermentation temperatures. Thermophilic fermentation led to 10-fold more accumulation of VFAs compared to mesophilic fermentation. α-glucosidase and protease had much higher activities in thermophilic reactor, especially protease. Illumina sequencing manifested that raising fermentation temperature increased the abundances of Clostridiaceae, Microthrixaceae and Thermotogaceae, which could facilitate either hydrolysis or acidification. Real-time PCR analysis demonstrated that under thermophilic condition the relative abundance of homoacetogens increased in batch tests and reached higher level at stable fermentation, whereas under mesophilic condition it only increased slightly in batch tests. Therefore, higher fermentation temperature increased the activities of key hydrolases, raised the proportions of bacteria involved in hydrolysis and acidification, and promoted the relative abundance of homoacetogens, which all resulted in higher VFAs production. Copyright © 2014 Elsevier Ltd. All rights reserved.