Modeling of the pyruvate production with Escherichia coli: comparison of mechanistic and neural networks-based models.

PubMed

Zelić, B; Bolf, N; Vasić-Racki, D

2006-06-01

Three different models: the unstructured mechanistic black-box model, the input-output neural network-based model and the externally recurrent neural network model were used to describe the pyruvate production process from glucose and acetate using the genetically modified Escherichia coli YYC202 ldhA::Kan strain. The experimental data were used from the recently described batch and fed-batch experiments [ Zelić B, Study of the process development for Escherichia coli-based pyruvate production. PhD Thesis, University of Zagreb, Faculty of Chemical Engineering and Technology, Zagreb, Croatia, July 2003. (In English); Zelić et al. Bioproc Biosyst Eng 26:249-258 (2004); Zelić et al. Eng Life Sci 3:299-305 (2003); Zelić et al Biotechnol Bioeng 85:638-646 (2004)]. The neural networks were built out of the experimental data obtained in the fed-batch pyruvate production experiments with the constant glucose feed rate. The model validation was performed using the experimental results obtained from the batch and fed-batch pyruvate production experiments with the constant acetate feed rate. Dynamics of the substrate and product concentration changes was estimated using two neural network-based models for biomass and pyruvate. It was shown that neural networks could be used for the modeling of complex microbial fermentation processes, even in conditions in which mechanistic unstructured models cannot be applied.

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.

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.

Integrated production of lactic acid and biomass on distillery stillage.

PubMed

Djukić-Vuković, Aleksandra P; Mojović, Ljiljana V; Vukašinović-Sekulić, Maja S; Nikolić, Svetlana B; Pejin, Jelena D

2013-09-01

The possibilities of parallel lactic acid and biomass production in batch and fed-batch fermentation on distillery stillage from bioethanol production were studied. The highest lactic acid yield and productivity of 92.3 % and 1.49 g L(-1) h(-1) were achieved in batch fermentation with initial sugar concentration of 55 g L(-1). A significant improvement of the process was achieved in fed-batch fermentation where the concentration of lactic acid was increased to 47.6 % and volumetric productivity for 21 % over the batch process. A high number of Lactobacillus rhamnosus ATCC 7469 viable cells of 10(9) CFU ml(-1) was attained at the end of fed-batch fermentation. The survival of 92.9 % of L. rhamnosus cells after 3 h of incubation at pH 2.5 validated that the fermentation media remained after lactic acid removal could be used as a biomass-enriched animal feed thus making an additional value to the process.

Improved Mannanase Production from Penicillium occitanis by Fed-Batch Fermentation Using Acacia Seeds

PubMed Central

Blibech, Monia; Ellouz Ghorbel, Raoudha; Chaari, Fatma; Dammak, Ilyes; Bhiri, Fatma; Neifar, Mohamed; Ellouz Chaabouni, Semia

2011-01-01

By applying a fed-batch strategy, production of Penicillium occitanis mannanases could be almost doubled as compared to a batch cultivation on acacia seeds (76 versus 41 U/mL). Also, a 10-fold increase of enzyme activities was observed from shake flask fermentation to the fed-batch fermentation. These production levels were 3-fold higher than those obtained on coconut meal. The high mannanase production using acacia seeds powder as inducer substrate showed the suitability of this culture process for industrial-scale development. PMID:23724314

Bioprocessing Data for the Production of Marine Enzymes

PubMed Central

Sarkar, Sreyashi; Pramanik, Arnab; Mitra, Anindita; Mukherjee, Joydeep

2010-01-01

This review is a synopsis of different bioprocess engineering approaches adopted for the production of marine enzymes. Three major modes of operation: batch, fed-batch and continuous have been used for production of enzymes (such as protease, chitinase, agarase, peroxidase) mainly from marine bacteria and fungi on a laboratory bioreactor and pilot plant scales. Submerged, immobilized and solid-state processes in batch mode were widely employed. The fed-batch process was also applied in several bioprocesses. Continuous processes with suspended cells as well as with immobilized cells have been used. Investigations in shake flasks were conducted with the prospect of large-scale processing in reactors. PMID:20479981

Ethanol fermentation integrated with PDMS composite membrane: An effective process.

PubMed

Fu, Chaohui; Cai, Di; Hu, Song; Miao, Qi; Wang, Yong; Qin, Peiyong; Wang, Zheng; Tan, Tianwei

2016-01-01

The polydimethylsiloxane (PDMS) membrane, prepared in water phase, was investigated in separation ethanol from model ethanol/water mixture and fermentation-pervaporation integrated process. Results showed that the PDMS membrane could effectively separate ethanol from model solution. When integrated with batch ethanol fermentation, the ethanol productivity was enhanced compared with conventional process. Fed-batch and continuous ethanol fermentation with pervaporation were also performed and studied. 396.2-663.7g/m(2)h and 332.4-548.1g/m(2)h of total flux with separation factor of 8.6-11.7 and 8-11.6, were generated in the fed-batch and continuous fermentation with pervaporation scenario, respectively. At the same time, high titre ethanol production of ∼417.2g/L and ∼446.3g/L were also achieved on the permeate side of membrane in the two scenarios, respectively. The integrated process was environmental friendly and energy saving, and has a promising perspective in long-terms operation. Copyright © 2015 Elsevier Ltd. All rights reserved.

Dynamic genome-scale metabolic modeling of the yeast Pichia pastoris.

PubMed

Saitua, Francisco; Torres, Paulina; Pérez-Correa, José Ricardo; Agosin, Eduardo

2017-02-21

Pichia pastoris shows physiological advantages in producing recombinant proteins, compared to other commonly used cell factories. This yeast is mostly grown in dynamic cultivation systems, where the cell's environment is continuously changing and many variables influence process productivity. In this context, a model capable of explaining and predicting cell behavior for the rational design of bioprocesses is highly desirable. Currently, there are five genome-scale metabolic reconstructions of P. pastoris which have been used to predict extracellular cell behavior in stationary conditions. In this work, we assembled a dynamic genome-scale metabolic model for glucose-limited, aerobic cultivations of Pichia pastoris. Starting from an initial model structure for batch and fed-batch cultures, we performed pre/post regression diagnostics to ensure that model parameters were identifiable, significant and sensitive. Once identified, the non-relevant ones were iteratively fixed until a priori robust modeling structures were found for each type of cultivation. Next, the robustness of these reduced structures was confirmed by calibrating the model with new datasets, where no sensitivity, identifiability or significance problems appeared in their parameters. Afterwards, the model was validated for the prediction of batch and fed-batch dynamics in the studied conditions. Lastly, the model was employed as a case study to analyze the metabolic flux distribution of a fed-batch culture and to unravel genetic and process engineering strategies to improve the production of recombinant Human Serum Albumin (HSA). Simulation of single knock-outs indicated that deviation of carbon towards cysteine and tryptophan formation improves HSA production. The deletion of methylene tetrahydrofolate dehydrogenase could increase the HSA volumetric productivity by 630%. Moreover, given specific bioprocess limitations and strain characteristics, the model suggests that implementation of a decreasing specific growth rate during the feed phase of a fed-batch culture results in a 25% increase of the volumetric productivity of the protein. In this work, we formulated a dynamic genome scale metabolic model of Pichia pastoris that yields realistic metabolic flux distributions throughout dynamic cultivations. The model can be calibrated with experimental data to rationally propose genetic and process engineering strategies to improve the performance of a P. pastoris strain of interest.

40 CFR 63.161 - Definitions.

Code of Federal Regulations, 2012 CFR

2012-07-01

... subpart that references this subpart. Batch process means a process in which the equipment is fed... generally emptied. Examples of industries that use batch processes include pharmaceutical production and pesticide production. Batch product-process equipment train means the collection of equipment (e.g...

40 CFR 63.161 - Definitions.

Code of Federal Regulations, 2013 CFR

2013-07-01

... subpart that references this subpart. Batch process means a process in which the equipment is fed... generally emptied. Examples of industries that use batch processes include pharmaceutical production and pesticide production. Batch product-process equipment train means the collection of equipment (e.g...

40 CFR 63.161 - Definitions.

Code of Federal Regulations, 2014 CFR

2014-07-01

... subpart that references this subpart. Batch process means a process in which the equipment is fed... generally emptied. Examples of industries that use batch processes include pharmaceutical production and pesticide production. Batch product-process equipment train means the collection of equipment (e.g...

40 CFR 63.161 - Definitions.

Code of Federal Regulations, 2011 CFR

2011-07-01

... subpart that references this subpart. Batch process means a process in which the equipment is fed... generally emptied. Examples of industries that use batch processes include pharmaceutical production and pesticide production. Batch product-process equipment train means the collection of equipment (e.g...

Kinetic modeling of multi-feed simultaneous saccharification and co-fermentation of pretreated birch to ethanol.

PubMed

Wang, Ruifei; Koppram, Rakesh; Olsson, Lisbeth; Franzén, Carl Johan

2014-11-01

Fed-batch simultaneous saccharification and fermentation (SSF) is a feasible option for bioethanol production from lignocellulosic raw materials at high substrate concentrations. In this work, a segregated kinetic model was developed for simulation of fed-batch simultaneous saccharification and co-fermentation (SSCF) of steam-pretreated birch, using substrate, enzymes and cell feeds. The model takes into account the dynamics of the cellulase-cellulose system and the cell population during SSCF, and the effects of pre-cultivation of yeast cells on fermentation performance. The model was cross-validated against experiments using different feed schemes. It could predict fermentation performance and explain observed differences between measured total yeast cells and dividing cells very well. The reproducibility of the experiments and the cell viability were significantly better in fed-batch than in batch SSCF at 15% and 20% total WIS contents. The model can be used for simulation of fed-batch SSCF and optimization of feed profiles. Copyright © 2014 Elsevier Ltd. All rights reserved.

Fumaric Acid Production from Alkali-Pretreated Corncob by Fed-Batch Simultaneous Saccharification and Fermentation Combined with Separated Hydrolysis and Fermentation at High Solids Loading.

PubMed

Li, Xin; Zhou, Jin; Ouyang, Shuiping; Ouyang, Jia; Yong, Qiang

2017-02-01

Production of fumaric acid from alkali-pretreated corncob (APC) at high solids loading was investigated using a combination of separated hydrolysis and fermentation (SHF) and fed-batch simultaneous saccharification and fermentation (SSF) by Rhizopus oryzae. Four different fermentation modes were tested to maximize fumaric acid concentration at high solids loading. The highest concentration of 41.32 g/L fumaric acid was obtained from 20 % (w/v) APC at 38 °C in the combined SHF and fed-batch SSF process, compared with 19.13 g/L fumaric acid in batch SSF alone. The results indicated that a combination of SHF and fed-batch SSF significantly improved production of fumaric acid from lignocellulose by R. oryzae than that achieved with batch SSF at high solids loading.

L-Lactic Acid Production by Lactobacillus rhamnosus ATCC 10863

PubMed Central

Senedese, Ana Lívia Chemeli; Maciel Filho, Rubens; Maciel, Maria Regina Wolf

2015-01-01

Lactic acid has been shown to have the most promising application in biomaterials as poly(lactic acid). L. rhamnosus ATCC 10863 that produces L-lactic acid was used to perform the fermentation and molasses was used as substrate. A solution containing 27.6 g/L of sucrose (main composition of molasses) and 3.0 g/L of yeast extract was prepared, considering the final volume of 3,571 mL (14.0% (v/v) inoculum). Batch and fed batch fermentations were performed with temperature of 43.4°C and pH of 5.0. At the fed batch, three molasses feed were applied at 12, 24, and 36 hours. Samples were taken every two hours and the amounts of lactic acid, sucrose, glucose, and fructose were determined by HPLC. The sucrose was barely consumed at both processes; otherwise the glucose and fructose were almost entirely consumed. 16.5 g/L of lactic acid was produced at batch and 22.0 g/L at fed batch. Considering that lactic acid was produced due to the low concentration of the well consumed sugars, the final amount was considerable. The cell growth was checked and no substrate inhibition was observed. A sucrose molasses hydrolysis is suggested to better avail the molasses fermentation with this strain, surely increasing the L-lactic acid. PMID:25922852

Conversion of cellulosic materials into glycolipid biosurfactants, mannosylerythritol lipids, by Pseudozyma spp. under SHF and SSF processes.

PubMed

Faria, Nuno Torres; Santos, Marisa; Ferreira, Carla; Marques, Susana; Ferreira, Frederico Castelo; Fonseca, César

2014-11-04

Mannosylerythritol lipids (MEL) are glycolipids with unique biosurfactant properties and are produced by Pseudozyma spp. from different substrates, preferably vegetable oils, but also sugars, glycerol or hydrocarbons. However, solvent intensive downstream processing and the relatively high prices of raw materials currently used for MEL production are drawbacks in its sustainable commercial deployment. The present work aims to demonstrate MEL production from cellulosic materials and investigate the requirements and consequences of combining commercial cellulolytic enzymes and Pseudozyma spp. under separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) processes. MEL was produced from cellulosic substrates, Avicel® as reference (>99% cellulose) and hydrothermally pretreated wheat straw, using commercial cellulolytic enzymes (Celluclast 1.5 L® and Novozyme 188®) and Pseudozyma antarctica PYCC 5048(T) or Pseudozyma aphidis PYCC 5535(T). The strategies included SHF, SSF and fed-batch SSF with pre-hydrolysis. While SSF was isothermal at 28°C, in SHF and fed-batch SSF, yeast fermentation was preceded by an enzymatic (pre-)hydrolysis step at 50°C for 48 h. Pseudozyma antarctica showed the highest MEL yields from both cellulosic substrates, reaching titres of 4.0 and 1.4 g/l by SHF of Avicel® and wheat straw (40 g/l glucan), respectively, using enzymes at low dosage (3.6 and 8.5 FPU/gglucan at 28°C and 50°C, respectively) with prior dialysis. Higher MEL titres were obtained by fed-batch SSF with pre-hydrolysis, reaching 4.5 and 2.5 g/l from Avicel® and wheat straw (80 g/l glucan), respectively. This work reports for the first time MEL production from cellulosic materials. The process was successfully performed through SHF, SSF or Fed-batch SSF, requiring, for maximal performance, dialysed commercial cellulolytic enzymes. The use of inexpensive lignocellulosic substrates associated to straightforward downstream processing from sugary broths is expected to have a great impact in the economy of MEL production for the biosurfactant market, inasmuch as low enzyme dosage is sufficient for good systems performance.

High Solid Fed-batch Butanol Fermentation with Simultaneous Product Recovery: Part II - Process Integration.

PubMed

Qureshi, Nasib; Klasson, K Thomas; Saha, Badal C; Liu, Siqing

2018-04-25

In these studies liquid hot water (LHW) pretreated and enzymatically hydrolyzed Sweet Sorghum Bagasse (SSB) hydrolyzates were fermented in a fed-batch reactor. As reported in the preceding paper, the culture was not able to ferment the hydrolyzate I in a batch process due to presence of high level of toxic chemicals, in particular acetic acid released from SSB during the hydrolytic process. To be able to ferment the hydrolyzate I obtained from 250 gL -1 SSB hydrolysis, a fed-batch reactor with in-situ butanol recovery was devised. The process was started with the hydrolyzate II and when good cell growth and vigorous fermentation were observed, the hydrolyzate I was slowly fed to the reactor. In this manner the culture was able to ferment all the sugars present in both the hydrolyzates to acetone butanol ethanol (ABE). In a control batch reactor in which ABE was produced from glucose, ABE productivity and yield of 0.42 gL -1 h -1 and 0.36 were obtained, respectively. In the fed-batch reactor fed with SSB hydrolyzates these productivity and yield values were 0.44 gL -1 h -1 and 0.45, respectively. ABE yield in the integrated system was high due to utilization of acetic acid to convert to ABE. In summary we were able to utilize both the hydrolyzates obtained from LHW pretreated and enzymatically hydrolyzed SSB (250 gL -1 ) and convert them to ABE. Complete fermentation was possible due to simultaneous recovery of ABE by vacuum. This article is protected by copyright. All rights reserved. © 2018 American Institute of Chemical Engineers.

Enhanced production and immunological characterization of recombinant West Nile virus envelope domain III protein.

PubMed

Tripathi, Nagesh K; Karothia, Divyanshi; Shrivastava, Ambuj; Banger, Swati; Kumar, Jyoti S

2018-05-13

West Nile virus (WNV) is an emerging mosquito-borne virus which is responsible for severe and fatal encephalitis in humans and for which there is no licensed vaccine or therapeutic available to prevent infection. The envelope domain III protein (EDIII) of WNV was over-expressed in Escherichia coli and purified using a two-step chromatography process which included immobilized metal affinity chromatography and ion exchange chromatography. E. coli cells were grown in a bioreactor to high density using batch and fed-batch cultivation. Wet biomass obtained after batch and fed-batch cultivation processes was 11.2 g and 84 g/L of culture respectively. Protein yield after affinity purification was 5.76 mg and 5.81 mg/g wet cell weight after batch and fed-batch processes respectively. The purified WNV EDIII elicited specific antibodies in rabbits, confirming its immunogenicity. Moreover, the antibodies were able to neutralize WNV in vitro. These results established that the refolded and purified WNV EDIII could be a potential vaccine candidate. Copyright © 2018 Elsevier B.V. All rights reserved.

Soft sensor modeling based on variable partition ensemble method for nonlinear batch processes

NASA Astrophysics Data System (ADS)

Wang, Li; Chen, Xiangguang; Yang, Kai; Jin, Huaiping

2017-01-01

Batch processes are always characterized by nonlinear and system uncertain properties, therefore, the conventional single model may be ill-suited. A local learning strategy soft sensor based on variable partition ensemble method is developed for the quality prediction of nonlinear and non-Gaussian batch processes. A set of input variable sets are obtained by bootstrapping and PMI criterion. Then, multiple local GPR models are developed based on each local input variable set. When a new test data is coming, the posterior probability of each best performance local model is estimated based on Bayesian inference and used to combine these local GPR models to get the final prediction result. The proposed soft sensor is demonstrated by applying to an industrial fed-batch chlortetracycline fermentation process.

Scale-up of industrial biodiesel production to 40 m(3) using a liquid lipase formulation.

PubMed

Price, Jason; Nordblad, Mathias; Martel, Hannah H; Chrabas, Brent; Wang, Huali; Nielsen, Per Munk; Woodley, John M

2016-08-01

In this work, we demonstrate the scale-up from an 80 L fed-batch scale to 40 m(3) along with the design of a 4 m(3) continuous process for enzymatic biodiesel production catalyzed by NS-40116 (a liquid formulation of a modified Thermomyces lanuginosus lipase). Based on the analysis of actual pilot plant data for the transesterification of used cooking oil and brown grease, we propose a method applying first order integral analysis to fed-batch data based on either the bound glycerol or free fatty acid content in the oil. This method greatly simplifies the modeling process and gives an indication of the effect of mixing at the various scales (80 L to 40 m(3) ) along with the prediction of the residence time needed to reach a desired conversion in a CSTR. Suitable process metrics reflecting commercial performance such as the reaction time, enzyme efficiency, and reactor productivity were evaluated for both the fed-batch and CSTR cases. Given similar operating conditions, the CSTR operation on average, has a reaction time which is 1.3 times greater than the fed-batch operation. We also showed how the process metrics can be used to quickly estimate the selling price of the enzyme. Assuming a biodiesel selling price of 0.6 USD/kg and a one-time use of the enzyme (0.1% (w/woil ) enzyme dosage); the enzyme can then be sold for 30 USD/kg which ensures that that the enzyme cost is not more than 5% of the biodiesel revenue. Biotechnol. Bioeng. 2016;113: 1719-1728. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Production of biomethane from palm oil mill effluent (POME) with fed batch system in beam-shaped digester

NASA Astrophysics Data System (ADS)

Aznury, Martha; Amin, Jaksen M.; Hasan, Abu; Himmatuliza, Astinesia

2017-05-01

Palm oil mill effluent (POME) is the biggest liquid waste which is produced from palm oil production. POME are containing organic matter, high levels of biological oxygen demand (BOD) and chemical oxygen demand (COD) were 28000 mg/L and 48000 mg/L. To reduce the levels of pollution caused by POME, is necessary to do stages of processing using a biological process that involves aerobic and anaerobic bacteria so that it can be utilized as a new product that has economic value, one is biogas. The processing into biogas in anaerobic performed by fed batch system. In the ratio between POME and activated microorganismes are 70:30%. The process of anaerobic fermentation in fed batch is done by time variation of the addition of the substrate. The mixture of POME and activated microorganismes were fermented for a month and then after one month substrates were added gradually as much as 1 liter into the digester with a variety of additional time are 1, 2, and 5 days. The interval of addition of the substrate give effect to the pH and the quantity of biogas produced. The highest increasing of the quantity of biomethane was 25.14 mol% at the time the addition of substrate every fifth day.

Optimization of fed-batch enzymatic hydrolysis from alkali-pretreated sugarcane bagasse for high-concentration sugar production.

PubMed

Gao, Yueshu; Xu, Jingliang; Yuan, Zhenhong; Zhang, Yu; Liu, Yunyun; Liang, Cuiyi

2014-09-01

Fed-batch enzymatic hydrolysis process from alkali-pretreated sugarcane bagasse was investigated to increase solids loading, produce high-concentration fermentable sugar and finally to reduce the cost of the production process. The optimal initial solids loading, feeding time and quantities were examined. The hydrolysis system was initiated with 12% (w/v) solids loading in flasks, where 7% fresh solids were fed consecutively at 6h, 12h, 24h to get a final solids loading of 33%. All the requested cellulase loading (10 FPU/g substrate) was added completely at the beginning of hydrolysis reaction. After 120 h of hydrolysis, the maximal concentrations of cellobiose, glucose and xylose obtained were 9.376 g/L, 129.50 g/L, 56.03 g/L, respectively. The final total glucan conversion rate attained to 60% from this fed-batch process. Copyright © 2014. Published by Elsevier Ltd.

Effects of the feeding ratio of food waste on fed-batch aerobic composting and its microbial community.

PubMed

Wang, Xiaojun; Pan, Songqing; Zhang, Zhaoji; Lin, Xiangyu; Zhang, Yuzhen; Chen, Shaohua

2017-01-01

To determine the suitable feeding ratio for fed-batch aerobic composting, four fermenters were operated by adding 0%, 5%, 10% or 15% of food waste every day. The results showed that the 5% and 10% treatments were able to maintain continuous thermophilic conditions, while the 15% treatment performed badly in regard to composting temperature, which was probably due to the negative effects of excessive moisture on microbial activity. As composting proceeded, both the 5% and the 10% treatments reached maturity and achieved weight losses of approximately 65%. High-throughput sequencing results indicated that Firmicutes, Proteobacteria, Bacteroidetes and Actinobacteria were the dominant phyla of the community structure. The communities sampled at the thermophilic phases had high similarity and relatively low diversity, while species diversity increased in the maturity phase. This study was devoted to optimizing the fed-batch composting process and assessing bacterial communities, both of which were supplied as a reference for practical application. Copyright © 2016 Elsevier Ltd. All rights reserved.

High-performance recombinant protein production with Escherichia coli in continuously operated cascades of stirred-tank reactors.

PubMed

Schmideder, Andreas; Weuster-Botz, Dirk

2017-07-01

The microbial expression of intracellular, recombinant proteins in continuous bioprocesses suffers from low product concentrations. Hence, a process for the intracellular production of photoactivatable mCherry with Escherichia coli in a continuously operated cascade of two stirred-tank reactors was established to separate biomass formation (first reactor) and protein expression (second reactor) spatially. Cascades of miniaturized stirred-tank reactors were implemented, which enable the 24-fold parallel characterization of cascade processes and the direct scale-up of results to the liter scale. With PAmCherry concentrations of 1.15 g L -1 cascades of stirred-tank reactors improved the process performance significantly compared to production processes in chemostats. In addition, an optimized fed-batch process was outperformed regarding space-time yield (149 mg L -1  h -1 ). This study implicates continuous cascade processes to be a promising alternative to fed-batch processes for microbial protein production and demonstrates that miniaturized stirred-tank reactors can reduce the timeline and costs for cascade process characterization.

Brewery and liquid manure wastewaters as potential feedstocks for microbial fuel cells: a performance study.

PubMed

Angosto, J M; Fernández-López, J A; Godínez, C

2015-01-01

This work aims at the comparison of the electrical and chemical performance of microbial fuel cells (MFCs) fed with several types of brewery and manure industrial wastewaters. Experiments were conducted in a single-cell MFC with the cathode exposed to air operated in batch and fed-batch modes. In fed-batch mode, after 4 days of operation, a standard MFC was refilled with crude wastewater to regenerate the biofilm and recreate initial feeding conditions. Brewery wastewater (CV1) mixed with pig-farm liquid manure (PU sample) gave the highest voltage (199.8 mV) and power density (340 mW/m3) outputs than non-mixed brewery waste water. Also, coulombic efficiency is much larger in the mixture (11%) than in the others (2-3%). However, in terms of chemical oxygen demand removal, the performance showed to be poorer (53%) for the mixed sample than in the pure brewery sample (93%). Fed-batch operation showed to be a good alternate for quasi-continuous operation, with equivalent electrical and chemical yields as compared with normal batchwise operation.

Exploiting the metabolism of PYC expressing HEK293 cells in fed-batch cultures.

PubMed

Vallée, Cédric; Durocher, Yves; Henry, Olivier

2014-01-01

The expression of recombinant yeast pyruvate carboxylase (PYC) in animal cell lines was shown in previous studies to reduce significantly the formation of waste metabolites, although it has translated into mixed results in terms of improved cellular growth and productivity. In this work, we demonstrate that the unique phenotype of PYC expressing cells can be exploited through the application of a dynamic fed-batch strategy and lead to significant process enhancements. Metabolically engineered HEK293 cells stably producing human recombinant IFNα2b and expressing the PYC enzyme were cultured in batch and fed-batch modes. Compared to parental cells, the maximum cell density in batch was increased 1.5-fold and the culture duration was extended by 2.5 days, but the product yield was only marginally increased. Further improvements were achieved by developing and implementing a dynamic fed-batch strategy using a concentrated feed solution. The feeding was based on an automatic control-loop to maintain a constant glucose concentration. This strategy led to a further 2-fold increase in maximum cell density (up to 10.7×10(6)cells/ml) and a final product titer of 160mg/l, representing nearly a 3-fold yield increase compared to the batch process with the parental cell clone. Copyright © 2013 Elsevier B.V. All rights reserved.

Effect of fed-batch vs. continuous mode of operation on microbial fuel cell performance treating biorefinery wastewater

DOE PAGES

Pannell, Tyler C.; Goud, R. Kannaiah; Schell, Daniel J.; ...

2016-05-01

Bioelectrochemical systems have been shown to treat low-value biorefinery streams while recovering energy, however, low current densities and anode conversion efficiencies (ACE) limit their application. A bioanode was developed via enrichment of electroactive biofilm under fed-batch and continuous feeding conditions using corn stover-derived waste stream. The continuously-fed MFC exhibited a current density of 5.8±0.06 A/m 2 and an ACE of 39%±4. The fed-batch MFC achieved a similar current density and an ACE of 19.2%, however, its performance dropped after 36 days of operation to 1.1 A/m 2 and 0.5%, respectively. In comparison, the ACE of the continuously-fed MFC remained stablemore » achieving an ACE of 30% ± 3 after 48 days of operation. An MFC treating a biorefinery stream post fuel separation achieved a current density of 10.7±0.1 A/m 2 and an ACE of 57% ± 9 at an organic loading of 12.5 g COD/L-day. Characterization of the microbial communities indicate higher abundance of Firmicutes and Proteobacteria and lower abundance of Bacteriodetes and a higher level of Geobacter spp. (1.4% vs. 0.2%) in continuously-fed MFC vs. fed-batch MFC. Finally, the results demonstrate that limiting substrate to the equivalent maximum current that the anode can generate, maintains MFC performance over a long term for high strength wastewaters, such as those generated in the biorefinery.« less

Effect of fed-batch vs. continuous mode of operation on microbial fuel cell performance treating biorefinery wastewater

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

Pannell, Tyler C.; Goud, R. Kannaiah; Schell, Daniel J.

Bioelectrochemical systems have been shown to treat low-value biorefinery streams while recovering energy, however, low current densities and anode conversion efficiencies (ACE) limit their application. A bioanode was developed via enrichment of electroactive biofilm under fed-batch and continuous feeding conditions using corn stover-derived waste stream. The continuously-fed MFC exhibited a current density of 5.8±0.06 A/m 2 and an ACE of 39%±4. The fed-batch MFC achieved a similar current density and an ACE of 19.2%, however, its performance dropped after 36 days of operation to 1.1 A/m 2 and 0.5%, respectively. In comparison, the ACE of the continuously-fed MFC remained stablemore » achieving an ACE of 30% ± 3 after 48 days of operation. An MFC treating a biorefinery stream post fuel separation achieved a current density of 10.7±0.1 A/m 2 and an ACE of 57% ± 9 at an organic loading of 12.5 g COD/L-day. Characterization of the microbial communities indicate higher abundance of Firmicutes and Proteobacteria and lower abundance of Bacteriodetes and a higher level of Geobacter spp. (1.4% vs. 0.2%) in continuously-fed MFC vs. fed-batch MFC. Finally, the results demonstrate that limiting substrate to the equivalent maximum current that the anode can generate, maintains MFC performance over a long term for high strength wastewaters, such as those generated in the biorefinery.« less

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.

miRNA profiling of high, low and non-producing CHO cells during biphasic fed-batch cultivation reveals process relevant targets for host cell engineering.

PubMed

Stiefel, Fabian; Fischer, Simon; Sczyrba, Alexander; Otte, Kerstin; Hesse, Friedemann

2016-05-10

Fed-batch cultivation of recombinant Chinese hamster ovary (CHO) cell lines is one of the most widely used production modes for commercial manufacturing of recombinant protein therapeutics. Furthermore, fed-batch cultivations are often conducted as biphasic processes where the culture temperature is decreased to maximize volumetric product yields. However, it remains to be elucidated which intracellular regulatory elements actually control the observed pro-productive phenotypes. Recently, several studies have revealed microRNAs (miRNAs) to be important molecular switches of cell phenotypes. In this study, we analyzed miRNA profiles of two different recombinant CHO cell lines (high and low producer), and compared them to a non-producing CHO DG44 host cell line during fed-batch cultivation at 37°C versus a temperature shift to 30°C. Taking advantage of next-generation sequencing combined with cluster, correlation and differential expression analyses, we could identify 89 different miRNAs, which were differentially expressed in the different cell lines and cultivation phases. Functional validation experiments using 19 validated target miRNAs confirmed that these miRNAs indeed induced changes in process relevant phenotypes. Furthermore, computational miRNA target prediction combined with functional clustering identified putative target genes and cellular pathways, which might be regulated by these miRNAs. This study systematically identified novel target miRNAs during different phases and conditions of a biphasic fed-batch production process and functionally evaluated their potential for host cell engineering. Copyright © 2016. Published by Elsevier B.V.

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.

Hybrid intelligent control of substrate feeding for industrial fed-batch chlortetracycline fermentation process.

PubMed

Jin, Huaiping; Chen, Xiangguang; Yang, Jianwen; Wu, Lei; Wang, Li

2014-11-01

The lack of accurate process models and reliable online sensors for substrate measurements poses significant challenges for controlling substrate feeding accurately, automatically and optimally in fed-batch fermentation industries. It is still a common practice to regulate the feeding rate based upon manual operations. To address this issue, a hybrid intelligent control method is proposed to enable automatic substrate feeding. The resulting control system consists of three modules: a presetting module for providing initial set-points; a predictive module for estimating substrate concentration online based on a new time interval-varying soft sensing algorithm; and a feedback compensator using expert rules. The effectiveness of the proposed approach is demonstrated through its successful applications to the industrial fed-batch chlortetracycline fermentation process. Copyright © 2014 ISA. Published by Elsevier Ltd. All rights reserved.

Feasibility of nitrification/denitrification in a sequencing batch biofilm reactor with liquid circulation applied to post-treatment.

PubMed

Andrade do Canto, Catarina Simone; Rodrigues, José Alberto Domingues; Ratusznei, Suzana Maria; Zaiat, Marcelo; Foresti, Eugênio

2008-02-01

An investigation was performed on the biological removal of ammonium nitrogen from synthetic wastewater by the simultaneous nitrification/denitrification (SND) process, using a sequencing batch biofilm reactor (SBBR). System behavior was analyzed as to the effects of sludge type used as inoculum (autotrophic/heterotrophic), wastewater feed strategy (batch/fed-batch) and aeration strategy (continuous/intermittent). The presence of an autotrophic aerobic sludge showed to be essential for nitrification startup, despite publications stating the existence of heterotrophic organisms capable of nitrifying organic and inorganic nitrogen compounds at low dissolved oxygen concentrations. As to feed strategy, batch operation (synthetic wastewater containing 100 mg COD/L and 50 mg N-NH(4)(+)/L) followed by fed-batch (synthetic wastewater with 100 mg COD/L) during a whole cycle seemed to be the most adequate, mainly during the denitrification phase. Regarding aeration strategy, an intermittent mode, with dissolved oxygen concentration of 2.0mg/L in the aeration phase, showed the best results. Under these optimal conditions, 97% of influent ammonium nitrogen (80% of total nitrogen) was removed at a rate of 86.5 mg N-NH(4)(+)/Ld. In the treated effluent only 0.2 mg N-NO(2)(-)/L,4.6 mg N-NO(3)(-)/L and 1.0 mg N-NH(4)(+)/L remained, demonstrating the potential viability of this process in post-treatment of wastewaters containing ammonium nitrogen.

Advanced control of dissolved oxygen concentration in fed batch cultures during recombinant protein production.

PubMed

Kuprijanov, A; Gnoth, S; Simutis, R; Lübbert, A

2009-02-01

Design and experimental validation of advanced pO(2) controllers for fermentation processes operated in the fed-batch mode are described. In most situations, the presented controllers are able to keep the pO(2) in fermentations for recombinant protein productions exactly on the desired value. The controllers are based on the gain-scheduling approach to parameter-adaptive proportional-integral controllers. In order to cope with the most often appearing distortions, the basic gain-scheduling feedback controller was complemented with a feedforward control component. This feedforward/feedback controller significantly improved pO(2) control. By means of numerical simulations, the controller behavior was tested and its parameters were determined. Validation runs were performed with three Escherichia coli strains producing different recombinant proteins. It is finally shown that the new controller leads to significant improvements in the signal-to-noise ratio of other key process variables and, thus, to a higher process quality.

Lactate production as representative of the fermentation potential of Corynebacterium glutamicum 2262 in a one-step process.

PubMed

Khuat, Hoang Bao Truc; Kaboré, Abdoul Karim; Olmos, Eric; Fick, Michel; Boudrant, Joseph; Goergen, Jean-Louis; Delaunay, Stéphane; Guedon, Emmanuel

2014-01-01

The fermentative properties of thermo-sensitive strain Corynebacterium glutamicum 2262 were investigated in processes coupling aerobic cell growth and the anaerobic fermentation phase. In particular, the influence of two modes of fermentation on the production of lactate, the fermentation product model, was studied. In both processes, lactate was produced in significant amount, 27 g/L in batch culture, and up to 55.8 g/L in fed-batch culture, but the specific production rate in the fed-batch culture was four times lower than that in the batch culture. Compared to other investigated fermentation processes, our strategy resulted in the highest yield of lactic acid from biomass. Lactate production by C. glutamicum 2262 thus revealed the capability of the strain to produce various fermentation products from pyruvate.

Fed-batch hydrolysate addition and cell separation by settling in high cell density lignocellulosic ethanol fermentations on AFEX™ corn stover in the Rapid Bioconversion with Integrated recycling Technology process.

PubMed

Sarks, Cory; Jin, Mingjie; Balan, Venkatesh; Dale, Bruce E

2017-09-01

The Rapid Bioconversion with Integrated recycling Technology (RaBIT) process uses enzyme and yeast recycling to improve cellulosic ethanol production economics. The previous versions of the RaBIT process exhibited decreased xylose consumption using cell recycle for a variety of different micro-organisms. Process changes were tested in an attempt to eliminate the xylose consumption decrease. Three different RaBIT process changes were evaluated in this work including (1) shortening the fermentation time, (2) fed-batch hydrolysate addition, and (3) selective cell recycling using a settling method. Shorting the RaBIT fermentation process to 11 h and introducing fed-batch hydrolysate addition eliminated any xylose consumption decrease over ten fermentation cycles; otherwise, decreased xylose consumption was apparent by the third cell recycle event. However, partial removal of yeast cells during recycle was not economical when compared to recycling all yeast cells.

Dynamic model of temperature impact on cell viability and major product formation during fed-batch and continuous ethanolic fermentation in Saccharomyces cerevisiae.

PubMed

Amillastre, Emilie; Aceves-Lara, César-Arturo; Uribelarrea, Jean-Louis; Alfenore, Sandrine; Guillouet, Stéphane E

2012-08-01

The impact of the temperature on an industrial yeast strain was investigated in very high ethanol performance fermentation fed-batch process within the range of 30-47 °C. As previously observed with a lab strain, decoupling between growth and glycerol formation occurred at temperature of 36 °C and higher. A dynamic model was proposed to describe the impact of the temperature on the total and viable biomass, ethanol and glycerol production. The model validation was implemented with experimental data sets from independent cultures under different temperatures, temperature variation profiles and cultivation modes. The proposed model fitted accurately the dynamic evolutions for products and biomass concentrations over a wide range of temperature profiles. R2 values were above 0.96 for ethanol and glycerol in most experiments. The best results were obtained at 37 °C in fed-batch and chemostat cultures. This dynamic model could be further used for optimizing and monitoring the ethanol fermentation at larger scale. Copyright © 2012 Elsevier Ltd. All rights reserved.

Combination of bioaugmentation and biostimulation for remediation of paddy soil contaminated with 2,4-dichlorophenoxyacetic acid.

PubMed

Yang, Zhiman; Xu, Xiaohui; Dai, Meng; Wang, Lin; Shi, Xiaoshuang; Guo, Rongbo

2018-04-22

The batch and fed-batch tests were performed to evaluate the efficiency of bioaugmentation in combination with biostimulation for remediation of paddy soil contaminated with 2,4-dichlorophenoxyacetic acid (2,4-D). 2,4-D degrading enrichments were used for bioaugmentation, and effluents prepared through biological hydrogen production process were used as substrate for biostimulation. The batch tests indicated that 2,4-D degradation depended on the enrichment/substrate ratio (E/S), where E/S of 0.03 showed an excellent performance. The fed-batch tests showed that biostimulation only led to an improvement in 2,4-D degradation, while the pattern of repeated augmentation of enrichments (FRA) together with biostimulation obviously improved degradation of 2,4-D, 2-chlorophenol (2-CP) and phenol. DNA-sequencing approach showed that the FRA pattern altered the bacterial community composition, and high removal of 2,4-D, 2-CP and phenol may be attributed to the acclimation and persistence of Thauera. The findings demonstrated the importance of the FRA pattern on remediation of paddy soil contaminated with 2,4-D. Copyright © 2018 Elsevier B.V. All rights reserved.

Kinetic characterization and fed-batch fermentation for maximal simultaneous production of esterase and protease from Lysinibacillus fusiformis AU01.

PubMed

Divakar, K; Suryia Prabha, M; Nandhinidevi, G; Gautam, P

2017-04-21

The simultaneous production of intracellular esterase and extracellular protease from the strain Lysinibacillus fusiformis AU01 was studied in detail. The production was performed both under batch and fed-batch modes. The maximum yield of intracellular esterase and protease was obtained under full oxygen saturation at the beginning of the fermentation. The data were fitted to the Luedeking-Piret model and it was shown that the enzyme (both esterase and protease) production was growth associated. A decrease in intracellular esterase and increase in the extracellular esterase were observed during late stationary phase. The appearance of intracellular proteins in extracellular media and decrease in viable cell count and biomass during late stationary phase confirmed that the presence of extracellular esterase is due to cell lysis. Even though the fed-batch fermentation with different feeding strategies showed improved productivity, feeding yeast extract under DO-stat fermentation conditions showed highest intracellular esterase and protease production. Under DO-stat fed-batch cultivation, maximum intracellular esterase activity of 820 × 10 3 U/L and extracellular protease activity of 172 × 10 3 U/L were obtained at the 16th hr. Intracellular esterase and extracellular protease production were increased fivefold and fourfold, respectively, when compared to batch fermentation performed under shake flask conditions.

Kinetics of sugars consumption and ethanol inhibition in carob pulp fermentation by Saccharomyces cerevisiae in batch and fed-batch cultures.

PubMed

Lima-Costa, Maria Emília; Tavares, Catarina; Raposo, Sara; Rodrigues, Brígida; Peinado, José M

2012-05-01

The waste materials from the carob processing industry are a potential resource for second-generation bioethanol production. These by-products are small carob kibbles with a high content of soluble sugars (45-50%). Batch and fed-batch Saccharomyces cerevisiae fermentations of high density sugar from carob pods were analyzed in terms of the kinetics of sugars consumption and ethanol inhibition. In all the batch runs, 90-95% of the total sugar was consumed and transformed into ethanol with a yield close to the theoretical maximum (0.47-0.50 g/g), and a final ethanol concentration of 100-110 g/l. In fed-batch runs, fresh carob extract was added when glucose had been consumed. This addition and the subsequent decrease of ethanol concentrations by dilution increased the final ethanol production up to 130 g/l. It seems that invertase activity and yeast tolerance to ethanol are the main factors to be controlled in carob fermentations. The efficiency of highly concentrated carob fermentation makes it a very promising process for use in a second-generation ethanol biorefinery.

Kinetic studies on batch cultivation of Trichoderma reesei and application to enhance cellulase production by fed-batch fermentation.

PubMed

Ma, Lijuan; Li, Chen; Yang, Zhenhua; Jia, Wendi; Zhang, Dongyuan; Chen, Shulin

2013-07-20

Reducing the production cost of cellulase as the key enzyme for cellulose hydrolysis to fermentable sugars remains a major challenge for biofuel production. Because of the complexity of cellulase production, kinetic modeling and mass balance calculation can be used as effective tools for process design and optimization. In this study, kinetic models for cell growth, substrate consumption and cellulase production in batch fermentation were developed, and then applied in fed-batch fermentation to enhance cellulase production. Inhibition effect of substrate was considered and a modified Luedeking-Piret model was developed for cellulase production and substrate consumption according to the growth characteristics of Trichoderma reesei. The model predictions fit well with the experimental data. Simulation results showed that higher initial substrate concentration led to decrease of cellulase production rate. Mass balance and kinetic simulation results were applied to determine the feeding strategy. Cellulase production and its corresponding productivity increased by 82.13% after employing the proper feeding strategy in fed-batch fermentation. This method combining mathematics and chemometrics by kinetic modeling and mass balance can not only improve cellulase fermentation process, but also help to better understand the cellulase fermentation process. The model development can also provide insight to other similar fermentation processes. Copyright © 2013 Elsevier B.V. All rights reserved.

Different cultivation methods to acclimatise ammonia-tolerant methanogenic consortia.

PubMed

Tian, Hailin; Fotidis, Ioannis A; Mancini, Enrico; Angelidaki, Irini

2017-05-01

Bioaugmentation with ammonia tolerant-methanogenic consortia was proposed as a solution to overcome ammonia inhibition during anaerobic digestion process recently. However, appropriate technology to generate ammonia tolerant methanogenic consortia is still lacking. In this study, three basic reactors (i.e. batch, fed-batch and continuous stirred-tank reactors (CSTR)) operated at mesophilic (37°C) and thermophilic (55°C) conditions were assessed, based on methane production efficiency, incubation time, TAN/FAN (total ammonium nitrogen/free ammonia nitrogen) levels and maximum methanogenic activity. Overall, fed-batch cultivation was clearly the most efficient method compared to batch and CSTR. Specifically, by saving incubation time up to 150%, fed-batch reactors were acclimatised to nearly 2-fold higher FAN levels with a 37%-153% methanogenic activity improvement, compared to batch method. Meanwhile, CSTR reactors were inhibited at lower ammonia levels. Finally, specific methanogenic activity test showed that hydrogenotrophic methanogens were more active than aceticlastic methanogens in all FAN levels above 540mgNH 3 -NL -1 . Copyright © 2017 Elsevier Ltd. All rights reserved.

An investigation into the preservation of microbial cell banks for α-amylase production during 5 l fed-batch Bacillus licheniformis fermentations.

PubMed

Hancocks, Nichola H; Thomas, Colin R; Stocks, Stuart M; Hewitt, Christopher J

2010-10-01

Fluorescent staining techniques were used for a systematic examination of methods used to cryopreserve microbial cell banks. The aim of cryopreservation here is to ensure subsequent reproducible fermentation performance rather than just post thaw viability. Bacillus licheniformis cell physiology post-thaw is dependent on the cryopreservant (either Tween 80, glycerol or dimethyl sulphoxide) and whilst this had a profound effect on the length of the lag phase, during subsequent 5 l fed-batch fermentations, it had little effect on maximum specific growth rate, final biomass concentration or α-amylase activity. Tween 80 not only protected the cells during freezing but also helped them recover post-thaw resulting in shorter process times.

Effect of moisture content on fed batch composting reactor of vegetable and fruit wastes.

PubMed

Jolanun, B; Tripetchkul, S; Chiemchaisri, C; Chaiprasert, P; Towprayoon, S

2005-03-01

Vegetable and fruit wastes mixed with sawdust were composted in a laboratory scale reactor by controlling the waste feeding rate at 21 kg m(-3) day(-1) and aeration rate at 10.6 l m(-3) min(-1). The effects of initial moisture content on organic matter degradation and process performance of fed batch composting were investigated. The absolute amount of removal, removal percentage, and removal rate of dry mass obtained were substantially different among the initial moisture contents. The rapid rise of moisture content and the lowest absolute amount of removal observed were achieved in the 50% condition. The initial moisture content yielding the largest absolute amount of removal in both feeding and curing stage was 30% whereas the removal percentage and rate constant of waste decomposition were highest in the 50% condition. Examined by traditional soil physics method, the moisture content at 50-55% was suitable for satisfying the degree of free air space (65-70%) of compost during the fed batch composting. Most degradable organic matter was mainly consumed in the feeding stage as indicated by a higher removal rate of dry mass in all cases. It is recommended that the initial moisture content of 30% and mode of aeration and agitation should be adopted for achieving practical fed batch composting of vegetable and fruit wastes. The study also demonstrated that the composting kinetics of vegetable and fruit wastes mixed with sawdust can be described by a first order model.

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.

Alcoholic fermentation with flocculant Saccharomyces cerevisiae in fed-batch process.

PubMed

Guidini, Carla Zanella; Marquez, Líbia Diniz Santos; de Almeida Silva, Helisângela; de Resende, Miriam Maria; Cardoso, Vicelma Luiz; Ribeiro, Eloízio Júlio

2014-02-01

Studies have been conducted on selecting yeast strains for use in fermentation for ethanol production to improve the performance of industrial plants and decrease production costs. In this paper, we study alcoholic fermentation in a fed-batch process using a Saccharomyces cerevisiae yeast strain with flocculant characteristics. Central composite design (CCD) was used to determine the optimal combination of the variables involved, with the sucrose concentration of 170 g/L, a cellular concentration in the inoculum of 40% (v/v), and a filling time of 6 h, which resulted in a 92.20% yield relative to the theoretical maximum yield, a productivity of 6.01 g/L h and a residual sucrose concentration of 44.33 g/L. With some changes in the process such as recirculation of medium during the fermentation process and increase in cellular concentration in the inoculum after use of the CCD was possible to reduce the residual sucrose concentration to 2.8 g/L in 9 h of fermentation and increase yield and productivity for 92.75% and 9.26 g/L h, respectively. A model was developed to describe the inhibition of alcoholic fermentation kinetics by the substrate and the product. The maximum specific growth rate was 0.103 h(-1), with K(I) and K(s) values of 109.86 and 30.24 g/L, respectively. The experimental results from the fed-batch reactor show a good fit with the proposed model, resulting in a maximum growth rate of 0.080 h(-1).

Efficient production of l-lactic acid from hydrolysate of Jerusalem artichoke with immobilized cells of Lactococcus lactis in fibrous bed bioreactors.

PubMed

Shi, Zhouming; Wei, Peilian; Zhu, Xiangcheng; Cai, Jin; Huang, Lei; Xu, Zhinan

2012-10-10

Hydrolysate of Jerusalem artichoke was applied for the production of l-lactic acid by immobilized Lactococcus lactis cells in a fibrous bed bioreactor system. Preliminary experiments had indicated that the high quality hydrolysate, which was derived from the 40 min acid treatment at 95 °C and pH 1.8, was sufficient to support the cell growth and synthesis of l-lactic acid. With the addition of 5 g/l yeast extract, the fermentative performance of free cell system was evidently improved. After the basal settlement of hydrolysate based fermentation, the batch mode and the fed-batch mode fermentation were carried out in the free cell system and the fibrous bed bioreactor system, respectively. In all cases the immobilized cells presented the superior ability to produce l-lactic acid. The comparison of batch mode and fed-batch mode also indicated that the growth-limiting feeding strategy could reduce the lag phase of fermentation process and enhance the production of l-lactic acid. The achieved maximum concentration of l-lactic acid was 142 g/l in the fed-batch mode. Subsequent repeated-batch fermentation of the fibrous bed bioreactor system had further exhibited the persistence and stability of this system for the high production of l-lactic acid in a long term. Our work suggested the great potential of the fibrous bed bioreactor system and hydrolysate of J. artichoke in the economical production of l-lactic acid at industrial scale. Copyright © 2012 Elsevier Inc. All rights reserved.

Bio-plasticizer production by hybrid acetone-butanol-ethanol fermentation with full cell catalysis of Candida sp. 99-125.

PubMed

Chen, Changjing; Cai, Di; Qin, Peiyong; Chen, Biqiang; Wang, Zheng; Tan, Tianwei

2018-06-01

Hybrid process that integrated fermentation, pervaporation and esterification was established aiming to improve the economic feasibility of the conventional acetone-butanol-ethanol (ABE) fermentation process. Candida sp 99-125 cells were used as full-cell catalyst. The feasibility of batch and fed-batch esterification using the ABE permeate of pervaporation (ranging from 286.9 g/L to 402.9 g/L) as substrate were compared. Valuable butyl oleate was produced along with ethyl oleate. For the batch esterification, due to severe inhibition of substrate to lipase, the yield of butyl oleate and ethyl oleate were only 24.9% and 3.3%, respectively. In contrast, 75% and 11.8% of butyl oleate and ethyl oleate were obtained, respectively, at the end of the fed-batch esterification. The novel integration process provides a promising strategy for in situ upgrading ABE products. Copyright © 2018 Elsevier Ltd. All rights reserved.

Switching the mode of sucrose utilization by Saccharomyces cerevisiae

PubMed Central

Badotti, Fernanda; Dário, Marcelo G; Alves, Sergio L; Cordioli, Maria Luiza A; Miletti, Luiz C; de Araujo, Pedro S; Stambuk, Boris U

2008-01-01

Background Overflow metabolism is an undesirable characteristic of aerobic cultures of Saccharomyces cerevisiae during biomass-directed processes. It results from elevated sugar consumption rates that cause a high substrate conversion to ethanol and other bi-products, severely affecting cell physiology, bioprocess performance, and biomass yields. Fed-batch culture, where sucrose consumption rates are controlled by the external addition of sugar aiming at its low concentrations in the fermentor, is the classical bioprocessing alternative to prevent sugar fermentation by yeasts. However, fed-batch fermentations present drawbacks that could be overcome by simpler batch cultures at relatively high (e.g. 20 g/L) initial sugar concentrations. In this study, a S. cerevisiae strain lacking invertase activity was engineered to transport sucrose into the cells through a low-affinity and low-capacity sucrose-H+ symport activity, and the growth kinetics and biomass yields on sucrose analyzed using simple batch cultures. Results We have deleted from the genome of a S. cerevisiae strain lacking invertase the high-affinity sucrose-H+ symporter encoded by the AGT1 gene. This strain could still grow efficiently on sucrose due to a low-affinity and low-capacity sucrose-H+ symport activity mediated by the MALx1 maltose permeases, and its further intracellular hydrolysis by cytoplasmic maltases. Although sucrose consumption by this engineered yeast strain was slower than with the parental yeast strain, the cells grew efficiently on sucrose due to an increased respiration of the carbon source. Consequently, this engineered yeast strain produced less ethanol and 1.5 to 2 times more biomass when cultivated in simple batch mode using 20 g/L sucrose as the carbon source. Conclusion Higher cell densities during batch cultures on 20 g/L sucrose were achieved by using a S. cerevisiae strain engineered in the sucrose uptake system. Such result was accomplished by effectively reducing sucrose uptake by the yeast cells, avoiding overflow metabolism, with the concomitant reduction in ethanol production. The use of this modified yeast strain in simpler batch culture mode can be a viable option to more complicated traditional sucrose-limited fed-batch cultures for biomass-directed processes of S. cerevisiae. PMID:18304329

Switching the mode of sucrose utilization by Saccharomyces cerevisiae.

PubMed

Badotti, Fernanda; Dário, Marcelo G; Alves, Sergio L; Cordioli, Maria Luiza A; Miletti, Luiz C; de Araujo, Pedro S; Stambuk, Boris U

2008-02-27

Overflow metabolism is an undesirable characteristic of aerobic cultures of Saccharomyces cerevisiae during biomass-directed processes. It results from elevated sugar consumption rates that cause a high substrate conversion to ethanol and other bi-products, severely affecting cell physiology, bioprocess performance, and biomass yields. Fed-batch culture, where sucrose consumption rates are controlled by the external addition of sugar aiming at its low concentrations in the fermentor, is the classical bioprocessing alternative to prevent sugar fermentation by yeasts. However, fed-batch fermentations present drawbacks that could be overcome by simpler batch cultures at relatively high (e.g. 20 g/L) initial sugar concentrations. In this study, a S. cerevisiae strain lacking invertase activity was engineered to transport sucrose into the cells through a low-affinity and low-capacity sucrose-H+ symport activity, and the growth kinetics and biomass yields on sucrose analyzed using simple batch cultures. We have deleted from the genome of a S. cerevisiae strain lacking invertase the high-affinity sucrose-H+ symporter encoded by the AGT1 gene. This strain could still grow efficiently on sucrose due to a low-affinity and low-capacity sucrose-H+ symport activity mediated by the MALx1 maltose permeases, and its further intracellular hydrolysis by cytoplasmic maltases. Although sucrose consumption by this engineered yeast strain was slower than with the parental yeast strain, the cells grew efficiently on sucrose due to an increased respiration of the carbon source. Consequently, this engineered yeast strain produced less ethanol and 1.5 to 2 times more biomass when cultivated in simple batch mode using 20 g/L sucrose as the carbon source. Higher cell densities during batch cultures on 20 g/L sucrose were achieved by using a S. cerevisiae strain engineered in the sucrose uptake system. Such result was accomplished by effectively reducing sucrose uptake by the yeast cells, avoiding overflow metabolism, with the concomitant reduction in ethanol production. The use of this modified yeast strain in simpler batch culture mode can be a viable option to more complicated traditional sucrose-limited fed-batch cultures for biomass-directed processes of S. cerevisiae.

Determination of model parameters for zinc (II) ion biosorption onto powdered waste sludge (PWS) in a fed-batch system.

PubMed

Kargi, Fikret; Cikla, Sinem

2007-12-01

Biosorption of zinc (II) ions onto pre-treated powdered waste sludge (PWS) was investigated using a completely mixed tank operating in fed-batch mode instead of an adsorption column. Experiments with variable feed flow rate (0.05-0.5 L h(-1)), feed Zn(II) ion concentrations (37.5-275 mg L(-1)) and amount of adsorbent (1-6 g PWS) were performed using fed-batch operation at pH 5 and room temperature (20-25 degrees C). Break-through curves describing variations of aqueous (effluent) zinc ion concentrations with time were determined for different operating conditions. Percent zinc removal from the aqueous phase decreased, but the biosorbed (solid phase) zinc ion concentration increased with increasing feed flow rate and zinc concentration. A modified Bohart-Adams equation was used to determine the biosorption capacity of PWS (q'(s)) and the rate constant (K) for zinc ion biosorption. Biosorption capacity (q'(s)=57.7 g Zn kg(-1) PWS) of PWS in fed-batch operation was found to be comparable with powdered activated carbon (PAC) in column operations. However, the adsorption rate constant (K=9.17 m(3) kg(-1) h(-1)) in fed-batch operation was an order of magnitude larger than those obtained in adsorption columns because of elimination of mass transfer limitations encountered in the column operations. Therefore, a completely mixed tank operated in fed-batch mode was proven to be more advantageous as compared to adsorption columns due to better contact between the phases yielding faster adsorption rates.

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.

Production of pullulan by a thermotolerant aureobasidium pullulans strain in non-stirred fed batch fermentation process.

PubMed

Singh, Ranjan; Gaur, Rajeeva; Tiwari, Soni; Gaur, Manogya Kumar

2012-07-01

Total 95 isolates of Aureobasidium pullulans were isolated from different flowers and leaves samples, out of which 11 thermotolerant strains produced pullulan. One thermotolerant non-melanin pullulan producing strain, designated as RG-5, produced highest pullulan (37.1±1.0 g/l) at 42(o)C, pH 5.5 in 48h of incubation with 3% sucrose and 0.5% ammonium sulphate in a non-stirred fed batch fermentor of 6 liters capacity. The two liters of initial volume of fermentation medium was further fed with the 2 liters in two successive batches at 5 h interval into the fermentor. The sterile air was supplied only for 10h at the rate of 0.5 vvm.

Model-based intensification of a fed-batch microbial process for the maximization of polyhydroxybutyrate (PHB) production rate.

PubMed

Penloglou, Giannis; Vasileiadou, Athina; Chatzidoukas, Christos; Kiparissides, Costas

2017-08-01

An integrated metabolic-polymerization-macroscopic model, describing the microbial production of polyhydroxybutyrate (PHB) in Azohydromonas lata bacteria, was developed and validated using a comprehensive series of experimental measurements. The model accounted for biomass growth, biopolymer accumulation, carbon and nitrogen sources utilization, oxygen mass transfer and uptake rates and average molecular weights of the accumulated PHB, produced under batch and fed-batch cultivation conditions. Model predictions were in excellent agreement with experimental measurements. The validated model was subsequently utilized to calculate optimal operating conditions and feeding policies for maximizing PHB productivity for desired PHB molecular properties. More specifically, two optimal fed-batch strategies were calculated and experimentally tested: (1) a nitrogen-limited fed-batch policy and (2) a nitrogen sufficient one. The calculated optimal operating policies resulted in a maximum PHB content (94% g/g) in the cultivated bacteria and a biopolymer productivity of 4.2 g/(l h), respectively. Moreover, it was demonstrated that different PHB grades with weight average molecular weights of up to 1513 kg/mol could be produced via the optimal selection of bioprocess operating conditions.

A novel process-based model of microbial growth: self-inhibition in Saccharomyces cerevisiae aerobic fed-batch cultures.

PubMed

Mazzoleni, Stefano; Landi, Carmine; Cartenì, Fabrizio; de Alteriis, Elisabetta; Giannino, Francesco; Paciello, Lucia; Parascandola, Palma

2015-07-30

Microbial population dynamics in bioreactors depend on both nutrients availability and changes in the growth environment. Research is still ongoing on the optimization of bioreactor yields focusing on the increase of the maximum achievable cell density. A new process-based model is proposed to describe the aerobic growth of Saccharomyces cerevisiae cultured on glucose as carbon and energy source. The model considers the main metabolic routes of glucose assimilation (fermentation to ethanol and respiration) and the occurrence of inhibition due to the accumulation of both ethanol and other self-produced toxic compounds in the medium. Model simulations reproduced data from classic and new experiments of yeast growth in batch and fed-batch cultures. Model and experimental results showed that the growth decline observed in prolonged fed-batch cultures had to be ascribed to self-produced inhibitory compounds other than ethanol. The presented results clarify the dynamics of microbial growth under different feeding conditions and highlight the relevance of the negative feedback by self-produced inhibitory compounds on the maximum cell densities achieved in a bioreactor.

Evaluation of enzymatic reactors for large-scale panose production.

PubMed

Fernandes, Fabiano A N; Rodrigues, Sueli

2007-07-01

Panose is a trisaccharide constituted by a maltose molecule bonded to a glucose molecule by an alpha-1,6-glycosidic bond. This trisaccharide has potential to be used in the food industry as a noncariogenic sweetener, as the oral flora does not ferment it. Panose can also be considered prebiotic for stimulating the growth of benefic microorganisms, such as lactobacillus and bifidobacteria, and for inhibiting the growth of undesired microorganisms such as E. coli and Salmonella. In this paper, the production of panose by enzymatic synthesis in a batch and a fed-batch reactor was optimized using a mathematical model developed to simulate the process. Results show that optimum production is obtained in a fed-batch process with an optimum production of 11.23 g/l h of panose, which is 51.5% higher than production with batch reactor.

Strategies to achieve high-solids enzymatic hydrolysis of dilute-acid pretreated corn stover.

PubMed

Geng, Wenhui; Jin, Yongcan; Jameel, Hasan; Park, Sunkyu

2015-01-01

Three strategies were presented to achieve high solids loading while maximizing carbohydrate conversion, which are fed-batch, splitting/thickening, and clarifier processes. Enzymatic hydrolysis was performed at water insoluble solids (WIS) of 15% using washed dilute-acid pretreated corn stover. The carbohydrate concentration increased from 31.8 to 99.3g/L when the insoluble solids content increased from 5% to 15% WIS, while the final carbohydrate conversion was decreased from 78.4% to 73.2%. For the fed-batch process, a carbohydrate conversion efficiency of 76.8% was achieved when solid was split into 60:20:20 ratio, with all enzymes added first. For the splitting/thickening process, a carbohydrate conversion of 76.5% was realized when the filtrate was recycled to simulate a steady-state process. Lastly, the clarifier process was evaluated and the highest carbohydrate conversion of 81.4% was achieved. All of these results suggests the possibility of enzymatic hydrolysis at high solids to make the overall conversion cost-competitive. Copyright © 2015 Elsevier Ltd. All rights reserved.

Fault detection and diagnosis in an industrial fed-batch cell culture process.

PubMed

Gunther, Jon C; Conner, Jeremy S; Seborg, Dale E

2007-01-01

A flexible process monitoring method was applied to industrial pilot plant cell culture data for the purpose of fault detection and diagnosis. Data from 23 batches, 20 normal operating conditions (NOC) and three abnormal, were available. A principal component analysis (PCA) model was constructed from 19 NOC batches, and the remaining NOC batch was used for model validation. Subsequently, the model was used to successfully detect (both offline and online) abnormal process conditions and to diagnose the root causes. This research demonstrates that data from a relatively small number of batches (approximately 20) can still be used to monitor for a wide range of process faults.

In vitro growth of Curcuma longa L. in response to five mineral elements and plant density in fed-batch culture systems.

PubMed

El-Hawaz, Rabia F; Bridges, William C; Adelberg, Jeffrey W

2015-01-01

Plant density was varied with P, Ca, Mg, and KNO3 in a multifactor experiment to improve Curcuma longa L. micropropagation, biomass and microrhizome development in fed-batch liquid culture. The experiment had two paired D-optimal designs, testing sucrose fed-batch and nutrient sucrose fed-batch techniques. When sucrose became depleted, volume was restored to 5% m/v sucrose in 200 ml of modified liquid MS medium by adding sucrose solutions. Similarly, nutrient sucrose fed-batch was restored to set points with double concentration of treatments' macronutrient and MS micronutrient solutions, along with sucrose solutions. Changes in the amounts of water and sucrose supplementations were driven by the interaction of P and KNO3 concentrations. Increasing P from 1.25 to 6.25 mM increased both multiplication and biomass. The multiplication ratio was greatest in the nutrient sucrose fed-batch technique with the highest level of P, 6 buds/vessel, and the lowest level of Ca and KNO3. The highest density (18 buds/vessel) produced the highest fresh biomass at the highest concentrations of KNO3 and P with nutrient sucrose fed-batch, and moderate Ca and Mg concentrations. However, maximal rhizome dry biomass required highest P, sucrose fed-batch, and a moderate plant density. Different media formulations and fed-batch techniques were identified to maximize the propagation and storage organ responses. A single experimental design was used to optimize these dual purposes.

In Vitro Growth of Curcuma longa L. in Response to Five Mineral Elements and Plant Density in Fed-Batch Culture Systems

PubMed Central

El-Hawaz, Rabia F.; Bridges, William C.; Adelberg, Jeffrey W.

2015-01-01

Plant density was varied with P, Ca, Mg, and KNO3 in a multifactor experiment to improve Curcuma longa L. micropropagation, biomass and microrhizome development in fed-batch liquid culture. The experiment had two paired D-optimal designs, testing sucrose fed-batch and nutrient sucrose fed-batch techniques. When sucrose became depleted, volume was restored to 5% m/v sucrose in 200 ml of modified liquid MS medium by adding sucrose solutions. Similarly, nutrient sucrose fed-batch was restored to set points with double concentration of treatments’ macronutrient and MS micronutrient solutions, along with sucrose solutions. Changes in the amounts of water and sucrose supplementations were driven by the interaction of P and KNO3 concentrations. Increasing P from 1.25 to 6.25 mM increased both multiplication and biomass. The multiplication ratio was greatest in the nutrient sucrose fed-batch technique with the highest level of P, 6 buds/vessel, and the lowest level of Ca and KNO3. The highest density (18 buds/vessel) produced the highest fresh biomass at the highest concentrations of KNO3 and P with nutrient sucrose fed-batch, and moderate Ca and Mg concentrations. However, maximal rhizome dry biomass required highest P, sucrose fed-batch, and a moderate plant density. Different media formulations and fed-batch techniques were identified to maximize the propagation and storage organ responses. A single experimental design was used to optimize these dual purposes. PMID:25830292

Estimation of the Maximum Theoretical Productivity of Fed-Batch Bioreactors

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

Bomble, Yannick J; St. John, Peter C; Crowley, Michael F

2017-10-18

A key step towards the development of an integrated biorefinery is the screening of economically viable processes, which depends sharply on the yields and productivities that can be achieved by an engineered microorganism. In this study, we extend an earlier method which used dynamic optimization to find the maximum theoretical productivity of batch cultures to explicitly include fed-batch bioreactors. In addition to optimizing the intracellular distribution of metabolites between cell growth and product formation, we calculate the optimal control trajectory of feed rate versus time. We further analyze how sensitive the productivity is to substrate uptake and growth parameters.

Modelling and properties of a nonlinear autonomous switching system in fed-batch culture of glycerol

NASA Astrophysics Data System (ADS)

Wang, Juan; Sun, Qingying; Feng, Enmin

2012-11-01

A nonlinear autonomous switching system is proposed to describe the coupled fed-batch fermentation with the pH as the feedback parameter. We prove the non-Zeno behaviors of the switching system and some basic properties of its solution, including the existence, uniqueness, boundedness and regularity. Numerical simulation is also carried out, which reveals that the proposed system can describe the factual fermentation process properly.

Principles and approach to developing mammalian cell culture media for high cell density perfusion process leveraging established fed-batch media.

PubMed

Lin, Henry; Leighty, Robert Woodrow; Godfrey, Scott; Wang, Samantha Boran

2017-07-01

Perfusion medium was successfully developed based on our fed-batch platform basal and feed media. A systematic development approach was undertaken by first optimizing the ratios of fed-batch basal and feed media followed by targeted removal of unnecessary and redundant components. With this reduction in components, the medium could then be further concentrated by 2× to increase medium depth. The medium osmolality was also optimized where we found ∼360 mOsm/kg was desirable resulting in a residual culture osmolality of ∼300 mOsm/kg for our cell lines. Further building on this, the amino acids Q, E, N, and D were rebalanced to reduce lactate and ammonium levels, and increase the cell-specific productivity without compromising on cell viability while leaving viable cell density largely unaffected. Further modifications were also made by increasing certain important vitamin and lipid concentrations, while eliminating other unnecessary vitamins. Overall, an effective perfusion medium was developed with all components remaining in the formulation understood to be important and their concentrations increased to improve medium depth. The critical cell-specific perfusion rate using this medium was then established for a cell line of interest to be 0.075 nL/cell-day yielding 1.2 g/L-day at steady state. This perfusion process was then successfully scaled up to a 100 L single-use bioreactor with an ATF6 demonstrating similar performance as a 2 L bioreactor with an ATF2. Large volume handling challenges in our fed-batch facility were overcome by developing a liquid medium version of the powder medium product contained in custom totes for plug-and-play use with the bioreactor. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:891-901, 2017. © 2017 American Institute of Chemical Engineers.

Production of pullulan by a thermotolerant aureobasidium pullulans strain in non-stirred fed batch fermentation process

PubMed Central

Singh, Ranjan; Gaur, Rajeeva; Tiwari, Soni; Gaur, Manogya Kumar

2012-01-01

Total 95 isolates of Aureobasidium pullulans were isolated from different flowers and leaves samples, out of which 11 thermotolerant strains produced pullulan. One thermotolerant non-melanin pullulan producing strain, designated as RG-5, produced highest pullulan (37.1±1.0 g/l) at 42oC, pH 5.5 in 48h of incubation with 3% sucrose and 0.5% ammonium sulphate in a non-stirred fed batch fermentor of 6 liters capacity. The two liters of initial volume of fermentation medium was further fed with the 2 liters in two successive batches at 5 h interval into the fermentor. The sterile air was supplied only for 10h at the rate of 0.5 vvm. PMID:24031927

Scale-up bioprocess development for production of the antibiotic valinomycin in Escherichia coli based on consistent fed-batch cultivations.

PubMed

Li, Jian; Jaitzig, Jennifer; Lu, Ping; Süssmuth, Roderich D; Neubauer, Peter

2015-06-12

Heterologous production of natural products in Escherichia coli has emerged as an attractive strategy to obtain molecules of interest. Although technically feasible most of them are still constrained to laboratory scale production. Therefore, it is necessary to develop reasonable scale-up strategies for bioprocesses aiming at the overproduction of targeted natural products under industrial scale conditions. To this end, we used the production of the antibiotic valinomycin in E. coli as a model system for scalable bioprocess development based on consistent fed-batch cultivations. In this work, the glucose limited fed-batch strategy based on pure mineral salt medium was used throughout all scales for valinomycin production. The optimal glucose feed rate was initially detected by the use of a biocatalytically controlled glucose release (EnBase® technology) in parallel cultivations in 24-well plates with continuous monitoring of pH and dissolved oxygen. These results were confirmed in shake flasks, where the accumulation of valinomycin was highest when the specific growth rate decreased below 0.1 h(-1). This correlation was also observed for high cell density fed-batch cultivations in a lab-scale bioreactor. The bioreactor fermentation produced valinomycin with titers of more than 2 mg L(-1) based on the feeding of a concentrated glucose solution. Valinomycin production was not affected by oscillating conditions (i.e. glucose and oxygen) in a scale-down two-compartment reactor, which could mimic similar situations in industrial bioreactors, suggesting that the process is very robust and a scaling of the process to a larger industrial scale appears a realistic scenario. Valinomycin production was scaled up from mL volumes to 10 L with consistent use of the fed-batch technology. This work presents a robust and reliable approach for scalable bioprocess development and represents an example for the consistent development of a process for a heterologously expressed natural product towards the industrial scale.

Research on On-Line Modeling of Fed-Batch Fermentation Process Based on v-SVR

NASA Astrophysics Data System (ADS)

Ma, Yongjun

The fermentation process is very complex and non-linear, many parameters are not easy to measure directly on line, soft sensor modeling is a good solution. This paper introduces v-support vector regression (v-SVR) for soft sensor modeling of fed-batch fermentation process. v-SVR is a novel type of learning machine. It can control the accuracy of fitness and prediction error by adjusting the parameter v. An on-line training algorithm is discussed in detail to reduce the training complexity of v-SVR. The experimental results show that v-SVR has low error rate and better generalization with appropriate v.

Data-driven monitoring for stochastic systems and its application on batch process

NASA Astrophysics Data System (ADS)

Yin, Shen; Ding, Steven X.; Haghani Abandan Sari, Adel; Hao, Haiyang

2013-07-01

Batch processes are characterised by a prescribed processing of raw materials into final products for a finite duration and play an important role in many industrial sectors due to the low-volume and high-value products. Process dynamics and stochastic disturbances are inherent characteristics of batch processes, which cause monitoring of batch processes a challenging problem in practice. To solve this problem, a subspace-aided data-driven approach is presented in this article for batch process monitoring. The advantages of the proposed approach lie in its simple form and its abilities to deal with stochastic disturbances and process dynamics existing in the process. The kernel density estimation, which serves as a non-parametric way of estimating the probability density function, is utilised for threshold calculation. An industrial benchmark of fed-batch penicillin production is finally utilised to verify the effectiveness of the proposed approach.

Improving lactate metabolism in an intensified CHO culture process: productivity and product quality considerations.

PubMed

Xu, Sen; Hoshan, Linda; Chen, Hao

2016-11-01

In this study, we discussed the development and optimization of an intensified CHO culture process, highlighting medium and control strategies to improve lactate metabolism. A few strategies, including supplementing glucose with other sugars (fructose, maltose, and galactose), controlling glucose level at <0.2 mM, and supplementing medium with copper sulfate, were found to be effective in reducing lactate accumulation. Among them, copper sulfate supplementation was found to be critical for process optimization when glucose was in excess. When copper sulfate was supplemented in the new process, two-fold increase in cell density (66.5 ± 8.4 × 10(6) cells/mL) and titer (11.9 ± 0.6 g/L) was achieved. Productivity and product quality attributes differences between batch, fed-batch, and concentrated fed-batch cultures were discussed. The importance of process and cell metabolism understanding when adapting the existing process to a new operational mode was demonstrated in the study.

A simple numerical model for predicting organic matter decomposition in a fed-batch composting operation.

PubMed

Nakasaki, Kiyohiko; Ohtaki, Akihito

2002-01-01

Using dog food as a model of the organic waste that comprises composting raw material, the degradation pattern of organic materials was examined by continuously measuring the quantity of CO2 evolved during the composting process in both batch and fed-batch operations. A simple numerical model was made on the basis of three suppositions for describing the organic matter decomposition in the batch operation. First, a certain quantity of carbon in the dog food was assumed to be recalcitrant to degradation in the composting reactor within the retention time allowed. Second, it was assumed that the decomposition rate of carbon is proportional to the quantity of easily degradable carbon, that is, the carbon recalcitrant to degradation was subtracted from the total carbon remaining in the dog food. Third, a certain lag time is assumed to occur before the start of active decomposition of organic matter in the dog food; this lag corresponds to the time required for microorganisms to proliferate and become active. It was then ascertained that the decomposition pattern for the organic matter in the dog food during the fed-batch operation could be predicted by the numerical model with the parameters obtained from the batch operation. This numerical model was modified so that the change in dry weight of composting materials could be obtained. The modified model was found suitable for describing the organic matter decomposition pattern in an actual fed-batch composting operation of the garbage obtained from a restaurant, approximately 10 kg d(-1) loading for 60 d.

The use of dissolved oxygen-controlled, fed-batch aerobic cultivation for recombinant protein subunit vaccine manufacturing.

PubMed

Farrell, Patrick; Sun, Jacob; Champagne, Paul-Philippe; Lau, Heron; Gao, Meg; Sun, Hong; Zeiser, Arno; D'Amore, Tony

2015-11-27

A simple "off-the-shelf" fed-batch approach to aerobic bacterial cultivation for recombinant protein subunit vaccine manufacturing is presented. In this approach, changes in the dissolved oxygen levels are used to adjust the nutrient feed rate (DO-stat), so that the desired dissolved oxygen level is maintained throughout cultivation. This enables high Escherichia coli cell densities and recombinant protein titers. When coupled to a kLa-matched scale-down model, process performance is shown to be consistent at the 2L, 20L, and 200L scales for two recombinant E. coli strains expressing different protein subunit vaccine candidates. Additionally, by mining historical DO-stat nutrient feeding data, a method to transition from DO-stat to a pre-determined feeding profile suitable for larger manufacturing scales without using feedback control is demonstrated at the 2L, 20L, and 200L scales. Copyright © 2015 Elsevier Ltd. All rights reserved.

Bioreactors for high cell density and continuous multi-stage cultivations: options for process intensification in cell culture-based viral vaccine production.

PubMed

Tapia, Felipe; Vázquez-Ramírez, Daniel; Genzel, Yvonne; Reichl, Udo

2016-03-01

With an increasing demand for efficacious, safe, and affordable vaccines for human and animal use, process intensification in cell culture-based viral vaccine production demands advanced process strategies to overcome the limitations of conventional batch cultivations. However, the use of fed-batch, perfusion, or continuous modes to drive processes at high cell density (HCD) and overextended operating times has so far been little explored in large-scale viral vaccine manufacturing. Also, possible reductions in cell-specific virus yields for HCD cultivations have been reported frequently. Taking into account that vaccine production is one of the most heavily regulated industries in the pharmaceutical sector with tough margins to meet, it is understandable that process intensification is being considered by both academia and industry as a next step toward more efficient viral vaccine production processes only recently. Compared to conventional batch processes, fed-batch and perfusion strategies could result in ten to a hundred times higher product yields. Both cultivation strategies can be implemented to achieve cell concentrations exceeding 10(7) cells/mL or even 10(8) cells/mL, while keeping low levels of metabolites that potentially inhibit cell growth and virus replication. The trend towards HCD processes is supported by development of GMP-compliant cultivation platforms, i.e., acoustic settlers, hollow fiber bioreactors, and hollow fiber-based perfusion systems including tangential flow filtration (TFF) or alternating tangential flow (ATF) technologies. In this review, these process modes are discussed in detail and compared with conventional batch processes based on productivity indicators such as space-time yield, cell concentration, and product titers. In addition, options for the production of viral vaccines in continuous multi-stage bioreactors such as two- and three-stage systems are addressed. While such systems have shown similar virus titers compared to batch cultivations, keeping high yields for extended production times is still a challenge. Overall, we demonstrate that process intensification of cell culture-based viral vaccine production can be realized by the consequent application of fed-batch, perfusion, and continuous systems with a significant increase in productivity. The potential for even further improvements is high, considering recent developments in establishment of new (designer) cell lines, better characterization of host cell metabolism, advances in media design, and the use of mathematical models as a tool for process optimization and control.

The operable modeling of simultaneous saccharification and fermentation of ethanol production from cellulose.

PubMed

Shen, Jiacheng; Agblevor, Foster A

2010-03-01

An operable batch model of simultaneous saccharification and fermentation (SSF) for ethanol production from cellulose has been developed. The model includes four ordinary differential equations that describe the changes of cellobiose, glucose, yeast, and ethanol concentrations with respect to time. These equations were used to simulate the experimental data of the four main components in the SSF process of ethanol production from microcrystalline cellulose (Avicel PH101). The model parameters at 95% confidence intervals were determined by a MATLAB program based on the batch experimental data of the SSF. Both experimental data and model simulations showed that the cell growth was the rate-controlling step at the initial period in a series of reactions of cellulose to ethanol, and later, the conversion of cellulose to cellobiose controlled the process. The batch model was extended to the continuous and fed-batch operating models. For the continuous operation in the SSF, the ethanol productivities increased with increasing dilution rate, until a maximum value was attained, and rapidly decreased as the dilution rate approached the washout point. The model also predicted a relatively high ethanol mass for the fed-batch operation than the batch operation.

Behavior of spores of Penicillium roquefortii during fed-batch bioconversion of octanoic acid into 2-heptanone

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

Larroche, C.; Besson, I.; Gros, J.B.

1994-09-05

The bioconversion of octanoic acid into 2-heptanone by spores of Penicillium roquefortii is performed using a fed-batch technique with pH control by addition of the liquid substrate itself. The early stage of this process takes place with a high bioconversion rate and high yield. These values then decrease as a result of germination and growth of the biocatalyst. An optimization strategy for the process would thus be to improve the characteristics of this first period, i.e., increase its duration and the reaction rate. An increase in duration is evidenced in two cases: (1) under oxygen limitation; and (2) when themore » spore content in the medium is less than 10[sup 7] spores/mL. These conditions give insufficient overall bioconversion rates; better optimization should be achieved without oxygen limitation and with high spore content. Characterization of the first period by material and bioenergetic balances suggests that an increase in the ethanol content of the medium, which acts as an energy source and a permeabilizer, and the use of a specific inhibitor of the Krebs cycle, may be a way to further improve the biocatalyst performance and stability.« less

Fed-batch coculture of Lactobacillus kefiranofaciens with Saccharomyces cerevisiae for effective production of kefiran.

PubMed

Tada, Shiori; Katakura, Yoshio; Ninomiya, Kazuaki; Shioya, Suteaki

2007-06-01

In a batch coculture of kefiran-producing lactic acid bacteria Lactobacillus kefiranofaciens and lactate-assimilating yeast Saccharomyces cerevisiae, lactate accumulation in the medium was observed, which inhibited kefiran production. To enhance kefiran productivity by preventing lactate accumulation, we conducted lactose-feeding batch operation with feedforward/feedback control during the coculture, so that the lactate production rate of L. kefiranofaciens was balanced with the lactate consumption rate of S. cerevisiae. The lactate concentration was maintained at less than 6 g l(-1) throughout the fed-batch coculture using a 5 l jar fermentor, although the concentration reached 33 g l(-1) in the batch coculture. Kefiran production was increased to 6.3 g in 102 h in the fed-batch coculture, whereas 4.5 g kefiran was produced in 97 h in the batch coculture. The kefiran yield on lactose basis was increased up to 0.033 g g(-1) in the fed-batch coculture, whereas that in the batch coculture was 0.027 g g(-1).

Change of Monascus pigment metabolism and secretion in different extractive fermentation process.

PubMed

Chen, Gong; Tang, Rui; Tian, Xiaofei; Qin, Peng; Wu, Zhenqiang

2017-06-01

Monascus pigments that were generally produced intracellularly from Monascus spp. are important natural colorants in food industry. In this study, change of pigment metabolism and secretion was investigated through fed-batch extractive fermentation and continuous extractive fermentation. The biomass, secreting rate of pigment and total pigment yield closely correlated with the activated time of extractive fermentation as well as the composition of feeding nutrients. Metal ions played a key role in both the cell growth and pigment metabolism. Nitrogen source was necessary for a high productivity of biomass but not for high pigment yield. Furthermore, fermentation period for the fed-batch extractive fermentation could be reduced by 18.75% with a nitrogen source free feeding medium. Through a 30-day continuous extractive fermentation, the average daily productivity for total pigments reached 74.9 AU day -1 with an increase by 32.6 and 296.3% compared to that in a 6-day conventional batch fermentation and a 16-day fed-batch extractive fermentation, respectively. At the meantime, proportions of extracellular pigments increased gradually from 2.7 to 71.3%, and yellow pigments gradually became dominated in both intracellular and extracellular pigments in the end of continuous extractive fermentation. This findings showed that either fed-batch or continuous extractive fermentation acted as a promising method in the efficient production of Monascus pigments.

The influence of sorbitol on the production of cellulases and xylanases in an airlift bioreactor.

PubMed

Ritter, Carla Eliana Todero; Fontana, Roselei Claudete; Camassola, Marli; da Silveira, Maurício Moura; Dillon, Aldo José Pinheiro

2013-11-01

The production of cellulases and xylanases by Penicillium echinulatum in an airlift bioreactor was evaluated. In batch production, we tested media with isolated or associated cellulose and sorbitol. In fed-batch production, we tested cellulose addition at two different times, 30 h and 48 h. Higher liquid circulation velocities in the downcomer were observed in sorbitol 10 g L(-1) medium. In batch production, higher FPA (filter paper activity) and endoglucanase activities were obtained with cellulose (7.5 g L(-1)) and sorbitol (2.5 g L(-1)), 1.0 U mL(-1) (120 h) and 6.4 U m L(-1) (100 h), respectively. For xylanases, the best production condition was cellulose 10 g L(-1), which achieved 5.5 U mL(-1) in 64 h. The fed-batch process was favorable for obtaining xylanases, but not for FPA and endoglucanases, suggesting that in the case of cellulases, the inducer must be added early in the process. Copyright © 2013 Elsevier Ltd. All rights reserved.

Anaerobic sequencing batch reactors for wastewater treatment: a developing technology.

PubMed

Zaiat, M; Rodrigues, J A; Ratusznei, S M; de Camargo, E F; Borzani, W

2001-01-01

This paper describes and discusses the main problems related to anaerobic batch and fed-batch processes for wastewater treatment. A critical analysis of the literature evaluated the industrial application viability and proposed alternatives to improve operation and control of this system. Two approaches were presented in order to make this anaerobic discontinuous process feasible for industrial application: (1) optimization of the operating procedures in reactors containing self-immobilized sludge as granules, and (2) design of bioreactors with inert support media for biomass immobilization.

Multi-stage high cell continuous fermentation for high productivity and titer.

PubMed

Chang, Ho Nam; Kim, Nag-Jong; Kang, Jongwon; Jeong, Chang Moon; Choi, Jin-dal-rae; Fei, Qiang; Kim, Byoung Jin; Kwon, Sunhoon; Lee, Sang Yup; Kim, Jungbae

2011-05-01

We carried out the first simulation on multi-stage continuous high cell density culture (MSC-HCDC) to show that the MSC-HCDC can achieve batch/fed-batch product titer with much higher productivity to the fed-batch productivity using published fermentation kinetics of lactic acid, penicillin and ethanol. The system under consideration consists of n-serially connected continuous stirred-tank reactors (CSTRs) with either hollow fiber cell recycling or cell immobilization for high cell-density culture. In each CSTR substrate supply and product removal are possible. Penicillin production is severely limited by glucose metabolite repression that requires multi-CSTR glucose feeding. An 8-stage C-HCDC lactic acid fermentation resulted in 212.9 g/L of titer and 10.6 g/L/h of productivity, corresponding to 101 and 429% of the comparable lactic acid fed-batch, respectively. The penicillin production model predicted 149% (0.085 g/L/h) of productivity in 8-stage C-HCDC with 40 g/L of cell density and 289% of productivity (0.165 g/L/h) in 7-stage C-HCDC with 60 g/L of cell density compared with referring batch cultivations. A 2-stage C-HCDC ethanol experimental run showed 107% titer and 257% productivity of the batch system having 88.8 g/L of titer and 3.7 g/L/h of productivity. MSC-HCDC can give much higher productivity than batch/fed-batch system, and yield a several percentage higher titer as well. The productivity ratio of MSC-HCDC over batch/fed-batch system is given as a multiplication of system dilution rate of MSC-HCDC and cycle time of batch/fed-batch system. We suggest MSC-HCDC as a new production platform for various fermentation products including monoclonal antibody.

Batch and fed-batch production of butyric acid by Clostridium butyricum ZJUCB

PubMed Central

He, Guo-qing; Kong, Qing; Chen, Qi-he; Ruan, Hui

2005-01-01

The production of butyric acid by Clostridium butyricum ZJUCB at various pH values was investigated. In order to study the effect of pH on cell growth, butyric acid biosynthesis and reducing sugar consumption, different cultivation pH values ranging from 6.0 to 7.5 were evaluated in 5-L bioreactor. In controlled pH batch fermentation, the optimum pH for cell growth and butyric acid production was 6.5 with a cell yield of 3.65 g/L and butyric acid yield of 12.25 g/L. Based on these results, this study then compared batch and fed-batch fermentation of butyric acid production at pH 6.5. Maximum value (16.74 g/L) of butyric acid concentration was obtained in fed-batch fermentation compared to 12.25 g/L in batch fermentation. It was concluded that cultivation under fed-batch fermentation mode could enhance butyric acid production significantly (P<0.01) by C. butyricum ZJUCB. PMID:16252341

The development of an industrial-scale fed-batch fermentation simulation.

PubMed

Goldrick, Stephen; Ştefan, Andrei; Lovett, David; Montague, Gary; Lennox, Barry

2015-01-10

This paper describes a simulation of an industrial-scale fed-batch fermentation that can be used as a benchmark in process systems analysis and control studies. The simulation was developed using a mechanistic model and validated using historical data collected from an industrial-scale penicillin fermentation process. Each batch was carried out in a 100,000 L bioreactor that used an industrial strain of Penicillium chrysogenum. The manipulated variables recorded during each batch were used as inputs to the simulator and the predicted outputs were then compared with the on-line and off-line measurements recorded in the real process. The simulator adapted a previously published structured model to describe the penicillin fermentation and extended it to include the main environmental effects of dissolved oxygen, viscosity, temperature, pH and dissolved carbon dioxide. In addition the effects of nitrogen and phenylacetic acid concentrations on the biomass and penicillin production rates were also included. The simulated model predictions of all the on-line and off-line process measurements, including the off-gas analysis, were in good agreement with the batch records. The simulator and industrial process data are available to download at www.industrialpenicillinsimulation.com and can be used to evaluate, study and improve on the current control strategy implemented on this facility. Crown Copyright © 2014. Published by Elsevier B.V. All rights reserved.

Development of a mathematical model for the growth associated Polyhydroxybutyrate fermentation by Azohydromonas australica and its use for the design of fed-batch cultivation strategies.

PubMed

Gahlawat, Geeta; Srivastava, Ashok K

2013-06-01

In the present investigation, batch cultivation of Azohydromonas australica DSM 1124 was carried out in a bioreactor for growth associated PHB production. The observed batch PHB production kinetics data was then used for the development of a mathematical model which adequately described the substrate limitation and inhibition during the cultivation. The statistical validity test demonstrated that the proposed mathematical model predictions were significant at 99% confidence level. The model was thereafter extrapolated to fed-batch to identify various nutrients feeding regimes during the bioreactor cultivation to improve the PHB accumulation. The distinct capability of the mathematical model to predict highly dynamic fed-batch cultivation strategies was demonstrated by experimental implementation of two fed-batch cultivation strategies. A significantly high PHB concentration of 22.65 g/L & an overall PHB content of 76% was achieved during constant feed rate fed-batch cultivation which is the highest PHB content reported so far using A. australica. Copyright © 2013 Elsevier Ltd. All rights reserved.

Physiological mechanism of the overproduction of ε-poly-L-lysine by acidic pH shock in fed-batch fermentation.

PubMed

Ren, Xi-Dong; Chen, Xu-Sheng; Tang, Lei; Zeng, Xin; Wang, Liang; Mao, Zhong-Gui

2015-11-01

The introduction of an environmental stress of acidic pH shock had successfully solved the common deficiency existed in ε-PL production, viz. the distinct decline of ε-PL productivity in the feeding phase of the fed-batch fermentation. To unravel the underlying mechanism, we comparatively studied the physiological changes of Streptomyces sp. M-Z18 during fed-batch fermentations with the pH shock strategy (PS) and pH non-shock strategy (PNS). Morphology investigation showed that pellet-shape change was negligible throughout both fermentations. In addition, the distribution of pellet size rarely changed in the PS, whereas pellet size and number decreased substantially with time in the PNS. This was consistent with the performances of ε-PL productivity in both strategies, demonstrating that morphology could be used as a predictor of ε-PL productivity during fed-batch fermentation. Furthermore, a second growth phase happened in the PS after pH shock, followed by the re-appearance of live mycelia in the dead core of the pellets. Meanwhile, mycelia respiration and key enzymes in the central metabolic and ε-PL biosynthetic pathways were overall strengthened until the end of the fed-batch fermentation. As a result, the physiological changes induced by the acidic pH shock have synergistically and permanently contributed to the stimulation of ε-PL productivity. However, this second growth phase and re-appearance of live mycelia were absent in the PNS. These results indicated that the introduction of a short-term suppression on mycelia physiological metabolism would guarantee the long-term high ε-PL productivity.

Electrochemical study of multi-electrode microbial fuel cells under fed-batch and continuous flow conditions

NASA Astrophysics Data System (ADS)

Ren, Lijiao; Ahn, Yongtae; Hou, Huijie; Zhang, Fang; Logan, Bruce E.

2014-07-01

Power production of four hydraulically connected microbial fuel cells (MFCs) was compared with the reactors operated using individual electrical circuits (individual), and when four anodes were wired together and connected to four cathodes all wired together (combined), in fed-batch or continuous flow conditions. Power production under these different conditions could not be made based on a single resistance, but instead required polarization tests to assess individual performance relative to the combined MFCs. Based on the power curves, power produced by the combined MFCs (2.12 ± 0.03 mW, 200 Ω) was the same as the summed power (2.13 mW, 50 Ω) produced by the four individual reactors in fed-batch mode. With continuous flow through the four MFCs, the maximum power (0.59 ± 0.01 mW) produced by the combined MFCs was slightly lower than the summed maximum power of the four individual reactors (0.68 ± 0.02 mW). There was a small parasitic current flow from adjacent anodes and cathodes, but overall performance was relatively unaffected. These findings demonstrate that optimal power production by reactors hydraulically and electrically connected can be predicted from performance by individual reactors.

Effect of Growth Conditions and Trehalose Content on Cryotolerance of Bakers' Yeast in Frozen Doughs

PubMed Central

Gélinas, Pierre; Fiset, Gisèle; LeDuy, Anh; Goulet, Jacques

1989-01-01

The cryotolerance in frozen doughs and in water suspensions of bakers' yeast (Saccharomyces cerevisiae) previously grown under various industrial conditions was evaluated on a laboratory scale. Fed-batch cultures were very superior to batch cultures, and strong aeration enhanced cryoresistance in both cases for freezing rates of 1 to 56°C min−1. Loss of cell viability in frozen dough or water was related to the duration of the dissolved-oxygen deficit during fed-batch growth. Strongly aerobic fed-batch cultures grown at a reduced average specific rate (μ = 0.088 h−1 compared with 0.117 h−1) also showed greater trehalose synthesis and improved frozen-dough stability. Insufficient aeration (dissolved-oxygen deficit) and lower growth temperature (20°C instead of 30°C) decreased both fed-batch-grown yeast cryoresistance and trehalose content. Although trehalose had a cryoprotective effect in S. cerevisiae, its effect was neutralized by even a momentary lack of excess dissolved oxygen in the fed-batch growth medium. PMID:16348024

Kinetics and modeling of hexavalent chromium reduction in Enterobacter cloacae

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

Yamamoto, Koji; Kato, Junichi; Yano, Takuo

1993-01-05

Kinetics of bacterial reduction of toxic hexavalent chromium (chromate: CrO[sub 4][sup [minus]2]) was investigated using batch and fed-batch cultures of Enterobacter cloacae strain HO1. In fed-batch cultures, the CrO[sub 4][sup [minus]2] feed was controlled on the basis of the rate of pH change. This control strategy has proven to be useful for avoiding toxic CrO[sub 3][sup [minus]2] overload. A simple mathematical model was developed to describe the bacterial process of CrO[sub 4][sup [minus]2] reduction. In this model, two types of bacterial cells were considered: induced, CrO[sub 4][sup [minus]2]-resistant cells and uninduced, sensitive ones. Only resistant cells were assumed to bemore » able to reduce CrO[sub 4][sup [minus]2]. These fundamental ideas were supported by the model predictions which well approximated all experimental data. In a simulation study, the model was also used to optimize fed-batch cultures, instead of lengthy and expensive laboratory experiments.« less

Continuous citric acid production in repeated-fed batch fermentation by Aspergillus niger immobilized on a new porous foam.

PubMed

Yu, Bin; Zhang, Xin; Sun, Wenjun; Xi, Xun; Zhao, Nan; Huang, Zichun; Ying, Zhuojun; Liu, Li; Liu, Dong; Niu, Huanqing; Wu, Jinglan; Zhuang, Wei; Zhu, Chenjie; Chen, Yong; Ying, Hanjie

2018-06-20

The efficiency of current methods for industrial production of citric acid is limited. To achieve continuous citric acid production with enhanced yield and reduced cost, immobilized fermentation was employed in an Aspergillus niger 831 repeated fed-batch fermentation system. We developed a new type of material (PAF201), which was used as a carrier for the novel adsorption immobilization system. Hydrophobicity, pore size and concentration of carriers were researched in A. niger immobilization. The efficiency of the A. niger immobilization process was analyzed by scanning electron microscopy. Then eight-cycle repeated fed-batch cultures for citric acid production were carried out over 600 h, which showed stable production with maximum citric acid concentrations and productivity levels of 162.7 g/L and 2.26 g L -1  h -1 , respectively. Compared with some other literatures about citric acid yield, PAF201 immobilization system is 11.3% higher than previous results. These results indicated that use of the new adsorption immobilization system could greatly improve citric acid productivity in repeated fed-batch fermentation. Moreover, these results could provide a guideline for A.niger or other filamentous fungi immobilization in industry. Copyright © 2018 Elsevier B.V. All rights reserved.

Production of branched-chain alcohols by recombinant Ralstonia eutropha in fed-batch cultivation

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

Fei, Q; Brigham, CJ; Lu, JN

Branched-chain alcohols are considered promising green energy sources due to their compatibility with existing infrastructure and their high energy density. We utilized a strain of Ralstonia eutropha capable of producing branched-chain alcohols and examined its production in flask cultures. In order to increase isobutanol and 3-methyl-1-butanol (isoamyl alcohol) productivity in the engineered strain, batch, fed-batch, and two-stage fed-batch cultures were carried out in this work. The effects of nitrogen source concentration on branched-chain alcohol production were investigated under four different initial concentrations in fermenters. A maximum 380 g m(-3) of branched-chain alcohol production was observed with 2 kg m(-3) initialmore » NH4Cl concentration in batch cultures. A pH-stat control strategy was utilized to investigate the optimum carbon source amount fed during fed-batch cultures for higher cell density. In cultures of R. eutropha strains that did not produce polyhydroxyalkanoate or branched-chain alcohols, a maximum cell dry weight of 36 kg m(-3) was observed using a fed-batch strategy, when 10 kg m(-3) carbon source was fed into culture medium. Finally, a total branched-chain alcohol titer of 790 g m(-3), the highest branched-chain alcohol yield of 0.03 g g(-1), and the maximum branched-chain alcohol productivity of 8.23 g m(-3) h(-1) were obtained from the engineered strain Re2410/pJL26 in a two-stage fed-batch culture system with pH-stat control. Isobutanol made up over 95% (mass fraction) of the total branched-chain alcohols titer produced in this study. (C) 2013 Published by Elsevier Ltd.« less

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.

Nisin production of Lactococcus lactis N8 with hemin-stimulated cell respiration in fed-batch fermentation system.

PubMed

Kördikanlıoğlu, Burcu; Şimşek, Ömer; Saris, Per E J

2015-01-01

In this study, nisin production of Lactococcus lactis N8 was optimized by independent variables of glucose, hemin and oxygen concentrations in fed-batch fermentation in which respiration of cells was stimulated with hemin. Response surface model was able to explain the changes of the nisin production of L. lactis N8 in fed-batch fermentation system with high fidelity (R(2) 98%) and insignificant lack of fit. Accordingly, the equation developed indicated the optimum parameters for glucose, hemin, and dissolved oxygen were 8 g L(-1) h(-1) , 3 μg mL(-1) and 40%, respectively. While 1711 IU mL(-1) nisin was produced by L. lactis N8 in control fed-batch fermentation, 5410 IU mL(-1) nisin production was achieved within the relevant optimum parameters where the respiration of cell was stimulated with hemin. Accordingly, nisin production was enhanced 3.1 fold in fed-batch fermentation using hemin. In conclusion the nisin production of L. lactis N8 was enhanced extensively as a result of increasing the biomass by stimulating the cell respiration with adding the hemin in the fed-batch fermentation. © 2015 American Institute of Chemical Engineers.

Removal and recovery of uranium(VI) by waste digested activated sludge in fed-batch stirred tank reactor.

PubMed

Jain, Rohan; Peräniemi, Sirpa; Jordan, Norbert; Vogel, Manja; Weiss, Stephan; Foerstendorf, Harald; Lakaniemi, Aino-Maija

2018-05-24

This study demonstrated the removal and recovery of uranium(VI) in a fed-batch stirred tank reactor (STR) using waste digested activated sludge (WDAS). The batch adsorption experiments showed that WDAS can adsorb 200 (±9.0) mg of uranium(VI) per g of WDAS. The maximum adsorption of uranium(VI) was achieved even at an acidic initial pH of 2.7 which increased to a pH of 4.0 in the equilibrium state. Desorption of uranium(VI) from WDAS was successfully demonstrated from the release of more than 95% of uranium(VI) using both acidic (0.5 M HCl) and alkaline (1.0 M Na 2 CO 3 ) eluents. Due to the fast kinetics of uranium(VI) adsorption onto WDAS, the fed-batch STR was successfully operated at a mixing time of 15 min. Twelve consecutive uranium(VI) adsorption steps with an average adsorption efficiency of 91.5% required only two desorption steps to elute more than 95% of uranium(VI) from WDAS. Uranium(VI) was shown to interact predominantly with the phosphoryl and carboxyl groups of the WDAS, as revealed by in situ infrared spectroscopy and time-resolved laser-induced fluorescence spectroscopy studies. This study provides a proof-of-concept of the use of fed-batch STR process based on WDAS for the removal and recovery of uranium(VI). Copyright © 2018 Elsevier Ltd. All rights reserved.

Citric acid production from hydrolysate of pretreated straw cellulose by Yarrowia lipolytica SWJ-1b using batch and fed-batch cultivation.

PubMed

Liu, Xiaoyan; Lv, Jinshun; Zhang, Tong; Deng, Yuanfang

2015-01-01

In this study, crude cellulase produced by Trichoderma reesei Rut-30 was used to hydrolyze pretreated straw. After the compositions of the hydrolysate of pretreated straw were optimized, the study showed that natural components of pretreated straw without addition of any other components such as (NH4)2SO4, KH2PO4, or Mg(2+) were suitable for citric acid production by Yarrowia lipolytica SWJ-1b, and the optimal ventilatory capacity was 10.0 L/min/L medium. Batch and fed-batch production of citric acid from the hydrolysate of pretreated straw by Yarrowia lipolytica SWJ-1b has been investigated. In the batch cultivation, 25.4 g/L and 26.7 g/L citric acid were yields from glucose and hydrolysate of straw cellulose, respectively, while the cultivation time was 120 hr. In the three-cycle fed-batch cultivation, citric acid (CA) production was increased to 42.4 g/L and the cultivation time was extended to 240 hr. However, iso-citric acid (ICA) yield in fed-batch cultivation (4.0 g/L) was similar to that during the batch cultivation (3.9 g/L), and only 1.6 g/L of reducing sugar was left in the medium at the end of fed-batch cultivation, suggesting that most of the added carbon was used in the cultivation.

Towards the Integration of Dark- and Photo-Fermentative Waste Treatment. 4. Repeated Batch Sequential Dark- and Photofermentation using Starch as Substrate

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

Laurinavichene, T. V.; Belokopytov, B. F.; Laurinavichius, K. S.

In this study we demonstrated the technical feasibility of a prolonged, sequential two-stage integrated process under a repeated batch mode of starch fermentation. In this durable scheme, the photobioreactor with purple bacteria in the second stage was fed directly with dark culture from the first stage without centrifugation, filtration, or sterilization (not demonstrated previously). After preliminary optimization, both the dark- and the photo-stages were performed under repeated batch modes with different process parameters. Continuous H{sub 2} production in this system was observed at a H{sub 2} yield of up to 1.4 and 3.9 mole mole{sup -1} hexose during the dark-more » and photo-stage, respectively (for a total of 5.3 mole mole{sup -1} hexose), and rates of 0.9 and 0.5 L L{sup -1} d{sup -1}, respectively. Prolonged repeated batch H{sub 2} production was maintained for up to 90 days in each stage and was rather stable under non-aseptic conditions. Potential for improvements in these results are discussed.« less

Nisin production in realkalized fed-batch cultures in whey with feeding with lactose- or glucose-containing substrates.

PubMed

Costas Malvido, Mónica; Alonso González, Elisa; Pérez Guerra, Nelson

2016-09-01

Nisin production by Lactococcus lactis CECT 539 was followed in batch cultures in whey supplemented with different concentrations of glucose and in two realkalized fed-batch fermentations in unsupplemented whey, which were fed, respectively, with concentrated solutions of lactose and glucose. In the batch fermentations, supplementation of whey with glucose inhibited both the growth and bacteriocin production. However, fed-batch cultures were characterized with high productions of biomass (1.34 and 1.51 g l(-1)) and nisin (50.6 and 60.3 BU ml(-1)) in comparison to the batch fermentations in unsupplemented whey (0.48 g l(-1) and 22.5 BU ml(-1)) and MRS broth (1.59 g l(-1) and 50.0 BU ml(-1)). In the two realkalized fed-batch fermentations, the increase in bacteriocin production parallels both the biomass production and pH drop generated in each realkalization and feeding cycle, suggesting that nisin was synthesized as a pH-dependent primary metabolite. A shift from homolactic to heterolactic fermentation was observed at the 108 h of incubation, and other metabolites (acetic acid and butane-2,3-diol) in addition to lactic acid accumulated in the medium. On the other hand, the feeding with glucose improved the efficiencies in glucose, nitrogen, and phosphorus consumption as compared to the batch cultures. The realkalized fed-batch fermentations showed to be an effective strategy to enhance nisin production in whey by using an appropriate feeding strategy to avoid the substrate inhibition.

Enhanced bioethanol production by fed-batch simultaneous saccharification and co-fermentation at high solid loading of Fenton reaction and sodium hydroxide sequentially pretreated sugarcane bagasse.

PubMed

Zhang, Teng; Zhu, Ming-Jun

2017-04-01

A study on the fed-batch simultaneous saccharification and co-fermentation (SSCF) of Fenton reaction combined with NaOH pretreated sugarcane bagasse (SCB) at a high solid loading of 10-30% (w/v) was investigated. Enzyme feeding mode, substrate feeding mode and combination of both were compared with the batch mode under respective solid loadings. Ethanol concentrations of above 80g/L were obtained in batch and enzyme feeding modes at a solid loading of 30% (w/v). Enzyme feeding mode was found to increase ethanol productivity and reduce enzyme loading to a value of 1.23g/L/h and 9FPU/g substrate, respectively. The present study provides an economically feasible process for high concentration bioethanol production. Copyright © 2017 Elsevier Ltd. All rights reserved.

Benchmarking of commercially available CHO cell culture media for antibody production.

PubMed

Reinhart, David; Damjanovic, Lukas; Kaisermayer, Christian; Kunert, Renate

2015-06-01

In this study, eight commercially available, chemically defined Chinese hamster ovary (CHO) cell culture media from different vendors were evaluated in batch culture using an IgG-producing CHO DG44 cell line as a model. Medium adaptation revealed that the occurrence of even small aggregates might be a good indicator of cell growth performance in subsequent high cell density cultures. Batch experiments confirmed that the culture medium has a significant impact on bioprocess performance, but high amino acid concentrations alone were not sufficient to ensure superior cell growth and high antibody production. However, some key amino acids that were limiting in most media could be identified. Unbalanced glucose and amino acids led to high cell-specific lactate and ammonium production rates. In some media, persistently high glucose concentrations probably induced the suppression of respiration and oxidative phosphorylation, known as Crabtree effect, which resulted in high cell-specific glycolysis rates along with a continuous and high lactate production. In additional experiments, two of the eight basal media were supplemented with feeds from two different manufacturers in six combinations, in order to understand the combined impact of media and feeds on cell metabolism in a CHO fed-batch process. Cell growth, nutrient consumption and metabolite production rates, antibody production, and IgG quality were evaluated in detail. Concentrated feed supplements boosted cell concentrations almost threefold and antibody titers up to sevenfold. Depending on the fed-batch strategy, fourfold higher peak cell concentrations and eightfold increased IgG titers (up to 5.8 g/L) were achieved. The glycolytic flux was remarkably similar among the fed-batches; however, substantially different specific lactate production rates were observed in the different media and feed combinations. Further analysis revealed that in addition to the feed additives, the basal medium can make a considerable contribution to the ammonium metabolism of the cells. The glycosylation of the recombinant antibody was influenced by the selection of basal medium and feeds. Differences of up to 50 % in the monogalacto-fucosylated (G1F) and high mannose fraction of the IgG were observed.

Use of response surface methodology in a fed-batch process for optimization of tricarboxylic acid cycle intermediates to achieve high levels of canthaxanthin from Dietzia natronolimnaea HS-1.

PubMed

Nasri Nasrabadi, Mohammad Reza; Razavi, Seyed Hadi

2010-04-01

In this work, we applied statistical experimental design to a fed-batch process for optimization of tricarboxylic acid cycle (TCA) intermediates in order to achieve high-level production of canthaxanthin from Dietzia natronolimnaea HS-1 cultured in beet molasses. A fractional factorial design (screening test) was first conducted on five TCA cycle intermediates. Out of the five TCA cycle intermediates investigated via screening tests, alfaketoglutarate, oxaloacetate and succinate were selected based on their statistically significant (P<0.05) and positive effects on canthaxanthin production. These significant factors were optimized by means of response surface methodology (RSM) in order to achieve high-level production of canthaxanthin. The experimental results of the RSM were fitted with a second-order polynomial equation by means of a multiple regression technique to identify the relationship between canthaxanthin production and the three TCA cycle intermediates. By means of this statistical design under a fed-batch process, the optimum conditions required to achieve the highest level of canthaxanthin (13172 + or - 25 microg l(-1)) were determined as follows: alfaketoglutarate, 9.69 mM; oxaloacetate, 8.68 mM; succinate, 8.51 mM. Copyright 2009 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Stochastic models to study the impact of mixing on a fed-batch culture of Saccharomyces cerevisiae.

PubMed

Delvigne, F; Lejeune, A; Destain, J; Thonart, P

2006-01-01

The mechanisms of interaction between microorganisms and their environment in a stirred bioreactor can be modeled by a stochastic approach. The procedure comprises two submodels: a classical stochastic model for the microbial cell circulation and a Markov chain model for the concentration gradient calculus. The advantage lies in the fact that the core of each submodel, i.e., the transition matrix (which contains the probabilities to shift from a perfectly mixed compartment to another in the bioreactor representation), is identical for the two cases. That means that both the particle circulation and fluid mixing process can be analyzed by use of the same modeling basis. This assumption has been validated by performing inert tracer (NaCl) and stained yeast cells dispersion experiments that have shown good agreement with simulation results. The stochastic model has been used to define a characteristic concentration profile experienced by the microorganisms during a fermentation test performed in a scale-down reactor. The concentration profiles obtained in this way can explain the scale-down effect in the case of a Saccharomyces cerevisiae fed-batch process. The simulation results are analyzed in order to give some explanations about the effect of the substrate fluctuation dynamics on S. cerevisiae.

Improved productivity of poly (3-hydroxybutyrate) (PHB) in thermophilic Chelatococcus daeguensis TAD1 using glycerol as the growth substrate in a fed-batch culture.

PubMed

Cui, Bin; Huang, Shaobin; Xu, Fuqian; Zhang, Ruijian; Zhang, Yongqing

2015-07-01

A particularly successful polyhydroxyalkanoate (PHA) in industrial applications is poly (3-hydroxybutyrate) (PHB). However, one of the major obstacles for wider application of PHB is the cost of its production and purification. Therefore, it is desirable to discover a method for producing PHB in large quantities at a competitive price. Glycerol is a cheap and widely used carbon source that can be applied in PHB production process. There are numerous advantages to operating fermentation at elevated temperatures; only several thermophilic bacteria are able to accumulate PHB when glycerol is the growth substrate. Here, we report on the possibility of increasing PHB production at low cost using thermophilic Chelatococcus daeguensis TAD1 when glycerol is the growth substrate in a fed-batch culture. We found that (1) excess glycerol inhibited PHB accumulation and (2) organic nitrogen sources, such as tryptone and yeast extract, promoted the growth of C. daeguensis TAD1. In the batch fermentation experiments, we found that using glycerol at low concentrations as the sole carbon source, along with the addition of mixed nitrate (NH4Cl, tryptone, and yeast extract), stimulated PHB accumulation in C. daeguensis TAD1. The results showed that the PHB productivity decreased in the following order: two-stage fed-batch fermentation > fed-batch fermentation > batch fermentation. In optimized culture conditions, a PHB amount of 17.4 g l(-1) was obtained using a two-stage feeding regimen, leading to a productivity rate of 0.434 g l(-1) h(-1), which is the highest productivity rate reported for PHB to date. This high PHB biosynthetic productivity could decrease the total production cost, allowing for further development of industrial applications of PHB.

Production of biomass and polysaccharides of Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum (W.Curt. :Fr.) P. Karst. (higher Basidiomycetes), by submerged cultivation.

PubMed

Habijanic, Jozica; Berovic, Marin; Boh, Bojana; Wraber, Branka; Petravic-Tominac, Vlatka

2013-01-01

Submerged batch and repeated fed-batch cultivation techniques were used for mycelia cultivation and polysaccharide production of the Lingzhi or Reishi medicinal mushroom Ganoderma lucidum. Although most publications use various Asiatic G. lucidum strains, the growth of the strain Ga.l 4 (Biotechnical Faculty Strain Collection, Ljubljana, Slovenia), originally isolated from the Slovenian forest, is much faster. The results between the batch and repeated fed-batch cultivation are compared with the polysaccharide production in batch cultivation. From the aspect of biomass production, the best results were obtained in repeated fed-batch after 44 days, where 12.4 g/L of dry fungal biomass was obtained.

A novel model-based control strategy for aerobic filamentous fungal fed-batch fermentation processes.

PubMed

Mears, Lisa; Stocks, Stuart M; Albaek, Mads O; Cassells, Benny; Sin, Gürkan; Gernaey, Krist V

2017-07-01

A novel model-based control strategy has been developed for filamentous fungal fed-batch fermentation processes. The system of interest is a pilot scale (550 L) filamentous fungus process operating at Novozymes A/S. In such processes, it is desirable to maximize the total product achieved in a batch in a defined process time. In order to achieve this goal, it is important to maximize both the product concentration, and also the total final mass in the fed-batch system. To this end, we describe the development of a control strategy which aims to achieve maximum tank fill, while avoiding oxygen limited conditions. This requires a two stage approach: (i) calculation of the tank start fill; and (ii) on-line control in order to maximize fill subject to oxygen transfer limitations. First, a mechanistic model was applied off-line in order to determine the appropriate start fill for processes with four different sets of process operating conditions for the stirrer speed, headspace pressure, and aeration rate. The start fills were tested with eight pilot scale experiments using a reference process operation. An on-line control strategy was then developed, utilizing the mechanistic model which is recursively updated using on-line measurements. The model was applied in order to predict the current system states, including the biomass concentration, and to simulate the expected future trajectory of the system until a specified end time. In this way, the desired feed rate is updated along the progress of the batch taking into account the oxygen mass transfer conditions and the expected future trajectory of the mass. The final results show that the target fill was achieved to within 5% under the maximum fill when tested using eight pilot scale batches, and over filling was avoided. The results were reproducible, unlike the reference experiments which show over 10% variation in the final tank fill, and this also includes over filling. The variance of the final tank fill is reduced by over 74%, meaning that it is possible to target the final maximum fill reproducibly. The product concentration achieved at a given set of process conditions was unaffected by the control strategy. Biotechnol. Bioeng. 2017;114: 1459-1468. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

Adaptation of the xylose fermenting yeast Saccharomyces cerevisiae F12 for improving ethanol production in different fed-batch SSF processes.

PubMed

Tomás-Pejó, E; Ballesteros, M; Oliva, J M; Olsson, L

2010-11-01

An efficient fermenting microorganism for bioethanol production from lignocellulose is highly tolerant to the inhibitors released during pretreatment and is able to ferment efficiently both glucose and xylose. In this study, directed evolution was employed to improve the xylose fermenting Saccharomyces cerevisiae F12 strain for bioethanol production at high substrate loading. Adapted and parental strains were compared with respect to xylose consumption and ethanol production. Adaptation led to an evolved strain more tolerant to the toxic compounds present in the medium. When using concentrated prehydrolysate from steam-pretreated wheat straw with high inhibitor concentration, an improvement of 65 and 20% in xylose consumption and final ethanol concentration, respectively, were achieved using the adapted strain. To address the need of high substrate loadings, fed-batch SSF experiments were performed and an ethanol concentration as high as 27.4 g/l (61% of the theoretical) was obtained with 11.25% (w/w) of water insoluble solids (WIS).

Effects of carbon brush anode size and loading on microbial fuel cell performance in batch and continuous mode

NASA Astrophysics Data System (ADS)

Lanas, Vanessa; Ahn, Yongtae; Logan, Bruce E.

2014-02-01

Larger scale microbial fuel cells (MFCs) require compact architectures to efficiently treat wastewater. We examined how anode-brush diameter, number of anodes, and electrode spacing affected the performance of the MFCs operated in fed-batch and continuous flow mode. All anodes were initially tested with the brush core set at the same distance from the cathode. In fed-batch mode, the configuration with three larger brushes (25 mm diameter) produced 80% more power (1240 mW m-2) than reactors with eight smaller brushes (8 mm) (690 mW m-2). The higher power production by the larger brushes was due to more negative and stable anode potentials than the smaller brushes. The same general result was obtained in continuous flow operation, although power densities were reduced. However, by moving the center of the smaller brushes closer to the cathode (from 16.5 to 8 mm), power substantially increased from 690 to 1030 mW m-2 in fed batch mode. In continuous flow mode, power increased from 280 to 1020 mW m-2, resulting in more power production from the smaller brushes than the larger brushes (540 mW m-2). These results show that multi-electrode MFCs can be optimized by selecting smaller anodes, placed as close as possible to the cathode.

Modeling and parameters identification of 2-keto-L-gulonic acid fed-batch fermentation.

PubMed

Wang, Tao; Sun, Jibin; Yuan, Jingqi

2015-04-01

This article presents a modeling approach for industrial 2-keto-L-gulonic acid (2-KGA) fed-batch fermentation by the mixed culture of Ketogulonicigenium vulgare (K. vulgare) and Bacillus megaterium (B. megaterium). A macrokinetic model of K. vulgare is constructed based on the simplified metabolic pathways. The reaction rates obtained from the macrokinetic model are then coupled into a bioreactor model such that the relationship between substrate feeding rates and the main state variables, e.g., the concentrations of the biomass, substrate and product, is constructed. A differential evolution algorithm using the Lozi map as the random number generator is utilized to perform the model parameters identification, with the industrial data of 2-KGA fed-batch fermentation. Validation results demonstrate that the model simulations of substrate and product concentrations are well in coincidence with the measurements. Furthermore, the model simulations of biomass concentrations reflect principally the growth kinetics of the two microbes in the mixed culture.

Development of a GMP Phase III purification process for VB4-845, an immunotoxin expressed in E. coli using high cell density fermentation.

PubMed

Premsukh, Arjune; Lavoie, Joelle M; Cizeau, Jeannick; Entwistle, Joycelyn; MacDonald, Glen C

2011-07-01

VB4-845 is a recombinant immunotoxin comprised of an anti-epithelial cell adhesion molecule (EpCAM) scFv fused to a truncated form of the bacterial toxin, Pseudomonas exotoxin A. VB4-845, purified from TB fed-batch fermentation, showed clinical efficacy when administered locally to treat non-muscle invasive bladder cancer (NMIBC) and squamous cell carcinomas of the head and neck (SCCHN). Here, we describe the implementation of an Escherichia coli high cell density (HCD) cultivation and purification process for VB4-845. HCD cultivation was a prerequisite for achieving higher yields necessary for Phase III clinical trials and commercialization. Using this process, the VB4-845 titer in the supernatant was increased by 30-fold over the original TB fed-batch cultivation. To obtain clinical grade material, a process involving a five-step column purification procedure was implemented and led to an overall recovery of ∼ 40%. VB4-845 purity of >97% was achieved after the first three columns following the removal of low-molecular weight product-related impurities and aggregates. Endotoxins were effectively separated from VB4-845 on the Q-columns and by washing the Ni-column with a detergent buffer while host cell proteins were removed using ceramic hydroxyapatite. Comparability studies demonstrated that the purified product from the Phase III process was identical to the Phase II reference standard produced using TB fed-batch fermentation. Copyright © 2011 Elsevier Inc. All rights reserved.

Conversion of H2 and CO2 to CH4 and acetate in fed-batch biogas reactors by mixed biogas community: a novel route for the power-to-gas concept.

PubMed

Szuhaj, Márk; Ács, Norbert; Tengölics, Roland; Bodor, Attila; Rákhely, Gábor; Kovács, Kornél L; Bagi, Zoltán

2016-01-01

Applications of the power-to-gas principle for the handling of surplus renewable electricity have been proposed. The feasibility of using hydrogenotrophic methanogens as CH4 generating catalysts has been demonstrated. Laboratory and scale-up experiments have corroborated the benefits of the CO2 mitigation via biotechnological conversion of H2 and CO2 to CH4. A major bottleneck in the process is the gas-liquid mass transfer of H2. Fed-batch reactor configuration was tested at mesophilic temperature in laboratory experiments in order to improve the contact time and H2 mass transfer between the gas and liquid phases. Effluent from an industrial biogas facility served as biocatalyst. The bicarbonate content of the effluent was depleted after some time, but the addition of stoichiometric CO2 sustained H2 conversion for an extended period of time and prevented a pH shift. The microbial community generated biogas from the added α-cellulose substrate with concomitant H2 conversion, but the organic substrate did not facilitate H2 consumption. Fed-batch operational mode allowed a fourfold increase in volumetric H2 load and a 6.5-fold augmentation of the CH4 formation rate relative to the CSTR reactor configuration. Acetate was the major by-product of the reaction. Fed-batch reactors significantly improve the efficiency of the biological power-to-gas process. Besides their storage function, biogas fermentation effluent reservoirs can serve as large-scale bio CH4 reactors. On the basis of this recognition, a novel concept is proposed, which merges biogas technology with other means of renewable electricity production for improved efficiency and sustainability.

Integrated continuous dissolution, refolding and tag removal of fusion proteins from inclusion bodies in a tubular reactor.

PubMed

Pan, Siqi; Zelger, Monika; Jungbauer, Alois; Hahn, Rainer

2014-09-20

An integrated continuous tubular reactor system was developed for processing an autoprotease expressed as inclusion bodies. The inclusion bodies were suspended and fed into the tubular reactor system for continuous dissolving, refolding and precipitation. During refolding, the dissolved autoprotease cleaves itself, separating the fusion tag from the target peptide. Subsequently, the cleaved fusion tag and any uncleaved autoprotease were precipitated out in the precipitation step. The processed exiting solution results in the purified soluble target peptide. Refolding and precipitation yields performed in the tubular reactor were similar to batch reactor and process was stable for at least 20 h. The authenticity of purified peptide was also verified by mass spectroscopy. Productivity (in mg/l/h and mg/h) calculated in the tubular process was twice and 1.5 times of the batch process, respectively. Although it is more complex to setup a tubular than a batch reactor, it offers faster mixing, higher productivity and better integration to other bioprocessing steps. With increasing interest of integrated continuous biomanufacturing, the use of tubular reactors in industrial settings offers clear advantages. Copyright © 2014 Elsevier B.V. All rights reserved.

Improvement of constraint-based flux estimation during L-phenylalanine production with Escherichia coli using targeted knock-out mutants.

PubMed

Weiner, Michael; Tröndle, Julia; Albermann, Christoph; Sprenger, Georg A; Weuster-Botz, Dirk

2014-07-01

Fed-batch production of the aromatic amino acid L-phenylalanine was studied with recombinant Escherichia coli strains on a 15 L-scale using glycerol as carbon source. Flux Variability Analysis (FVA) was applied for intracellular flux estimation to obtain an insight into intracellular flux distribution during L-phenylalanine production. Variability analysis revealed great flux uncertainties in the central carbon metabolism, especially concerning malate consumption. Due to these results two recombinant strains were genetically engineered differing in the ability of malate degradation and anaplerotic reactions (E. coli FUS4.11 ΔmaeA pF81kan and E. coli FUS4.11 ΔmaeA ΔmaeB pF81kan). Applying these malic enzyme knock-out mutants in the standardized L-phenylalanine production process resulted in almost identical process performances (e.g., L-phenylalanine concentration, production rate and byproduct formation). This clearly highlighted great redundancies in central metabolism in E. coli. Uncertainties of intracellular flux estimations by constraint-based analyses during fed-batch production of L-phenylalanine were drastically reduced by application of the malic enzyme knock-out mutants. © 2014 Wiley Periodicals, Inc.

Very High Density of Chinese Hamster Ovary Cells in Perfusion by Alternating Tangential Flow or Tangential Flow Filtration in WAVE Bioreactor™—Part II: Applications for Antibody Production and Cryopreservation

PubMed Central

Clincke, Marie-Françoise; Mölleryd, Carin; Samani, Puneeth K; Lindskog, Eva; Fäldt, Eric; Walsh, Kieron; Chotteau, Véronique

2013-01-01

A high cell density perfusion process of monoclonal antibody (MAb) producing Chinese hamster ovary (CHO) cells was developed in disposable WAVE Bioreactor™ using external hollow fiber (HF) filter as cell separation device. Tangential flow filtration (TFF) and alternating tangential flow (ATF) systems were compared and process applications of high cell density perfusion were studied here: MAb production and cryopreservation. Operations by perfusion using microfiltration (MF) or ultrafiltration (UF) with ATF or TFF and by fed-batch were compared. Cell densities higher than 108 cells/mL were obtained using UF TFF or UF ATF. The cells produced comparable amounts of MAb in perfusion by ATF or TFF, MF or UF. MAbs were partially retained by the MF using ATF or TFF but more severely using TFF. Consequently, MAbs were lost when cell broth was discarded from the bioreactor in the daily bleeds. The MAb cell-specific productivity was comparable at cell densities up to 1.3 × 108 cells/mL in perfusion and was comparable or lower in fed-batch. After 12 days, six times more MAbs were harvested using perfusion by ATF or TFF with MF or UF, compared to fed-batch and 28× more in a 1-month perfusion at 108 cells/mL density. Pumping at a recirculation rate up to 2.75 L/min did not damage the cells with the present TFF settings with HF short circuited. Cell cryopreservation at 0.5 × 108 and 108 cells/mL was performed using cells from a perfusion run at 108 cells/mL density. Cell resuscitation was very successful, showing that this system was a reliable process for cell bank manufacturing. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:768–777, 2013 PMID:23436783

Comparison of semi-batch vs. continuously fed anaerobic bioreactors for the treatment of a high-strength, solids-rich pumpkin-processing wastewater.

PubMed

del Agua, Isabel; Usack, Joseph G; Angenent, Largus T

2015-01-01

The objective of this work was to compare two different high-rate anaerobic bioreactor configurations--the anaerobic sequencing batch reactor (ASBR) and the upflow anaerobic solid removal (UASR) reactor--for the treatment of a solid-rich organic wastewater with a high strength. The two, 4.5-L reactors were operated in parallel for close to 100 days under mesophilic conditions (37°C) with non-granular biomass by feeding a pumpkin wastewater with ∼4% solids. The organic loading rate of pumpkin wastewater was increased periodically to a maximum of 8 g COD L(-1) d(-1) by shortening the hydraulic retention time to 5.3 days. Compositional analysis of pumpkin wastewater revealed deficiencies in the trace metal cobalt and alkalinity. With supplementation, the ASBR outperformed the UASR reactor with total chemical oxygen demand (COD) removal efficiencies of 64% and 53%, respectively, achieving a methane yield of 0.27 and 0.20 L CH4 g(-1) COD fed to the ASBR and UASR, respectively. The better performance realized with the ASBR and this specific wastewater was attributed to its semi-batch, dynamic operating conditions rather than the continuous operating conditions of the UASR reactor.

Rational development of solid dispersions via hot-melt extrusion using screening, material characterization, and numeric simulation tools.

PubMed

Zecevic, Damir E; Wagner, Karl G

2013-07-01

Effective and predictive small-scale selection tools are inevitable during the development of a solubility enhanced drug product. For hot-melt extrusion, this selection process can start with a microscale performance evaluation on a hot-stage microscope (HSM). A batch size of 400 mg can provide sufficient materials to assess the drug product attributes such as solid-state properties, solubility enhancement, and physical stability as well as process related attributes such as processing temperature in a twin-screw extruder (TSE). Prototype formulations will then be fed into a 5 mm TSE (~1-2 g) to confirm performance from the HSM under additional shear stress. Small stress stability testing might be performed with these samples or a larger batch (20-40 g) made by 9 or 12 mm TSE. Simultaneously, numeric process simulations are performed using process data as well as rheological and thermal properties of the formulations. Further scale up work to 16 and 18 mm TSE confirmed and refined the simulation model. Thus, at the end of the laboratory-scale development, not only the clinical trial supply could be manufactured, but also one can form a sound risk assessment to support further scale up even without decades of process experience. Copyright © 2013 Wiley Periodicals, Inc.

Comparison of batch cultivation strategies for cost-effective biomass production of Micractinium inermum NLP-F014 using a blended wastewater medium.

PubMed

Park, Seonghwan; Kim, Jeongmi; Park, Younghyun; Son, Suyoung; Cho, Sunja; Kim, Changwon; Lee, Taeho

2017-06-01

Two competitive strategies, fed-batch and sequencing-batch cultivation, were compared in cost-effective biomass production of a high lipid microalgae, Micractinium inermum NLP-F014 using a blended wastewater medium. For fed-batch cultivations, additional nutrient was supplemented at day 2 (FB1) or consecutively added at day 2 and 4 (FB2). Through inoculum size test, 1.0g-DCWL -1 was selected for the sequencing-batch cultivation (SB) where about 65% of culture was replaced with fresh medium every 2days. Both fed-batch cultivations showed the maximum biomass productivity of 0.95g-DCWL -1 d -1 , while average biomass productivity in SB was slightly higher as 0.96±0.08g-DCWL -1 d -1 . Furthermore, remained concentrations of organics (426mg-CODL -1 ), total nitrogen (15.4mg-NL -1 ) and phosphorus (0.6mg-PL -1 ) in SB were much lower than those of fed-batch conditions. The results suggested that SB could be a promising strategy to cultivate M. inermum NLP-F014 with the blended wastewater medium. Copyright © 2017 Elsevier Ltd. All rights reserved.

Modeling of Fusarium redolens Dzf2 mycelial growth kinetics and optimal fed-batch fermentation for beauvericin production.

PubMed

Xu, Li-Jian; Liu, Yuan-Shuai; Zhou, Li-Gang; Wu, Jian-Yong

2011-09-01

Beauvericin (BEA) is a cyclic hexadepsipeptide mycotoxin with notable phytotoxic and insecticidal activities. Fusarium redolens Dzf2 is a highly BEA-producing fungus isolated from a medicinal plant. The aim of the current study was to develop a simple and valid kinetic model for F. redolens Dzf2 mycelial growth and the optimal fed-batch operation for efficient BEA production. A modified Monod model with substrate (glucose) and product (BEA) inhibition was constructed based on the culture characteristics of F. redolens Dzf2 mycelia in a liquid medium. Model parameters were derived by simulation of the experimental data from batch culture. The model fitted closely with the experimental data over 20-50 g l(-1) glucose concentration range in batch fermentation. The kinetic model together with the stoichiometric relationships for biomass, substrate and product was applied to predict the optimal feeding scheme for fed-batch fermentation, leading to 54% higher BEA yield (299 mg l(-1)) than in the batch culture (194 mg l(-1)). The modified Monod model incorporating substrate and product inhibition was proven adequate for describing the growth kinetics of F. redolens Dzf2 mycelial culture at suitable but not excessive initial glucose levels in batch and fed-batch cultures.

Optimal quality control of bakers' yeast fed-batch culture using population dynamics.

PubMed

Dairaku, K; Izumoto, E; Morikawa, H; Shioya, S; Takamatsu, T

1982-12-01

An optimal quality control policy for the overall specific growth rate of bakers' yeast, which maximizes the fermentative activity in the making of bread, was obtained by direct searching based on the mathematical model proposed previously. The mathematical model had described the age distribution of bakers' yeast which had an essential relationship to the ability of fermentation in the making of bread. The mathematical model is a simple aging model with two periods: Nonbudding and budding. Based on the result obtained by direct searching, the quality control of bakers' yeast fed-batch culture was performed and confirmed to be experimentally valid.

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.

A Simple Method to Reduce both Lactic Acid and Ammonium Production in Industrial Animal Cell Culture

PubMed Central

Freund, Nathaniel W.; Croughan, Matthew S.

2018-01-01

Fed-batch animal cell culture is the most common method for commercial production of recombinant proteins. However, higher cell densities in these platforms are still limited due to factors such as excessive ammonium production, lactic acid production, nutrient limitation, and/or hyperosmotic stress related to nutrient feeds and base additions to control pH. To partly overcome these factors, we investigated a simple method to reduce both ammonium and lactic acid production—termed Lactate Supplementation and Adaptation (LSA) technology—through the use of CHO cells adapted to a lactate-supplemented medium. Using this simple method, we achieved a reduction of nearly 100% in lactic acid production with a simultaneous 50% reduction in ammonium production in batch shaker flasks cultures. In subsequent fed-batch bioreactor cultures, lactic acid production and base addition were both reduced eight-fold. Viable cell densities of 35 million cells per mL and integral viable cell days of 273 million cell-days per mL were achieved, both among the highest currently reported for a fed-batch animal cell culture. Investigating the benefits of LSA technology in animal cell culture is worthy of further consideration and may lead to process conditions more favorable for advanced industrial applications. PMID:29382079

A Simple Method to Reduce both Lactic Acid and Ammonium Production in Industrial Animal Cell Culture.

PubMed

Freund, Nathaniel W; Croughan, Matthew S

2018-01-28

Fed-batch animal cell culture is the most common method for commercial production of recombinant proteins. However, higher cell densities in these platforms are still limited due to factors such as excessive ammonium production, lactic acid production, nutrient limitation, and/or hyperosmotic stress related to nutrient feeds and base additions to control pH. To partly overcome these factors, we investigated a simple method to reduce both ammonium and lactic acid production-termed Lactate Supplementation and Adaptation (LSA) technology-through the use of CHO cells adapted to a lactate-supplemented medium. Using this simple method, we achieved a reduction of nearly 100% in lactic acid production with a simultaneous 50% reduction in ammonium production in batch shaker flasks cultures. In subsequent fed-batch bioreactor cultures, lactic acid production and base addition were both reduced eight-fold. Viable cell densities of 35 million cells per mL and integral viable cell days of 273 million cell-days per mL were achieved, both among the highest currently reported for a fed-batch animal cell culture. Investigating the benefits of LSA technology in animal cell culture is worthy of further consideration and may lead to process conditions more favorable for advanced industrial applications.

A novel fed-batch based strategy for enhancing cell-density and recombinant cyprosin B production in bioreactors.

PubMed

Sampaio, P N; Pais, M S; Fonseca, L P

2014-12-01

Nowadays, the dairy industry is continuously looking for new and more efficient clotting enzymes to create innovative products. Cyprosin B is a plant aspartic protease characterized by clotting activity that was previously cloned in Saccharomyces cerevisiae BJ1991 strain. The production of recombinant cyprosin B by a batch and fed-batch culture was compared using glucose and galactose as carbon sources. The strategy for fed-batch cultivation involved two steps: in the first batch phase, the culture medium presented glucose 1 % (w/v) and galactose 0.5 % (w/v), while in the feed step the culture medium was constituted by 5 % (w/v) galactose with the aim to minimize the GAL7 promoter repression. Based on fed-batch, in comparison to batch growth, an increase in biomass (6.6-fold), protein concentration (59 %) and cyprosin B activity (91 %) was achieved. The recombinant cyprosin B was purified by a single hydrophobic chromatography, presenting a specific activity of 6 × 10(4) U·mg(-1), corresponding to a purification degree of 12.5-fold and a recovery yield of 16.4 %. The SDS-PAGE analysis showed that recovery procedure is suitable for achieving the purified recombinant cyprosin B. The results show that the recombinant cyprosin B production can be improved based on two distinct steps during the fed-batch, presenting that this strategy, associated with a simplified purification procedure, could be applied to large-scale production, constituting a new and efficient alternative for animal and fungal enzymes widely used in cheese making.

Enhanced production of GDP-L-fucose by overexpression of NADPH regenerator in recombinant Escherichia coli.

PubMed

Lee, Won-Heong; Chin, Young-Wook; Han, Nam Soo; Kim, Myoung-Dong; Seo, Jin-Ho

2011-08-01

Biosynthesis of guanosine 5'-diphosphate-L-fucose (GDP-L-fucose) requires NADPH as a reducing cofactor. In this study, endogenous NADPH regenerating enzymes such as glucose-6-phosphate dehydrogenase (G6PDH), isocitrate dehydrogenase (Icd), and NADP(+)-dependent malate dehydrogenase (MaeB) were overexpressed to increase GDP-L-fucose production in recombinant Escherichia coli. The effects of overexpression of each NADPH regenerating enzyme on GDP-L-fucose production were investigated in a series of batch and fed-batch fermentations. Batch fermentations showed that overexpression of G6PDH was the most effective for GDP-L-fucose production. However, GDP-L-fucose production was not enhanced by overexpression of G6PDH in the glucose-limited fed-batch fermentation. Hence, a glucose feeding strategy was optimized to enhance GDP-L-fucose production. Fed-batch fermentation with a pH-stat feeding mode for sufficient supply of glucose significantly enhanced GDP-L-fucose production compared with glucose-limited fed-batch fermentation. A maximum GDP-L-fucose concentration of 235.2 ± 3.3 mg l(-1), corresponding to a 21% enhancement in the GDP-L-fucose production compared with the control strain overexpressing GDP-L-fucose biosynthetic enzymes only, was achieved in the pH-stat fed-batch fermentation of the recombinant E. coli overexpressing G6PDH. It was concluded that sufficient glucose supply and efficient NADPH regeneration are crucial for NADPH-dependent GDP-L-fucose production in recombinant E. coli.

Xylose induces cellulase production in Thermoascus aurantiacus.

PubMed

Schuerg, Timo; Prahl, Jan-Philip; Gabriel, Raphael; Harth, Simon; Tachea, Firehiwot; Chen, Chyi-Shin; Miller, Matthew; Masson, Fabrice; He, Qian; Brown, Sarah; Mirshiaghi, Mona; Liang, Ling; Tom, Lauren M; Tanjore, Deepti; Sun, Ning; Pray, Todd R; Singer, Steven W

2017-01-01

Lignocellulosic biomass is an important resource for renewable production of biofuels and bioproducts. Enzymes that deconstruct this biomass are critical for the viability of biomass-based biofuel production processes. Current commercial enzyme mixtures have limited thermotolerance. Thermophilic fungi may provide enzyme mixtures with greater thermal stability leading to more robust processes. Understanding the induction of biomass-deconstructing enzymes in thermophilic fungi will provide the foundation for strategies to construct hyper-production strains. Induction of cellulases using xylan was demonstrated during cultivation of the thermophilic fungus Thermoascus aurantiacus . Simulated fed-batch conditions with xylose induced comparable levels of cellulases. These fed-batch conditions were adapted to produce enzymes in 2 and 19 L bioreactors using xylose and xylose-rich hydrolysate from dilute acid pretreatment of corn stover. Enzymes from T. aurantiacus that were produced in the xylose-fed bioreactor demonstrated comparable performance in the saccharification of deacetylated, dilute acid-pretreated corn stover when compared to a commercial enzyme mixture at 50 °C. The T. aurantiacus enzymes retained this activity at of 60 °C while the commercial enzyme mixture was largely inactivated. Xylose induces both cellulase and xylanase production in T. aurantiacus and was used to produce enzymes at up to the 19 L bioreactor scale. The demonstration of induction by xylose-rich hydrolysate and saccharification of deacetylated, dilute acid-pretreated corn stover suggests a scenario to couple biomass pretreatment with onsite enzyme production in a biorefinery. This work further demonstrates the potential for T. aurantiacus as a thermophilic platform for cellulase development.

Degradation of the unbiodegradable particulate fraction (XU) from different activated sludges during batch digestion tests at ambient temperature.

PubMed

Habermacher, Jonathan; Benetti, Antonio Domingues; Derlon, Nicolas; Morgenroth, Eberhard

2016-07-01

One strategy for the management of excess sludge in small wastewater treatment plants (WWTPs) consists in minimizing the excess sludge production by operating the WWTP at very long solids retention times (SRTs > 30 days). A number of recent studies have suggested that sludge minimization at very long SRT results from the degradation of the unbiodegradable particulate fraction (XU) (influent unbiodegradable compounds and endogenous decay products). But the biodegradability of the unbiodegradable particulate fraction has only been evaluated during batch digestion test performed at ambient temperature with sludge fed with synthetic wastewaters. It is not clear to what extent observations made for sludge fed with synthetic influents can be transposed to sludge fed with real influent. The current study thus focused on evaluating the biodegradability of the unbiodegradable particulate fraction for sludge fed with real wastewater. Batch digestion tests (400 days, ambient temperature) were conducted with three different sludges fed with either synthetic or real influents and exposed to aerobic or intermittent aeration conditions. Our results indicate that volatile suspended solids (VSS) decreased even after complete decay of the active biomass (i.e., after 30 days of aerobic batch digestion) indicating that the unbiodegradable particulate fraction is biodegradable. However, very low degradation rates of the unbiodegradable particulate fraction were monitored after day 30 of digestion (0.7-1.7·10(-3) d(-1)). These values were in the lower range of previously published values for synthetic wastewaters (1-7.5·10(-3) d(-1)). The low values determined in our study indicate that the rate could decrease over time or that sludge composition influences the degradability of the unbiodegradable particulate fraction. But our results also demonstrate that extracellular polymeric substances (EPS) have a minor impact on the biodegradability of the unbiodegradable particulate fraction. Overall bound EPS were indeed biodegradable under all conditions and thus did not accumulate in the unbiodegradable particulate fraction. Different bound EPS pools (e.g., cation bound EPS) were associated with specific degradation behaviors. Besides improved mechanistic understanding of sludge degradation processes, our results have implications for the development of decentralized wastewater treatment technologies with on-site reduction of excess sludge. Copyright © 2016 Elsevier Ltd. All rights reserved.

Application of enzymatic apple pomace hydrolysate to production of 2,3-butanediol by alkaliphilic Bacillus licheniformis NCIMB 8059.

PubMed

Białkowska, Aneta M; Gromek, Ewa; Krysiak, Joanna; Sikora, Barbara; Kalinowska, Halina; Jędrzejczak-Krzepkowska, Marzena; Kubik, Celina; Lang, Siegmund; Schütt, Fokko; Turkiewicz, Marianna

2015-12-01

2,3-Butanediol (2,3-BD) synthesis by a nonpathogenic bacterium Bacillus licheniformis NCIMB 8059 from enzymatic hydrolysate of depectinized apple pomace and its blend with glucose was studied. In shake flasks, the maximum diol concentration in fed-batch fermentations was 113 g/L (in 163 h, from the hydrolysate, feedings with glucose) while in batch processes it was around 27 g/L (in 32 h, from the hydrolysate and glucose blend). Fed-batch fermentations in the 0.75 and 30 L fermenters yielded 87.71 g/L 2,3-BD in 160 h, and 72.39 g/L 2,3-BD in 94 h, respectively (from the hydrolysate and glucose blend, feedings with glucose). The hydrolysate of apple pomace, which was for the first time used for microbial 2,3-BD production is not only a source of sugars but also essential minerals.

Production of nattokinase by batch and fed-batch culture of Bacillus subtilis.

PubMed

Cho, Young-Han; Song, Jae Yong; Kim, Kyung Mi; Kim, Mi Kyoung; Lee, In Young; Kim, Sang Bum; Kim, Hyeon Shup; Han, Nam Soo; Lee, Bong Hee; Kim, Beom Soo

2010-09-30

Nattokinase was produced by batch and fed-batch culture of Bacillus subtilis in flask and fermentor. Effect of supplementing complex media (peptone, yeast extract, or tryptone) was investigated on the production of nattokinase. In flask culture, the highest cell growth and nattokinase activity were obtained with 50 g/L of peptone supplementation. In this condition, nattokinase activity was 630 unit/ml at 12 h. In batch culture of B. subtilis in fermentor, the highest nattokinase activity of 3400 unit/ml was obtained at 10h with 50 g/L of peptone supplementation. From the batch kinetics data, it was shown that nattokinase production was growth-associated and culture should be harvested before stationary phase for maximum nattokinase production. In fed-batch culture of B. subtilis using pH-stat feeding strategy, cell growth (optical density monitored at 600 nm) increased to ca. 100 at 22 h, which was 2.5 times higher than that in batch culture. The highest nattokinase activity was 7100 unit/ml at 19 h, which was also 2.1 times higher than that in batch culture. Copyright 2010 Elsevier B.V. All rights reserved.

Acetone-butanol-ethanol (ABE) fermentation using Clostridium acetobutylicum XY16 and in situ recovery by PDMS/ceramic composite membrane.

PubMed

Wu, Hao; Chen, Xiao-Peng; Liu, Gong-Ping; Jiang, Min; Guo, Ting; Jin, Wan-Qin; Wei, Ping; Zhu, Da-Wei

2012-09-01

PDMS/ceramic composite membrane was directly integrated with acetone-butanol-ethanol (ABE) fermentation using Clostridium acetobutylicum XY16 at 37 °C and in situ removing ABE from fermentation broth. The membrane was integrated with batch fermentation, and approximately 46 % solvent was extracted. The solvent in permeates was 118 g/L, and solvent productivity was 0.303 g/(L/h), which was approximately 33 % higher compared with the batch fermentation without in situ recovery. The fed-batch fermentation with in situ recovery by pervaporation continued for more than 200 h, 61 % solvent was extracted, and the solvent in penetration was 96.2 g/L. The total flux ranged from 0.338 to 0.847 kg/(m(2)/h) and the separation factor of butanol ranged from 5.1 to 27.1 in this process. The membrane was fouled by the active fermentation broth, nevertheless the separation performances were partially recovered by offline membrane cleaning, and the solvent productivity was increased to 0.252 g/(L/h), which was 19 % higher compared with that in situ recovery process without membrane cleaning.

Development of a chemically defined platform fed-batch culture media for monoclonal antibody-producing CHO cell lines with optimized choline content.

PubMed

Kuwae, Shinobu; Miyakawa, Ichiko; Doi, Tomohiro

2018-01-11

A chemically defined platform basal medium and feed media were developed using a single Chinese hamster ovary (CHO) cell line that produces a monoclonal antibody (mAb). Cell line A, which showed a peak viable cell density of 5.9 × 10 6  cells/mL and a final mAb titer of 0.5 g/L in batch culture, was selected for the platform media development. Stoichiometrically balanced feed media were developed using glucose as an indicator of cell metabolism to determine the feed rates of all other nutrients. A fed-batch culture of cell line A using the platform fed-batch medium yielded a 6.4 g/L mAb titer, which was 12-fold higher than that of the batch culture. To examine the applicability of the platform basal medium and feed media, three other cell lines (A16, B, and C) that produce mAbs were cultured using the platform fed-batch medium, and they yielded mAb titers of 8.4, 3.3, and 6.2 g/L, respectively. The peak viable cell densities of the three cell lines ranged from 1.3 × 10 7 to 1.8 × 10 7  cells/mL. These results show that the nutritionally balanced fed-batch medium and feeds worked well for other cell lines. During the medium development, we found that choline limitation caused a lower cell viability, a lower mAb titer, a higher mAb aggregate content, and a higher mannose-5 content. The optimal choline chloride to glucose ratio for the CHO cell fed-batch culture was determined. Our platform basal medium and feed media will shorten the medium-development time for mAb-producing cell lines.

Scaling-up vaccine production: implementation aspects of a biomass growth observer and controller.

PubMed

Soons, Zita I T A; van den IJssel, Jan; van der Pol, Leo A; van Straten, Gerrit; van Boxtel, Anton J B

2009-04-01

This study considers two aspects of the implementation of a biomass growth observer and specific growth rate controller in scale-up from small- to pilot-scale bioreactors towards a feasible bulk production process for whole-cell vaccine against whooping cough. The first is the calculation of the oxygen uptake rate, the starting point for online monitoring and control of biomass growth, taking into account the dynamics in the gas-phase. Mixing effects and delays are caused by amongst others the headspace and tubing to the analyzer. These gas phase dynamics are modelled using knowledge of the system in order to reconstruct oxygen consumption. The second aspect is to evaluate performance of the monitoring and control system with the required modifications of the oxygen consumption calculation on pilot-scale. In pilot-scale fed-batch cultivation good monitoring and control performance is obtained enabling a doubled concentration of bulk vaccine compared to standard batch production.

Optimization of high solids fed-batch saccharification of sugarcane bagasse based on system viscosity changes.

PubMed

Liu, Yunyun; Xu, Jingliang; Zhang, Yu; Yuan, Zhenhong; Xie, Jun

2015-10-10

Viscosity trends in alkali-pretreated sugarcane bagasse (SCB) slurries undergoing high solids fed-batch enzymatic hydrolysis were measured for a range of solids loading from 15% to 36%. Solids liquefaction times were related to system viscosity changes. The viscosity decreased quickly for low solids loading, and increased with increasing solids content. Fed-batch hydrolysis was initiated with 15% solids loading, and an additional 8%, 7% and 6% were successively added after the system viscosity decreased to stable values to achieve a final solids content of 36%. Two enzyme-adding modes with 8.5FPU/g solid were investigated. The batch mode with all enzyme being added at the beginning of the reaction produced the highest yields, with approximately 231.7g/L total sugars and 134.9g/L glucose being obtained after 96h with nearly 60% of the final glucan conversion rate. This finding indicates that under the right conditions, the fed-batch strategy might be a plausible way to produce high sugars under high solids. Copyright © 2015 Elsevier B.V. All rights reserved.

Lipid Content and Cryotolerance of Bakers' Yeast in Frozen Doughs †

PubMed Central

Gélinas, Pierre; Fiset, Gisèle; Willemot, Claude; Goulet, Jacques

1991-01-01

The relationship between lipid content and tolerance to freezing at −50°C was studied in Saccharomyces cerevisiae grown under batch or fed-batch mode and various aeration and temperature conditions. A higher free-sterol-to-phospholipid ratio as well as higher free sterol and phospholipid contents correlated with the superior cryoresistance in dough or in water of the fed-batch-grown compared with the batch-grown cells. For both growth modes, the presence of excess dissolved oxygen in the culture medium greatly improved yeast cryoresistance and trehalose content (P. Gélinas, G. Fiset, A. LeDuy, and J. Goulet, Appl. Environ. Microbiol. 26:2453-2459, 1989) without significantly changing the lipid profile. Under the batch or fed-batch modes, no correlation was found between the cryotolerance of bakers' yeast and the total cellular lipid content, the total sterol content, the phospholipid unsaturation index, the phosphate or crude protein content, or the yeast cell morphology (volume and roundness). PMID:16348412

Online automatic tuning and control for fed-batch cultivation

PubMed Central

van Straten, Gerrit; van der Pol, Leo A.; van Boxtel, Anton J. B.

2007-01-01

Performance of controllers applied in biotechnological production is often below expectation. Online automatic tuning has the capability to improve control performance by adjusting control parameters. This work presents automatic tuning approaches for model reference specific growth rate control during fed-batch cultivation. The approaches are direct methods that use the error between observed specific growth rate and its set point; systematic perturbations of the cultivation are not necessary. Two automatic tuning methods proved to be efficient, in which the adaptation rate is based on a combination of the error, squared error and integral error. These methods are relatively simple and robust against disturbances, parameter uncertainties, and initialization errors. Application of the specific growth rate controller yields a stable system. The controller and automatic tuning methods are qualified by simulations and laboratory experiments with Bordetella pertussis. PMID:18157554

Heat shock protein 27 overexpression in CHO cells modulates apoptosis pathways and delays activation of caspases to improve recombinant monoclonal antibody titre in fed-batch bioreactors.

PubMed

Tan, Janice G L; Lee, Yih Yean; Wang, Tianhua; Yap, Miranda G S; Tan, Tin Wee; Ng, Say Kong

2015-05-01

CHO cells are major production hosts for recombinant biologics including the rapidly expanding recombinant monoclonal antibodies (mAbs). Heat shock protein 27 (HSP27) expression was observed to be down-regulated towards the late-exponential and stationary phase of CHO fed-batch bioreactor cultures, whereas HSP27 was found to be highly expressed in human pathological cells and reported to have anti-apoptotic functions. These phenotypes suggest that overexpression of HSP27 is a potential cell line engineering strategy for improving robustness of CHO cells. In this work, HSP27 was stably overexpressed in CHO cells producing recombinant mAb and the effects of HSP27 on cell growth, volumetric production titer and product quality were assessed. Concomitantly, HSP27 anti-apoptosis functions in CHO cells were investigated. Stably transfected clones cultured in fed-batch bioreactors displayed 2.2-fold higher peak viable cell density, delayed loss of culture viability by two days and 2.3-fold increase in mAb titer without affecting the N-glycosylation profile, as compared to clones stably transfected with the vector backbone. Co-immunoprecipitation studies revealed HSP27 interactions with Akt, pro-caspase 3 and Daxx and caspase activity profiling showed delayed increase in caspase 2, 3, 8 and 9 activities. These results suggest that HSP27 modulates apoptosis signaling pathways and delays caspase activities to improve performance of CHO fed-batch bioreactor cultures. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Bioconversion of Agricultural Waste to Ethanol by SSF Using Recombinant Cellulase from Clostridium thermocellum

PubMed Central

Mutreja, Ruchi; Das, Debasish; Goyal, Dinesh; Goyal, Arun

2011-01-01

The effect of different pretreatment methods, temperature, and enzyme concentration on ethanol production from 8 lignocellulosic agrowaste by simultaneous saccharification and fermentation (SSF) using recombinant cellulase and Saccharomyces cerevisiae were studied. Recombinant cellulase was isolated from E. coli BL21 cells transformed with CtLic26A-Cel5-CBM11 full-length gene from Clostridium thermocellum and produced in both batch and fed-batch processes. The maximum cell OD and specific activity in batch mode were 1.6 and 1.91 U/mg, respectively, whereas in the fed-batch mode, maximum cell OD and specific activity were 3.8 and 3.5 U/mg, respectively, displaying a 2-fold increase. Eight substrates, Syzygium cumini (jamun), Azadirachta indica (neem), Saracens indica (asoka), bambusa dendrocalmus (bamboo), Populas nigra (poplar), Achnatherum hymenoides (wild grass), Eucalyptus marginata (eucalyptus), and Mangifera indica (mango), were subjected to SSF. Of three pretreatments, acid, alkali, and steam explosion, acid pretreatment Syzygium cumini (Jamun) at 30°C gave maximum ethanol yield of 1.42 g/L. PMID:21811671

Bioconversion of Agricultural Waste to Ethanol by SSF Using Recombinant Cellulase from Clostridium thermocellum.

PubMed

Mutreja, Ruchi; Das, Debasish; Goyal, Dinesh; Goyal, Arun

2011-01-01

The effect of different pretreatment methods, temperature, and enzyme concentration on ethanol production from 8 lignocellulosic agrowaste by simultaneous saccharification and fermentation (SSF) using recombinant cellulase and Saccharomyces cerevisiae were studied. Recombinant cellulase was isolated from E. coli BL21 cells transformed with CtLic26A-Cel5-CBM11 full-length gene from Clostridium thermocellum and produced in both batch and fed-batch processes. The maximum cell OD and specific activity in batch mode were 1.6 and 1.91 U/mg, respectively, whereas in the fed-batch mode, maximum cell OD and specific activity were 3.8 and 3.5 U/mg, respectively, displaying a 2-fold increase. Eight substrates, Syzygium cumini (jamun), Azadirachta indica (neem), Saracens indica (asoka), bambusa dendrocalmus (bamboo), Populas nigra (poplar), Achnatherum hymenoides (wild grass), Eucalyptus marginata (eucalyptus), and Mangifera indica (mango), were subjected to SSF. Of three pretreatments, acid, alkali, and steam explosion, acid pretreatment Syzygium cumini (Jamun) at 30°C gave maximum ethanol yield of 1.42 g/L.

Optimization of fed-batch fermentation for xylitol production by Candida tropicalis.

PubMed

Kim, J-H; Han, K-C; Koh, Y-H; Ryu, Y-W; Seo, J-H

2002-07-01

Xylitol, a functional sweetener, was produced from xylose by biological conversion using Candida tropicalis ATCC 13803. Based on a two-substrate fermentation using glucose for cell growth and xylose for xylitol production, fed-batch fermentations were undertaken to increase the final xylitol concentration. The effects of xylose and xylitol on xylitol production rate were studied to determine the optimum concentrations for fed-batch fermentation. Xylose concentration in the medium (100 g l(-1)) and less than 200 g l(-1) total xylose plus xylitol concentration were determined as optimum for maximum xylitol production rate and xylitol yield. Increasing the concentrations of xylose and xylitol decreased the rate and yield of xylitol production and the specific cell growth rate, probably because of an increase in osmotic stress that would interfere with xylose transport, xylitol flux to secretion to cell metabolism. The feeding rate of xylose solution during the fed-batch mode of operation was determined by using the mass balance equations and kinetic parameters involved in the equations in order to increase final xylitol concentration without affecting xylitol and productivity. The optimized fed-batch fermentation resulted in 187 g l(-1) xylitol concentration, 0.75 g xylitol g xylose(-1) xylitol yield and 3.9 g xylitol l(-1) h(-1) volumetric productivity.

Phosphoenolpyruvate Transporter Enables Targeted Perturbation During Metabolic Analysis of L-Phenylalanine Production With Escherichia coli.

PubMed

Tröndle, Julia; Albermann, Christoph; Weiner, Michael; Sprenger, Georg A; Weuster-Botz, Dirk

2018-05-01

Usually perturbation of the metabolism of cells by addition of substrates is applied for metabolic analysis of production organisms, but perturbation studies are restricted to the endogenous substrates of the cells under study. The goal of this study is to overcome this limitation by making phosphoenolpyruvate (PEP) available for perturbation studies with Escherichia coli producing L-phenylalanine. A production strain overexpressing a PEP-transporter variant (UhpT-D388C) is applied in a standardized fed-batch production-process on a 42 L-scale. Four parallel short-term perturbation experiments of 20 min are performed with glucose and glycerol as fed-batch carbon sources after rapid media transition of cells from the production-process. PEP is added after 9 min and is immediately consumed by the cells with up to 1.5 mmol g CDW -1  h -1 . L-phenylalanine production rates increased by up to 200% after addition of PEP. This clearly indicates an intracellular PEP-limitation in the L-phenylalanine production strain under study. Thus, it is shown that overexpressing specific transporters for analytical reasons makes exogenous substrates available as perturbation substrates for metabolic analyses of cells sampled from production-processes and thereby allows a very targeted perturbation of whole-cell metabolism. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Characteristics of Corn Stover Pretreated with Liquid Hot Water and Fed-Batch Semi-Simultaneous Saccharification and Fermentation for Bioethanol Production

PubMed Central

Li, Xuezhi; Lu, Jie; Zhao, Jian; Qu, Yinbo

2014-01-01

Corn stover is a promising feedstock for bioethanol production because of its abundant availability in China. To obtain higher ethanol concentration and higher ethanol yield, liquid hot water (LHW) pretreatment and fed-batch semi-simultaneous saccharification and fermentation (S-SSF) were used to enhance the enzymatic digestibility of corn stover and improve bioconversion of cellulose to ethanol. The results show that solid residues from LHW pretreatment of corn stover can be effectively converted into ethanol at severity factors ranging from 3.95 to 4.54, and the highest amount of xylan removed was approximately 89%. The ethanol concentrations of 38.4 g/L and 39.4 g/L as well as ethanol yields of 78.6% and 79.7% at severity factors of 3.95 and 4.54, respectively, were obtained by fed-batch S-SSF in an optimum conditions (initial substrate consistency of 10%, and 6.1% solid residues added into system at the prehydrolysis time of 6 h). The changes in surface morphological structure, specific surface area, pore volume and diameter of corn stover subjected to LHW process were also analyzed for interpreting the possible improvement mechanism. PMID:24763192

Recombinant production of biologically active giant grouper (Epinephelus lanceolatus) growth hormone from inclusion bodies of Escherichia coli by fed-batch culture.

PubMed

Chung, Wen-Jen; Huang, Chi-Lung; Gong, Hong-Yi; Ou, Tsung-Yin; Hsu, Jue-Liang; Hu, Shao-Yang

2015-06-01

Growth hormone (GH) performs important roles in regulating somatic growth, reproduction, osmoregulation, metabolism and immunity in teleosts, and thus, it has attracted substantial attention in the field of aquaculture application. Herein, giant grouper GH (ggGH) cDNA was cloned into the pET28a vector and expressed in Shuffle® T7 Competent Escherichia coli. Recombinant N-terminal 6× His-tagged ggGH was produced mainly in insoluble inclusion bodies; the recombinant ggGH content reached 20% of total protein. For large-scale ggGH production, high-cell density E. coli culture was achieved via fed-batch culture with pH-stat. After 30h of cultivation, a cell concentration of 41.1g/l dry cell weight with over 95% plasmid stability was reached. Maximal ggGH production (4.0g/l; 22% total protein) was achieved via mid-log phase induction. Various centrifugal forces, buffer pHs and urea concentrations were optimized for isolation and solubilization of ggGH from inclusion bodies. Hydrophobic interactions and ionic interactions were the major forces in ggGH inclusion body formation. Complete ggGH inclusion body solubilization was obtained in PBS buffer at pH 12 containing 3M urea. Through a simple purification process including Ni-NTA affinity chromatography and refolding, 5.7mg of ggGH was obtained from 10ml of fed-batch culture (45% recovery). The sequence and secondary structure of the purified ggGH were confirmed by LC-MS/MS mass spectrometry and circular dichroism analysis. The cell proliferation-promoting activity was confirmed in HepG2, ZFL and GF-1 cells with the WST-1 colorimetric bioassay. Copyright © 2015 Elsevier Inc. All rights reserved.

SLUDGE BATCH 4 BASELINE MELT RATE FURNACE AND SLURRY-FED MELT RATE FURNACE TESTS WITH FRITS 418 AND 510 (U)

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

Smith, M. E.; Jones, T. M.; Miller, D. H.

Several Slurry-Fed Melt Rate Furnace (SMRF) tests with earlier projections of the Sludge Batch 4 (SB4) composition have been performed.1,2 The first SB4 SMRF test used Frits 418 and 320, however it was found after the test that the REDuction/OXidation (REDOX) correlation at that time did not have the proper oxidation state for manganese. Because the manganese level in the SB4 sludge was higher than previous sludge batches tested, the impact of the higher manganese oxidation state was greater. The glasses were highly oxidized and very foamy, and therefore the results were inconclusive. After resolving this REDOX issue, Frits 418,more » 425, and 503 were tested in the SMRF with the updated baseline SB4 projection. Based on dry-fed Melt Rate Furnace (MRF) tests and the above mentioned SMRF tests, two previous frit recommendations were made by the Savannah River National Laboratory (SRNL) for processing of SB4 in the Defense Waste Processing Facility (DWPF). The first was Frit 503 based on the June 2006 composition projections.3 The recommendation was changed to Frit 418 as a result of the October 2006 composition projections (after the Tank 40 decant was implemented as part of the preparation plan). However, the start of SB4 processing was delayed due to the control room consolidation outage and the repair of the valve box in the Tank 51 to Tank 40 transfer line. These delays resulted in changes to the projected SB4 composition. Due to the slight change in composition and based on preliminary dry-fed MRF testing, SRNL believed that Frit 510 would increase throughput in processing SB4 in DWPF. Frit 418, which was used in processing Sludge Batch 3 (SB3), was a viable candidate and available in DWPF. Therefore, it was used during the initial SB4 processing. Due to the potential for higher melt rates with Frit 510, SMRF tests with the latest SB4 composition (1298 canisters) and Frits 510 and 418 were performed at a targeted waste loading (WL) of 35%. The '1298 canisters' describes the number of equivalent canisters that would be produced from the beginning of the current contract period before SB3 is blended with SB4. The melt rate for the SMRF SB4/Frit 510 test was 14.6 grams/minute. Due to cold cap mounding problems with the SMRF SB4/Frit 418 feed at 50 weight % solids that prevented a melt rate determination, this feed was diluted to 45 weight % solids. The melt rate for this diluted feed was 8.9 grams/minute. A correction factor of 1.2 for estimating the melt rate at 50 weight % solids from 45 weight % solids test results (based on previous SMRF testing5) was then used to estimate a melt rate of 10.7 grams/minute for SB4/Frit 418 at 50 weight % solids. Therefore, the use of Frit 510 versus Frit 418 with SB4 resulted in a higher melt rate (14.6 versus an estimated 10.7 grams/minute). For reference, a previous SMRF test with SB3/Frit 418 feed at 35% waste loading and 50 weight % solids resulted in a melt rate of 14.1 grams/minute. Therefore, depending on the actual feed rheology, the use of Frit 510 with SB4 could result in similar melt rates as experienced with SB3/Frit 418 feed in the DWPF.« less

Process for concentrated biomass saccharification

DOEpatents

Hennessey, Susan M.; Seapan, Mayis; Elander, Richard T.; Tucker, Melvin P.

2010-10-05

Processes for saccharification of pretreated biomass to obtain high concentrations of fermentable sugars are provided. Specifically, a process was developed that uses a fed batch approach with particle size reduction to provide a high dry weight of biomass content enzymatic saccharification reaction, which produces a high sugars concentration hydrolysate, using a low cost reactor system.

Cocoa residues as viable biomass for renewable energy production through anaerobic digestion.

PubMed

Acosta, Nayaret; De Vrieze, Jo; Sandoval, Verónica; Sinche, Danny; Wierinck, Isabella; Rabaey, Korneel

2018-05-31

The aim of this work was to evaluate the bioenergy potential of cocoa residue via anaerobic digestion. Batch and fed-batch lab-scale reactors were operated under low and high solids conditions. In the batch tests, 59 ± 4% of Chemical Oxygen Demand (COD) was recovered as methane. This corresponded with an average methane yield of 174 (wet) and 193 (dry) L kg -1 volatile solids fed, whereas a series of fed-batch reactors produced 70 ± 24 (wet) and 107 ± 39 (dry) L CH 4  kg -1 volatile solids fed during stable conditions. A case study was developed for canton Balao (Ecuador) based on our experimental data, operational estimates and available cocoa waste in the area. Annually, 8341 MWh could be produced, meeting 88% of the current electricity demand in Balao. This case study proves the potential for cocoa waste as a source of renewable energy in rural areas. Copyright © 2018 Elsevier Ltd. All rights reserved.

[Enhanced ε-poly-L-lysine production through pH regulation and organic nitrogen addition in fed-batch fermentation].

PubMed

Sun, Qixing; Chen, Xusheng; Ren, Xidong; Zheng, Gencheng; Mao, Zhonggui

2015-05-01

During the production of ε-poly-L-lysine (ε-PL) in fed-batch fermentation, the decline of ε-PL synthesis often occurs at middle or late phase of the fermentation. To solve the problem, we adopted two strategies, namely pH shift and feeding yeast extract, to improve the productivity of ε-PL. ε-PL productivity in fermentation by pH shift and feeding yeast extract achieved 4.62 g/(L x d) and 5.16 g/(L x d), which were increased by 27.3% and 42.2% compared with the control ε-PL fed-batch fermentation, respectively. Meanwhile, ε-PL production enhanced 36.95 g/L and 41.32 g/L in 192 h with these two strategies, increased by 27.4% and 42.48% compared to the control, respectively. ε-PL production could be improved at middle or late phase of fed-batch fermentation by pH shift or feeding yeast extract.

Xylose induces cellulase production in Thermoascus aurantiacus

DOE PAGES

Schuerg, Timo; Prahl, Jan -Philip; Gabriel, Raphael; ...

2017-11-15

Lignocellulosic biomass is an important resource for renewable production of biofuels and bioproducts. Enzymes that deconstruct this biomass are critical for the viability of biomass-based biofuel production processes. Current commercial enzyme mixtures have limited thermotolerance. Thermophilic fungi may provide enzyme mixtures with greater thermal stability leading to more robust processes. Understanding the induction of biomass-deconstructing enzymes in thermophilic fungi will provide the foundation for strategies to construct hyper-production strains. Induction of cellulases using xylan was demonstrated during cultivation of the thermophilic fungus Thermoascus aurantiacus. Simulated fed-batch conditions with xylose induced comparable levels of cellulases. These fed-batch conditions were adapted tomore » produce enzymes in 2 and 19 L bioreactors using xylose and xylose-rich hydrolysate from dilute acid pretreatment of corn stover. Enzymes from T. aurantiacus that were produced in the xylose-fed bioreactor demonstrated comparable performance in the saccharification of deacetylated, dilute acid-pretreated corn stover when compared to a commercial enzyme mixture at 50 °C. The T. aurantiacus enzymes retained this activity at of 60 °C while the commercial enzyme mixture was largely inactivated. CXylose induces both cellulase and xylanase production in T. aurantiacus and was used to produce enzymes at up to the 19 L bioreactor scale. The demonstration of induction by xylose-rich hydrolysate and saccharification of deacetylated, dilute acid-pretreated corn stover suggests a scenario to couple biomass pretreatment with onsite enzyme production in a biorefinery. This work further demonstrates the potential for T. aurantiacus as a thermophilic platform for cellulase development.« less

Xylose induces cellulase production in Thermoascus aurantiacus

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

Schuerg, Timo; Prahl, Jan -Philip; Gabriel, Raphael

Lignocellulosic biomass is an important resource for renewable production of biofuels and bioproducts. Enzymes that deconstruct this biomass are critical for the viability of biomass-based biofuel production processes. Current commercial enzyme mixtures have limited thermotolerance. Thermophilic fungi may provide enzyme mixtures with greater thermal stability leading to more robust processes. Understanding the induction of biomass-deconstructing enzymes in thermophilic fungi will provide the foundation for strategies to construct hyper-production strains. Induction of cellulases using xylan was demonstrated during cultivation of the thermophilic fungus Thermoascus aurantiacus. Simulated fed-batch conditions with xylose induced comparable levels of cellulases. These fed-batch conditions were adapted tomore » produce enzymes in 2 and 19 L bioreactors using xylose and xylose-rich hydrolysate from dilute acid pretreatment of corn stover. Enzymes from T. aurantiacus that were produced in the xylose-fed bioreactor demonstrated comparable performance in the saccharification of deacetylated, dilute acid-pretreated corn stover when compared to a commercial enzyme mixture at 50 °C. The T. aurantiacus enzymes retained this activity at of 60 °C while the commercial enzyme mixture was largely inactivated. CXylose induces both cellulase and xylanase production in T. aurantiacus and was used to produce enzymes at up to the 19 L bioreactor scale. The demonstration of induction by xylose-rich hydrolysate and saccharification of deacetylated, dilute acid-pretreated corn stover suggests a scenario to couple biomass pretreatment with onsite enzyme production in a biorefinery. This work further demonstrates the potential for T. aurantiacus as a thermophilic platform for cellulase development.« less

Acetate production from whey lactose using co-immobilized cells of homolactic and homoacetic bacteria in a fibrous-bed bioreactor

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

Huang, Y.; Yang, S.T.

1998-11-20

Acetate was produced from whey lactose in batch and fed-batch fermentations using co-immobilized cells of Clostridium formicoaceticum and Lactococcus lactis. The cells were immobilized in a spirally wound fibrous sheet packed in a 0.45-L column reactor, with liquid circulated through a 5-L stirred-tank fermentor. Industrial-grade nitrogen sources, including corn steep liquor, casein hydrolysate, and yeast hydrolysate, were studied as inexpensive nutrient supplements to whey permeate and acid whey. Supplementation with either 2.5% (v/v) corn steep liquor or 1.5 g/L casein hydrolysate was adequate for the cocultured fermentation. The overall acetic acid yield from lactose was 0.9 g/g, and the productivitymore » was 0.25 g/(L h). Both lactate and acetate at high concentrations inhibited the homoacetic fermentation. To overcome these inhibitions, fed-batch fermentations were used to keep lactate concentration low and to adapt cells to high-concentration acetate. The final acetate concentration obtained in the fed-batch fermentations were used to keep lactate concentration low and to adapt cells to high-concentration acetate. The final acetate concentration obtained in the fed-batch fermentation was 75 g/L, which was the highest acetate concentration ever produced by C. formicoaceticum. Even at this high acetate concentration, the overall productivity was 0.18 g/(L h) based on the total medium volume and 1.23 g/(L h) based on the fibrous-bed reactor volume. The cells isolated from the fibrous-bed bioreactor at the end of this study were more tolerant to acetic acid than the original culture used to seed the bioreactor, indicating that adaptation and natural selection of acetate-tolerant strains occurred. This cocultured fermentation process could be used to produce a low-cost acetate deicer from whey permeate and acid whey.« less

Batch process fault detection and identification based on discriminant global preserving kernel slow feature analysis.

PubMed

Zhang, Hanyuan; Tian, Xuemin; Deng, Xiaogang; Cao, Yuping

2018-05-16

As an attractive nonlinear dynamic data analysis tool, global preserving kernel slow feature analysis (GKSFA) has achieved great success in extracting the high nonlinearity and inherently time-varying dynamics of batch process. However, GKSFA is an unsupervised feature extraction method and lacks the ability to utilize batch process class label information, which may not offer the most effective means for dealing with batch process monitoring. To overcome this problem, we propose a novel batch process monitoring method based on the modified GKSFA, referred to as discriminant global preserving kernel slow feature analysis (DGKSFA), by closely integrating discriminant analysis and GKSFA. The proposed DGKSFA method can extract discriminant feature of batch process as well as preserve global and local geometrical structure information of observed data. For the purpose of fault detection, a monitoring statistic is constructed based on the distance between the optimal kernel feature vectors of test data and normal data. To tackle the challenging issue of nonlinear fault variable identification, a new nonlinear contribution plot method is also developed to help identifying the fault variable after a fault is detected, which is derived from the idea of variable pseudo-sample trajectory projection in DGKSFA nonlinear biplot. Simulation results conducted on a numerical nonlinear dynamic system and the benchmark fed-batch penicillin fermentation process demonstrate that the proposed process monitoring and fault diagnosis approach can effectively detect fault and distinguish fault variables from normal variables. Copyright © 2018 ISA. Published by Elsevier Ltd. All rights reserved.

Fed-batch culture of Escherichia coli for L-valine production based on in silico flux response analysis.

PubMed

Park, Jin Hwan; Kim, Tae Yong; Lee, Kwang Ho; Lee, Sang Yup

2011-04-01

We have previously reported the development of a 100% genetically defined engineered Escherichia coli strain capable of producing L-valine from glucose with a high yield of 0.38 g L-valine per gram glucose (0.58 mol L-valine per mol glucose) by batch culture. Here we report a systems biological strategy of employing flux response analysis in bioprocess development using L-valine production by fed-batch culture as an example. Through the systems-level analysis, the source of ATP was found to be important for efficient L-valine production. There existed a trade-off between L-valine production and biomass formation, which was optimized for the most efficient L-valine production. Furthermore, acetic acid feeding strategy was optimized based on flux response analysis. The final fed-batch cultivation strategy allowed production of 32.3 g/L L-valine, the highest concentration reported for E. coli. This approach of employing systems-level analysis of metabolic fluxes in developing fed-batch cultivation strategy would also be applicable in developing strategies for the efficient production of other bioproducts. Copyright © 2010 Wiley Periodicals, Inc.

Optimization of the microbial synthesis of dihydroxyacetone from glycerol with Gluconobacter oxydans.

PubMed

Hekmat, D; Bauer, R; Fricke, J

2003-12-01

An optimized repeated-fed-batch fermentation process for the synthesis of dihydroxyacetone (DHA) from glycerol utilizing Gluconobacter oxydans is presented. Cleaning, sterilization, and inoculation procedures could be reduced significantly compared to the conventional fed-batch process. A stringent requirement was that the product concentration was kept below a critical threshold level at all times in order to avoid irreversible product inhibition of the cells. On the basis of experimentally validated model calculations, a threshold value of about 60 kg x m(-3) DHA was obtained. The innovative bioreactor system consisted of a stirred tank reactor combined with a packed trickle-bed column. In the packed column, active cells could be retained by in situ immobilization on a hydrophilized Ralu-ring carrier material. Within 17 days, the productivity of the process could be increased by 75% to about 2.8 kg x m(-3) h(-1). However, it was observed that the maximum achievable productivity had not been reached yet.

A simple method to determine evaporation and compensate for liquid losses in small-scale cell culture systems.

PubMed

Wiegmann, Vincent; Martinez, Cristina Bernal; Baganz, Frank

2018-04-24

Establish a method to indirectly measure evaporation in microwell-based cell culture systems and show that the proposed method allows compensating for liquid losses in fed-batch processes. A correlation between evaporation and the concentration of Na + was found (R 2  = 0.95) when using the 24-well-based miniature bioreactor system (micro-Matrix) for a batch culture with GS-CHO. Based on these results, a method was developed to counteract evaporation with periodic water additions based on measurements of the Na + concentration. Implementation of this method resulted in a reduction of the relative liquid loss after 15 days of a fed-batch cultivation from 36.7 ± 6.7% without volume corrections to 6.9 ± 6.5% with volume corrections. A procedure was established to indirectly measure evaporation through a correlation with the level of Na + ions in solution and deriving a simple formula to account for liquid losses.

Sol-gel immobilization as a suitable technique for enhancement of α-amylase activity of Aspergillus oryzae PP.

PubMed

Evstatieva, Yana; Yordanova, Mariya; Chernev, Georgi; Ruseva, Yanislava; Nikolova, Dilyana

2014-07-04

Bioencapsulation of microbial cells in silica-based matrices has proved to be a good strategy to enhance the biosynthetic capabilities and viability of bioproducers. In the present study, mycelium and pellet cultures of strain Aspergillus oryzae PP were successfully immobilized in sol-gel hybrid matrices composed of tetraethylorthosilicate as an inorganic precursor, 5% (w/v) starch and 10 or 15% (w/v) polyethylene oxide, or 10% (w/v) calcium alginate as organic compounds. Biosynthetic activity of immobilized cultures was investigated by batch and fed-batch cultivation and the obtained results of 3042.04 IU cm -3 were comparable with the enzyme activity of the free cell culture. Immobilized cultures retained their viability and biosynthetic capabilities up to the 744th h during fed-batch fermentation processes. Consequently, sol-gel encapsulation in hybrid matrices could be considered as a promising technique for immobilization of Aspergillus oryzae PP in order to increase the α-amylase production.

Amino acid and glucose metabolism in fed-batch CHO cell culture affects antibody production and glycosylation.

PubMed

Fan, Yuzhou; Jimenez Del Val, Ioscani; Müller, Christian; Wagtberg Sen, Jette; Rasmussen, Søren Kofoed; Kontoravdi, Cleo; Weilguny, Dietmar; Andersen, Mikael Rørdam

2015-03-01

Fed-batch Chinese hamster ovary (CHO) cell culture is the most commonly used process for IgG production in the biopharmaceutical industry. Amino acid and glucose consumption, cell growth, metabolism, antibody titer, and N-glycosylation patterns are always the major concerns during upstream process optimization, especially media optimization. Gaining knowledge on their interrelations could provide insight for obtaining higher immunoglobulin G (IgG) titer and better controlling glycosylation-related product quality. In this work, different fed-batch processes with two chemically defined proprietary media and feeds were studied using two IgG-producing cell lines. Our results indicate that the balance of glucose and amino acid concentration in the culture is important for cell growth, IgG titer and N-glycosylation. Accordingly, the ideal fate of glucose and amino acids in the culture could be mainly towards energy and recombinant product, respectively. Accumulation of by-products such as NH4(+) and lactate as a consequence of unbalanced nutrient supply to cell activities inhibits cell growth. The levels of Leu and Arg in the culture, which relate to cell growth and IgG productivity, need to be well controlled. Amino acids with the highest consumption rates correlate with the most abundant amino acids present in the produced IgG, and thus require sufficient availability during culture. Case-by-case analysis is necessary for understanding the effect of media and process optimization on glycosylation. We found that in certain cases the presence of Man5 glycan can be linked to limitation of UDP-GlcNAc biosynthesis as a result of insufficient extracellular Gln. However, under different culture conditions, high Man5 levels can also result from low α-1,3-mannosyl-glycoprotein 2-β-N-acetylglucosaminyltransferase (GnTI) and UDP-GlcNAc transporter activities, which may be attributed to high level of NH4+ in the cell culture. Furthermore, galactosylation of the mAb Fc glycans was found to be limited by UDP-Gal biosynthesis, which was observed to be both cell line and cultivation condition-dependent. Extracellular glucose and glutamine concentrations and uptake rates were positively correlated with intracellular UDP-Gal availability. All these findings are important for optimization of fed-batch culture for improving IgG production and directing glycosylation quality. © 2014 Wiley Periodicals, Inc.

40 CFR Table 2 to Subpart Cccc of... - Requirements for Performance Tests

Code of Federal Regulations, 2012 CFR

2012-07-01

... As stated in § 63.2161, if you demonstrate compliance by monitoring brew ethanol, you must comply with the requirements for performance tests in the following table: [Brew Ethanol Monitoring Only] For each fed-batch fermenter for which compliance is determined by monitoring brew ethanol concentration...

40 CFR Table 2 to Subpart Cccc of... - Requirements for Performance Tests

Code of Federal Regulations, 2014 CFR

2014-07-01

... As stated in § 63.2161, if you demonstrate compliance by monitoring brew ethanol, you must comply with the requirements for performance tests in the following table: [Brew Ethanol Monitoring Only] For each fed-batch fermenter for which compliance is determined by monitoring brew ethanol concentration...

40 CFR Table 2 to Subpart Cccc of... - Requirements for Performance Tests

Code of Federal Regulations, 2013 CFR

2013-07-01

... As stated in § 63.2161, if you demonstrate compliance by monitoring brew ethanol, you must comply with the requirements for performance tests in the following table: [Brew Ethanol Monitoring Only] For each fed-batch fermenter for which compliance is determined by monitoring brew ethanol concentration...

Propionic acid production from corn stover hydrolysate by Propionibacterium acidipropionici

DOE PAGES

Wang, Xiaoqing; Salvachua, Davinia; Sanchez i Nogue, Violeta; ...

2017-08-17

The production of value-added chemicals alongside biofuels from lignocellulosic hydrolysates is critical for developing economically viable biorefineries. Here, the production of propionic acid (PA), a potential building block for C3-based chemicals, from corn stover hydrolysate is investigated using the native PA-producing bacterium Propionibacterium acidipropionici. A wide range of culture conditions and process parameters were examined and experimentally optimized to maximize titer, rate, and yield of PA. The effect of gas sparging during fermentation was first examined, and N 2 was found to exhibit improved performance over CO 2. Subsequently, the effects of different hydrolysate concentrations, nitrogen sources, and neutralization agentsmore » were investigated. One of the best combinations found during batch experiments used yeast extract (YE) as the primary nitrogen source and NH 4OH for pH control. This combination enabled PA titers of 30.8 g/L with a productivity of 0.40 g/L h from 76.8 g/L biomass sugars, while successfully minimizing lactic acid production. Due to the economic significance of downstream separations, increasing titers using fed-batch fermentation was examined by changing both feeding media and strategy. Continuous feeding of hydrolysate was found to be superior to pulsed feeding and combined with high YE concentrations increased PA titers to 62.7 g/L and improved the simultaneous utilization of different biomass sugars. Additionally, applying high YE supplementation maintains the lactic acid concentration below 4 g/L for the duration of the fermentation. Finally, with the aim of increasing productivity, high cell density fed-batch fermentations were conducted. PA titers increased to 64.7 g/L with a productivity of 2.35 g/L h for the batch stage and 0.77 g/L h for the overall process. These results highlight the importance of media and fermentation strategy to improve PA production. Altogether, this work demonstrates the feasibility of producing PA from corn stover hydrolysate.« less

Propionic acid production from corn stover hydrolysate by Propionibacterium acidipropionici

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

Wang, Xiaoqing; Salvachua, Davinia; Sanchez i Nogue, Violeta

The production of value-added chemicals alongside biofuels from lignocellulosic hydrolysates is critical for developing economically viable biorefineries. Here, the production of propionic acid (PA), a potential building block for C3-based chemicals, from corn stover hydrolysate is investigated using the native PA-producing bacterium Propionibacterium acidipropionici. A wide range of culture conditions and process parameters were examined and experimentally optimized to maximize titer, rate, and yield of PA. The effect of gas sparging during fermentation was first examined, and N 2 was found to exhibit improved performance over CO 2. Subsequently, the effects of different hydrolysate concentrations, nitrogen sources, and neutralization agentsmore » were investigated. One of the best combinations found during batch experiments used yeast extract (YE) as the primary nitrogen source and NH 4OH for pH control. This combination enabled PA titers of 30.8 g/L with a productivity of 0.40 g/L h from 76.8 g/L biomass sugars, while successfully minimizing lactic acid production. Due to the economic significance of downstream separations, increasing titers using fed-batch fermentation was examined by changing both feeding media and strategy. Continuous feeding of hydrolysate was found to be superior to pulsed feeding and combined with high YE concentrations increased PA titers to 62.7 g/L and improved the simultaneous utilization of different biomass sugars. Additionally, applying high YE supplementation maintains the lactic acid concentration below 4 g/L for the duration of the fermentation. Finally, with the aim of increasing productivity, high cell density fed-batch fermentations were conducted. PA titers increased to 64.7 g/L with a productivity of 2.35 g/L h for the batch stage and 0.77 g/L h for the overall process. These results highlight the importance of media and fermentation strategy to improve PA production. Altogether, this work demonstrates the feasibility of producing PA from corn stover hydrolysate.« less

Biotransformation of sweet lime pulp waste into high-quality nanocellulose with an excellent productivity using Komagataeibacter europaeus SGP37 under static intermittent fed-batch cultivation.

PubMed

Dubey, Swati; Singh, Jyoti; Singh, R P

2018-01-01

Herein, sweet lime pulp waste (SLPW) was utilized as a low- or no-cost feedstock for the production of bacterial nanocellulose (BNC) alone and in amalgamation with other nutritional supplements by the isolate K. europaeus SGP37 under static batch and static intermittent fed-batch cultivation. The highest yield (26.2±1.50gL -1 ) was obtained in the hot water extract of SLPW supplemented with the components of HS medium, which got further boosted to 38±0.85gL -1 as the cultivation strategy was shifted from static batch to static intermittent fed-batch. BNC obtained from various SLPW medium was similar or even superior to that obtained with standard HS medium in terms of its physicochemical properties. The production yields of BNC thus obtained are significantly higher and fit well in terms of industrial scale production. Copyright © 2017 Elsevier Ltd. All rights reserved.

High cell density fed-batch fermentations for lipase production: feeding strategies and oxygen transfer.

PubMed

Salehmin, M N I; Annuar, M S M; Chisti, Y

2013-11-01

This review is focused on the production of microbial lipases by high cell density fermentation. Lipases are among the most widely used of the enzyme catalysts. Although lipases are produced by animals and plants, industrial lipases are sourced almost exclusively from microorganisms. Many of the commercial lipases are produced using recombinant species. Microbial lipases are mostly produced by batch and fed-batch fermentation. Lipases are generally secreted by the cell into the extracellular environment. Thus, a crude preparation of lipases can be obtained by removing the microbial cells from the fermentation broth. This crude cell-free broth may be further concentrated and used as is, or lipases may be purified from it to various levels. For many large volume applications, lipases must be produced at extremely low cost. High cell density fermentation is a promising method for low-cost production: it allows a high concentration of the biomass and the enzyme to be attained rapidly and this eases the downstream recovery of the enzyme. High density fermentation enhances enzyme productivity compared with the traditional submerged culture batch fermentation. In production of enzymes, a high cell density is generally achieved through fed-batch operation, not through perfusion culture which is cumbersome. The feeding strategies used in fed-batch fermentations for producing lipases and the implications of these strategies are discussed. Most lipase-producing microbial fermentations require oxygen. Oxygen transfer in such fermentations is discussed.

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.

Intact cell mass spectrometry as a progress tracking tool for batch and fed-batch fermentation processes.

PubMed

Helmel, Michaela; Marchetti-Deschmann, Martina; Raus, Martin; Posch, Andreas E; Herwig, Christoph; Šebela, Marek; Allmaier, Günter

2015-02-01

Penicillin production during a fermentation process using industrial strains of Penicillium chrysogenum is a research topic permanently discussed since the accidental discovery of the antibiotic. Intact cell mass spectrometry (ICMS) can be a fast and novel monitoring tool for the fermentation progress during penicillin V production in a nearly real-time fashion. This method is already used for the characterization of microorganisms and the differentiation of fungal strains; therefore, the application of ICMS to samples directly harvested from a fermenter is a promising possibility to get fast information about the progress of fungal growth. After the optimization of the ICMS method to penicillin V fermentation broth samples, the obtained ICMS data were evaluated by hierarchical cluster analysis or an in-house software solution written especially for ICMS data comparison. Growth stages of a batch and fed-batch fermentation of Penicillium chrysogenum are differentiated by one of those statistical approaches. The application of two matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) instruments in the linear positive ion mode from different vendors demonstrated the universal applicability of the developed ICMS method. The base for a fast and easy-to-use method for monitoring the fermentation progress of P. chrysogenum is created with this ICMS method developed especially for fermentation broth samples. Copyright © 2014 Elsevier Inc. All rights reserved.

Ethanol Production from Traditional and Emerging Raw Materials

NASA Astrophysics Data System (ADS)

Rudolf, Andreas; Karhumaa, Kaisa; Hahn-Hägerdal, Bärbel

The ethanol industry of today utilizes raw materials rich in saccharides, such as sugar cane or sugar beets, and raw materials rich in starch, such as corn and wheat. The concern about supply of liquid transportation fuels, which has brought the crude oil price above 100 /barrel during 2006, together with the concern about global warming, have turned the interest towards large-scale ethanol production from lignocellulosic materials, such as agriculture and forestry residues. Baker's yeast Saccharomyces cerevisiae is the preferred fermenting microorganism for ethanol production because of its superior and well-documented industrial performance. Extensive work has been made to genetically improve S. cerevisiae to enable fermentation of lignocellulosic raw materials. Ethanolic fermentation processes are conducted in batch, fed-batch, or continuous mode, with or without cell recycling, the relative merit of which will be discussed.

Bioprocess development for the production of mouse-human chimeric anti-epidermal growth factor receptor vIII antibody C12 by suspension culture of recombinant Chinese hamster ovary cells.

PubMed

Hu, Suwen; Deng, Lei; Wang, Huamao; Zhuang, Yingping; Chu, Ju; Zhang, Siliang; Li, Zhonghai; Guo, Meijin

2011-05-01

The mouse-human chimeric anti-epidermal growth factor receptor vIII (EGFRvIII) antibody C12 is a promising candidate for the diagnosis of hepatocellular carcinoma (HCC). In this study, 3 processes were successfully developed to produce C12 by cultivation of recombinant Chinese hamster ovary (CHO-DG44) cells in serum-free medium. The effect of inoculum density was evaluated in batch cultures of shaker flasks to obtain the optimal inoculum density of 5 × 10(5) cells/mL. Then, the basic metabolic characteristics of CHO-C12 cells were studied in stirred bioreactor batch cultures. The results showed that the limiting concentrations of glucose and glutamine were 6 and 1 mM, respectively. The culture process consumed significant amounts of aspartate, glutamate, asparagine, serine, isoleucine, leucine, and lysine. Aspartate, glutamate, asparagine, and serine were particularly exhausted in the early growth stage, thus limiting cell growth and antibody synthesis. Based on these findings, fed-batch and perfusion processes in the bioreactor were successfully developed with a balanced amino acid feed strategy. Fed-batch and especially perfusion culture effectively maintained high cell viability to prolong the culture process. Furthermore, perfusion cultures maximized the efficiency of nutrient utilization; the mean yield coefficient of antibody to consumed glucose was 44.72 mg/g and the mean yield coefficient of glutamine to antibody was 721.40 mg/g. Finally, in small-scale bioreactor culture, the highest total amount of C12 antibody (1,854 mg) was realized in perfusion cultures. Therefore, perfusion culture appears to be the optimal process for small-scale production of C12 antibody by rCHO-C12 cells.

An optimized fed-batch culture strategy integrated with a one-step fermentation improves L-lactic acid production by Rhizopus oryzae.

PubMed

Fu, Yongqian; Sun, Xiaolong; Zhu, Huayue; Jiang, Ru; Luo, Xi; Yin, Longfei

2018-05-21

In previous work, we proposed a novel modified one-step fermentation fed-batch strategy to efficiently generate L-lactic acid (L-LA) using Rhizopus oryzae. In this study, to further enhance efficiency of L-LA production through one-step fermentation in fed-batch cultures, we systematically investigated the initial peptone- and glucose-feeding approaches, including different initial peptone and glucose concentrations and maintained residual glucose levels. Based on the results of this study, culturing R. oryzae with initial peptone and glucose concentrations of 3.0 and 50.0 g/l, respectively, using a fed-batch strategy is an effective approach of producing L-LA through one-step fermentation. Changing the residual glucose had no obvious effect on the generation of L-LA. We determined the maximum LA production and productivity to be 162 g/l and 6.23 g/(l·h), respectively, during the acid production stage. Compared to our previous work, there was almost no change in L-LA production or yield; however, the productivity of L-LA increased by 14.3%.

The digester modification for biogas production from palm oil mill effluent by Fed-batch

NASA Astrophysics Data System (ADS)

Aznury, M.; Amin, J. M.; Hasan, A.; Harsyah, A.

2018-03-01

The purpose of this research is to biogas production in the digester modification equipment by Fed-batch of the palm oil mill effluent (POME) to determine the quality of POME after a treatment and the concentration of biogas that is formed every 24 hours within 10 days. The raw materials used are POME from PT Mitra Ogan, Tbk. In the initial stage is sedimentation process in the first digester tank at a flow rate 6 liters/minute and then observing the retention time of 24 hours. POME flowed into the second digester tank for fermentation process with the addition of active microbes seed every 24 hours to produce biogas. After the fermentation process is complete, POME flowed to third digester tank for water treatment stage before being released into the environment. COD content test values obtained after processing are 766, 362 and 350 mg/L, approximately. While the BOD value is 212.75; 125 and 110.9 mg/L, approximately. Biogas production for 10 days fermentation are 10.88% methane, 19.2% oxygen and 75.83% nitrogen, approximately.

Process model comparison and transferability across bioreactor scales and modes of operation for a mammalian cell bioprocess.

PubMed

Craven, Stephen; Shirsat, Nishikant; Whelan, Jessica; Glennon, Brian

2013-01-01

A Monod kinetic model, logistic equation model, and statistical regression model were developed for a Chinese hamster ovary cell bioprocess operated under three different modes of operation (batch, bolus fed-batch, and continuous fed-batch) and grown on two different bioreactor scales (3 L bench-top and 15 L pilot-scale). The Monod kinetic model was developed for all modes of operation under study and predicted cell density, glucose glutamine, lactate, and ammonia concentrations well for the bioprocess. However, it was computationally demanding due to the large number of parameters necessary to produce a good model fit. The transferability of the Monod kinetic model structure and parameter set across bioreactor scales and modes of operation was investigated and a parameter sensitivity analysis performed. The experimentally determined parameters had the greatest influence on model performance. They changed with scale and mode of operation, but were easily calculated. The remaining parameters, which were fitted using a differential evolutionary algorithm, were not as crucial. Logistic equation and statistical regression models were investigated as alternatives to the Monod kinetic model. They were less computationally intensive to develop due to the absence of a large parameter set. However, modeling of the nutrient and metabolite concentrations proved to be troublesome due to the logistic equation model structure and the inability of both models to incorporate a feed. The complexity, computational load, and effort required for model development has to be balanced with the necessary level of model sophistication when choosing which model type to develop for a particular application. Copyright © 2012 American Institute of Chemical Engineers (AIChE).

Significantly enhanced production of acarbose in fed-batch fermentation with the addition of S-adenosylmethionine.

PubMed

Sun, Li-Hui; Li, Ming-Gang; Wang, Yuan-Shan; Zheng, Yu-Guo

2012-06-01

Acarbose, a pseudo-oligosaccharide, is widely used clinically in therapies for non-insulin-dependent diabetes. In the present study, S-adenosylmethionine (SAM) was added to selected media in order to investigate its effect on acarbose fermentation by Actinoplanes utahensis ZJB- 08196. Acarbose titer was seen to increase markedly when concentrations of SAM were added over a period of time. The effects of glucose and maltose on the production of acarbose were investigated in both batch and fed-batch fermentation. Optimal acarbose production was observed at relatively low glucose levels and high maltose levels. Based on these results, a further fed-batch experiment was designed so as to enhance the production of acarbose. Fed-batch fermentation was carried out at an initial glucose level of 10 g/l and an initial maltose level of 60 g/l. Then, 12 h post inoculation, 100 micromol/l SAM was added. In addition, 8 g/l of glucose was added every 24 h, and 20 g/l of maltose was added at 96 h. By way of this novel feeding strategy, the maximum titer of acarbose achieved was 6,113 mg/l at 192 h. To our knowledge, the production level of acarbose achieved in this study is the highest ever reported.

40 CFR Table 2 to Subpart Cccc of... - Requirements for Performance Tests

Code of Federal Regulations, 2011 CFR

2011-07-01

... stated in § 63.2161, if you demonstrate compliance by monitoring brew ethanol, you must comply with the requirements for performance tests in the following table: [Brew Ethanol Monitoring Only] For each fed-batch fermenter for which compliance is determined by monitoring brew ethanol concentration and calculating VOC...

Nocardia globerula NHB-2 nitrilase catalysed biotransformation of 4-cyanopyridine to isonicotinic acid

PubMed Central

2012-01-01

Isonicotinic acid (INA) is an important pyridine derivative used in the manufacture of isoniazid (antituberculosatic drug) and other pharmaceutically important drugs. Nitrilase catalysed processes for the synthesis of pharmaceutically important acids from their corresponding nitriles are promising alternative over the cumbersome, hazardous, and energy demanding chemical processes. Nitrilase of Nocardia globerula NHB-2 (NitNHB2) is expressed in presence of isobutyronitrile in the growth medium (1.0% glucose, 0.5% peptone, 0.3% beef extract, and 0.1 % yeast extract, pH 7.5). NitNHB2 hydrolyses 4-cyanopyridine (4-CP) to INA without accumulation of isonicotinamide, which is common in the reaction catalysed via fungal nitrilases. The NitNHB2 suffers from substrate inhibition effect and hydrolysing activity up to 250 mM 4-CP was recorded. Complete conversion of 200 mM 4-CP to INA was achieved in 40 min using resting cell concentration corresponding to 10 U mL-1 nitrilase activity in the reaction. Substrate inhibition effect in the fed batch reaction (200 mM substrate feed/40min) led to formation of only 729 mM INA. In a fed batch reaction (100 mM 4-CP/20min), substrate inhibition effect was encountered after 7th feed and a total of 958 mM INA was produced in 400 min. The fed batch reaction scaled up to 1 L and 100% hydrolysis of 700 mM of 4-CP to INA at 35°C achieved in 140 min. The rate of INA production was 21.1 g h-1 mgDCW-1. This is the fastest biotransformation process ever reported for INA production with time and space productivity of 36 g L-1 h-1 using a bacterial nitrilase. PMID:22537922

Quantitative modeling of viable cell density, cell size, intracellular conductivity, and membrane capacitance in batch and fed-batch CHO processes using dielectric spectroscopy.

PubMed

Opel, Cary F; Li, Jincai; Amanullah, Ashraf

2010-01-01

Dielectric spectroscopy was used to analyze typical batch and fed-batch CHO cell culture processes. Three methods of analysis (linear modeling, Cole-Cole modeling, and partial least squares regression), were used to correlate the spectroscopic data with routine biomass measurements [viable packed cell volume, viable cell concentration (VCC), cell size, and oxygen uptake rate (OUR)]. All three models predicted offline biomass measurements accurately during the growth phase of the cultures. However, during the stationary and decline phases of the cultures, the models decreased in accuracy to varying degrees. Offline cell radius measurements were unsuccessfully used to correct for the deviations from the linear model, indicating that physiological changes affecting permittivity were occurring. The beta-dispersion was analyzed using the Cole-Cole distribution parameters Deltaepsilon (magnitude of the permittivity drop), f(c) (critical frequency), and alpha (Cole-Cole parameter). Furthermore, the dielectric parameters static internal conductivity (sigma(i)) and membrane capacitance per area (C(m)) were calculated for the cultures. Finally, the relationship between permittivity, OUR, and VCC was examined, demonstrating how the definition of viability is critical when analyzing biomass online. The results indicate that the common assumptions of constant size and dielectric properties used in dielectric analysis are not always valid during later phases of cell culture processes. The findings also demonstrate that dielectric spectroscopy, while not a substitute for VCC, is a complementary measurement of viable biomass, providing useful auxiliary information about the physiological state of a culture. (c) 2010 American Institute of Chemical Engineers

Production of the potential sweetener 5-ketofructose from fructose in fed-batch cultivation with Gluconobacter oxydans.

PubMed

Herweg, Elena; Schöpping, Marie; Rohr, Katja; Siemen, Anna; Frank, Oliver; Hofmann, Thomas; Deppenmeier, Uwe; Büchs, Jochen

2018-07-01

Sweeteners improve the dietary properties of many foods. A candidate for a new natural sweetener is 5-ketofructose. In this study a fed-batch process for the production of 5-ketofructose was developed. A Gluconobacter oxydans strain overexpressing a fructose dehydrogenase from G. japonicus was used and the sensory properties of 5-ketofructose were analyzed. The compound showed an identical sweet taste quality as fructose and a similar intrinsic sweet threshold concentration of 16.4 mmol/L. The production of 5-ketofructose was characterized online by monitoring of the respiration activity in shake flasks. Pulsed and continuous fructose feeding was realized in 2 L stirred tank reactors and maximum fructose consumption rates were determined. 5-Ketofructose concentrations of up to 489 g/L, product yields up to 0.98 g 5-KF /g fructose and space time yields up to 8.2 g/L/h were reached highlighting the potential of the presented process. Copyright © 2018 Elsevier Ltd. All rights reserved.

Ethanol production from lignocellulosic byproducts of olive oil extraction.

PubMed

Ballesteros, I; Oliva, J M; Saez, F; Ballesteros, M

2001-01-01

The recent implementation of a new two-step centrifugation process for extracting olive oil in Spain has substantially reduced water consumption, thereby eliminating oil mill wastewater. However, a new high sugar content residue is still generated. In this work the two fractions present in the residue (olive pulp and fragmented stones) were assayed as substrate for ethanol production by the simultaneous saccharification and fermentation (SSF) process. Pretreatment of fragmented olive stones by sulfuric acid-catalyzed steam explosion was the most effective treatment for increasing enzymatic digestibility; however, a pretreatment step was not necessary to bioconvert the olive pulp into ethanol. The olive pulp and fragmented olive stones were tested by the SSF process using a fed-batch procedure. By adding the pulp three times at 24-h intervals, 76% of the theoretical SSF yield was obtained. Experiments with fed-batch pretreated olive stones provided SSF yields significantly lower than those obtained at standard SSF procedure. The preferred SSF conditions to obtain ethanol from olives stones (61% of theoretical yield) were 10% substrate and addition of cellulases at 15 filter paper units/g of substrate.

Pilot scale repeated fed-batch fermentation processes of the wine yeast Dekkera bruxellensis for mass production of resveratrol from Polygonum cuspidatum.

PubMed

Kuo, Hsiao-Ping; Wang, Reuben; Lin, Yi-Sheng; Lai, Jinn-Tsyy; Lo, Yi-Chen; Huang, Shyue-Tsong

2017-11-01

Resveratrol has long been used as an ingredient in functional foods. Currently, Polygonum cuspidatum extract is the greatest natural source for resveratrol because of high concentrations of glycosidic-linked resveratrol. Thus, developing a cost-effective procedure to hydrolyze glucoside could substantially enhance resveratrol production from P. cuspidatum. This study selected Dekkera bruxellensis from several microorganisms based on its bioconversion and enzyme-specific activities. We demonstrated that the cells could be reused at least nine times while maintaining an average of 180.67U/L β-glucosidase activity. The average resveratrol bioconversion efficiency within five rounds of repeated usage was 108.77±0.88%. This process worked effectively when the volume was increased to 1200L, a volume at which approximately 35mgL -1 h -1 resveratrol per round was produced. This repeated fed-batch bioconversion process for resveratrol production is comparable to enzyme or cell immobilization strategies in terms of reusing cycles, but without incurring additional costs for immobilization. Copyright © 2017 Elsevier Ltd. All rights reserved.

ANAMMOX-like performances for nitrogen removal from ammonium-sulfate-rich wastewater in an anaerobic sequencing batch reactor.

PubMed

Prachakittikul, Pensiri; Wantawin, Chalermraj; Noophan, Pongsak Lek; Boonapatcharoen, Nimaradee

2016-01-01

Ammonium removal by the ANaerobic AMonium OXidation (ANAMMOX) process was observed through the Sulfate-Reducing Ammonium Oxidation (SRAO) process. The same concentration of ammonium (100 mg N L(-1)) was applied to two anaerobic sequencing batch reactors (AnSBRs) that were inoculated with the same activated sludge from the Vermicelli wastewater treatment process, while nitrite was fed in ANAMMOX and sulfate in SRAO reactors. In SRAO-AnSBR, in substrates that were fed with a ratio of NH4(+)/SO4(2-) at 1:0.4 ± 0.03, a hydraulic retention time (HRT) of 48 h and without sludge draining, the Ammonium Removal Rate (ARR) was 0.02 ± 0.01 kg N m(-3).d(-1). Adding specific ANAMMOX substrates to SRAO-AnSBR sludge in batch tests results in specific ammonium and nitrite removal rates of 0.198 and 0.139 g N g(-1) VSS.d, respectively, indicating that the ANAMMOX activity contributes to the removal of ammonium in the SRAO process using the nitrite that is produced from SRAO. Nevertheless, the inability of ANAMMOX to utilize sulfate to oxidize ammonium was also investigated in batch tests by augmenting enriched ANAMMOX culture in SRAO-AnSBR sludge and without nitrite supply. The time course of sulfate in a 24-hour cycle of SRAO-AnSBR showed an increase in sulfate after 6 h. For enriched SRAO culture, the uptake molar ratio of NH4(+)/SO4(2-) at 8 hours in a batch test was 1:0.82 lower than the value of 1:0.20 ± 0.09 as obtained in an SRAO-AnSBR effluent, while the stoichiometric ratio of 1:0.5 that includes the ANAMMOX reaction was in this range. After a longer operation of more than 2 years without sludge draining, the accumulation of sulfate and the reduction of ammonium removal were observed, probably due to the gradual increase in the sulfur denitrification rate and the competitive use of nitrite with ANAMMOX. The 16S rRNA gene PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) and PCR cloning analyses resulted in the detection of the ANAMMOX bacterium (Candidatus Brocadia sinica JPN1) Desulfacinum subterraneum belonging to the genus Desulfacinum and bacteria that are involved in sulfur metabolism (Pseudomonas aeruginosa strain SBTPe-001 and Paracoccus denitrificans strain IAM12479) in SRAO-AnSBR.

Ultrasound pretreatment for enhanced biogas production from olive mill wastewater.

PubMed

Oz, Nilgun Ayman; Uzun, Alev Cagla

2015-01-01

This study investigates applicability of low frequency ultrasound technology to olive mill wastewaters (OMWs) as a pretreatment step prior to anaerobic batch reactors to improve biogas production and methane yield. OMWs originating from three phase processes are characterized with high organic content and complex nature. The treatment of the wastewater is problematic and alternative treatment options should be investigated. In the first part of the study, OMW samples were subjected to ultrasound at a frequency of 20kHz with applied powers varying between 50 and 100W under temperature controlled conditions for different time periods in order to determine the most effective sonication conditions. The level of organic matter solubilization at ultrasound experiments was assessed by calculating the ratio of soluble chemical oxygen demand/total chemical oxygen demand (SCOD/TCOD). The results revealed that the optimum ultrasonic condition for diluted OMW is 20kHz, 0.4W/mL for 10min. The application of ultrasound to OMW increased SCOD/TCOD ratio from 0.59 to 0.79. Statistical analysis (Friedman's tests) show that ultrasound was significantly effective on diluted OMW (p<0.05) in terms of SCOD parameter, but not for raw OMW (p>0.05). For raw OMW, this increase has been found to be limited due to high concentration of suspended solids (SS). In the second part of the study, biogas and methane production rates of anaerobic batch reactor fed with the ultrasound pretreated OMW samples were compared with the results of control reactor fed with untreated OMW in order to determine the effect of sonication. A nonparametric statistical procedure, Mann-Whitney U test, was used to compare biogas and methane production from anaerobic batch reactors for control and ultrasound pretreated samples. Results showed that application of low frequency ultrasound to OMW significantly improved both biogas and methane production in anaerobic batch reactor fed with the wastewater (p<0.05). Anaerobic batch reactor fed with ultrasound pretreated diluted OMW produced approximately 20% more biogas and methane compared with the untreated one (control reactor). The overall results indicated that low frequency ultrasound pretreatment increased soluble COD in OMW and subsequently biogas production. Copyright © 2014 Elsevier B.V. All rights reserved.

Production of acetone butanol (AB) from liquefied corn starch, a commercial substrate, using Clostridium beijerinckii coupled with product recovery by gas stripping.

PubMed

Ezeji, Thaddeus C; Qureshi, Nasib; Blaschek, Hans P

2007-12-01

A potential industrial substrate (liquefied corn starch; LCS) has been employed for successful acetone butanol ethanol (ABE) production. Fermentation of LCS (60 g l(-1)) in a batch process resulted in the production of 18.4 g l(-1) ABE, comparable to glucose: yeast extract based medium (control experiment, 18.6 g l(-1) ABE). A batch fermentation of LCS integrated with product recovery resulted in 92% utilization of sugars present in the feed. When ABE was recovered by gas stripping (to relieve inhibition) from the fed-batch reactor fed with saccharified liquefied cornstarch (SLCS), 81.3 g l(-1) ABE was produced compared to 18.6 g l(-1) (control). In this integrated system, 225.8 g l(-1) SLCS sugar (487 % of control) was consumed. In the absence of product removal, it is not possible for C. beijerinckii BA101 to utilize more than 46 g l(-1) glucose. A combination of fermentation of this novel substrate (LCS) to butanol together with product recovery by gas stripping may economically benefit this fermentation.

Use of Model-Based Nutrient Feeding for Improved Production of Artemisinin by Hairy Roots of Artemisia Annua in a Modified Stirred Tank Bioreactor.

PubMed

Patra, Nivedita; Srivastava, Ashok K

2015-09-01

Artemisinin has been indicated to be a potent drug for the cure of malaria. Batch growth and artemisinin production kinetics of hairy root cultures of Artemisia annua were studied under shake flask conditions which resulted in accumulation of 12.49 g/L biomass and 0.27 mg/g artemisinin. Using the kinetic data, a mathematical model was identified to understand and optimize the system behavior. The developed model was then extrapolated to design nutrient feeding strategies during fed-batch cultivation for enhanced production of artemisinin. In one of the fed-batch cultivation, sucrose (37 g/L) feeding was done at a constant feed rate of 0.1 L/day during 10-15 days, which led to improved artemisinin accumulation of 0.77 mg/g. The second strategy of fed-batch hairy root cultivation involved maintenance of pseudo-steady state sucrose concentration (20.8 g/L) during 10-15 days which resulted in artemisinin accumulation of 0.99 mg/g. Fed-batch cultivation (with the maintenance of pseudo-steady state of substrate) of Artemisia annua hairy roots was, thereafter, implemented in bioreactor cultivation, which featured artemisinin accumulation of 1.0 mg/g artemisinin in 16 days of cultivation. This is the highest reported artemisinin yield by hairy root cultivation in a bioreactor.

Ethanol production using whole plant biomass of Jerusalem artichoke by Kluyveromyces marxianus CBS1555.

PubMed

Kim, Seonghun; Park, Jang Min; Kim, Chul Ho

2013-03-01

Jerusalem artichoke is a low-requirement sugar crop containing cellulose and hemicellulose in the stalk and a high content of inulin in the tuber. However, the lignocellulosic component in Jerusalem artichoke stalk reduces the fermentability of the whole plant for efficient bioethanol production. In this study, Jerusalem artichoke stalk was pretreated sequentially with dilute acid and alkali, and then hydrolyzed enzymatically. During enzymatic hydrolysis, approximately 88 % of the glucan and xylan were converted to glucose and xylose, respectively. Batch and fed-batch simultaneous saccharification and fermentation of both pretreated stalk and tuber by Kluyveromyces marxianus CBS1555 were effectively performed, yielding 29.1 and 70.2 g/L ethanol, respectively. In fed-batch fermentation, ethanol productivity was 0.255 g ethanol per gram of dry Jerusalem artichoke biomass, or 0.361 g ethanol per gram of glucose, with a 0.924 g/L/h ethanol productivity. These results show that combining the tuber and the stalk hydrolysate is a useful strategy for whole biomass utilization in effective bioethanol fermentation from Jerusalem artichoke.

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.

Optimization of glutathione production in batch and fed-batch cultures by the wild-type and recombinant strains of the methylotrophic yeast Hansenula polymorpha DL-1

PubMed Central

2011-01-01

Background Tripeptide glutathione (gamma-glutamyl-L-cysteinyl-glycine) is the most abundant non-protein thiol that protects cells from metabolic and oxidative stresses and is widely used as medicine, food additives and in cosmetic industry. The methylotrophic yeast Hansenula polymorpha is regarded as a rich source of glutathione due to the role of this thiol in detoxifications of key intermediates of methanol metabolism. Cellular and extracellular glutathione production of H. polymorpha DL-1 in the wild type and recombinant strains which overexpress genes of glutathione biosynthesis (GSH2) and its precursor cysteine (MET4) was studied. Results Glutathione producing capacity of H. polymorpha DL-1 depending on parameters of cultivation (dissolved oxygen tension, pH, stirrer speed), carbon substrate (glucose, methanol) and type of overexpressed genes of glutathione and its precursor biosynthesis during batch and fed-batch fermentations were studied. Under optimized conditions of glucose fed-batch cultivation, the glutathione productivity of the engineered strains was increased from ~900 up to ~ 2300 mg of Total Intracellular Glutathione (TIG) or GSH+GSSGin, per liter of culture medium. Meantime, methanol fed-batch cultivation of one of the recombinant strains allowed achieving the extracellular glutathione productivity up to 250 mg of Total Extracellular Glutathione (TEG) or GSH+GSSGex, per liter of the culture medium. Conclusions H. polymorpha is an competitive glutathione producer as compared to other known yeast and bacteria strains (Saccharomyces cerevisiae, Candida utilis, Escherichia coli, Lactococcus lactis etc.) with good perspectives for further improvement especially for production of extracellular form of glutathione. PMID:21255454

AnSBBR applied to organic matter and sulfate removal: interaction effect between feed strategy and COD/sulfate ratio.

PubMed

Friedl, Gregor F; Mockaitis, Gustavo; Rodrigues, José A D; Ratusznei, Suzana M; Zaiat, Marcelo; Foresti, Eugênio

2009-10-01

A mechanically stirred anaerobic sequencing batch reactor containing anaerobic biomass immobilized on polyurethane foam cubes, treating low-strength synthetic wastewater (500 mg COD L(-1)), was operated under different operational conditions to assess the removal of organic matter and sulfate. These conditions were related to fill time, defined by the following feed strategies: batch mode of 10 min, fed-batch mode of 3 h and fed-batch mode of 6 h, and COD/[SO(4)(2-)] ratios of 1.34, 0.67, and 0.34 defined by organic matter concentration of 500 mg COD L(-1) and sulfate concentrations of 373, 746, and 1,493 mg SO(4)(2-) L(-1) in the influent. Thus, nine assays were performed to investigate the influence of each of these parameters, as well as the interaction effect, on the performance of the system. The reactor operated with agitation of 400 rpm, total volume of 4.0 L, and treated 2.0 L synthetic wastewater in 8-h cycles at 30 +/- 1 degrees C. During all assays, the reactor showed operational stability in relation to the monitored variables such as COD, sulfate, sulfide, sulfite, volatile acids, bicarbonate alkalinity, and solids, thus demonstrating the potential to apply this technology to the combined removal of organic matter and sulfate. In general, the results showed that the 3-h fed-batch operation with a COD/[SO(4)(2-)] ratio of 0.34 presented the best conditions for organic matter removal (89%). The best efficiency for sulfate removal (71%) was accomplished during the assay with a COD/[SO(4)(2-)] ratio of 1.34 and a fill time of 6 h. It was also observed that as fill time and sulfate concentration in the influent increased, the ratio between removed sulfate load and removed organic load also increased. However, it should be pointed out that the aim of this study was not to optimize the removal of organic matter and sulfate, but rather to analyze the behavior of the reactor during the different feed strategies and applied COD/[SO(4)(2-)] ratios, and mainly to analyze the interaction effect, an aspect that has not yet been explored in the literature for batch reactors.

Effects of pH and Temperature on Recombinant Manganese Peroxidase Production and Stability

NASA Astrophysics Data System (ADS)

Jiang, Fei; Kongsaeree, Puapong; Schilke, Karl; Lajoie, Curtis; Kelly, Christine

The enzyme manganese peroxidase (MnP) is produced by numerous white-rot fungi to overcome biomass recalcitrance caused by lignin. MnP acts directly on lignin and increases access of the woody structure to synergistic wood-degrading enzymes such as cellulases and xylanases. Recombinant MnP (rMnP) can be produced in the yeast Pichia pastoris αMnP1-1 in fed-batch fermentations. The effects of pH and temperature on recombinant manganese peroxidase (rMnP) production by P. pastoris αMnP1-1 were investigated in shake flask and fed-batch fermentations. The optimum pH and temperature for a standardized fed-batch fermentation process for rMnP production in P. pastoris ctMnP1-1 were determined to be pH 6 and 30 °C, respectively. P. pastoris αMnP1-1 constitutively expresses the manganese peroxidase (mnp1) complementary DNA from Phanerochaete chrysosporium, and the rMnP has similar kinetic characteristics and pH activity and stability ranges as the wild-type MnP (wtMnP). Cultivation of P. chrysosporium mycelia in stationary flasks for production of heme peroxidases is commonly conducted at low pH (pH 4.2). However, shake flask and fed-batch fermentation experiments with P. pastoris αMnP1-1 demonstrated that rMnP production is highest at pH 6, with rMnP concentrations in the medium declining rapidly at pH less than 5.5, although cell growth rates were similar from pH 4-7. Investigations of the cause of low rMnP production at low pH were consistent with the hypothesis that intracellular proteases are released from dead and lysed yeast cells during the fermentation that are active against rMnP at pH less than 5.5.

40 CFR 63.1036 - Alternative means of emission limitation: Batch processes.

Code of Federal Regulations, 2010 CFR

2010-07-01

... SOURCE CATEGORIES (CONTINUED) National Emission Standards for Equipment Leaks-Control Level 2 Standards... leaks. The owner or operator may switch among the alternatives provided the change is documented as... shall be pressure-tested for leaks before regulated material is first fed to the equipment and the...

40 CFR 65.117 - Alternative means of emission limitation: Batch processes.

Code of Federal Regulations, 2010 CFR

2010-07-01

... (CONTINUED) AIR PROGRAMS (CONTINUED) CONSOLIDATED FEDERAL AIR RULE Equipment Leaks § 65.117 Alternative means... section provide the options of pressure testing or monitoring the equipment for leaks. The owner or... pressure-tested for leaks before regulated material is first fed to the equipment and the equipment is...

Process development for a recombinant Chinese hamster ovary (CHO) cell line utilizing a metal induced and amplified metallothionein expression system.

PubMed

Huang, Edwin P; Marquis, Christopher P; Gray, Peter P

2004-11-20

The suspension Chinese Hamster Ovary cell line, 13-10-302, utilizing the metallothionein (MT) expression system producing recombinant human growth hormone (hGH) was studied in a serum-free and cadmium-free medium at different fermentation scales and modes of operation. Initial experiments were carried out to optimize the concentration of metal addition to induce the MT promoter. Subsequently, the cultivation of the 13-10-302 cell line was scaled up from spinner flasks into bioreactors, and the cultivation duration was extended with fed-batch and perfusion strategies utilizing 180 microM zinc to induce the promoter controlling expression of recombinant hGH. It was shown that a fed-batch process could increase the maximum cell numbers twofold, from 3.3 to 6.3 x 10(6) cell/mL, over those obtained in normal batch fermentations, and this coupled with extended fermentation times resulted in a fourfold increase in final hGH titer, from 135 +/- 15 to 670 +/- 70 mg/L at a specific productivity q(hGH) value of 12 pg cell(-1)d(-1). The addition of sodium butyrate increased the specific productivity of hGH in cells to a value of approximately 48 pg cell(-1)d(-1), resulting in a final hGH titer of over a gram per liter during fed-batch runs. A BioSep acoustic cell recycler was used to retain the cells in the bioreactor during perfusion operation. It was necessary to maintain the specific feeding rates (SFR) above a value of 0.2 vvd/(10(6) cell/mL) to maintain the viability and productivity of the 13-10-302 cells; under these conditions the viable cell number increased to over 10(7) cell/mL and resulted in a volumetric productivity of over 120 mg(hGH) L(-1)d(-1). Process development described in this work demonstrates cultivation at various scales and sustained high levels of productivity under cadmium free condition in a CHO cell line utilizing an inducible metallothionein expression system. (c) 2004 Wiley Periodicals, Inc

Supplementation of serum free media with HT is not sufficient to restore growth properties of DHFR-/- cells in fed-batch processes - Implications for designing novel CHO-based expression platforms.

PubMed

Florin, Lore; Lipske, Carolin; Becker, Eric; Kaufmann, Hitto

2011-04-10

DHFR-deficient CHO cells are the most commonly used host cells in the biopharmaceutical industry and over the years, individual substrains have evolved, some have been engineered with improved properties and platform technologies have been designed around them. Unexpectedly, we have observed that different DHFR-deficient CHO cells show only poor growth in fed-batch cultures even in HT supplemented medium, whereas antibody producer cells derived from these hosts achieved least 2-3 fold higher peak cell densities. Using a set of different expression vectors, we were able to show that this impaired growth performance was not due to the selection procedure possibly favouring fast growing clones, but a direct consequence of DHFR deficiency. Re-introduction of the DHFR gene reproducibly restored the growth phenotype to the level of wild-type CHO cells or even beyond which seemed to be dose-dependent. The requirement for a functional DHFR gene to achieve optimal growth under production conditions has direct implications for cell line generation since it suggests that changing to a selection system other than DHFR would require another CHO host which - especially for transgenic CHO strains and tailor-suited process platforms - this could mean significant investments and potential changes in product quality. In these cases, DHFR engineering of the current CHO-DG44 or DuxB11-based host could be an attractive alternative. Copyright © 2011 Elsevier B.V. All rights reserved.

Proteome analysis to assess physiological changes in Escherichia coli grown under glucose-limited fed-batch conditions.

PubMed

Raman, Babu; Nandakumar, M P; Muthuvijayan, Vignesh; Marten, Mark R

2005-11-05

Proteome analysis was used to compare global protein expression changes in Escherichia coli fermentation between exponential and glucose-limited fed-batch phase. Two-dimensional gel electrophoresis and MALDI-TOF mass spectrometry were used to separate and identify 49 proteins showing >2-fold difference in expression. Proteins upregulated during exponential phase include ribonucleotide biosynthesis enzymes and ribosomal recycling factor. Proteins upregulated during fed-batch phase include those involved in high-affinity glucose uptake, transport and degradation of alternate carbon sources and TCA cycle, suggesting an enhanced role of the cycle under glucose- and energy-limited conditions. We report the upregulation of several putative proteins (ytfQ, ygiS, ynaF, yggX, yfeX), not identified in any previous study under carbon-limited conditions. Copyright (c) 2005 Wiley Periodicals, Inc.

High rate psychrophilic anaerobic digestion of high solids (35%) dairy manure in sequence batch reactor.

PubMed

Saady, Noori M Cata; Massé, Daniel I

2015-06-01

Zero liquid discharge is increasingly adopted as an objective for waste treatment process. The objective of this study was to increase the feed total solids (TS) and the organic loading rate (OLR) fed to a novel psychrophilic (20°C) dry anaerobic digestion (PDAD). Duplicate laboratory-scale bioreactors were fed cow feces and wheat straw (35% TS in feed) at OLR of 6.0 g TCOD kg(-1) inoculum d(-1) during long-term operation (147 days consisting of 7 successive cycles). An overall average specific methane yield (SMY) of 151.8±7.9 N L CH4 kg(-1) VS fed with an averaged volatile solids removal of 42.4±4.3% were obtained at a volatile solids-based inoculum-to-substrate ratio (ISR) of 2.13±0.2. The operation was stable as indicated by biogas and VFAs profiles and the results were reproducible in successive cycles; a maximum SMY of 163.3±5.7 N L CH4 kg(-1) VS fed was obtained. Hydrolysis was the reaction limiting step. High rate PDAD of 35% TS dairy manure is possible in sequential batch reactor within 21 days treatment cycle length. Crown Copyright © 2015. Published by Elsevier Ltd. All rights reserved.

Cellulase production by Penicillium funiculosum and its application in the hydrolysis of sugar cane bagasse for second generation ethanol production by fed batch operation.

PubMed

Maeda, Roberto Nobuyuki; Barcelos, Carolina Araújo; Santa Anna, Lídia Maria Melo; Pereira, Nei

2013-01-10

This study aimed to produce a cellulase blend and to evaluate its application in a simultaneous saccharification and fermentation (SSF) process for second generation ethanol production from sugar cane bagasse. The sugar cane bagasse was subjected to pretreatments (diluted acid and alkaline), as for disorganizing the ligocellulosic complex, and making the cellulose component more amenable to enzymatic hydrolysis. The residual solid fraction was named sugar cane bagasse partially delignified cellulignin (PDC), and was used for enzyme production and ethanol fermentation. The enzyme production was performed in a bioreactor with two inoculum concentrations (5 and 10% v/v). The fermentation inoculated with higher inoculum size reduced the time for maximum enzyme production (from 72 to 48). The enzyme extract was concentrated using tangential ultrafiltration in hollow fiber membranes, and the produced cellulase blend was evaluated for its stability at 37 °C, operation temperature of the simultaneous SSF process, and at 50 °C, optimum temperature of cellulase blend activity. The cellulolytic preparation was stable for at least 300 h at both 37 °C and 50 °C. The ethanol production was carried out by PDC fed-batch SSF process, using the onsite cellulase blend. The feeding strategy circumvented the classic problems of diffusion limitations by diminishing the presence of a high solid:liquid ratio at any time, resulting in high ethanol concentration at the end of the process (100 g/L), which corresponded to a fermentation efficiency of 78% of the maximum obtainable theoretically. The experimental results led to the ratio of 380 L of ethanol per ton of sugar cane bagasse PDC. Copyright © 2012 Elsevier B.V. All rights reserved.

A strategy for the highly efficient production of docosahexaenoic acid by Aurantiochytrium limacinum SR21 using glucose and glycerol as the mixed carbon sources.

PubMed

Li, Jing; Liu, Ruijie; Chang, Guifang; Li, Xiangyu; Chang, Ming; Liu, Yuanfa; Jin, Qingzhe; Wang, Xingguo

2015-02-01

Glucose and glycerol are useful carbon sources for the cultivation of Aurantiochytrium limacinum SR21. Glucose facilitates rapid growth and lipid synthesis, and glycerol promotes the accumulation of docosahexaenoic acid (DHA) in A. limacinum SR21. To improve the DHA productivity of A. limacinum SR21, shake flask and fed-batch cultures were performed using glucose and glycerol as mixed carbon sources (MCSs). Along with optimization of the MCSs, the best DHA yield and productivity (32.36 g/L and 337.1 mg/L/h) were obtained via fed-batch fermentation with maintenance of a constant air supply. The DHA productivity was 15.24% higher than that obtained using glucose as single carbon source (SCS). This study presents a highly efficient and economic strategy for the production of DHA by A. limacinum SR21. Copyright © 2014 Elsevier Ltd. All rights reserved.

Progressing batch hydrolysis process

DOEpatents

Wright, J.D.

1985-01-10

A progressive batch hydrolysis process is disclosed for producing sugar from a lignocellulosic feedstock. It comprises passing a stream of dilute acid serially through a plurality of percolation hydrolysis reactors charged with feed stock, at a flow rate, temperature and pressure sufficient to substantially convert all the cellulose component of the feed stock to glucose. The cooled dilute acid stream containing glucose, after exiting the last percolation hydrolysis reactor, serially fed through a plurality of pre-hydrolysis percolation reactors, charged with said feedstock, at a flow rate, temperature and pressure sufficient to substantially convert all the hemicellulose component of said feedstock to glucose. The dilute acid stream containing glucose is cooled after it exits the last prehydrolysis reactor.

40 CFR 63.2192 - What definitions apply to this subpart?

Code of Federal Regulations, 2010 CFR

2010-07-01

... follows: Batch means a single fermentation cycle in a single fermentation vessel (fermenter). Batch... exhaust. This correlation is specific to each fed-batch fermentation stage and is established while... additives during fermentation in the vessel. In contrast, carbohydrates and additives are added to “set...

Fed-batch strategy for enhancing cell growth and C-phycocyanin production of Arthrospira (Spirulina) platensis under phototrophic cultivation.

PubMed

Xie, Youping; Jin, Yiwen; Zeng, Xianhai; Chen, Jianfeng; Lu, Yinghua; Jing, Keju

2015-03-01

The C-phycocyanin generated in blue-green algae Arthrospira platensis is gaining commercial interest due to its nutrition and healthcare value. In this study, the light intensity and initial biomass concentration were manipulated to improve cell growth and C-phycocyanin production of A.platensis in batch cultivation. The results show that low light intensity and high initial biomass concentration led to increased C-phycocyanin accumulation. The best C-phycocyanin productivity occurred when light intensity and initial biomass concentration were 300μmol/m(2)/s and 0.24g/L, respectively. The fed-batch cultivation proved to be an effective strategy to further enhance C-phycocyanin production of A.platensis. The results indicate that C-phycocyanin accumulation not only requires nitrogen-sufficient condition, but also needs other nutrients. The highest C-phycocyanin content (16.1%), production (1034mg/L) and productivity (94.8mg/L/d) were obtained when using fed-batch strategy with 5mM medium feeding. Copyright © 2014 Elsevier Ltd. All rights reserved.

Bacteriocin Production with Lactobacillus amylovorus DCE 471 Is Improved and Stabilized by Fed-Batch Fermentation

PubMed Central

Callewaert, Raf; De Vuyst, Luc

2000-01-01

Amylovorin L471 is a small, heat-stable, and hydrophobic bacteriocin produced by Lactobacillus amylovorus DCE 471. The nutritional requirements for amylovorin L471 production were studied with fed-batch fermentations. A twofold increase in bacteriocin titer was obtained when substrate addition was controlled by the acidification rate of the culture, compared with the titers reached with constant substrate addition or pH-controlled batch cultures carried out under the same conditions. An interesting feature of fed-batch cultures observed under certain culture conditions (constant feed rate) is the apparent stabilization of bacteriocin activity after obtaining maximum production. Finally, a mathematical model was set up to simulate cell growth, glucose and complex nitrogen source consumption, and lactic acid and bacteriocin production kinetics. The model showed that bacterial growth was dependent on both the energy and the complex nitrogen source. Bacteriocin production was growth associated, with a simultaneous bacteriocin adsorption on the producer cells dependent on the lactic acid accumulated and hence the viability of the cells. Both bacteriocin production and adsorption were inhibited by high concentrations of the complex nitrogen source. PMID:10653724

Improved production of an enzyme that hydrolyses raw yam starch by Penicillium sp. S-22 using fed-batch fermentation.

PubMed

Sun, Hai-Yan; Ge, Xiang-Yang; Zhang, Wei-Guo

2006-11-01

A newly isolated strain, Penicillium sp. S-22, was used to produce an enzyme that hydrolyses raw yam starch [raw yam starch digesting enzyme (RYSDE)]. The enzyme activity and overall enzyme productivity were respectively 16 U/ml and 0.19 U/ml h in the batch culture. The enzyme activity increased to 85 U/ml by feeding of partially hydrolyzed raw yam starch. When a mixture containing partially hydrolyzed raw yam starch and peptone was fed by a pH-stat strategy, the enzyme activity reached 366 U/ml, 23-fold of that obtained in the batch culture, and the overall productivity reached 3.4 U/ml h, which was 18-fold of that in the batch culture.

Effects of cell culture conditions on antibody N-linked glycosylation--what affects high mannose 5 glycoform.

PubMed

Pacis, Efren; Yu, Marcella; Autsen, Jennifer; Bayer, Robert; Li, Feng

2011-10-01

The glycosylation profile of therapeutic antibodies is routinely analyzed throughout development to monitor the impact of process parameters and to ensure consistency, efficacy, and safety for clinical and commercial batches of therapeutic products. In this study, unusually high levels of the mannose-5 (Man5) glycoform were observed during the early development of a therapeutic antibody produced from a Chinese hamster ovary (CHO) cell line, model cell line A. Follow up studies indicated that the antibody Man5 level was increased throughout the course of cell culture production as a result of increasing cell culture medium osmolality levels and extending culture duration. With model cell line A, Man5 glycosylation increased more than twofold from 12% to 28% in the fed-batch process through a combination of high basal and feed media osmolality and increased run duration. The osmolality and culture duration effects were also observed for four other CHO antibody producing cell lines by adding NaCl in both basal and feed media and extending the culture duration of the cell culture process. Moreover, reduction of Man5 level from model cell line A was achieved by supplementing MnCl2 at appropriate concentrations. To further understand the role of glycosyltransferases in Man5 level, N-acetylglucosaminyltransferase I GnT-I mRNA levels at different osmolality conditions were measured. It has been hypothesized that specific enzyme activity in the glycosylation pathway could have been altered in this fed-batch process. Copyright © 2011 Wiley Periodicals, Inc.

Lactic acid production from lime-treated wheat straw by Bacillus coagulans: neutralization of acid by fed-batch addition of alkaline substrate

PubMed Central

Maas, Ronald H. W.; Bakker, Robert R.; Jansen, Mickel L. A.; Visser, Diana; de Jong, Ed; Eggink, Gerrit

2008-01-01

Conventional processes for lignocellulose-to-organic acid conversion requires pretreatment, enzymatic hydrolysis, and microbial fermentation. In this study, lime-treated wheat straw was hydrolyzed and fermented simultaneously to lactic acid by an enzyme preparation and Bacillus coagulans DSM 2314. Decrease in pH because of lactic acid formation was partially adjusted by automatic addition of the alkaline substrate. After 55 h of incubation, the polymeric glucan, xylan, and arabinan present in the lime-treated straw were hydrolyzed for 55%, 75%, and 80%, respectively. Lactic acid (40.7 g/l) indicated a fermentation efficiency of 81% and a chiral l(+)-lactic acid purity of 97.2%. In total, 711 g lactic acid was produced out of 2,706 g lime-treated straw, representing 43% of the overall theoretical maximum yield. Approximately half of the lactic acid produced was neutralized by fed-batch feeding of lime-treated straw, whereas the remaining half was neutralized during the batch phase with a Ca(OH)2 suspension. Of the lime added during the pretreatment of straw, 61% was used for the neutralization of lactic acid. This is the first demonstration of a process having a combined alkaline pretreatment of lignocellulosic biomass and pH control in fermentation resulting in a significant saving of lime consumption and avoiding the necessity to recycle lime. PMID:18247027

Dual utilization of NADPH and NADH cofactors enhances xylitol production in engineered Saccharomyces cerevisiae.

PubMed

Jo, Jung-Hyun; Oh, Sun-Young; Lee, Hyeun-Soo; Park, Yong-Cheol; Seo, Jin-Ho

2015-12-01

Xylitol, a natural sweetener, can be produced by hydrogenation of xylose in hemicelluloses. In microbial processes, utilization of only NADPH cofactor limited commercialization of xylitol biosynthesis. To overcome this drawback, Saccharomyces cerevisiae D452-2 was engineered to express two types of xylose reductase (XR) with either NADPH-dependence or NADH-preference. Engineered S. cerevisiae DWM expressing both the XRs exhibited higher xylitol productivity than the yeast strain expressing NADPH-dependent XR only (DWW) in both batch and glucose-limited fed-batch cultures. Furthermore, the coexpression of S. cerevisiae ZWF1 and ACS1 genes in the DWM strain increased intracellular concentrations of NADPH and NADH and improved maximum xylitol productivity by 17%, relative to that for the DWM strain. Finally, the optimized fed-batch fermentation of S. cerevisiae DWM-ZWF1-ACS1 resulted in 196.2 g/L xylitol concentration, 4.27 g/L h productivity and almost the theoretical yield. Expression of the two types of XR utilizing both NADPH and NADH is a promising strategy to meet the industrial demands for microbial xylitol production. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Translational arrest due to cytoplasmic redox stress delays adaptation to growth on methanol and heterologous protein expression in a typical fed-batch culture of Pichia pastoris.

PubMed

Edwards-Jones, Bryn; Aw, Rochelle; Barton, Geraint R; Tredwell, Gregory D; Bundy, Jacob G; Leak, David J

2015-01-01

We have followed a typical fed-batch induction regime for heterologous protein production under the control of the AOX1 promoter using both microarray and metabolomic analysis. The genetic constructs involved 1 and 3 copies of the TRY1 gene, encoding human trypsinogen. In small-scale laboratory cultures, expression of the 3 copy-number construct induced the unfolded protein response (UPR) sufficiently that titres of extracellular trypsinogen were lower in the 3-copy construct than with the 1-copy construct. In the fed-batch-culture, a similar pattern was observed, with higher expression from the 1-copy construct, but in this case there was no significant induction of UPR with the 3-copy strain. Analysis of the microarray and metabolomic information indicates that the 3-copy strain was undergoing cytoplasmic redox stress at the point of induction with methanol. In this Crabtree-negative yeast, this redox stress appeared to delay the adaptation to growth on methanol and supressed heterologous protein production, probably due to a block in translation. Although redox imbalance as a result of artificially imposed hypoxia has previously been described, this is the first time that it has been characterised as a result of a transient metabolic imbalance and shown to involve a stress response which can lead to translational arrest. Without detailed analysis of the underlying processes it could easily have been mis-interpreted as secretion stress, transmitted through the UPR.

Enhanced poly(L-malic acid) production from pretreated cane molasses by Aureobasidium pullulans in fed-batch fermentation.

PubMed

Xia, Jun; Xu, Jiaxing; Hu, Lei; Liu, Xiaoyan

2016-11-16

Poly(L-malic acid) (PMA) is a natural polyester with many attractive properties for biomedical application. However, the cost of PMA production is high when glucose is used as a carbon source. To solve this problem, cane molasses as a low-cost feedstock was applied for the production of PMA. Six pretreatment methods were applied to cane molasses before fermentation. Pretreatment with combined tricalcium phosphate, potassium ferrocyanide, and sulfuric acid (TPFSA) removed significant amounts of metal ions from cane molasses. The PMA concentration increased from 5.4 g/L (untreated molasses) to 36.9 g/L (TPFSA-pretreated molasses) after fermentation in shake flasks. A fed-batch fermentation strategy was then developed. In this method, TPFSA-pretreated cane molasses solution was continuously fed into the fermentor to maintain the total sugar concentration at 20 g/L. This technique generated approximately 95.4 g/L PMA with a productivity of 0.57 g/L/hr. The present study indicated that fed-batch fermentation using pretreated cane molasses is a feasible technique for producing high amounts of PMA.

Design of two-column batch-to-batch recirculation to enhance performance in ion-exchange chromatography.

PubMed

Persson, Oliver; Andersson, Niklas; Nilsson, Bernt

2018-01-05

Preparative liquid chromatography is a separation technique widely used in the manufacturing of fine chemicals and pharmaceuticals. A major drawback of traditional single-column batch chromatography step is the trade-off between product purity and process performance. Recirculation of impure product can be utilized to make the trade-off more favorable. The aim of the present study was to investigate the usage of a two-column batch-to-batch recirculation process step to increase the performance compared to single-column batch chromatography at a high purity requirement. The separation of a ternary protein mixture on ion-exchange chromatography columns was used to evaluate the proposed process. The investigation used modelling and simulation of the process step, experimental validation and optimization of the simulated process. In the presented case the yield increases from 45.4% to 93.6% and the productivity increases 3.4 times compared to the performance of a batch run for a nominal case. A rapid concentration build-up product can be seen during the first cycles, before the process reaches a cyclic steady-state with reoccurring concentration profiles. The optimization of the simulation model predicts that the recirculated salt can be used as a flying start of the elution, which would enhance the process performance. The proposed process is more complex than a batch process, but may improve the separation performance, especially while operating at cyclic steady-state. The recirculation of impure fractions reduces the product losses and ensures separation of product to a high degree of purity. Copyright © 2017 Elsevier B.V. All rights reserved.

Ultraviolet Light (UV) Inactivation of Porcine Parvovirus in Liquid Plasma and Effect of UV Irradiated Spray Dried Porcine Plasma on Performance of Weaned Pigs

PubMed Central

Polo, Javier; Rodríguez, Carmen; Ródenas, Jesús; Russell, Louis E.; Campbell, Joy M.; Crenshaw, Joe D.; Torrallardona, David; Pujols, Joan

2015-01-01

A novel ultraviolet light irradiation (UV-C, 254 nm) process was designed as an additional safety feature for manufacturing of spray dried porcine plasma (SDPP). In Exp. 1, three 10-L batches of bovine plasma were inoculated with 105.2±0.12 tissue culture infectious dose 50 (TCID50) of porcine parvovirus (PPV) per mL of plasma and subjected to UV-C ranging from 0 to 9180 J/L. No viable PPV was detected in bovine plasma by micro-titer assay in SK6 cell culture after UV-C at 2295 J/L. In Exp. 2, porcine plasma was subjected to UV-C (3672 J/L), then spray dried and mixed in complete mash diets. Diets were a control without SDPP (Control), UV-C SDPP either at 3% (UVSDPP3) or 6% (UVSDPP6) and non-UV-C SDPP at 3% (SDPP3) or 6% (SDPP6). Diets were fed ad libitum to 320 weaned pigs (26 d of age; 16 pens/diet; 4 pigs/pen) for 14 d after weaning and a common diet was fed d 15 to 28. During d 0 to 14, pigs fed UVSDPP3, UVSDPP6, or SDPP6 had higher (P < 0.05) weight gain and feed intake than control. During d 0 to 28, pigs fed UVSDPP3 and UVSDPP6 had higher (P < 0.05) weight gain and feed intake than control and SDPP3, and SDPP6 had higher (P < 0.05) feed intake than control. Also, pigs fed UVSDPP had higher (P < 0.05) weight gain than pigs fed SDPP. In conclusion, UV-C inactivated PPV in liquid plasma and UVSDPP used in pig feed had no detrimental effects on pig performance. PMID:26171968

Performance and fat quality of heavy pigs fed maize differing in linoleic acid content.

PubMed

Della Casa, G; Bochicchio, D; Faeti, V; Marchetto, G; Poletti, E; Rossi, A; Panciroli, A; Mordenti, A L; Brogna, N

2010-01-01

Maize shows wide differences in linoleic acid due both to total lipid content and to fatty acid profile. Therefore, diets containing the same high maize percentage (up to 55%) can differ in linoleic acid content and lead to subcutaneous fats of differing suitability for raw ham curing. Two trials were performed on heavy pigs; in the first, 60 pigs (body weight 48.7+/-5.1 kg) were fed three diets made using three maize batches differing in linoleic acid due to different total lipid content, in the second trial, 40 pigs (live weight 70.4+/-3.4 kg) were fed two diets made using two maize batches differing in linoleic acid due to their fatty acid profile. Pigs were slaughtered at 170 kg of live weight. In both trials, the growth and slaughtering performance did not differ. In the first trial the three diets lead to a different content of linoleic acid both in subcutaneous (low linoleic vs medium linoleic vs high linoleic P0.01) and intramuscular fat (low linoleic vs high linoleic P0.05). In the second trial different linoleic acid content was observed for subcutaneous fat (P0.01) but not for intramuscular fat. To formulate diets for heavy pigs, it is crucial to know the linoleic acid content of the maize used, because differences of only 0.3% can lead to significant differences in fatty acids composition of depot fats.

Evaluation of an anaerobic digestion system for processing CELSS crop residues for resource recovery

NASA Astrophysics Data System (ADS)

Strayer, R. F.; Finger, B. W.; Alazraki, M. P.

1997-01-01

Three bioreactors, connected in series, were used to process CELSS potato residues for recovery of resources. The first stage was an anaerobic digestor (8 L working volume; cow rumen contents inoculum; fed-batch; 8 day retention time; feed rate 25 gdw day^-1) that converted 33% of feed (dry weight loss) to CO_2 and ``volatile fatty acids'' (vfa, 83:8:8 mmolar ratio acetic:propionic:butyric). High nitrate-N in the potato residue feed was absent in the anaerobic effluent, with a high portion converted to NH_4^+-N and the remainder unaccounted and probably lost to denitrification and NH_4^+ volatilization. Liquid anaerobic effluent was fed to an aerobic, yeast biomass production vessel (2 L volume; Candida ingens inoculum; batch [pellicle] growth; 2 day retention time) where the VFAs and some NH_4^+-N were converted into yeast biomass. Yeast yields accounted for up to 8% of potato residue fed into the anaerobic bioreactor. The third bioreactor (0.5 L liquid working volume; commercial nitrifier inoculum; packed-bed biofilm; continuous yeast effluent feed; recirculating; constant volume; 2 day hydraulic retention time) was used to convert successfully the remaining NH_4^+-N into nitrate-N (preferred form of N for CELSS crop production) and to remove the remaining degradable soluble organic carbon. Effluents from the last two stages were used for partial replenishment of minerals for hydroponic potato production.

Evaluation of an Anaerobic Digestion System for Processing CELSS Crop Residues for Resource Recovery

NASA Technical Reports Server (NTRS)

Strayer, R. F.; Finger, B. W.; Alazraki, M. P.

1997-01-01

Three bioreactors, connected in series, were used to process CELSS potato residues for recovery of resources. The first stage was an anaerobic digestor (8 L working volume; cow rumen contents inoculum; fed-batch; 8 day retention time; feed rate 25 gdw/day) that converted 33% of feed (dry weight loss) to CO2 and "volatile fatty acids" (vfa, 83:8:8 mmolar ratio acetic:propionic:butyric). High nitrate-N in the potato residue feed was absent in the anaerobic effluent, with a high portion converted to NH4(+)-N and the remainder unaccounted and probably lost to denitrification and NH4(+) volatilization. Liquid anaerobic effluent was fed to an aerobic, yeast biomass production vessel (2 L volume; Candida ingens inoculum; batch [pellicle] growth; 2 day retention time) where the VFAs and some NH4(+)-N were converted into yeast biomass. Yeast yields accounted for up to 8% of potato residue fed into the anaerobic bioreactor. The third bioreactor (0.5 L liquid working volume; commercial nitrifier inoculum; packed-bed biofilm; continuous yeast effluent feed; recirculating; constant volume; 2 day hydraulic retention time) was used to convert successfully the remaining NH4(+)-N into nitrate-N (preferred form of N for CELSS crop production) and to remove the remaining degradable soluble organic carbon. Effluents from the last two stages were used for partial replenishment of minerals for hydroponic potato production.

Fermentation strategy for second generation ethanol production from sugarcane bagasse hydrolyzate by Spathaspora passalidarum and Scheffersomyces stipitis.

PubMed

Nakanishi, Simone C; Soares, Lauren B; Biazi, Luiz Eduardo; Nascimento, Viviane M; Costa, Aline C; Rocha, George Jackson M; Ienczak, Jaciane L

2017-10-01

Alcoholic fermentation of released sugars in pretreatment and enzymatic hydrolysis of biomass is a central feature for second generation ethanol (E2G) production. Saccharomyces cerevisiae used industrially in the production of first generation ethanol (E1G) convert sucrose, fructose, and glucose into ethanol. However, these yeasts have no ability to ferment pentose (xylose). Therefore, the present work has focused on E2G production by Scheffersomyces stipitis and Spathaspora passalidarum. The fermentation strategy with high pitch, cell recycle, fed-batch mode, and temperature decrease for each batch were performed in a hydrolyzate obtained from a pretreatment at 130°C with NaOH solution (1.5% w/v) added with 0.15% (w/w) of anthraquinone (AQ) and followed by enzymatic hydrolysis. The process strategy has increased volumetric productivity from 0.35 to 0.38 g · L -1  · h -1 (first to third batch) for S. stipitis and from 0.38 to 0.81 g · L -1  · h -1 for S. passalidarum (first to fourth batch). Mass balance for the process proposed in this work showed the production of 177.33 kg ethanol/ton of sugar cane bagasse for S. passalidarum compared to 124.13 kg ethanol/ton of sugar cane bagasse for S. stipitis fermentation. The strategy proposed in this work can be considered as a promising strategy in the production of second generation ethanol. Biotechnol. Bioeng. 2017;114: 2211-2221. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

The Long Non-Coding RNA Transcriptome Landscape in CHO Cells Under Batch and Fed-Batch Conditions.

PubMed

Vito, Davide; Smales, C Mark

2018-05-21

The role of non-coding RNAs in determining growth, productivity and recombinant product quality attributes in Chinese hamster ovary (CHO) cells has received much attention in recent years, exemplified by studies into microRNAs in particular. However, other classes of non-coding RNAs have received less attention. One such class are the non-coding RNAs known collectively as long non-coding RNAs (lncRNAs). We have undertaken the first landscape analysis of the lncRNA transcriptome in CHO using a mouse based microarray that also provided for the surveillance of the coding transcriptome. We report on those lncRNAs present in a model host CHO cell line under batch and fed-batch conditions on two different days and relate the expression of different lncRNAs to each other. We demonstrate that the mouse microarray was suitable for the detection and analysis of thousands of CHO lncRNAs and validated a number of these by qRT-PCR. We then further analysed the data to identify those lncRNAs whose expression changed the most between growth and stationary phases of culture or between batch and fed-batch culture to identify potential lncRNA targets for further functional studies with regard to their role in controlling growth of CHO cells. We discuss the implications for the publication of this rich dataset and how this may be used by the community. This article is protected by copyright. All rights reserved.

D-lactic acid production by Sporolactobacillus inulinus Y2-8 immobilized in fibrous bed bioreactor using corn flour hydrolyzate.

PubMed

Zhao, Ting; Liu, Dong; Ren, Hengfei; Shi, Xinchi; Zhao, Nan; Chen, Yong; Ying, Hanjie

2014-12-28

In this study, a fibrous bed bioreactor (FBB) was used for D-lactic acid (D-LA) production by Sporolactobacillus inulinus Y2-8. Corn flour hydrolyzed with α-amylase and saccharifying enzyme was used as a cost-efficient and nutrient-rich substrate for D-LA production. A maximal starch conversion rate of 93.78% was obtained. The optimum pH for D-LA production was determined to be 6.5. Ammonia water was determined to be an ideal neutralizing agent, which improved the D-LA production and purification processes. Batch fermentation and fedbatch fermentation, with both free cells and immobilized cells, were compared to highlight the advantages of FBB fermentation. In batch mode, the D-LA production rate of FBB fermentation was 1.62 g/l/h, which was 37.29% higher than that of free-cell fermentation, and the D-LA optical purities of the two fermentation methods were above 99.00%. In fed-batch mode, the maximum D-LA concentration attained by FBB fermentation was 218.8 g/l, which was 37.67% higher than that of free-cell fermentation. Repeated-batch fermentation was performed to determine the long-term performance of the FBB system, and the data indicated that the average D-LA production rate was 1.62 g/l/h and the average yield was 0.98 g/g. Thus, hydrolyzed corn flour fermented by S. inulinus Y2-8 in a FBB may be used for improving D-LA fermentation by using ammonia water as the neutralizing agent.

Performance and membrane fouling of a step-fed submerged membrane sequencing batch reactor treating swine biogas digestion slurry.

PubMed

Han, Zhiying; Chen, Shixia; Lin, Xiaochang; Yu, Hongjun; Duan, Li'an; Ye, Zhangying; Jia, Yanbo; Zhu, Songming; Liu, Dezhao

2018-01-02

To identify the performance of step-fed submerged membrane sequencing batch reactor (SMSBR) treating swine biogas digestion slurry and to explore the correlation between microbial metabolites and membrane fouling within this novel reactor, a lab-scale step-fed SMSBR was operated under nitrogen loading rate of 0.026, 0.052 and 0.062 g NH 4 + -N (gVSS·d) -1 . Results show that the total removal efficiencies for NH 4 + -N, total nitrogen and chemical oxygen demand in the reactor (>94%, >89% and >97%, respectively) were high during the whole experiment. However, the cycle removal efficiency of NH 4 + -N decreased significantly when the nitrogen loading rate was increased to 0.062 g NH 4 + -N (gVSS·d) -1 . The total removal efficiency of total phosphorus in the step-fed SMSBR was generally higher than 75%, though large fluctuations were observed during the experiments. In addition, the concentrations of microbial metabolites, i.e., soluble microbial products (SMP) and extracellular polymeric substances (EPS) from activated sludge increased as nitrogen loading rate increased, both showing quadratic equation correlations with viscosity of the mixed liquid in the step-fed SMSBR (both R 2 > 0.90). EPS content was higher than SMP content, while protein (PN) was detected as the main component in both SMP and EPS. EPS PN was found to be well correlated with transmembrane pressure, membrane flux and the total membrane fouling resistance. Furthermore, the three-dimensional excitation-emission matrix fluorescence spectroscopy results suggested the tryptophan-like protein as one of the main contributors to the membrane fouling. Overall, this study showed that the step-fed SMSBR could be used to treat swine digestion slurry at nitrogen loading rate of 0.052 g NH 4 + -N (gVSS·d) -1 , and the control strategy of membrane fouling should be developed based on reducing the tryptophan-like PN in EPS.

Hydrogen production from formic acid in pH-stat fed-batch operation for direct supply to fuel cell.

PubMed

Shin, Jong-Hwan; Yoon, Jong Hyun; Lee, Seung Hoon; Park, Tai Hyun

2010-01-01

Enterobacter asburiae SNU-1 harvested after cultivation was used as a whole cell biocatalyst, for the production of hydrogen. Formic acid was efficiently converted to hydrogen using the harvested cells with an initial hydrogen production rate and total hydrogen production of 491 ml/l/h and 6668 ml/l, respectively, when 1 g/l of whole cell enzyme was used. Moreover, new pH-stat fed-batch operation was conducted, and total hydrogen production was 1.4 times higher than that of batch operation. For practical application, bio-hydrogen produced from formic acid using harvested cells was directly applied to PEMFC for power generation.

Effects of pelleting, extrusion, or extrusion and pelleting on energy and nutrient digestibility in diets containing different levels of fiber and fed to growing pigs.

PubMed

Rojas, O J; Vinyeta, E; Stein, H H

2016-05-01

An experiment was conducted to determine effects of pelleting, extrusion, and extrusion and pelleting on energy and nutrient digestibility in diets containing low, medium, or high concentrations of fiber. Three diets were formulated: 1) the low-fiber diet contained corn and soybean meal; 2) the medium-fiber diet contained corn, soybean meal, and 25% distillers dried grains with solubles (DDGS); and 3) the high-fiber diet contained corn, soybean meal, 25% DDGS, and 20% soybean hulls. Each diet was divided into 4 batches after mixing. One batch was not further processed and was fed in a meal form, one batch was pelleted at 85°C, one batch was extruded at 115°C using a single-screw extruder, and one batch was extruded at 115°C and then pelleted at 85°C. Thus, 12 different diets were produced. Twenty-four growing pigs (26.5 ± 1.5 kg initial BW) had a T-cannula installed in the distal ileum and were allotted to the 12 diets in a split-plot design with 8 pigs allotted to the low-fiber diets, the medium-fiber diets, and the high-fiber diets, respectively. Diets were fed to the pigs during four 14-d periods. Within each type of diet, the 8 pigs were fed the diets produced using the 4 processing technologies. Therefore, there were 8 replicate pigs per diet. Pigs were adjusted to their diets for 14 d before the experiment was initiated. Each of the four 14-d periods consisted of 5 d for adaptation, 5 d of fecal collection according to the marker to marker approach, and ileal digesta were collected on d 13 and 14. Results indicated that pelleting, extrusion, or extrusion and pelleting improved ( < 0.05) the apparent ileal digestibility of starch and most indispensable AA. In most cases, there were no differences between the pelleted, the extruded, and the extruded and pelleted diets. The apparent total tract digestibility of GE was also improved ( < 0.05) by pelleting and by the combination of extrusion and pelleting. The ME of pelleted diets was greater ( < 0.05) than that of meal diets for the low- and medium-fiber diets, but this was not the case for high-fiber diets (interaction, < 0.05). Medium- and high-fiber diets that were extruded had greater ME ( < 0.05) than meal diets, but that was not the case for low-fiber diets. These data indicate that energy utilization may be improved by pelleting or extrusion or by a combination of the 2 technologies, but the response seems to be greater for extrusion in diets that are relatively high in fiber.

Capacity planning for batch and perfusion bioprocesses across multiple biopharmaceutical facilities.

PubMed

Siganporia, Cyrus C; Ghosh, Soumitra; Daszkowski, Thomas; Papageorgiou, Lazaros G; Farid, Suzanne S

2014-01-01

Production planning for biopharmaceutical portfolios becomes more complex when products switch between fed-batch and continuous perfusion culture processes. This article describes the development of a discrete-time mixed integer linear programming (MILP) model to optimize capacity plans for multiple biopharmaceutical products, with either batch or perfusion bioprocesses, across multiple facilities to meet quarterly demands. The model comprised specific features to account for products with fed-batch or perfusion culture processes such as sequence-dependent changeover times, continuous culture constraints, and decoupled upstream and downstream operations that permit independent scheduling of each. Strategic inventory levels were accounted for by applying cost penalties when they were not met. A rolling time horizon methodology was utilized in conjunction with the MILP model and was shown to obtain solutions with greater optimality in less computational time than the full-scale model. The model was applied to an industrial case study to illustrate how the framework aids decisions regarding outsourcing capacity to third party manufacturers or building new facilities. The impact of variations on key parameters such as demand or titres on the optimal production plans and costs was captured. The analysis identified the critical ratio of in-house to contract manufacturing organization (CMO) manufacturing costs that led the optimization results to favor building a future facility over using a CMO. The tool predicted that if titres were higher than expected then the optimal solution would allocate more production to in-house facilities, where manufacturing costs were lower. Utilization graphs indicated when capacity expansion should be considered. © 2014 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers.

Capacity Planning for Batch and Perfusion Bioprocesses Across Multiple Biopharmaceutical Facilities

PubMed Central

Siganporia, Cyrus C; Ghosh, Soumitra; Daszkowski, Thomas; Papageorgiou, Lazaros G; Farid, Suzanne S

2014-01-01

Production planning for biopharmaceutical portfolios becomes more complex when products switch between fed-batch and continuous perfusion culture processes. This article describes the development of a discrete-time mixed integer linear programming (MILP) model to optimize capacity plans for multiple biopharmaceutical products, with either batch or perfusion bioprocesses, across multiple facilities to meet quarterly demands. The model comprised specific features to account for products with fed-batch or perfusion culture processes such as sequence-dependent changeover times, continuous culture constraints, and decoupled upstream and downstream operations that permit independent scheduling of each. Strategic inventory levels were accounted for by applying cost penalties when they were not met. A rolling time horizon methodology was utilized in conjunction with the MILP model and was shown to obtain solutions with greater optimality in less computational time than the full-scale model. The model was applied to an industrial case study to illustrate how the framework aids decisions regarding outsourcing capacity to third party manufacturers or building new facilities. The impact of variations on key parameters such as demand or titres on the optimal production plans and costs was captured. The analysis identified the critical ratio of in-house to contract manufacturing organization (CMO) manufacturing costs that led the optimization results to favor building a future facility over using a CMO. The tool predicted that if titres were higher than expected then the optimal solution would allocate more production to in-house facilities, where manufacturing costs were lower. Utilization graphs indicated when capacity expansion should be considered. © 2013 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers Biotechnol. Prog., 30:594–606, 2014 PMID:24376262

Microbial production of polyhydroxybutyrate with tailor-made properties: an integrated modelling approach and experimental validation.

PubMed

Penloglou, Giannis; Chatzidoukas, Christos; Kiparissides, Costas

2012-01-01

The microbial production of polyhydroxybutyrate (PHB) is a complex process in which the final quantity and quality of the PHB depend on a large number of process operating variables. Consequently, the design and optimal dynamic operation of a microbial process for the efficient production of PHB with tailor-made molecular properties is an extremely interesting problem. The present study investigates how key process operating variables (i.e., nutritional and aeration conditions) affect the biomass production rate and the PHB accumulation in the cells and its associated molecular weight distribution. A combined metabolic/polymerization/macroscopic modelling approach, relating the process performance and product quality with the process variables, was developed and validated using an extensive series of experiments and measurements. The model predicts the dynamic evolution of the biomass growth, the polymer accumulation, the consumption of carbon and nitrogen sources and the average molecular weights of the PHB in a bioreactor, under batch and fed-batch operating conditions. The proposed integrated model was used for the model-based optimization of the production of PHB with tailor-made molecular properties in Azohydromonas lata bacteria. The process optimization led to a high intracellular PHB accumulation (up to 95% g of PHB per g of DCW) and the production of different grades (i.e., different molecular weight distributions) of PHB. Copyright © 2011 Elsevier Inc. All rights reserved.

Quick generation of Raman spectroscopy based in-process glucose control to influence biopharmaceutical protein product quality during mammalian cell culture.

PubMed

Berry, Brandon N; Dobrowsky, Terrence M; Timson, Rebecca C; Kshirsagar, Rashmi; Ryll, Thomas; Wiltberger, Kelly

2016-01-01

Mitigating risks to biotherapeutic protein production processes and products has driven the development of targeted process analytical technology (PAT); however implementing PAT during development without significantly increasing program timelines can be difficult. The development of a monoclonal antibody expressed in a Chinese hamster ovary (CHO) cell line via fed-batch processing presented an opportunity to demonstrate capabilities of altering percent glycated protein product. Glycation is caused by pseudo-first order, non-enzymatic reaction of a reducing sugar with an amino group. Glucose is the highest concentration reducing sugar in the chemically defined media (CDM), thus a strategy controlling glucose in the production bioreactor was developed utilizing Raman spectroscopy for feedback control. Raman regions for glucose were determined by spiking studies in water and CDM. Calibration spectra were collected during 8 bench scale batches designed to capture a wide glucose concentration space. Finally, a PLS model capable of translating Raman spectra to glucose concentration was built using the calibration spectra and spiking study regions. Bolus feeding in mammalian cell culture results in wide glucose concentration ranges. Here we describe the development of process automation enabling glucose setpoint control. Glucose-free nutrient feed was fed daily, however glucose stock solution was fed as needed according to online Raman measurements. Two feedback control conditions were executed where glucose was controlled at constant low concentration or decreased stepwise throughout. Glycation was reduced from ∼9% to 4% using a low target concentration but was not reduced in the stepwise condition as compared to the historical bolus glucose feeding regimen. © 2015 American Institute of Chemical Engineers.

Model-Based Nutrient Feeding Strategies for the Increased Production of Polyhydroxybutyrate (PHB) by Alcaligenes latus.

PubMed

Gahlawat, Geeta; Srivastava, Ashok K

2017-10-01

Polyhydroxyalkanoates (PHAs) are biodegradable polymers which are considered as an effective alternative for conventional plastics due to their mechanical properties similar to the latter. However, the widespread use of these polymers is still hampered due to their higher cost of production as compared to plastics. The production cost could be overcome by obtaining high yields and productivity. The goal of the present research was to enhance the yield of polyhydroxybutyrate (PHB) with the help of two simple fed-batch cultivation strategies. In the present study, average batch kinetic and substrate limitation/inhibition study data of Alcaligenes latus was used for the development of PHB model which was then adopted for designing various off-line nutrient feeding strategies to enhance PHB accumulation. The predictive ability of the model was validated by experimental implementation of two fed-batch strategies. One such dynamic strategy of fed-batch cultivation under pseudo-steady state with respect to nitrogen and simultaneous carbon feeding strategy resulted in significantly high biomass and PHB concentration of 39.17 g/L and 29.64 g/L, respectively. This feeding strategy demonstrated a high PHB productivity and PHB content of 0.6 g/L h and 75%, respectively, which were remarkably high in comparison to batch cultivation. The mathematical model can also be employed for designing various other nutrient feeding strategies.

High bioethanol titre from Manihot glaziovii through fed-batch simultaneous saccharification and fermentation in Automatic Gas Potential Test System.

PubMed

Moshi, Anselm P; Crespo, Carla F; Badshah, Malik; Hosea, Kenneth M M; Mshandete, Anthony Manoni; Mattiasson, Bo

2014-03-01

A process for the production of high bioethanol titre was established through fed-batch and simultaneous saccharification and fermentation (FB-SSF) of wild, non-edible cassava Manihot glaziovii. FB-SSF allowed fermentation of up to 390g/L of starch-derived glucose achieving high bioethanol concentration of up to 190g/L (24% v/v) with yields of around 94% of the theoretical value. The wild cassava M. glaziovii starch is hydrolysable with a low dosage of amylolytic enzymes (0.1-0.15% v/w, Termamyl® and AMG®). The Automatic Gas Potential Test System (AMPTS) was adapted to yeast ethanol fermentation and demonstrated to be an accurate, reliable and flexible device for studying the kinetics of yeast in SSF and FB-SSF. The bioethanol derived stoichiometrically from the CO2 registered in the AMPTS software correlated positively with samples analysed by HPLC (R(2)=0.99). Copyright © 2013 Elsevier Ltd. All rights reserved.

A rational approach to improving productivity in recombinant Pichia pastoris fermentation.

PubMed

d'Anjou, M C; Daugulis, A J

2001-01-05

A Mut(S) Pichia pastoris strain that had been genetically modified to produce and secrete sea raven antifreeze protein was used as a model system to demonstrate the implementation of a rational, model-based approach to improve process productivity. A set of glycerol/methanol mixed-feed continuous stirred-tank reactor (CSTR) experiments was performed at the 5-L scale to characterize the relationship between the specific growth rate and the cell yield on methanol, the specific methanol consumption rate, the specific recombinant protein formation rate, and the productivity based on secreted protein levels. The range of dilution rates studied was 0. 01 to 0.10 h(-1), and the residual methanol concentration was kept constant at approximately 2 g/L (below the inhibitory level). With the assumption that the cell yield on glycerol was constant, the cell yield on methanol increased from approximately 0.5 to 1.5 over the range studied. A maximum specific methanol consumption rate of 20 mg/g. h was achieved at a dilution rate of 0.06 h(-1). The specific product formation rate and the volumetric productivity based on product continued to increase over the range of dilution rates studied, and the maximum values were 0.06 mg/g. h and 1.7 mg/L. h, respectively. Therefore, no evidence of repression by glycerol was observed over this range, and operating at the highest dilution rate studied maximized productivity. Fed-batch mass balance equations, based on Monod-type kinetics and parameters derived from data collected during the CSTR work, were then used to predict cell growth and recombinant protein production and to develop an exponential feeding strategy using two carbon sources. Two exponential fed-batch fermentations were conducted according to the predicted feeding strategy at specific growth rates of 0.03 h(-1) and 0.07 h(-1) to verify the accuracy of the model. Cell growth was accurately predicted in both fed-batch runs; however, the model underestimated recombinant product concentration. The overall volumetric productivity of both runs was approximately 2.2 mg/L. h, representing a tenfold increase in the productivity compared with a heuristic feeding strategy. Copyright 2001 John Wiley & Sons, Inc.

Metabolic Control in Mammalian Fed-Batch Cell Cultures for Reduced Lactic Acid Accumulation and Improved Process Robustness

PubMed Central

Konakovsky, Viktor; Clemens, Christoph; Müller, Markus Michael; Bechmann, Jan; Berger, Martina; Schlatter, Stefan; Herwig, Christoph

2016-01-01

Biomass and cell-specific metabolic rates usually change dynamically over time, making the “feed according to need” strategy difficult to realize in a commercial fed-batch process. We here demonstrate a novel feeding strategy which is designed to hold a particular metabolic state in a fed-batch process by adaptive feeding in real time. The feed rate is calculated with a transferable biomass model based on capacitance, which changes the nutrient flow stoichiometrically in real time. A limited glucose environment was used to confine the cell in a particular metabolic state. In order to cope with uncertainty, two strategies were tested to change the adaptive feed rate and prevent starvation while in limitation: (i) inline pH and online glucose concentration measurement or (ii) inline pH alone, which was shown to be sufficient for the problem statement. In this contribution, we achieved metabolic control within a defined target range. The direct benefit was two-fold: the lactic acid profile was improved and pH could be kept stable. Multivariate Data Analysis (MVDA) has shown that pH influenced lactic acid production or consumption in historical data sets. We demonstrate that a low pH (around 6.8) is not required for our strategy, as glucose availability is already limiting the flux. On the contrary, we boosted glycolytic flux in glucose limitation by setting the pH to 7.4. This new approach led to a yield of lactic acid/glucose (Y L/G) around zero for the whole process time and high titers in our labs. We hypothesize that a higher carbon flux, resulting from a higher pH, may lead to more cells which produce more product. The relevance of this work aims at feeding mammalian cell cultures safely in limitation with a desired metabolic flux range. This resulted in extremely stable, low glucose levels, very robust pH profiles without acid/base interventions and a metabolic state in which lactic acid was consumed instead of being produced from day 1. With this contribution, we wish to extend the basic repertoire of available process control strategies, which will open up new avenues in automation technology and radically improve process robustness in both process development and manufacturing. PMID:28952567

Expression of a mutated SPT15 gene in Saccharomyces cerevisiae enhances both cell growth and ethanol production in microaerobic batch, fed-batch, and simultaneous saccharification and fermentations.

PubMed

Seong, Yeong-Je; Park, Haeseong; Yang, Jungwoo; Kim, Soo-Jung; Choi, Wonja; Kim, Kyoung Heon; Park, Yong-Cheol

2017-05-01

The SPT15 gene encodes a Saccharomyces cerevisiae TATA-binding protein, which is able to globally control the transcription levels of various metabolic and regulatory genes. In this study, a SPT15 gene mutant (S42N, S78R, S163P, and I212N) was expressed in S. cerevisiae BY4741 (BSPT15-M3), of which effects on fermentative yeast properties were evaluated in a series of culture types. By applying different nitrogen sources and air supply conditions in batch culture, organic nitrogen sources and microaerobic condition were decided to be more favorable for both cell growth and ethanol production of the BSPT15-M3 strain than the control S. cerevisiae BY4741 strain expressing the SPT15 gene (BSPT15wt). Microaerobic fed-batch cultures of BSPT15-M3 with glucose shock in the presence of high ethanol content resulted in a 9.5-13.4% higher glucose consumption rate and ethanol productivity than those for the BSPT15wt strain. In addition, BSPT15-M3 showed 4.5 and 3.9% increases in ethanol productivity from cassava hydrolysates and corn starch in simultaneous saccharification and fermentation processes, respectively. It was concluded that overexpression of the mutated SPT15 gene would be a potent strategy to develop robust S. cerevisiae strains with enhanced cell growth and ethanol production abilities.

Methanogenic community shifts during the transition from sewage mono-digestion to co-digestion of grass biomass.

PubMed

Hardegen, Justus; Latorre-Pérez, Adriel; Vilanova, Cristina; Günther, Thomas; Porcar, Manuel; Luschnig, Olaf; Simeonov, Claudia; Abendroth, Christian

2018-06-06

In this work, liquid and solid fractions of grass biomass were used as co-substrates for anaerobic co-digestion of sewage sludge. The input of grass biomass was increased gradually, and the underlying methanogenic microbiome was assessed by means of microscopy-based cell counting and full-length 16S rRNA gene high-throughput sequencing, proving for the first time the suitability of nanopore-based portable sequencers as a monitoring tool for anaerobic digestion systems. In both cases co-fermentation resulted in an increased number of bacteria and methanogenic archaea. Interestingly, the microbial communities were highly different between solid and liquid-fed batches. Liquid-fed batches developed a more stable microbiome, enriched in Methanosarcina spp., and resulted in higher methanogenic yield. In contrast, solid-fed batches were highly unstable at higher substrate concentrations, and kept Methanosaeta spp. - typically associated to sewage sludge - as the majoritary methanogenic archaea. Copyright © 2018 Elsevier Ltd. All rights reserved.

Improved docosahexaenoic acid production in Aurantiochytrium by glucose limited pH-auxostat fed-batch cultivation.

PubMed

Janthanomsuk, Panyawut; Verduyn, Cornelis; Chauvatcharin, Somchai

2015-11-01

Fed-batch, pH auxostat cultivation of the docosahexaenoic acid (DHA)-producing microorganism Aurantiochytrium B072 was performed to obtain high cell density and record high productivity of both total fatty acid (TFA) and DHA. Using glucose feeding by carbon excess (C-excess) and by C-limitation at various feeding rates (70%, 50% or 20% of C-excess), high biomass density was obtained and DHA/TFA content (w/w) was improved from 30% to 37% with a 50% glucose feed rate when compared with C-excess. To understand the biochemistry behind these improvements, lipogenic enzyme assays and in silico metabolic flux calculations were used and revealed that enzyme activity and C-fluxes to TFA were reduced with C-limited feeding but that the carbon flux to the polyketide synthase pathway increased relative to the fatty acid synthase pathway. As a result, a new strategy to improve the DHA to TFA content while maintaining relatively high DHA productivity is proposed. Copyright © 2015 Elsevier Ltd. All rights reserved.

Improving industrial full-scale production of baker's yeast by optimizing aeration control.

PubMed

Blanco, Carlos A; Rayo, Julia; Giralda, José M

2008-01-01

This work analyzes the control of optimum dissolved oxygen of an industrial fed-batch procedure in which baker's yeast (Saccharomyces cerevisiae) is grown under aerobic conditions. Sugar oxidative metabolism was controlled by monitoring aeration, molasses flows, and yeast concentration in the propagator along the later stage of the propagation, and keeping pH and temperature under controlled conditions. A large number of fed-batch growth experiments were performed in the tank for a period of 16 h, for each of the 3 manufactured commercial products. For optimization and control of cultivations, the growth and metabolite formation were quantified through measurement of specific growth and ethanol concentration. Data were adjusted to a model of multiple lineal regression, and correlations representing dissolved oxygen as a function of aeration, molasses, yeast concentration in the broth, temperature, and pH were obtained. The actual influence of each variable was consistent with the mathematical model, further justified by significant levels of each variable, and optimum aeration profile during the yeast propagation.

Translational Arrest Due to Cytoplasmic Redox Stress Delays Adaptation to Growth on Methanol and Heterologous Protein Expression in a Typical Fed-Batch Culture of Pichia pastoris

PubMed Central

Edwards-Jones, Bryn; Aw, Rochelle; Barton, Geraint R.; Tredwell, Gregory D.; Bundy, Jacob G.; Leak, David J.

2015-01-01

Results We have followed a typical fed-batch induction regime for heterologous protein production under the control of the AOX1 promoter using both microarray and metabolomic analysis. The genetic constructs involved 1 and 3 copies of the TRY1 gene, encoding human trypsinogen. In small-scale laboratory cultures, expression of the 3 copy-number construct induced the unfolded protein response (UPR) sufficiently that titres of extracellular trypsinogen were lower in the 3-copy construct than with the 1-copy construct. In the fed-batch-culture, a similar pattern was observed, with higher expression from the 1-copy construct, but in this case there was no significant induction of UPR with the 3-copy strain. Analysis of the microarray and metabolomic information indicates that the 3-copy strain was undergoing cytoplasmic redox stress at the point of induction with methanol. In this Crabtree-negative yeast, this redox stress appeared to delay the adaptation to growth on methanol and supressed heterologous protein production, probably due to a block in translation. Conclusion Although redox imbalance as a result of artificially imposed hypoxia has previously been described, this is the first time that it has been characterised as a result of a transient metabolic imbalance and shown to involve a stress response which can lead to translational arrest. Without detailed analysis of the underlying processes it could easily have been mis-interpreted as secretion stress, transmitted through the UPR. PMID:25785713

Growth kinetics of the photosynthetic bacterium Chlorobium thiosulfatophilum in a fed-batch reactor.

PubMed

Kim, B W; Chang, H N; Kim, I K; Lee, K S

1992-08-01

Hydrogen sulfide dissolved in water can be converted to elementary sulfur or sulfate by the photosynthetic bacterium Chlorobium thiosulfatophilum. Substrate inhibition occurred at sulfide concentrations above 5.7 mM. Light inhibition was found at average light intensities of 40,000 lux in a sulfide concentration of 5 mM, where no substrate inhibition occurred. Light intensity, the most important growth parameter, was attenuated through both scattering by sulfur particles and absorption by the cells. Average cell and sulfur particle sizes were 1.1 and 9.4 microm, respectively. Cells contributed 10 times as much to the turbidity as sulfur particles of the same weight concentration. The light attenuation factor was mathematically modeled, considering both the absorption and scattering effects based on the Beer-Lambert law and the Rayleigh theory, which were introduced to the cell growth model. Optimal operational conditions relating feed rate vs. light intensity were obtained to suppress the accumulation of sulfate and sulfide and save light energy for 2- and 4-L fed-batch reactors. Light intensity should be greater for the same performance (H(2)S removal rate/unit cell concentration) in larger reactors due to the scaleup effect on light transmission. Knowledge of appropriate growth kinetics in photosynthetic fed-batch reactors was essential to increase feed rate and light intensity and therefore cell growth. A mathematical model was developed that describes the cell growth by considering the light attenuation factor due to scattering and absorption and the crowding effect of the cells. This model was in good agreement with the experimental results. (c) 1992 John Wiley & Sons, Inc.

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.

Antibiotic free selection for the high level biosynthesis of a silk-elastin-like protein

PubMed Central

Barroca, Mário; Rodrigues, Paulo; Sobral, Rómulo; Costa, M. Manuela R.; Chaves, Susana R.; Machado, Raul; Casal, Margarida; Collins, Tony

2016-01-01

Silk-elastin-like proteins (SELPs) are a family of genetically engineered recombinant protein polymers exhibiting mechanical and biological properties suited for a wide range of applications in the biomedicine and materials fields. They are being explored as the next generation of biomaterials but low productivities and use of antibiotics during production undermine their economic viability and safety. We have developed an industrially relevant, scalable, fed-batch process for the high level production of a novel SELP in E. coli in which the commonly used antibiotic selection marker of the expression vector is exchanged for a post segregational suicide system, the separate-component-stabilisation system (SCS). SCS significantly augments SELP productivity but also enhances the product safety profile and reduces process costs by eliminating the use of antibiotics. Plasmid content increased following induction but no significant differences in plasmid levels were discerned when using SCS or the antibiotic selection markers under the controlled fed-batch conditions employed. It is suggested that the absence of competing plasmid-free cells improves host cell viability and enables increased productivity with SCS. With the process developed, 12.8 g L−1 purified SELP was obtained, this is the highest SELP productivity reported to date and clearly demonstrates the commercial viability of these promising polymers. PMID:27982135

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.

Economic process to co-produce poly(ε-l-lysine) and poly(l-diaminopropionic acid) by a pH and dissolved oxygen control strategy.

PubMed

Xu, Zhaoxian; Feng, Xiaohai; Sun, Zhuzhen; Cao, Changhong; Li, Sha; Xu, Zheng; Xu, Zongqi; Bo, Fangfang; Xu, Hong

2015-01-01

This study tended to apply biorefinery of indigenous microbes to the fermentation of target-product generation through a novel control strategy. A novel strategy for co-producing two valuable homopoly(amino acid)s, poly(ε-l-lysine) (ε-PL) and poly(l-diaminopropionic acid) (PDAP), was developed by controlling pH and dissolved oxygen concentrations in Streptomyces albulus PD-1 fermentation. The production of ε-PL and PDAP got 29.4 and 9.6gL(-1), respectively, via fed-batch cultivation in a 5L bioreactor. What is more, the highest production yield (21.8%) of similar production systems was achieved by using this novel strategy. To consider the economic-feasibility, large-scale production in a 1t fermentor was also implemented, which would increase the gross profit of 54,243.5USD from one fed-batch bioprocess. This type of fermentation, which produces multiple commercial products from a unified process is attractive, because it will improve the utilization rate of raw materials, enhance production value and enrich product variety. Copyright © 2015 Elsevier Ltd. All rights reserved.

Hybrid modeling of microbial exopolysaccharide (EPS) production: The case of Enterobacter A47.

PubMed

Marques, Rodolfo; von Stosch, Moritz; Portela, Rui M C; Torres, Cristiana A V; Antunes, Sílvia; Freitas, Filomena; Reis, Maria A M; Oliveira, Rui

2017-03-20

Enterobacter A47 is a bacterium that produces high amounts of a fucose-rich exopolysaccharide (EPS) from glycerol residue of the biodiesel industry. The fed-batch process is characterized by complex non-linear dynamics with highly viscous pseudo-plastic rheology due to the accumulation of EPS in the culture medium. In this paper, we study hybrid modeling as a methodology to increase the predictive power of models for EPS production optimization. We compare six hybrid structures that explore different levels of knowledge-based and machine-learning model components. Knowledge-based components consist of macroscopic material balances, Monod type kinetics, cardinal temperature and pH (CTP) dependency and power-law viscosity models. Unknown dependencies are set to be identified by a feedforward artificial neural network (ANN). A semiparametric identification schema is applied resorting to a data set of 13 independent fed-batch experiments. A parsimonious hybrid model was identified that describes the dynamics of the 13 experiments with the same parameterization. The final model is specific to Enterobacter A47 but can be easily extended to other microbial EPS processes. Copyright © 2017 Elsevier B.V. All rights reserved.

Environmentally Friendly Production of D(-) Lactic Acid by Sporolactobacillus nakayamae: Investigation of Fermentation Parameters and Fed-Batch Strategies.

PubMed

Michelz Beitel, Susan; Fontes Coelho, Luciana; Sass, Daiane Cristina; Contiero, Jonas

2017-01-01

The interest in the production of lactic acid has increased due to its wide range of applications. In the present study, the variables that affect fermentative D(-) lactic acid production were investigated: neutralizing agents, pH, temperature, inoculum percentage, agitation, and concentration of the medium components. An experimental design was applied to determine the optimal concentrations of the medium components and fermentation was studied using different feeding strategies. High production (122.41 g/L) and productivity (3.65 g/L·h) were efficiently achieved by Sporolactobacillus nakayamae in 54 h using a multipulse fed-batch technique with an initial medium containing 35 g/L of yeast extract (byproduct of alcohol production), 60 g/L of crystallized sugar, and 7.5 mL/L of salts. The fermentation process was conducted at 35°C and pH 6.0 controlled by NaOH with a 20% volume of inoculum and agitation at 125 rpm. The production of a high optically pure concentration of D(-) lactic acid combined with an environmentally friendly NaOH-based process demonstrates that S. nakayamae is a promising strain for D(-) lactic acid production.

High-titer lactic acid production from NaOH-pretreated corn stover by Bacillus coagulans LA204 using fed-batch simultaneous saccharification and fermentation under non-sterile condition.

PubMed

Hu, Jinlong; Zhang, Zhenting; Lin, Yanxu; Zhao, Shumiao; Mei, Yuxia; Liang, Yunxiang; Peng, Nan

2015-04-01

Lactic acid (LA) is an important chemical with various industrial applications. Non-food feedstock is commercially attractive for use in LA production; however, efficient LA fermentation from lignocellulosic biomass resulting in both high yield and titer faces technical obstacles. In this study, the thermophilic bacterium Bacillus coagulans LA204 demonstrated considerable ability to ferment glucose, xylose, and cellobiose to LA. Importantly, LA204 produces LA from several NaOH-pretreated agro stovers, with remarkably high yields through simultaneous saccharification and fermentation (SSF). A fed-batch SSF process conducted at 50°C and pH 6.0, using a cellulase concentration of 30 FPU (filter paper unit)/g stover and 10 g/L yeast extract in a 5-L bioreactor, was developed to produce LA from 14.4% (w/w) NaOH-pretreated non-sterile corn stover. LA titer, yield, and average productivity reached 97.59 g/L, 0.68 g/g stover, and 1.63 g/L/h, respectively. This study presents a feasible process for lignocellulosic LA production from abundant agro stovers. Copyright © 2015 Elsevier Ltd. All rights reserved.

Comparative performances of microbial capacitive deionization cell and microbial fuel cell fed with produced water from the Bakken shale.

PubMed

Shrestha, Namita; Chilkoor, Govinda; Wilder, Joseph; Ren, Zhiyong Jason; Gadhamshetty, Venkataramana

2018-06-01

This study evaluates and compares the performance of microbial fuel cells (MFCs) and microbial capacitive deionization cells (MCDCs) fed with wastewater produced from the Bakken shale. The produced water was characterized by high levels of dissolved solids and chemical oxygen demand (COD). Two-compartment MFCs and three-compartment MCDCs were evaluated under batch-fed mode using mixed microbial consortia in the anode, ferricyanide in the cathode, and produced water as the electrolyte in the anode and capacitive deionization units. COD removal in the MFCs was 88%, while that in the MCDCs was limited to 76%. The lower performance of the MCDCs was due to the large impedance (6600 Ω cm 2 ) compared with the MFCs (870 Ω cm 2 ). However, the MCDCs achieved two-fold higher removal of dissolved solids. Both the MFCs and MCDCs suffered from a higher impedance induced by fouling in the latter stages of the operation. Copyright © 2018 Elsevier B.V. All rights reserved.

Enhancement of n-butanol production by in situ butanol removal using permeating-heating-gas stripping in acetone-butanol-ethanol fermentation.

PubMed

Chen, Yong; Ren, Hengfei; Liu, Dong; Zhao, Ting; Shi, Xinchi; Cheng, Hao; Zhao, Nan; Li, Zhenjian; Li, Bingbing; Niu, Huanqing; Zhuang, Wei; Xie, Jingjing; Chen, Xiaochun; Wu, Jinglan; Ying, Hanjie

2014-07-01

Butanol recovery from acetone-butanol-ethanol (ABE) fed-batch fermentation using permeating-heating-gas was determined in this study. Fermentation was performed with Clostridium acetobutylicum B3 in a fibrous bed bioreactor and permeating-heating-gas stripping was used to eliminate substrate and product inhibition, which normally restrict ABE production and sugar utilization to below 20 g/L and 60 g/L, respectively. In batch fermentation (without permeating-heating-gas stripping), C. acetobutylicum B3 utilized 60 g/L glucose and produced 19.9 g/L ABE and 12 g/L butanol, while in the integrated process 290 g/L glucose was utilized and 106.27 g/L ABE and 66.09 g/L butanol were produced. The intermittent gas stripping process generated a highly concentrated condensate containing approximately 15% (w/v) butanol, 4% (w/v) acetone, a small amount of ethanol (<1%), and almost no acids, resulting in a highly concentrated butanol solution [∼ 70% (w/v)] after phase separation. Butanol removal by permeating-heating-gas stripping has potential for commercial ABE production. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

Operational strategies, monitoring and control of heterologous protein production in the methylotrophic yeast Pichia pastoris under different promoters: A review

PubMed Central

Cos, Oriol; Ramón, Ramón; Montesinos, José Luis; Valero, Francisco

2006-01-01

The methylotrophic yeast Pichia pastoris has been widely reported as a suitable expression system for heterologous protein production. The use of different phenotypes under PAOX promoter, other alternative promoters, culture medium, and operational strategies with the objective to maximize either yield or productivity of the heterologous protein, but also to obtain a repetitive product batch to batch to get a robust process for the final industrial application have been reported. Medium composition, kinetics growth, fermentation operational strategies from fed-batch to continuous cultures using different phenotypes with the most common PAOX promoter and other novel promoters (GAP, FLD, ICL), the use of mixed substrates, on-line monitoring of the key fermentation parameters (methanol) and control algorithms applied to the bioprocess are reviewed and discussed in detail. PMID:16600031

Production of exopolysaccharides by Acinetobacter strains in a controlled fed-batch fermentation process using soap stock oil (SSO) as carbon source.

PubMed

Shabtai, Y

1990-04-01

The production of two extracellular capsular heteropolysaccharides by two different Acinetobacter strains has been studied in separate controlled fermentation processes with a view to their industrial applications as specific dispersing agents. The first, emulsan, is an extracellular polyanionic amphipathic heteropolysaccharide (MW 10(6) D) made by A. calcoaceticus RAG-1. It forms and stabilizes oil in water emulsions. The other, biodispersan (PS-A2), is another extracellular zwitterionic heteropolysaccharide (MW 51 kD) made by A. calcoaceticus A2. This polysaccharide disperses big solid limestone granules forming micron-size water suspension. Both polysaccharides are synthesized within the cells, exported to their outer surface to form an extracellular cell-associated capsule and released subsequently into the growth medium. The polymers were produced in a computer-controlled fed-batch intensively aerated fermentation process. A commercially available and cheap fatty acids mixture (soap stock oil) served as the carbon source, and was fed in coordination with the required nitrogen. The coordinated feed of carbon and nitrogen was operated on the basis of two metabolic correlations: The first correlation related the cell protein produced and the ammonium nitrogen consumed with the outcoming coeffients of 24 and 21 mM NH3/g protein for the emulsan and the biodispersan fermentations respectively. The second correlation linked the consumption of the fatty acids with that of the nitrogen source dictating the appropriate C/N ratio of the feed into the operating fermentor. These ratios were 7.7 g C/g N for the emulsan fermentation and 8.5 gC/g N in the case of the biodispersan production process.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparison of Different Strategies for Selection/Adaptation of Mixed Microbial Cultures Able to Ferment Crude Glycerol Derived from Second-Generation Biodiesel.

PubMed

Varrone, C; Heggeset, T M B; Le, S B; Haugen, T; Markussen, S; Skiadas, I V; Gavala, H N

2015-01-01

Objective of this study was the selection and adaptation of mixed microbial cultures (MMCs), able to ferment crude glycerol generated from animal fat-based biodiesel and produce building-blocks and green chemicals. Various adaptation strategies have been investigated for the enrichment of suitable and stable MMC, trying to overcome inhibition problems and enhance substrate degradation efficiency, as well as generation of soluble fermentation products. Repeated transfers in small batches and fed-batch conditions have been applied, comparing the use of different inoculum, growth media, and Kinetic Control. The adaptation of activated sludge inoculum was performed successfully and continued unhindered for several months. The best results showed a substrate degradation efficiency of almost 100% (about 10 g/L glycerol in 21 h) and different dominant metabolic products were obtained, depending on the selection strategy (mainly 1,3-propanediol, ethanol, or butyrate). On the other hand, anaerobic sludge exhibited inactivation after a few transfers. To circumvent this problem, fed-batch mode was used as an alternative adaptation strategy, which led to effective substrate degradation and high 1,3-propanediol and butyrate production. Changes in microbial composition were monitored by means of Next Generation Sequencing, revealing a dominance of glycerol consuming species, such as Clostridium, Klebsiella, and Escherichia.

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.

Batch and fed-batch cultivation for excretive production of human epidermal growth factor (hEGF) with recombinant E. Coli K12 system.

PubMed

Wang, J; Chen, J; Xu, R; Xu, Z

2008-01-01

Batch and fed-batch production of recombinant human epidermal growth factor (hEGF) was studied in an E. coli secretary expression system. By using MMBL medium containing 5 g/L glucose, controlling the temperature at 32 degrees C and maintaining the dissolved oxgen level over 20% saturation, a high yield of hEGF (32 mg/L) was obtained after an 18 hr batch cultivation with 0.2 mM IPTG induction at mid-log phase. Three different glucose feeding strategies were employed to further improve hEGF productivity in a bench top fermentor. Compared with the batch results, hEGF yield was improved up to 25.5% or 28.1%, respectively by intermittent or pH-stat glucose feeding, and up to 150% improvement of hEGF production was achieved by constant feeding of 200 g/L glucose solution at a rate of 0.11 mL/min. The effects of further combined feeding with other medium components and inducer on hEGF yield were also examined in the benchtop fermentor. This work is very helpful to further improve the productivity of extracellular hEGF in the recombinant E. coli system.

Generally applicable fed-batch culture concept based on the detection of metabolic state by on-line balancing.

PubMed

Jobé, Anna Marya; Herwig, Christoph; Surzyn, Martin; Walker, Bernhard; Marison, Ian; von Stockar, Urs

2003-06-20

In many microorganisms, flux limitations in oxidative metabolism lead to the formation of overflow metabolites even under fully aerobic conditions. This can be avoided if the specific growth rate is controlled at a low enough value. This is usually accomplished by controlling the substrate feeding profile in a fed-batch process. The present work proposes a control concept which is based on the on-line detection of metabolic state by on-line calculation of mass and elemental balances. The advantages of this method are: 1) the check of measurement consistency based on all of the available measurements, 2) the minimum requirement of a priori knowledge of metabolism, and 3) the exclusive use of simple and established on-line techniques which do not require direct measurement of the metabolite in question. The control concept has been linked to a simple adaptive controller and applied to fed-batch cultures of S. cerevisiae and E. coli, organisms which express different overflow metabolites, ethanol and acetic acid, respectively. Oxidative and oxidoreductive states of S. cerevisiae and E. coli cultures were detected with high precision. As demonstrated by the formation of acetic acid in E. coli cultures, metabolic states could be correctly distinguished for systems for which traditional methods, such as respiratory quotient (RQ), are insensitive. Hence, it could be shown that the control concept allowed avoidance of overflow metabolite formation and operation at maximum oxidative biomass productivity and oxidative conversion of substrate into biomass. Based on mass and elemental balances, the proposed method additionally provides a richness of additional information, such as yield coefficients and estimation of concentrations and specific conversion rates. These data certainly help the operator to additionally evaluate the state of the process on-line. Copyright 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 82: 627-639, 2003.

The impact of pH inhomogeneities on CHO cell physiology and fed-batch process performance - two-compartment scale-down modelling and intracellular pH excursion.

PubMed

Brunner, Matthias; Braun, Philipp; Doppler, Philipp; Posch, Christoph; Behrens, Dirk; Herwig, Christoph; Fricke, Jens

2017-07-01

Due to high mixing times and base addition from top of the vessel, pH inhomogeneities are most likely to occur during large-scale mammalian processes. The goal of this study was to set-up a scale-down model of a 10-12 m 3 stirred tank bioreactor and to investigate the effect of pH perturbations on CHO cell physiology and process performance. Short-term changes in extracellular pH are hypothesized to affect intracellular pH and thus cell physiology. Therefore, batch fermentations, including pH shifts to 9.0 and 7.8, in regular one-compartment systems are conducted. The short-term adaption of the cells intracellular pH are showed an immediate increase due to elevated extracellular pH. With this basis of fundamental knowledge, a two-compartment system is established which is capable of simulating defined pH inhomogeneities. In contrast to state-of-the-art literature, the scale-down model is included parameters (e.g. volume of the inhomogeneous zone) as they might occur during large-scale processes. pH inhomogeneity studies in the two-compartment system are performed with simulation of temporary pH zones of pH 9.0. The specific growth rate especially during the exponential growth phase is strongly affected resulting in a decreased maximum viable cell density and final product titer. The gathered results indicate that even short-term exposure of cells to elevated pH values during large-scale processes can affect cell physiology and overall process performance. In particular, it could be shown for the first time that pH perturbations, which might occur during the early process phase, have to be considered in scale-down models of mammalian processes. Copyright © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Heterotrophic growth and lipid accumulation of Chlorella protothecoides in whey permeate, a dairy by-product stream, for biofuel production.

PubMed

Espinosa-Gonzalez, Isabel; Parashar, Archana; Bressler, David C

2014-03-01

This study proposes a novel alternative for the utilization of whey permeate, a by-product stream from the dairy industry, as the feedstock for the biomass and lipid production of the microalgae Chlorella protothecoides. Glucose and galactose from the pre-hydrolyzed whey permeate were used as main carbon sources in a base mineral media for establishing batch and fed batch cultures. Batch cultures reached a biomass production of 9.1±0.2g/L with a total lipid accumulation of 42.0±6.6% (dry weight basis), while in the fed batch cultures 17.2±1.3g/L of biomass with 20.5±0.3% lipid accumulation (dry weight basis) were obtained. A third strategy for the direct utilization of whey permeate was investigated by simultaneous saccharification and fermentation (SSF), wherein, 7.3±1.3g/L of biomass with 49.9±3.3% lipid accumulation (dry weight basis) was obtained in batch mode using immobilized enzyme. Copyright © 2013 Elsevier Ltd. All rights reserved.

Ultrasound pre-treatment for anaerobic digestion improvement.

PubMed

Pérez-Elvira, S; Fdz-Polanco, M; Plaza, F I; Garralón, G; Fdz-Polanco, F

2009-01-01

Prior research indicates that ultrasounds can be used in batch reactors as pre-treatment before anaerobic digestion, but the specific energy required at laboratory-scale is too high. This work evaluates both the continuous ultrasound device performance (efficiency and solubilisation) and the operation of anaerobic digesters continuously fed with sonicated sludge, and presents energy balance considerations. The results of sludge solubilisation after the sonication treatment indicate that, applying identical specific energy, it is better to increase the power than the residence time. Working with secondary sludge, batch biodegradability tests show that by applying 30 kWh/m3 of sludge, it is possible to increase biogas production by 42%. Data from continuous pilot-scale anaerobic reactors (V=100 L) indicate that operating with a conventional HRT=20 d, a reactor fed with pre-treated sludge increases the volatile solids removal and the biogas production by 25 and 37% respectively. Operating with HRT=15 d, the removal efficiency is similar to the obtained with a reactor fed with non-hydrolysed sludge at HTR=20 d, although the specific biogas productivity per volume of reactor is higher for the pretreated sludge. Regarding the energy balance, although for laboratory-scale devices it is negative, full-scale suppliers state a net generation of 3-10 kW per kW of energy used.

A comparison of the energy use of in situ product recovery techniques for the Acetone Butanol Ethanol fermentation.

PubMed

Outram, Victoria; Lalander, Carl-Axel; Lee, Jonathan G M; Davis, E Timothy; Harvey, Adam P

2016-11-01

The productivity of the Acetone Butanol Ethanol (ABE) fermentation can be significantly increased by application of various in situ product recovery (ISPR) techniques. There are numerous technically viable processes, but it is not clear which is the most economically viable in practice. There is little available information about the energy requirements and economics of ISPR for the ABE fermentation. This work compares various ISPR techniques based on UniSim process simulations of the ABE fermentation. The simulations provide information on the process energy and separation efficiency, which is fed into an economic assessment. Perstraction was the only technique to reduce the energy demand below that of a batch process, by approximately 5%. Perstraction also had the highest profit increase over a batch process, by 175%. However, perstraction is an immature technology, so would need significant development before being integrated to an industrial process. Copyright © 2016 Elsevier Ltd. All rights reserved.

Acetone-butanol-ethanol from sweet sorghum juice by an immobilized fermentation-gas stripping integration process.

PubMed

Cai, Di; Wang, Yong; Chen, Changjing; Qin, Peiyong; Miao, Qi; Zhang, Changwei; Li, Ping; Tan, Tianwei

2016-07-01

In this study, sweet sorghum juice (SSJ) was used as the substrate in a simplified ABE fermentation-gas stripping integration process without nutrients supplementation. The sweet sorghum bagasse (SSB) after squeezing the fermentable juice was used as the immobilized carrier. The results indicated that the productivity of ABE fermentation process was improved by gas stripping integration. A total 24g/L of ABE solvents was obtained from 59.6g/L of initial sugar after 80h of fermentation with gas stripping. Then, long-term of fed-batch fermentation with continuous gas stripping was further performed. 112.9g/L of butanol, 44.1g/L of acetone, 9.5g/L of ethanol (total 166.5g/L of ABE) was produced in overall 312h of fermentation. At the same time, concentrated ABE product was obtained in the condensate of gas stripping. Copyright © 2016 Elsevier Ltd. All rights reserved.

Combined substrate, enzyme and yeast feed in simultaneous saccharification and fermentation allow bioethanol production from pretreated spruce biomass at high solids loadings

PubMed Central

2014-01-01

Background Economically feasible cellulosic ethanol production requires that the process can be operated at high solid loadings, which currently imparts technical challenges including inefficient mixing leading to heat and mass transfer limitations and high concentrations of inhibitory compounds hindering microbial activity during simultaneous saccharification and fermentation (SSF) process. Consequently, there is a need to develop cost effective processes overcoming the challenges when working at high solid loadings. Results In this study we have modified the yeast cultivation procedure and designed a SSF process to address some of the challenges at high water insoluble solids (WIS) content. The slurry of non-detoxified pretreated spruce when used in a batch SSF at 19% (w/w) WIS was found to be inhibitory to Saccharomyces cerevisiae Thermosacc that produced 2 g l-1 of ethanol. In order to reduce the inhibitory effect, the non-washed solid fraction containing reduced amount of inhibitors compared to the slurry was used in the SSF. Further, the cells were cultivated in the liquid fraction of pretreated spruce in a continuous culture wherein the outflow of cell suspension was used as cell feed to the SSF reactor in order to maintain the metabolic state of the cell. Enhanced cell viability was observed with cell, enzyme and substrate feed in a SSF producing 40 g l-1 ethanol after 96 h corresponding to 53% of theoretical yield based on available hexose sugars compared to 28 g l-1 ethanol in SSF with enzyme and substrate feed but no cell feed resulting in 37% of theoretical yield at a high solids loading of 20% (w/w) WIS content. The fed-batch SSF also significantly eased the mixing, which is usually challenging in batch SSF at high solids loading. Conclusions A simple modification of the cell cultivation procedure together with a combination of yeast, enzyme and substrate feed in a fed-batch SSF process, made it possible to operate at high solids loadings in a conventional bioreactor. The proposed process strategy significantly increased the yeast cell viability and overall ethanol yield. It was also possible to obtain 4% (w/v) ethanol concentration, which is a minimum requirement for an economical distillation process. PMID:24713027

Combined substrate, enzyme and yeast feed in simultaneous saccharification and fermentation allow bioethanol production from pretreated spruce biomass at high solids loadings.

PubMed

Koppram, Rakesh; Olsson, Lisbeth

2014-04-08

Economically feasible cellulosic ethanol production requires that the process can be operated at high solid loadings, which currently imparts technical challenges including inefficient mixing leading to heat and mass transfer limitations and high concentrations of inhibitory compounds hindering microbial activity during simultaneous saccharification and fermentation (SSF) process. Consequently, there is a need to develop cost effective processes overcoming the challenges when working at high solid loadings. In this study we have modified the yeast cultivation procedure and designed a SSF process to address some of the challenges at high water insoluble solids (WIS) content. The slurry of non-detoxified pretreated spruce when used in a batch SSF at 19% (w/w) WIS was found to be inhibitory to Saccharomyces cerevisiae Thermosacc that produced 2 g l-1 of ethanol. In order to reduce the inhibitory effect, the non-washed solid fraction containing reduced amount of inhibitors compared to the slurry was used in the SSF. Further, the cells were cultivated in the liquid fraction of pretreated spruce in a continuous culture wherein the outflow of cell suspension was used as cell feed to the SSF reactor in order to maintain the metabolic state of the cell. Enhanced cell viability was observed with cell, enzyme and substrate feed in a SSF producing 40 g l-1 ethanol after 96 h corresponding to 53% of theoretical yield based on available hexose sugars compared to 28 g l-1 ethanol in SSF with enzyme and substrate feed but no cell feed resulting in 37% of theoretical yield at a high solids loading of 20% (w/w) WIS content. The fed-batch SSF also significantly eased the mixing, which is usually challenging in batch SSF at high solids loading. A simple modification of the cell cultivation procedure together with a combination of yeast, enzyme and substrate feed in a fed-batch SSF process, made it possible to operate at high solids loadings in a conventional bioreactor. The proposed process strategy significantly increased the yeast cell viability and overall ethanol yield. It was also possible to obtain 4% (w/v) ethanol concentration, which is a minimum requirement for an economical distillation process.

Direct analysis in real time mass spectrometry, a process analytical technology tool for real-time process monitoring in botanical drug manufacturing.

PubMed

Wang, Lu; Zeng, Shanshan; Chen, Teng; Qu, Haibin

2014-03-01

A promising process analytical technology (PAT) tool has been introduced for batch processes monitoring. Direct analysis in real time mass spectrometry (DART-MS), a means of rapid fingerprint analysis, was applied to a percolation process with multi-constituent substances for an anti-cancer botanical preparation. Fifteen batches were carried out, including ten normal operations and five abnormal batches with artificial variations. The obtained multivariate data were analyzed by a multi-way partial least squares (MPLS) model. Control trajectories were derived from eight normal batches, and the qualification was tested by R(2) and Q(2). Accuracy and diagnosis capability of the batch model were then validated by the remaining batches. Assisted with high performance liquid chromatography (HPLC) determination, process faults were explained by corresponding variable contributions. Furthermore, a batch level model was developed to compare and assess the model performance. The present study has demonstrated that DART-MS is very promising in process monitoring in botanical manufacturing. Compared with general PAT tools, DART-MS offers a particular account on effective compositions and can be potentially used to improve batch quality and process consistency of samples in complex matrices. Copyright © 2014 Elsevier B.V. All rights reserved.

Enhancement of ε-poly-L-lysine synthesis in Streptomyces by exogenous glutathione.

PubMed

Yan, Peng; Sun, Haoben; Lu, Pengqi; Liu, Haili; Tang, Lei

2018-01-01

Our previous work indicated that the vigor of Streptomyces decreased at the later stage of ε-poly-L-lysine (ε-PL) fermentation. In this study, we observed that the level of reactive oxygen species (ROS) in vivo increased sharply after 24 h, and the addition of an antioxidant glutathione (GSH) before this increase in ROS stimulated ε-PL synthesis in shake-flask fermentation. The enhancement of ε-PL production by GSH was further verified in batch and fed-batch fermentations. On a 5-l fermenter scale, the highest increasement was 68.8% in batch fermentation and the highest ε-PL level was 46.5 g l - 1 in fed-batch fermentation. The RT-qPCR analysis showed that the transcriptional level of the catalase gene was down-regulated, and the decrease in cell activity was alleviated by the addition of GSH. The results revealed that exogenous antioxidant might maintain the cell vigor by reducing the excess ROS which provided a novel approach to regulate ε-PL synthesis.

Optimization of the yield of dark microaerobic production of hydrogen from lactate by Rhodopseudomonas palustris.

PubMed

Lazaro, Carolina Zampol; Hitit, Zeynep Yilmazer; Hallenbeck, Patrick C

2017-12-01

Hydrogen yields of dark fermentation are limited due to the need to also produce reduced side products, and photofermentation, an alternative, is limited by the need for light. A relatively new strategy, dark microaerobic fermentation, could potentially overcome both these constraints. Here, application of this strategy demonstrated for the first time significant hydrogen production from lactate by a single organism in the dark. Response surface methodology (RSM) was used to optimize substrate and oxygen concentration as well as inoculum using both (1) regular batch and (2) O 2 fed batch cultures. The highest hydrogen yield (HY) was observed under regular batch (1.4±0.1molH 2 /mollactate) and the highest hydrogen production (HP) (173.5µmolH 2 ) was achieved using O 2 fed batch. This study has provided proof of principal for the ability of microaerobic fermentation to drive thermodynamically difficult reactions, such as the conversion of lactate to hydrogen. Copyright © 2017 Elsevier Ltd. All rights reserved.

Plasmid fermentation process for DNA immunization applications.

PubMed

Carnes, Aaron E; Williams, James A

2014-01-01

Plasmid DNA for immunization applications must be of the highest purity and quality. The ability of downstream purification to efficiently produce a pure final product is directly influenced by the performance of the upstream fermentation process. While several clinical manufacturing facilities already have validated fermentation processes in place to manufacture plasmid DNA for use in humans, a simple and inexpensive laboratory-scale fermentation process can be valuable for in-house production of plasmid DNA for use in animal efficacy studies. This chapter describes a simple fed-batch fermentation process for producing bacterial cell paste enriched with high-quality plasmid DNA. A constant feeding strategy results in a medium cell density culture with continuously increasing plasmid amplification towards the end of the process. Cell banking and seed culture preparation protocols, which can dramatically influence final product yield and quality, are also described. These protocols are suitable for production of research-grade plasmid DNA at the 100 mg-to-1.5 g scale from a typical 10 L laboratory benchtop fermentor.

Benzoate-induced stress enhances xylitol yield in aerobic fed-batch culture of Candida mogii TISTR 5892.

PubMed

Wannawilai, Siwaporn; Sirisansaneeyakul, Sarote; Chisti, Yusuf

2015-01-20

Production of the natural sweetener xylitol from xylose via the yeast Candida mogii TISTR 5892 was compared with and without the growth inhibitor sodium benzoate in the culture medium. Sodium benzoate proved to be an uncompetitive inhibitor in relatively poorly oxygenated shake flask aerobic cultures. In a better controlled aerobic environment of a bioreactor, the role of sodium benzoate could equally well be described as competitive, uncompetitive or noncompetitive inhibitor of growth. In intermittent fed-batch fermentations under highly aerobic conditions, the presence of sodium benzoate at 0.15gL(-1) clearly enhanced the xylitol titer relative to the control culture without the sodium benzoate. The final xylitol concentration and the average xylitol yield on xylose were nearly 50gL(-1) and 0.57gg(-1), respectively, in the presence of sodium benzoate. Both these values were substantially higher than reported for the same fermentation under microaerobic conditions. Therefore, a fed-batch aerobic fermentation in the presence of sodium benzoate is promising for xylitol production using C. mogii. Copyright © 2014 Elsevier B.V. All rights reserved.

Fed-batch production of green coconut hydrolysates for high-gravity second-generation bioethanol fermentation with cellulosic yeast.

PubMed

Soares, Jimmy; Demeke, Mekonnen M; Van de Velde, Miet; Foulquié-Moreno, Maria R; Kerstens, Dorien; Sels, Bert F; Verplaetse, Alex; Fernandes, Antonio Alberto Ribeiro; Thevelein, Johan M; Fernandes, Patricia Machado Bueno

2017-11-01

The residual biomass obtained from the production of Cocos nucifera L. (coconut) is a potential source of feedstock for bioethanol production. Even though coconut hydrolysates for ethanol production have previously been obtained, high-solid loads to obtain high sugar and ethanol levels remain a challenge. We investigated the use of a fed-batch regime in the production of sugar-rich hydrolysates from the green coconut fruit and its mesocarp. Fermentation of the hydrolysates obtained from green coconut or its mesocarp, containing 8.4 and 9.7% (w/v) sugar, resulted in 3.8 and 4.3% (v/v) ethanol, respectively. However, green coconut hydrolysate showed a prolonged fermentation lag phase. The inhibitor profile suggested that fatty acids and acetic acid were the main fermentation inhibitors. Therefore, a fed-batch regime with mild alkaline pretreatment followed by saccharification, is presented as a strategy for fermentation of such challenging biomass hydrolysates, even though further improvement of yeast inhibitor tolerance is also needed. Copyright © 2017 Elsevier Ltd. All rights reserved.

Sucrose fed-batch strategy enhanced biomass, polysaccharide, and ganoderic acids production in fermentation of Ganoderma lucidum 5.26.

PubMed

Wei, Zhen-hua; Liu, Lianliang; Guo, Xiao-feng; Li, Yan-jun; Hou, Bao-chao; Fan, Qiu-ling; Wang, Kai-xiang; Luo, Yingdi; Zhong, Jian-jiang

2016-01-01

Ganoderma, as a Chinese traditional medicine, has multiple bioactivities. However, industrial production was limited due to low yield during Ganoderma fermentation. In this work, sucrose was found to greatly enhance intracellular polysaccharide (IPS) content and specific extracellular polysaccharide (EPS) production rate. The mechanism was studied by analyzing the activities of enzymes related to polysaccharide biosynthesis. The results revealed that sucrose regulated the activities of phosphoglucomutase and phosphoglucose isomerase. When glucose and sucrose mixture was used as carbon source, biomass, polysaccharide and ganoderic acids (GAs) production was greatly enhanced. A sucrose fed-batch strategy was developed in 10-L bioreactor, and was scaled up to 300-L bioreactor. The biomass, EPS and IPS production was 25.5, 2.9 and 4.8 g/L, respectively, which was the highest biomass and IPS production in pilot scale. This study provides information for further understanding the regulation mechanism of Ganoderma polysaccharide biosynthesis. It demonstrates that sucrose fed-batch is a useful strategy for enhancing Ganoderma biomass, polysaccharide and GAs production.

Efficient production of butyric acid from Jerusalem artichoke by immobilized Clostridium tyrobutyricum in a fibrous-bed bioreactor.

PubMed

Huang, Jin; Cai, Jin; Wang, Jin; Zhu, Xiangcheng; Huang, Lei; Yang, Shang-Tian; Xu, Zhinan

2011-02-01

Butyric acid is an important specialty chemical with wide industrial applications. The feasible large-scale fermentation for the economical production of butyric acid requires low-cost substrate and efficient process. In the present study, butyric acid production by immobilized Clostridium tyrobutyricum was successfully performed in a fibrous-bed bioreactor using Jerusalem artichoke as the substrate. Repeated-batch fermentation was carried out to produce butyric acid with a high butyrate yield (0.44 g/g), high productivity (2.75 g/L/h) and a butyrate concentration of 27.5 g/L. Furthermore, fed-batch fermentation using sulfuric acid pretreated Jerusalem artichoke hydrolysate resulted in a high butyric acid concentration of 60.4 g/L, with the yield of 0.38 g/g and the selectivity of ∼ 85.1 (85.1g butyric acid/g acetic acid). Thus, the production of butyric acid from Jerusalem artichoke on a commercial scale could be achieved based on the system developed in this work. Copyright © 2010. Published by Elsevier Ltd.

Fed-batch control based upon the measurement of intracellular NADH

NASA Technical Reports Server (NTRS)

Armiger, W. B.; Lee, J. F.; Montalvo, L. M.; Forro, J. R.

1987-01-01

A series of experiments demonstrating that on-line measurements of intracellular NADH by culture fluorescence can be used to monitor and control the fermentation process are described. A distinct advantage of intercellular NADH measurements over other monitoring techniques such as pH and dissolved oxygen is that it directly measures real time events occurring within the cell rather than changes in the environment. When coupled with other measurement parameters, it can provide a finer degree of sophistication in process control.

Effective alkaline metal-catalyzed oxidative delignification of hybrid poplar

DOE PAGES

Bhalla, Aditya; Bansal, Namita; Stoklosa, Ryan J.; ...

2016-02-09

Background: Strategies to improve copper-catalyzed alkaline hydrogen peroxide (Cu-AHP) pretreatment of hybrid poplar were investigated. These improvements included a combination of increasing hydrolysis yields, while simultaneously decreasing process inputs through (i) more efficient utilization of H 2O 2 and (ii) the addition of an alkaline extraction step prior to the metal-catalyzed AHP pretreatment. We hypothesized that utilizing this improved process could substantially lower the chemical inputs needed during pretreatment. Results: Hybrid poplar was pretreated utilizing a modified process in which an alkaline extraction step was incorporated prior to the Cu-AHP treatment step and H 2O 2 was added batch-wise overmore » the course of 10 h. Our results revealed that the alkaline pre-extraction step improved both lignin and xylan solubilization, which ultimately led to improved glucose (86 %) and xylose (95 %) yields following enzymatic hydrolysis. An increase in the lignin solubilization was also observed with fed-batch H 2O 2 addition relative to batch-only addition, which again resulted in increased glucose and xylose yields (77 and 93 % versus 63 and 74 %, respectively). Importantly, combining these strategies led to significantly improved sugar yields (96 % glucose and 94 % xylose) following enzymatic hydrolysis. In addition, we found that we could substantially lower the chemical inputs (enzyme, H 2O 2, and catalyst), while still maintaining high product yields utilizing the improved Cu-AHP process. This pretreatment also provided a relatively pure lignin stream consisting of ≥90 % Klason lignin and only 3 % xylan and 2 % ash following precipitation. Two-dimensional heteronuclear single-quantum coherence (2D HSQC) NMR and size-exclusion chromatography demonstrated that the solubilized lignin was high molecular weight (Mw ≈ 22,000 Da) and only slightly oxidized relative to lignin from untreated poplar. In conclusion: This study demonstrated that the fed-batch, two-stage Cu-AHP pretreatment process was effective in pretreating hybrid poplar for its conversion into fermentable sugars. Results showed sugar yields near the theoretical maximum were achieved from enzymatically hydrolyzed hybrid poplar by incorporating an alkaline extraction step prior to pretreatment and by efficiently utilizing H 2O 2 during the Cu-AHP process. Significantly, this study reports high sugar yields from woody biomass treated with an AHP pretreatment under mild reaction conditions.« less

Enhanced butyric acid tolerance and bioproduction by Clostridium tyrobutyricum immobilized in a fibrous bed bioreactor.

PubMed

Jiang, Ling; Wang, Jufang; Liang, Shizhong; Cai, Jin; Xu, Zhinan; Cen, Peilin; Yang, Shangtian; Li, Shuang

2011-01-01

Repeated fed-batch fermentation of glucose by Clostridium tyrobutyricum immobilized in a fibrous bed bioreactor (FBB) was successfully employed to produce butyric acid at a high final concentration as well as to adapt a butyric-acid-tolerant strain. At the end of the eighth fed-batch fermentation, the butyric acid concentration reached 86.9 ± 2.17 g/L, which to our knowledge is the highest butyric acid concentration ever produced in the traditional fermentation process. To understand the mechanism and factors contributing to the improved butyric acid production and enhanced acid tolerance, adapted strains were harvested from the FBB and characterized for their physiological properties, including specific growth rate, acid-forming enzymes, intracellular pH, membrane-bound ATPase and cell morphology. Compared with the original culture used to seed the bioreactor, the adapted culture showed significantly reduced inhibition effects of butyric acid on specific growth rate, cellular activities of butyric-acid-forming enzyme phosphotransbutyrylase (PTB) and ATPase, together with elevated intracellular pH, and elongated rod morphology. © 2010 Wiley Periodicals, Inc.

Life-cycle and cost of goods assessment of fed-batch and perfusion-based manufacturing processes for mAbs.

PubMed

Bunnak, Phumthep; Allmendinger, Richard; Ramasamy, Sri V; Lettieri, Paola; Titchener-Hooker, Nigel J

2016-09-01

Life-cycle assessment (LCA) is an environmental assessment tool that quantifies the environmental impact associated with a product or a process (e.g., water consumption, energy requirements, and solid waste generation). While LCA is a standard approach in many commercial industries, its application has not been exploited widely in the bioprocessing sector. To contribute toward the design of more cost-efficient, robust and environmentally-friendly manufacturing process for monoclonal antibodies (mAbs), a framework consisting of an LCA and economic analysis combined with a sensitivity analysis of manufacturing process parameters and a production scale-up study is presented. The efficiency of the framework is demonstrated using a comparative study of the two most commonly used upstream configurations for mAb manufacture, namely fed-batch (FB) and perfusion-based processes. Results obtained by the framework are presented using a range of visualization tools, and indicate that a standard perfusion process (with a pooling duration of 4 days) has similar cost of goods than a FB process but a larger environmental footprint because it consumed 35% more water, demanded 17% more energy, and emitted 17% more CO 2 than the FB process. Water consumption was the most important impact category, especially when scaling-up the processes, as energy was required to produce process water and water-for-injection, while CO 2 was emitted from energy generation. The sensitivity analysis revealed that the perfusion process can be made more environmentally-friendly than the FB process if the pooling duration is extended to 8 days. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1324-1335, 2016. © 2016 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers.

Effect of cycle time on polyhydroxybutyrate (PHB) production in aerobic mixed cultures.

PubMed

Ozdemir, Sebnem; Akman, Dilek; Cirik, Kevser; Cinar, Ozer

2014-03-01

The aim of this study was to investigate the effect of cycle time on polyhydroxybutyrate (PHB) production under aerobic dynamic feeding system. The acetate-fed feast and famine sequencing batch reactor was used to enrich PHB accumulating microorganism. Sequencing batch reactor (SBR) was operated in four different cycle times (12, 8, 4, and 2 h) fed with a synthetic wastewater. The system performance was determined by monitoring total dissolved organic carbon, dissolved oxygen, oxidation-reduction potential, and PHB concentration. In this study, under steady-state conditions, the feast period of the SBR was found to allow the PHB storage while a certain part of stored PHB was used for continued growth in famine period. The percentage PHB storages by aerobic microorganism were at 16, 18, 42, and 55% for the 12, 8, 4, and 2-h cycle times, respectively. The PHB storage was increased as the length of the cycle time was decreased, and the ratio of the feast compared to the total cycle length was increased from around 13 to 33% for the 12 and 2-h cycle times, respectively.

Enhanced kefiran production by mixed culture of Lactobacillus kefiranofaciens and Saccharomyces cerevisiae.

PubMed

Cheirsilp, Benjamas; Shimizu, Hiroshi; Shioya, Suteaki

2003-01-09

In a batch mixed culture of Lactobacillus kefiranofaciens and Saccharomyces cerevisiae, which could assimilate lactic acid, cell growth and kefiran production rates of L. kefiranofaciens significantly increased, compared with those in pure cultures. The kefiran production rate was 36 mg l(-1) h(-1) in the mixed culture under the anaerobic condition, which was greater than that in the pure culture (24 mg l(-1) h(-1)). Under the aerobic condition, a more intensive interaction between these two strains was observed and higher kefiran production rate (44 mg l(-1) h(-1)) was obtained compared with that under the anaerobic condition. Kefiran production was further enhanced by an addition of fresh medium in the fed-batch mixed culture. In the fed-batch mixed culture, a final kefiran concentration of 5.41 g l(-1) was achieved at 87 h, thereby attaining the highest productivity at 62 mg l(-1) h(-1). Simulation study considered the reduction of lactic acid in pure culture was performed to estimate the additional effect of coculture with S. cerevisiae. Slightly higher cell growth and kefiran production rates in the mixed culture than those expected from pure culture by simulation were observed. These results suggest that coculture of L. kefiranofaciens and S. cerevisiae not only reduces the lactic acid concentration by consumption but also stimulates cell growth and kefiran production of L. kefiranofaciens.

Bioaugmentation for Groundwater Remediation

DTIC Science & Technology

2010-02-01

continually grown and harvested , in fed batch mode where a culture is grown in a vessel and harvested and then another culture begun, or in a hybrid of...the two whereby the culture is maintained in a fermentor until a volume of culture is harvested and then replaced with fresh medium. The primary...producing cultures for bioaugmentation is a semi- continuous process whereby the culture is maintained in the fermentor until needed and then harvested

Expression and purification of ELP-intein-tagged target proteins in high cell density E. coli fermentation.

PubMed

Fong, Baley A; Wood, David W

2010-10-19

Elastin-like polypeptides (ELPs) are useful tools that can be used to non-chromatographically purify proteins. When paired with self-cleaving inteins, they can be used as economical self-cleaving purification tags. However, ELPs and ELP-tagged target proteins have been traditionally expressed using highly enriched media in shake flask cultures, which are generally not amenable to scale-up. In this work, we describe the high cell-density expression of self-cleaving ELP-tagged targets in a supplemented minimal medium at a 2.5 liter fermentation scale, with increased yields and purity compared to traditional shake flask cultures. This demonstration of ELP expression in supplemented minimal media is juxtaposed to previous expression of ELP tags in extract-based rich media. We also describe several sets of fed-batch conditions and their impact on ELP expression and growth medium cost. By using fed batch E. coli fermentation at high cell density, ELP-intein-tagged proteins can be expressed and purified at high yield with low cost. Further, the impact of media components and fermentation design can significantly impact the overall process cost, particularly at large scale. This work thus demonstrates an important advances in the scale up of self-cleaving ELP tag-mediated processes.

Expression and purification of ELP-intein-tagged target proteins in high cell density E. coli fermentation

PubMed Central

2010-01-01

Background Elastin-like polypeptides (ELPs) are useful tools that can be used to non-chromatographically purify proteins. When paired with self-cleaving inteins, they can be used as economical self-cleaving purification tags. However, ELPs and ELP-tagged target proteins have been traditionally expressed using highly enriched media in shake flask cultures, which are generally not amenable to scale-up. Results In this work, we describe the high cell-density expression of self-cleaving ELP-tagged targets in a supplemented minimal medium at a 2.5 liter fermentation scale, with increased yields and purity compared to traditional shake flask cultures. This demonstration of ELP expression in supplemented minimal media is juxtaposed to previous expression of ELP tags in extract-based rich media. We also describe several sets of fed-batch conditions and their impact on ELP expression and growth medium cost. Conclusions By using fed batch E. coli fermentation at high cell density, ELP-intein-tagged proteins can be expressed and purified at high yield with low cost. Further, the impact of media components and fermentation design can significantly impact the overall process cost, particularly at large scale. This work thus demonstrates an important advances in the scale up of self-cleaving ELP tag-mediated processes. PMID:20959011

Environmentally Friendly Production of D(−) Lactic Acid by Sporolactobacillus nakayamae: Investigation of Fermentation Parameters and Fed-Batch Strategies

PubMed Central

Michelz Beitel, Susan; Fontes Coelho, Luciana; Sass, Daiane Cristina

2017-01-01

The interest in the production of lactic acid has increased due to its wide range of applications. In the present study, the variables that affect fermentative D(−) lactic acid production were investigated: neutralizing agents, pH, temperature, inoculum percentage, agitation, and concentration of the medium components. An experimental design was applied to determine the optimal concentrations of the medium components and fermentation was studied using different feeding strategies. High production (122.41 g/L) and productivity (3.65 g/L·h) were efficiently achieved by Sporolactobacillus nakayamae in 54 h using a multipulse fed-batch technique with an initial medium containing 35 g/L of yeast extract (byproduct of alcohol production), 60 g/L of crystallized sugar, and 7.5 mL/L of salts. The fermentation process was conducted at 35°C and pH 6.0 controlled by NaOH with a 20% volume of inoculum and agitation at 125 rpm. The production of a high optically pure concentration of D(−) lactic acid combined with an environmentally friendly NaOH-based process demonstrates that S. nakayamae is a promising strain for D(−) lactic acid production. PMID:29081803

Sludge reduction by uncoupling metabolism: SBR tests with para-nitrophenol and a commercial uncoupler.

PubMed

Zuriaga-Agustí, E; Mendoza-Roca, J A; Bes-Piá, A; Alonso-Molina, J L; Amorós-Muñoz, I

2016-11-01

Nowadays cost reduction is a very important issue in wastewater treatment plants. One way, is to minimize the sludge production. Microorganisms break down the organic matter into inorganic compounds through catabolism. Uncoupling metabolism is a method which promote catabolism reactions instead of anabolism ones, where adenosine triphosphate synthesis is inhibited. In this work, the influence of the addition of para-nitrophenol and a commercial reagent to a sequencing batch reactor (SBR) on sludge production and process performance has been analyzed. Three laboratory SBRs were operated in parallel to compare the effect of the addition of both reagents with a control reactor. SBRs were fed with synthetic wastewater and were operated with the same conditions. Results showed that sludge production was slightly reduced for the tested para-nitrophenol concentrations (20 and 25 mg/L) and for a LODOred dose of 1 mL/day. Biological process performance was not influenced and high COD removals were achieved. Copyright © 2016 Elsevier Ltd. All rights reserved.

40 CFR Table 1 to Subpart Cccc of... - Emission Limitations

Code of Federal Regulations, 2013 CFR

2013-07-01

...) National Emission Standards for Hazardous Air Pollutants: Manufacturing of Nutritional Yeast Pt. 63, Subpt... comply with the emission limitations in the following table: For each fed-batch fermenter producing yeast... duration of a batch.b. The emission limitation does not apply during the production of specialty yeast. ...

40 CFR Table 1 to Subpart Cccc of... - Emission Limitations

Code of Federal Regulations, 2014 CFR

2014-07-01

...) National Emission Standards for Hazardous Air Pollutants: Manufacturing of Nutritional Yeast Pt. 63, Subpt... comply with the emission limitations in the following table: For each fed-batch fermenter producing yeast... duration of a batch.b. The emission limitation does not apply during the production of specialty yeast. ...

40 CFR Table 1 to Subpart Cccc of... - Emission Limitations

Code of Federal Regulations, 2012 CFR

2012-07-01

...) National Emission Standards for Hazardous Air Pollutants: Manufacturing of Nutritional Yeast Pt. 63, Subpt... comply with the emission limitations in the following table: For each fed-batch fermenter producing yeast... duration of a batch.b. The emission limitation does not apply during the production of specialty yeast. ...

Multi-cycle operation of enhanced biological phosphorus removal (EBPR) with different carbon sources under high temperature.

PubMed

Shen, Nan; Chen, Yun; Zhou, Yan

2017-05-01

Many studies reported that it is challenging to apply enhanced biological phosphorus removal (EBPR) process at high temperature. Glycogen accumulating organisms (GAOs) could easily gain their dominance over poly-phosphate accumulating organisms (PAOs) when the operating temperature was in the range of 25 °C-30 °C. However, a few successful EBPR processes operated at high temperature have been reported recently. This study aimed to have an in-depth understanding on the impact of feeding strategy and carbon source types on EBPR performance in tropical climate. P-removal performance of two EBPR systems was monitored through tracking effluent quality and cyclic studies. The results confirmed that EBPR was successfully obtained and maintained at high temperature with a multi-cycle strategy. More stable performance was observed with acetate as the sole carbon source compared to propionate. Stoichiometric ratios of phosphorus and carbon transformation during both anaerobic and aerobic phases were higher at high temperature than low temperature (20±1 °C) except anaerobic PHA/C ratios within most of the sub-cycles. Furthermore, the fractions of PHA and glycogen in biomass were lower compared with one-cycle pulse feed operation. The microbial community structure was more stable in acetate-fed sequencing batch reactor (C2-SBR) than that in propionate-fed reactor (C3-SBR). Accumulibacter Clade IIC was found to be highly abundant in both reactors. Copyright © 2017 Elsevier Ltd. All rights reserved.

Artisanal cheese

USDA-ARS?s Scientific Manuscript database

Artisanal cheese, which is handmade in small batches, differs from mass-produced cheese because of the milk and procedures used. Artisanal cheese is made from the milk of pasture-fed cows, sheep, or goats instead of conventionally-fed cows, and is affected by plants eaten, stage of lactation, and s...

The Conductivity and pH Values of Dispersions of Nanospheres for Targeted Drug Delivery in the Course of Forced Equilibrium Dialysis.

PubMed

Musiał, Witold; Pluta, Janusz; Byrski, Tomasz; Valh, Julija V

2015-01-01

In the available literature, the problem of pH and conductivity in FED is evaluated separately, and limited mainly to the final purity of the synthesized polymer. In this study data from conductivity and pH measurements were evaluated in the context of the structure of the macromolecule. The aim of the study was to evaluate the conductivity and pH of dispersions of nanospheres synthesized with the use of N-isopropyl acrylamide (NIPA) as the main monomer, N,N'-methylenebisacrylamide (MBA) as the cross-linker and acrylic acid (AcA) as the anionic comonomer during the purification of dispersions via forced equilibrium dialysis (FED). Six batches of nanospheres were obtained in the process of surfactant free precipitation polymerization (SFPP) under inert nitrogen. The conductivity and pH of the dispersions of nanospheres were measured at the beginning of FED and after finishing that process. The conductivity in the systems being studied decreased significantly in the process of FED. The initial values of conductivity ranged from 736.85±8.13 μS×cm(-1) to 1048.90±67.53 μS×cm(-1) After 10 days, when the systems being assessed gained stability in terms of conductivity level, the values of conductivity were between 4.29±0.01 μS×cm(-1) and 33.56±0.04 μS×cm(-1). The pH values inreased significantly after FED. The resulting pH was between 6.92±0.07 and 8.21±0.07, while the initial values were between 3.42±0.23 μS×cm(-1) and 4.30±0.22 μS×cm(-1). Conductivity and pH measurements performed during purification via FED provide important information on the composition of the resulting nanospheres, including the functional groups embedded in the structure of the polymer in the course of the synthesis, as well as the purity of the structures. The presence of a cross-linker and acidic comonomer in the poly-N-isopropyl acrylamide (polyNIPA) macromolecule may be confirmed by both the pH and the conductivity measurements.

Enhanced recombinant factor VII expression in Chinese hamster ovary cells by optimizing signal peptides and fed-batch medium.

PubMed

Peng, Lin; Yu, Xiao; Li, Chengyuan; Cai, Yanfei; Chen, Yun; He, Yang; Yang, Jianfeng; Jin, Jian; Li, Huazhong

2016-04-01

Signal peptides play an important role in directing and efficiently transporting secretory proteins to their proper locations in the endoplasmic reticulum of mammalian cells. The aim of this study was to enhance the expression of recombinant coagulation factor VII (rFVII) in CHO cells by optimizing the signal peptides and type of fed-batch culture medium used. Five sub-clones (O2, I3, H3, G2 and M3) with different signal peptide were selected by western blot (WB) analysis and used for suspension culture. We compared rFVII expression levels of 5 sub-clones and found that the highest rFVII expression level was obtained with the IgK signal peptide instead of Ori, the native signal peptide of rFVII. The high protein expression of rFVII with signal peptide IgK was mirrored by a high transcription level during suspension culture. After analyzing culture and feed media, the combination of M4 and F4 media yielded the highest rFVII expression of 20 mg/L during a 10-day suspension culture. After analyzing cell density and cell cycle, CHO cells feeding by F4 had a similar percentage of cells in G0/G1 and a higher cell density compared to F2 and F3. This may be the reason for high rFVII expression in M4+F4. In summary, rFVII expression was successfully enhanced by optimizing the signal peptide and fed-batch medium used in CHO suspension culture. Our data may be used to improve the production of other therapeutic proteins in fed-batch culture.

A novel in situ gas stripping-pervaporation process integrated with acetone-butanol-ethanol fermentation for hyper n-butanol production.

PubMed

Xue, Chuang; Liu, Fangfang; Xu, Mengmeng; Zhao, Jingbo; Chen, Lijie; Ren, Jiangang; Bai, Fengwu; Yang, Shang-Tian

2016-01-01

Butanol is considered as an advanced biofuel, the development of which is restricted by the intensive energy consumption of product recovery. A novel two-stage gas stripping-pervaporation process integrated with acetone-butanol-ethanol (ABE) fermentation was developed for butanol recovery, with gas stripping as the first-stage and pervaporation as the second-stage using the carbon nanotubes (CNTs) filled polydimethylsiloxane (PDMS) mixed matrix membrane (MMM). Compared to batch fermentation without butanol recovery, more ABE (27.5 g/L acetone, 75.5 g/L butanol, 7.0 g/L ethanol vs. 7.9 g/L acetone, 16.2 g/L butanol, 1.4 g/L ethanol) were produced in the fed-batch fermentation, with a higher butanol productivity (0.34 g/L · h vs. 0.30 g/L · h) due to reduced butanol inhibition by butanol recovery. The first-stage gas stripping produced a condensate containing 155.6 g/L butanol (199.9 g/L ABE), which after phase separation formed an organic phase containing 610.8 g/L butanol (656.1 g/L ABE) and an aqueous phase containing 85.6 g/L butanol (129.7 g/L ABE). Fed with the aqueous phase of the condensate from first-stage gas stripping, the second-stage pervaporation using the CNTs-PDMS MMM produced a condensate containing 441.7 g/L butanol (593.2 g/L ABE), which after mixing with the organic phase from gas stripping gave a highly concentrated product containing 521.3 g/L butanol (622.9 g/L ABE). The outstanding performance of CNTs-PDMS MMM can be attributed to the hydrophobic CNTs giving an alternative route for mass transport through the inner tubes or along the smooth surface of CNTs. This gas stripping-pervaporation process with less contaminated risk is thus effective in increasing butanol production and reducing energy consumption. © 2015 Wiley Periodicals, Inc.

A Model-based B2B (Batch to Batch) Control for An Industrial Batch Polymerization Process

NASA Astrophysics Data System (ADS)

Ogawa, Morimasa

This paper describes overview of a model-based B2B (batch to batch) control for an industrial batch polymerization process. In order to control the reaction temperature precisely, several methods based on the rigorous process dynamics model are employed at all design stage of the B2B control, such as modeling and parameter estimation of the reaction kinetics which is one of the important part of the process dynamics model. The designed B2B control consists of the gain scheduled I-PD/II2-PD control (I-PD with double integral control), the feed-forward compensation at the batch start time, and the model adaptation utilizing the results of the last batch operation. Throughout the actual batch operations, the B2B control provides superior control performance compared with that of conventional control methods.

Toward intensifying design of experiments in upstream bioprocess development: An industrial Escherichia coli feasibility study.

PubMed

von Stosch, Moritz; Hamelink, Jan-Martijn; Oliveira, Rui

2016-09-01

In this study, step variations in temperature, pH, and carbon substrate feeding rate were performed within five high cell density Escherichia coli fermentations to assess whether intraexperiment step changes, can principally be used to exploit the process operation space in a design of experiment manner. A dynamic process modeling approach was adopted to determine parameter interactions. A bioreactor model was integrated with an artificial neural network that describes biomass and product formation rates as function of varied fed-batch fermentation conditions for heterologous protein production. A model reliability measure was introduced to assess in which process region the model can be expected to predict process states accurately. It was found that the model could accurately predict process states of multiple fermentations performed at fixed conditions within the determined validity domain. The results suggest that intraexperimental variations of process conditions could be used to reduce the number of experiments by a factor, which in limit would be equivalent to the number of intraexperimental variations per experiment. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1343-1352, 2016. © 2016 American Institute of Chemical Engineers.

Optimal bioprocess design through a gene regulatory network - growth kinetic hybrid model: Towards Replacing Monod kinetics.

PubMed

Tsipa, Argyro; Koutinas, Michalis; Usaku, Chonlatep; Mantalaris, Athanasios

2018-05-02

Currently, design and optimisation of biotechnological bioprocesses is performed either through exhaustive experimentation and/or with the use of empirical, unstructured growth kinetics models. Whereas, elaborate systems biology approaches have been recently explored, mixed-substrate utilisation is predominantly ignored despite its significance in enhancing bioprocess performance. Herein, bioprocess optimisation for an industrially-relevant bioremediation process involving a mixture of highly toxic substrates, m-xylene and toluene, was achieved through application of a novel experimental-modelling gene regulatory network - growth kinetic (GRN-GK) hybrid framework. The GRN model described the TOL and ortho-cleavage pathways in Pseudomonas putida mt-2 and captured the transcriptional kinetics expression patterns of the promoters. The GRN model informed the formulation of the growth kinetics model replacing the empirical and unstructured Monod kinetics. The GRN-GK framework's predictive capability and potential as a systematic optimal bioprocess design tool, was demonstrated by effectively predicting bioprocess performance, which was in agreement with experimental values, when compared to four commonly used models that deviated significantly from the experimental values. Significantly, a fed-batch biodegradation process was designed and optimised through the model-based control of TOL Pr promoter expression resulting in 61% and 60% enhanced pollutant removal and biomass formation, respectively, compared to the batch process. This provides strong evidence of model-based bioprocess optimisation at the gene level, rendering the GRN-GK framework as a novel and applicable approach to optimal bioprocess design. Finally, model analysis using global sensitivity analysis (GSA) suggests an alternative, systematic approach for model-driven strain modification for synthetic biology and metabolic engineering applications. Copyright © 2018. Published by Elsevier Inc.

Current Development in Treatment and Hydrogen Energy Conversion of Organic Solid Waste

NASA Astrophysics Data System (ADS)

Shin, Hang-Sik

2008-02-01

This manuscript summarized current developments on continuous hydrogen production technologies researched in Korea advanced institute of science and technology (KAIST). Long-term continuous pilot-scale operation of hydrogen producing processes fed with non-sterile food waste exhibited successful results. Experimental findings obtained by the optimization processes of growth environments for hydrogen producing bacteria, the development of high-rate hydrogen producing strategies, and the feasibility tests for real field application could contribute to the progress of fermentative hydrogen production technologies. Three major technologies such as controlling dilution rate depending on the progress of acidogenesis, maintaining solid retention time independently from hydraulic retention time, and decreasing hydrogen partial pressure by carbon dioxide sparging could enhance hydrogen production using anaerobic leaching beds reactors and anaerobic sequencing batch reactors. These findings could contribute to stable, reliable and effective performances of pilot-scale reactors treating organic wastes.

Production of nattokinase by high cell density fed-batch culture of Bacillus subtilis.

PubMed

Kwon, Eun-Yeong; Kim, Kyung Mi; Kim, Mi Kyoung; Lee, In Young; Kim, Beom Soo

2011-09-01

Bacillus subtilis was cultivated to high cell density for nattokinase production by pH-stat fed-batch culture. A concentrated mixture solution of glucose and peptone was automatically added by acid-supplying pump when culture pH rose above high limit. Effect of the ratio of glucose to peptone in feeding solution was investigated on cell growth and nattokinase production by changing the ratio from 0.2 to 5 g glucose/g peptone. The highest cell concentration was 77 g/L when the ratio was 0.2 g glucose/g peptone. Cell concentration decreased with increasing the ratio of glucose to peptone in feeding solution, while the optimum condition existed for nattokinase production. The highest nattokinase activity was 14,500 unit/mL at a ratio of 0.33 g glucose/g peptone, which was 4.3 times higher than that in batch culture.

High lactic acid and fructose production via Mn2+-mediated conversion of inulin by Lactobacillus paracasei.

PubMed

Petrov, Kaloyan; Popova, Luiza; Petrova, Penka

2017-06-01

Lactobacillus paracasei DSM 23505 is able to produce high amounts of lactic acid (LA) by simultaneous saccharification and fermentation (SSF) of inulin. Aiming to obtain the highest possible amounts of LA and fructose, the present study is devoted to evaluate the impact of bivalent metal ions on the process of inulin conversion. It was shown that Mn 2+ strongly increases the activity of the purified key enzyme β-fructosidase. In vivo, batch fermentation kinetics revealed that the high Mn 2+ concentrations accelerated inulin hydrolysis by raise of the inulinase activity, and increased sugars conversion to LA through enhancement of the whole glycolytic flux. The highest LA concentration and yield were reached by addition of 15 mM Mn 2+ -151 g/L (corresponding to 40% increase) and 0.83 g/g, respectively. However, the relative quantification by real-time reverse transcription assay showed that the presence of Mn 2+ decreases the expression levels of fosE gene encoding β-fructosidase. Contrariwise, the full exclusion of metal ions resulted in fosE gene expression enhancement, blocked fructose transport, and hindered fructose conversion thus leading to huge fructose accumulation. During fed-batch with optimized medium and fermentation parameters, the fructose content reached 35.9% (w/v), achieving yield of 467 g fructose from 675 g inulin containing chicory flour powder (0.69 g/g). LA received in course of the batch fermentation and fructose gained by the fed-batch are the highest amounts ever obtained from inulin, thus disclosing the key role of Mn 2+ as a powerful tool to guide inulin conversion to targeted bio-chemicals.

Production of carotenoids and lipids by Rhodococcus opacus PD630 in batch and fed-batch culture.

PubMed

Thanapimmetha, Anusith; Suwaleerat, Tharatron; Saisriyoot, Maythee; Chisti, Yusuf; Srinophakun, Penjit

2017-01-01

Production of carotenoids by Rhodococcus opacus PD630 is reported. A modified mineral salt medium formulated with glycerol as an inexpensive carbon source was used for the fermentation. Ammonium acetate was the nitrogen source. A dry cell mass concentration of nearly 5.4 g/L could be produced in shake flasks with a carotenoid concentration of 0.54 mg/L. In batch culture in a 5 L bioreactor, without pH control, the maximum dry biomass concentration was ~30 % lower than in shake flasks and the carotenoids concentration was 0.09 mg/L. Both the biomass concentration and the carotenoids concentration could be raised using a fed-batch operation with a feed mixture of ammonium acetate and acetic acid. With this strategy, the final biomass concentration was 8.2 g/L and the carotenoids concentration was 0.20 mg/L in a 10-day fermentation. A control of pH proved to be unnecessary for maximizing the production of carotenoids in this fermentation.

Microbial conversion of synthetic and food waste-derived volatile fatty acids to lipids.

PubMed

Vajpeyi, Shashwat; Chandran, Kartik

2015-01-01

Lipid accumulation in the oleaginous yeast Cryptococcus albidus was evaluated using mixtures of volatile fatty acids (VFA) as substrates. In general, batch growth under nitrogen limitation led to higher lipid accumulation using synthetic VFA. During batch growth, an initial COD:N ratio of 25:1mg COD:mg N led to maximum intracellular lipid accumulation (28.3 ± 0.7% g/g dry cell weight), which is the maximum reported for C. albidus using VFA as the carbon source, without compromising growth kinetics. At this feed COD:N ratio, chemostat cultures fed with synthetic VFA yielded statistically similar intracellular lipid content as batch cultures (29.9 ± 1.9%, g/g). However, batch cultures fed with VFA produced from the fermentation of food waste, yielded a lower lipid content (14.9 ± 0.1%, g/g). The lipid composition obtained with synthetic and food-waste-derived VFA was similar to commercial biodiesel feedstock. We therefore demonstrate the feasibility of linking biochemical waste treatment and biofuel production using VFA as key intermediates. Copyright © 2015 Elsevier Ltd. All rights reserved.

Nutrient availability of different batches of wheat distillers dried grains with solubles with and without exogenous enzymes for broiler chickens.

PubMed

Whiting, I M; Pirgozliev, V; Rose, S P; Wilson, J; Amerah, A M; Ivanova, S G; Staykova, G P; Oluwatosin, O O; Oso, A O

2017-03-01

Wheat distillers' dried grains with solubles (DDGS) are being used increasingly in the poultry feed industry but their nutritional value is variable. The aim of this experiment was to examine the effect of batch to batch variation of wheat DDGS produced by the same manufacturer on the growth performance, dietary N corrected apparent metabolizable energy (AMEn), energy conversion ratio (ECR), total tract dry matter retention (DMR), nitrogen retention (NR) and fat digestibility (FD) coefficients when fed to broilers in complete diets with and without enzyme supplementation. Six UK wheat DDGS samples, produced by a single manufacturer, were used in a broiler experiment. Six diets containing 150 g/kg of each selected wheat DDGS sample were mixed. Each diet was then split into two batches and one of them was supplemented with commercial enzyme preparation, providing 1220 units xylanase and 152 units of β-glucanase/kg diet, resulting in 12 experimental diets. Each diet was fed ad libitum to five pens of two male Ross 308 broilers from 7 to 21 d old. Enzyme supplementation improved dietary AMEn, DMR, NR (P < 0.001) and FD (P < 0.05) compared to non-supplemented diets. There was DDGS sample by enzyme interaction (P < 0.05) on daily weight gain and ECR. The results suggest that the variability in AMEn of DDGS samples produced from a single manufacturer is greater than expected compared to the variability of whole wheat samples but substantially lower than expected from wheat DDGS samples from different EU manufacturers. This experiment has shown that the variation in feeding value of wheat DDGS may be explained by the variability in polysaccharide contents. © 2016 Poultry Science Association Inc.

Scheduling algorithm for flow shop with two batch-processing machines and arbitrary job sizes

NASA Astrophysics Data System (ADS)

Cheng, Bayi; Yang, Shanlin; Hu, Xiaoxuan; Li, Kai

2014-03-01

This article considers the problem of scheduling two batch-processing machines in flow shop where the jobs have arbitrary sizes and the machines have limited capacity. The jobs are processed in batches and the total size of jobs in each batch cannot exceed the machine capacity. Once a batch is being processed, no interruption is allowed until all the jobs in it are completed. The problem of minimising makespan is NP-hard in the strong sense. First, we present a mathematical model of the problem using integer programme. We show the scale of feasible solutions of the problem and provide optimality properties. Then, we propose a polynomial time algorithm with running time in O(nlogn). The jobs are first assigned in feasible batches and then scheduled on machines. For the general case, we prove that the proposed algorithm has a performance guarantee of 4. For the special case where the processing times of each job on the two machines satisfy p 1 j = ap 2 j , the performance guarantee is ? for a > 0.

Application of quality by design principles to the development and technology transfer of a major process improvement for the manufacture of a recombinant protein.

PubMed

Looby, Mairead; Ibarra, Neysi; Pierce, James J; Buckley, Kevin; O'Donovan, Eimear; Heenan, Mary; Moran, Enda; Farid, Suzanne S; Baganz, Frank

2011-01-01

This study describes the application of quality by design (QbD) principles to the development and implementation of a major manufacturing process improvement for a commercially distributed therapeutic protein produced in Chinese hamster ovary cell culture. The intent of this article is to focus on QbD concepts, and provide guidance and understanding on how the various components combine together to deliver a robust process in keeping with the principles of QbD. A fed-batch production culture and a virus inactivation step are described as representative examples of upstream and downstream unit operations that were characterized. A systematic approach incorporating QbD principles was applied to both unit operations, involving risk assessment of potential process failure points, small-scale model qualification, design and execution of experiments, definition of operating parameter ranges and process validation acceptance criteria followed by manufacturing-scale implementation and process validation. Statistical experimental designs were applied to the execution of process characterization studies evaluating the impact of operating parameters on product quality attributes and process performance parameters. Data from process characterization experiments were used to define the proven acceptable range and classification of operating parameters for each unit operation. Analysis of variance and Monte Carlo simulation methods were used to assess the appropriateness of process design spaces. Successful implementation and validation of the process in the manufacturing facility and the subsequent manufacture of hundreds of batches of this therapeutic protein verifies the approaches taken as a suitable model for the development, scale-up and operation of any biopharmaceutical manufacturing process. Copyright © 2011 American Institute of Chemical Engineers (AIChE).

Impact of organic loading rate on the performance of psychrophilic dry anaerobic digestion of dairy manure and wheat straw: long-term operation.

PubMed

Saady, Noori M Cata; Massé, Daniel I

2015-04-01

Development of efficient processes for valorising animal wastes would be a major advancement in cold-climate regions. This paper reports the results of long term (315 days experiment) of novel psychrophilic (20°C) dry anaerobic digestion (PDAD) of cow feces and wheat straw in laboratory scale sequence batch reactor operated at increasing organic loading rate. The PDAD process fed with a mixture of feces and straw (TS of 27%) over a treatment cycle length of 21 days at organic loading rate (OLR) 4.0, 5.0 and 6.0 g TCOD kg(-1) inoculum d(-1) (of 2.9 ± 0.1, 3.7 ± 0.1, and 4.4 ± 0.1g VS kg(-1) inoculum d(-1), respectively) resulted in average specific methane yield (SMY) of 187.3 ± 18.1, 163.6 ± 39.5, 150.8 ± 32.9 N L CH4 kg(-1)VS fed, respectively. PDAD of cow feces and wheat straw is possible with VS-based inoculum-to-substrate ratio of 1.4 at OLR of 6.0 g TCOD kg(-1) inoculum d(-1). Hydrolysis was the limiting step reaction. Crown Copyright © 2015. Published by Elsevier Ltd. All rights reserved.

Enhanced lipid production by Rhodosporidium toruloides using different fed-batch feeding strategies with lignocellulosic hydrolysate as the sole carbon source

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

Fei, Qiang; O'Brien, Marykate; Nelson, Robert

Industrial biotechnology that is able to provide environmentally friendly bio-based products has attracted more attention in replacing petroleum-based industries. Currently, most of the carbon sources used for fermentation-based bioprocesses are obtained from agricultural commodities that are used as foodstuff for human beings. Lignocellulose-derived sugars as the non-food, green, and sustainable alternative carbon sources have great potential to avoid this dilemma for producing the renewable, bio-based hydrocarbon fuel precursors, such as microbial lipid. Efficient bioconversion of lignocellulose-based sugars into lipids is one of the critical parameters for industrial application. Therefore, the fed-batch cultivation, which is a common method used in industrialmore » applications, was investigated to achieve a high cell density culture along with high lipid yield and productivity. In this study, several fed-batch strategies were explored to improve lipid production using lignocellulosic hydrolysates derived from corn stover. Compared to the batch culture giving a lipid yield of 0.19 g/g, the dissolved-oxygen-stat feeding mode increased the lipid yield to 0.23 g/g and the lipid productivity to 0.33 g/L/h. The pulse feeding mode further improved lipid productivity to 0.35 g/L/h and the yield to 0.24 g/g. However, the highest lipid yield (0.29 g/g) and productivity (0.4 g/L/h) were achieved using an automated online sugar control feeding mode, which gave a dry cell weight of 54 g/L and lipid content of 59 % (w/w). The major fatty acids of the lipid derived from lignocellulosic hydrolysates were predominately palmitic acid and oleic acid, which are similar to those of conventional oilseed plants. Our results suggest that the fed-batch feeding strategy can strongly influence the lipid production. Lastly, the online sugar control feeding mode was the most appealing strategy for high cell density, lipid yield, and lipid productivity using lignocellulosic hydrolysates as the sole carbon source.« less

Enhanced lipid production by Rhodosporidium toruloides using different fed-batch feeding strategies with lignocellulosic hydrolysate as the sole carbon source

DOE PAGES

Fei, Qiang; O'Brien, Marykate; Nelson, Robert; ...

2016-06-23

Industrial biotechnology that is able to provide environmentally friendly bio-based products has attracted more attention in replacing petroleum-based industries. Currently, most of the carbon sources used for fermentation-based bioprocesses are obtained from agricultural commodities that are used as foodstuff for human beings. Lignocellulose-derived sugars as the non-food, green, and sustainable alternative carbon sources have great potential to avoid this dilemma for producing the renewable, bio-based hydrocarbon fuel precursors, such as microbial lipid. Efficient bioconversion of lignocellulose-based sugars into lipids is one of the critical parameters for industrial application. Therefore, the fed-batch cultivation, which is a common method used in industrialmore » applications, was investigated to achieve a high cell density culture along with high lipid yield and productivity. In this study, several fed-batch strategies were explored to improve lipid production using lignocellulosic hydrolysates derived from corn stover. Compared to the batch culture giving a lipid yield of 0.19 g/g, the dissolved-oxygen-stat feeding mode increased the lipid yield to 0.23 g/g and the lipid productivity to 0.33 g/L/h. The pulse feeding mode further improved lipid productivity to 0.35 g/L/h and the yield to 0.24 g/g. However, the highest lipid yield (0.29 g/g) and productivity (0.4 g/L/h) were achieved using an automated online sugar control feeding mode, which gave a dry cell weight of 54 g/L and lipid content of 59 % (w/w). The major fatty acids of the lipid derived from lignocellulosic hydrolysates were predominately palmitic acid and oleic acid, which are similar to those of conventional oilseed plants. Our results suggest that the fed-batch feeding strategy can strongly influence the lipid production. Lastly, the online sugar control feeding mode was the most appealing strategy for high cell density, lipid yield, and lipid productivity using lignocellulosic hydrolysates as the sole carbon source.« less

Online optimal experimental re-design in robotic parallel fed-batch cultivation facilities.

PubMed

Cruz Bournazou, M N; Barz, T; Nickel, D B; Lopez Cárdenas, D C; Glauche, F; Knepper, A; Neubauer, P

2017-03-01

We present an integrated framework for the online optimal experimental re-design applied to parallel nonlinear dynamic processes that aims to precisely estimate the parameter set of macro kinetic growth models with minimal experimental effort. This provides a systematic solution for rapid validation of a specific model to new strains, mutants, or products. In biosciences, this is especially important as model identification is a long and laborious process which is continuing to limit the use of mathematical modeling in this field. The strength of this approach is demonstrated by fitting a macro-kinetic differential equation model for Escherichia coli fed-batch processes after 6 h of cultivation. The system includes two fully-automated liquid handling robots; one containing eight mini-bioreactors and another used for automated at-line analyses, which allows for the immediate use of the available data in the modeling environment. As a result, the experiment can be continually re-designed while the cultivations are running using the information generated by periodical parameter estimations. The advantages of an online re-computation of the optimal experiment are proven by a 50-fold lower average coefficient of variation on the parameter estimates compared to the sequential method (4.83% instead of 235.86%). The success obtained in such a complex system is a further step towards a more efficient computer aided bioprocess development. Biotechnol. Bioeng. 2017;114: 610-619. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Comprehensive assessment of the L-lysine production process from fermentation of sugarcane molasses.

PubMed

Anaya-Reza, Omar; Lopez-Arenas, Teresa

2017-07-01

L-Lysine is an essential amino acid that can be produced by chemical processes from fossil raw materials, as well as by microbial fermentation, the latter being a more efficient and environmentally friendly procedure. In this work, the production process of L-lysine-HCl is studied using a systematic approach based on modeling and simulation, which supports decision making in the early stage of process design. The study considers two analysis stages: first, the dynamic analysis of the fermentation reactor, where the conversion of sugars from sugarcane molasses to L-lysine with a strain of Corynebacterium glutamicum is carried out. In this stage, the operation mode (either batch or fed batch) and operating conditions of the fermentation reactor are defined to reach the maximum technical criteria. Afterwards, the second analysis stage relates to the industrial production process of L-lysine-HCl, where the fermentation reactor, upstream processing, and downstream processing are included. In this stage, the influence of key parameters on the overall process performance is scrutinized through the evaluation of several technical, economic, and environmental criteria, to determine a profitable and sustainable design of the L-lysine production process. The main results show how the operating conditions, process design, and selection of evaluation criteria can influence in the conceptual design. The best plant design shows maximum product yield (0.31 g L-lysine/g glucose) and productivity (1.99 g/L/h), achieving 26.5% return on investment (ROI) with a payback period (PBP) of 3.8 years, decreasing water and energy consumption, and with a low potential environmental impact (PEI) index.

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.

Effect of four processed animal proteins in the diet on digestibility and performance in laying hens.

PubMed

van Krimpen, M M; Veldkamp, T; Binnendijk, G P; de Veer, R

2010-12-01

An experiment was performed to investigate the effect of animal vs. vegetable protein sources in the diet of laying hens on the development of hen performance. A diet containing protein sources of only vegetable origin was compared with 4 diets, each containing 1 of 4 processed animal proteins (PAP). Two PAP (Daka-58 and Sonac-60) were classified as meat meals, and the remaining 2 (Daka-40 and Sonac-50) were classified as meat and bone meals. First, fecal digestibility of nutrients in the PAP was determined in Lohmann Brown layers. Hens (n = 132) were housed in 22 cages (6 hens/cage) and allotted to 5 dietary treatments. In the PAP diets (4 replicates/treatment), 100 g/kg of CP of animal origin was added, replacing soybean meal and corn (Zea mays) in the basal diet (6 replicates/treatment). The PAP sources differed largely in chemical composition and digestibility coefficients. Energy content (AME(n)) varied from 1,817 (Daka-40) to 3,107 kcal/kg (Sonac-60), and digestible lysine varied from 15.4 (Daka-40) to 28.3 g/kg (Sonac-50). Subsequently, the effect of a control diet (without PAP) vs. 4 PAP diets (50 g/kg of CP of animal origin from the same batches as used in the digestibility study) on performance was determined. All diets were isocaloric (AME(n) = 2,825 kcal/kg) and isonitrogenous (digestible lysine = 6.8 g/kg). Hens were housed in 40 floor pens (12 hens/pen, 8 pens/treatment) from 20 to 40 wk of age. Feed intake levels of the hens fed the meat and bone meal diets were reduced compared with those of hens fed the meat meal diets, whereas the feed intake level of hens fed the control diet was intermediate. Laying hen performance differed between treatments, being was most favorable for the Sonac-50 treatment and most adverse for the Daka-40 treatment. Differences in laying hen performance seemed to be related partly to differences in feed intake and corresponding amino acid intake.

High cell density cultivation and recombinant protein production with Escherichia coli in a rocking-motion-type bioreactor

PubMed Central

2010-01-01

Background Single-use rocking-motion-type bag bioreactors provide advantages compared to standard stirred tank bioreactors by decreased contamination risks, reduction of cleaning and sterilization time, lower investment costs, and simple and cheaper validation. Currently, they are widely used for cell cultures although their use for small and medium scale production of recombinant proteins with microbial hosts might be very attractive. However, the utilization of rocking- or wave-induced motion-type bioreactors for fast growing aerobic microbes is limited because of their lower oxygen mass transfer rate. A conventional approach to reduce the oxygen demand of a culture is the fed-batch technology. New developments, such as the BIOSTAT® CultiBag RM system pave the way for applying advanced fed-batch control strategies also in rocking-motion-type bioreactors. Alternatively, internal substrate delivery systems such as EnBase® Flo provide an opportunity for adopting simple to use fed-batch-type strategies to shaken cultures. Here, we investigate the possibilities which both strategies offer in view of high cell density cultivation of E. coli and recombinant protein production. Results Cultivation of E. coli in the BIOSTAT® CultiBag RM system in a conventional batch mode without control yielded an optical density (OD600) of 3 to 4 which is comparable to shake flasks. The culture runs into oxygen limitation. In a glucose limited fed-batch culture with an exponential feed and oxygen pulsing, the culture grew fully aerobically to an OD600 of 60 (20 g L-1 cell dry weight). By the use of an internal controlled glucose delivery system, EnBase® Flo, OD600 of 30 (10 g L-1 cell dry weight) is obtained without the demand of computer controlled external nutrient supply. EnBase® Flo also worked well in the CultiBag RM system with a recombinant E. coli RB791 strain expressing a heterologous alcohol dehydrogenase (ADH) to very high levels, indicating that the enzyme based feed supply strategy functions well for recombinant protein production also in a rocking-motion-type bioreactor. Conclusions Rocking-motion-type bioreactors may provide an interesting alternative to standard cultivation in bioreactors for cultivation of bacteria and recombinant protein production. The BIOSTAT® Cultibag RM system with the single-use sensors and advanced control system paves the way for the fed-batch technology also to rocking-motion-type bioreactors. It is possible to reach cell densities which are far above shake flasks and typical for stirred tank reactors with the improved oxygen transfer rate. For more simple applications the EnBase® Flo method offers an easy and robust solution for rocking-motion-systems which do not have such advanced control possibilities. PMID:20509968

Harvesting microalgal biomass using a magnetically induced membrane vibration (MMV) system: filtration performance and energy consumption.

PubMed

Bilad, M R; Discart, V; Vandamme, D; Foubert, I; Muylaert, K; Vankelecom, Ivo F J

2013-06-01

This study was performed to investigate the effectiveness of submerged microfiltration to harvest both a marine diatom Phaeodactylum tricornutum and a Chlorella vulgaris in a recently developed magnetically induced membrane vibrating (MMV) system. We assess the filtration performance by conducting the improved flux step method (IFM), fed-batch concentration filtrations and membrane fouling autopsy using two lab-made membranes with different porosity. The full-scale energy consumption was also estimated. Overall results suggest that the MMV offers a good fouling control and the process was proven to be economically attractive. By combining the membrane filtration (15× concentration) with centrifugation to reach a final concentration of 25% w/v, the energy consumption to harvest P. tricornutum and C. vulgaris was, respectively, as low as 0.84 and 0.77kWh/m(3), corresponding to 1.46 and 1.39 kWh/kg of the harvested biomass. Copyright © 2013 Elsevier Ltd. All rights reserved.

Semi-industrial scale (30 m3) fed-batch fermentation for the production of D-lactate by Escherichia coli strain HBUT-D15.

PubMed

Fu, Xiangmin; Wang, Yongze; Wang, Jinhua; Garza, Erin; Manow, Ryan; Zhou, Shengde

2017-02-01

D(-)-lactic acid is needed for manufacturing of stereo-complex poly-lactic acid polymer. Large scale D-lactic acid fermentation, however, has yet to be demonstrated. A genetically engineered Escherichia coli strain, HBUT-D, was adaptively evolved in a 15% calcium lactate medium for improved lactate tolerance. The resulting strain, HBUT-D15, was tested at a lab scale (7 L) by fed-batch fermentation with up to 200 g L -1 of glucose, producing 184-191 g L -1 of D-lactic acid, with a volumetric productivity of 4.38 g L -1  h -1 , a yield of 92%, and an optical purity of 99.9%. The HBUT-D15 was then evaluated at a semi-industrial scale (30 m 3 ) via fed-batch fermentation with up to 160 g L -1 of glucose, producing 146-150 g L -1 of D-lactic acid, with a volumetric productivity of 3.95-4.29 g L -1  h -1 , a yield of 91-94%, and an optical purity of 99.8%. These results are comparable to that of current industrial scale L(+)-lactic acid fermentation.

Integrated ecotechnology approach towards treatment of complex wastewater with simultaneous bioenergy production.

PubMed

Hemalatha, Manupati; Sravan, J Shanthi; Yeruva, Dileep Kumar; Venkata Mohan, S

2017-10-01

Sequential integration of three stage diverse biological processes was studied by exploiting the individual process advantage towards enhanced treatment of complex chemical based wastewater. A successful attempt to integrate sequence batch reactor (SBR) with bioelectrochemical treatment (BET) and finally with microalgae treatment was studied. The sequential integration has showed individual substrate degradation (COD) of 55% in SBR, 49% in BET and 56% in microalgae, accounting for a consolidated treatment efficiency of 90%. Nitrates removal efficiency of 25% was observed in SBR, 31% in BET and 44% in microalgae, with a total efficiency of 72%. The SBR treated effluents fed to BET with the electrode intervention showed TDS removal. BET exhibited relatively higher process performance than SBR. The integration approach significantly overcame the individual process limitations along with value addition as biomass (1.75g/L), carbohydrates (640mg/g), lipids (15%) and bioelectricity. The study resulted in providing a strategy of combining SBR as pretreatment step to BET process and finally polishing with microalgae cultivation achieving the benefits of enhanced wastewater treatment along with value addition. Copyright © 2017 Elsevier Ltd. All rights reserved.

Production of capsular polysaccharide from Escherichia coli K4 for biotechnological applications.

PubMed

Cimini, Donatella; Restaino, Odile Francesca; Catapano, Angela; De Rosa, Mario; Schiraldi, Chiara

2010-02-01

The production of industrially relevant microbial polysaccharides has recently gained much interest. The capsular polysaccharide of Escherichia coli K4 is almost identical to chondroitin, a commercially valuable biopolymer that is so far obtained from animal tissues entailing complex and expensive extraction procedures. In the present study, the production of capsular polysaccharide by E. coli K4 was investigated taking into consideration a potential industrial application. Strain physiology was first characterized in shake flask experiments to determine the optimal culture conditions for the growth of the microorganism and correlate it to polysaccharide production. Results show that the concentration of carbon source greatly affects polysaccharide production, while the complex nitrogen source is mainly responsible for the build up of biomass. Small-scale batch processes were performed to further evaluate the effect of the initial carbon source concentration and of growth temperatures on polysaccharide production, finally leading to the establishment of the medium to use in following fermentation experiments on a bigger scale. The fed-batch strategy next developed on a 2-L reactor resulted in a maximum cell density of 56 g(cww)/L and a titre of capsular polysaccharide equal to 1.4 g/L, approximately ten- and fivefold higher than results obtained in shake flask and 2-L batch experiments, respectively. The release kinetics of K4 polysaccharide into the medium were also explored to gain insight into the mechanisms underlying a complex aspect of the strain physiology.

Evaluation of the in vivo disintegration of solid dosage forms of a bile acid sequestrant in dogs using gamma-scintigraphy and correlation to in vitro disintegration.

PubMed

Hussain, Munir A; Chang, Rong-Kun; Sandefer, Erik; Page, Richard C; Digenis, George A

2003-03-01

[corrected] To evaluate the in vivo disintegration behavior of tablets and capsules of a bile acid sequestrant, DMP 504, in beagle dogs and to assess the significance of the in vitro disintegration of the dosage forms on subsequent in vivo behavior in order to draw possible in vitro-in vivo correlations. Tablet and capsule formulations of a bile acid sequestrant, DMP 504, were formulated with samarium oxide and neutron activated to produce radioactive 53Sm to noninvasively evaluate their in vivo behavior in beagle dogs by gamma-scintigraphy. A four-way crossover design was completed (n = 4) in which (a) tablets from two different batches were administered under the fasted condition and manufactured using different lots of drug substance where one batch exhibited relatively faster in vitro disintegration time (30 min) than the other tablet batch, which resulted in slower disintegration (45 min), (b) a capsule formulation was administered to fasted beagles, and (c) the tablet having slower in vitro disintegration was also administered in the fed state, and its in vivo disintegration was compared to that observed in the fasted state. Tablets manufactured using a lot of DMP 504 having relatively fast in vitro disintegration (approximately 30 min) resulted in relatively rapid in vivo disintegration time (15 min) in the fasted condition. This in vivo disintegration time was comparable to the in vivo disintegration of the capsules (17 min) even though the in vitro capsule disintegration time was considerably faster (2 min). Tablets prepared using a drug substance that provided a longer in vitro disintegration time (approximately 45 min) resulted in a slower in vivo disintegration (63 min). There was no difference observed in the in vivo disintegration behavior in fasted and fed dogs for the tablets that provided slower in vitro disintegration. In vivo disintegration of tablets of the bile acid sequestrant DMP 504 correlated with in vitro disintegration times. Gamma-Scintigraphy continues to be a good tool to use during early stages of product development to investigate in vivo performance of dosage forms. The results of this study provided evidence that the physical chemical specifications of the drug substance may not always be indicative of in vitro or in vivo performance of tablet dosage form, even when formulation and process are not changed.

Stable thermophilic anaerobic digestion of dissolved air flotation (DAF) sludge by co-digestion with swine manure.

PubMed

Creamer, K S; Chen, Y; Williams, C M; Cheng, J J

2010-05-01

Environmentally sound treatment of by-products in a value-adding process is an ongoing challenge in animal agriculture. The sludge produced as a result of the dissolved air flotation (DAF) wastewater treatment process in swine processing facilities is one such low-value residue. The objective of this study was to determine the fundamental performance parameters for thermophilic anaerobic digestion of DAF sludge. Testing in a semi-continuous stirred tank reactor and in batch reactors was conducted to determine the kinetics of degradation and biogas yield. Stable operation could not be achieved using pure DAF sludge as a substrate, possibly due to inhibition by long-chain fatty acids or to nutrient deficiencies. However, in a 1:1 ratio (w/w, dry basis) with swine manure, operation was both stable and productive. In the semi-continuous stirred reactor at 54.5 degrees Celsius, a hydraulic residence time of 10 days, and an organic loading rate of 4.68 gVS/day/L, the methane production rate was 2.19 L/L/day and the specific methane production rate was 0.47 L/gVS (fed). Maximum specific methanogenic activity (SMA) in batch testing was 0.15 mmoles CH(4) h(-1) gVS(-1) at a substrate concentration of 6.9 gVS L(-1). Higher substrate concentrations cause an initial lag in methane production, possibly due to long-chain fatty acid or nitrogen inhibition. Copyright 2009 Elsevier Ltd. All rights reserved.

Improving succinic acid production by Actinobacillus succinogenes from raw industrial carob pods.

PubMed

Carvalho, Margarida; Roca, Christophe; Reis, Maria A M

2016-10-01

Carob pods are an inexpensive by-product of locust bean gum industry that can be used as renewable feedstock for bio-based succinic acid. Here, for the first time, unprocessed raw carob pods were used to extract a highly enriched sugar solution, afterwards used as substrate to produce succinic acid using Actinobacillus succinogenes. Batch fermentations containing 30g/L sugars resulted in a production rate of 1.67gSA/L.h and a yield of 0.39gSA/g sugars. Taking advantage of A. succinogenes' metabolism, uncoupling cell growth from succinic acid production, a fed-batch mode was implemented to increase succinic acid yield and reduce by-products formation. This strategy resulted in a succinic acid yield of 0.94gSA/g sugars, the highest yield reported in the literature for fed-batch and continuous experiments, while maintaining by-products at residual values. Results demonstrate that raw carob pods are a highly efficient feedstock for bio-based succinic acid production. Copyright © 2016 Elsevier Ltd. All rights reserved.

Anaerobic co-digestion of sugarcane press mud with vinasse on methane yield.

PubMed

López González, Lisbet Mailin; Pereda Reyes, Ileana; Romero Romero, Osvaldo

2017-10-01

The conversion efficiency of high solids waste digestion as sugarcane press mud (P) may be limited due to hydrolysis step. The option of co-digestion with vinasse, main liquid waste generated from ethanol production, was investigated under batch regime at mesophilic conditions (37.5±1°C) and the best mixture was evaluated under semicontinuous regime in stirred-tank reactors. The maximum values for methane yield in batch tests were for V 75 /P 25 and V 50 /P 50 mixtures (on basis of the chemical oxygen demand (COD) percentage added in the mixture), with an average value of 246NmL CH 4 g -1 COD fed , which was 13% higher than that of press mud alone. A highest methane production rate of 69.6NmL CH 4 g -1 COD fed -1 d -1 was obtained for the mixtureV 75 /P 25 . During the experiment carried out in CSTR reactors, the organic loading rate (OLR) was increased from 0.5 up to 2.2gVSL -1 d -1 . Methane yields of 365L CH 4 kg -1 VS and biogas productivities of 1.6LL -1 were obtained in co-digestion, which was 64% higher in comparison to mono-digestion. The performance of the process in mono-digestion was less stable than in co-digestion, with a significant fall of methane yield to 1.8kgVSm -3 d -1 , and a partial inhibition of the methanogenic archaeas when the OLR was increased up to 2.2kgVSm -3 d -1 . The co-digestion of vinasse with press mud is a good option for the treatment of streams at the alcohol-sugar industry. Copyright © 2017 Elsevier Ltd. All rights reserved.

Development of hyper osmotic resistant CHO host cells for enhanced antibody production.

PubMed

Kamachi, Yasuharu; Omasa, Takeshi

2018-04-01

Cell culture platform processes are generally employed to shorten the duration of new product development. A fed-batch process with continuous feeding is a conventional platform process for monoclonal antibody production using Chinese hamster ovary (CHO) cells. To establish a simplified platform process, the feeding method can be changed from continuous feed to bolus feed. However, this change induces a rapid increase of osmolality by the bolus addition of nutrients. The increased osmolality suppresses cell culture growth, and the final product concentration is decreased. In this study, osmotic resistant CHO host cells were developed to attain a high product concentration. To establish hyper osmotic resistant CHO host cells, CHO-S host cells were passaged long-term in a hyper osmotic basal medium. There were marked differences in cell growth of the original and established host cells under iso- (328 mOsm/kg) or hyper-osmolality (over 450 mOsm/kg) conditions. Cell growth of the original CHO host cells was markedly decreased by the induction of osmotic stress, whereas cell growth of the hyper osmotic resistant CHO host cells was not affected. The maximum viable cell concentration of hyper osmotic resistant CHO host cells was 132% of CHO-S host cells after the induction of osmotic stress. Moreover, the hyper osmotic resistant characteristic of established CHO host cells was maintained even after seven passages in iso-osmolality basal medium. The use of hyper osmotic resistance CHO host cells to create a monoclonal antibody production cell line might be a new approach to increase final antibody concentrations with a fed-batch process. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Improvement of mammalian cell culture performance through surfactant enabled concentrated feed media.

PubMed

Hossler, Patrick; McDermott, Sean; Racicot, Christopher; Fann, John C H

2013-01-01

The design of basal and feed media in mammalian cell culture is paramount towards ensuring acceptable upstream process performance in various operation modes, especially fed-batch culture. Mammalian cell culture media designs have evolved from the classical formulations designed by Eagle and Ham, to today's formulations designed from continuous improvement and statistical frameworks. Feed media is especially important for ensuring robust cell growth, productivity, and ensuring the product quality of recombinant therapeutics are within acceptable ranges. Numerous studies have highlighted the benefit of various media designs, supplements, and feed addition strategies towards the resulting cell culture process. In this work we highlight the use of a top-down level approach towards feed media design enabled by the use of select surfactants for the targeted enrichment of a chemically defined feed media. The use of the enriched media was able to improve product titers at g/L levels, without adversely impacting the growth of multiple Chinese Hamster Ovary cell lines or the product quality of multiple recombinant antibodies. © 2013 American Institute of Chemical Engineers.

Online intelligent controllers for an enzyme recovery plant: design methodology and performance.

PubMed

Leite, M S; Fujiki, T L; Silva, F V; Fileti, A M F

2010-12-27

This paper focuses on the development of intelligent controllers for use in a process of enzyme recovery from pineapple rind. The proteolytic enzyme bromelain (EC 3.4.22.4) is precipitated with alcohol at low temperature in a fed-batch jacketed tank. Temperature control is crucial to avoid irreversible protein denaturation. Fuzzy or neural controllers offer a way of implementing solutions that cover dynamic and nonlinear processes. The design methodology and a comparative study on the performance of fuzzy-PI, neurofuzzy, and neural network intelligent controllers are presented. To tune the fuzzy PI Mamdani controller, various universes of discourse, rule bases, and membership function support sets were tested. A neurofuzzy inference system (ANFIS), based on Takagi-Sugeno rules, and a model predictive controller, based on neural modeling, were developed and tested as well. Using a Fieldbus network architecture, a coolant variable speed pump was driven by the controllers. The experimental results show the effectiveness of fuzzy controllers in comparison to the neural predictive control. The fuzzy PI controller exhibited a reduced error parameter (ITAE), lower power consumption, and better recovery of enzyme activity.

Online Intelligent Controllers for an Enzyme Recovery Plant: Design Methodology and Performance

PubMed Central

Leite, M. S.; Fujiki, T. L.; Silva, F. V.; Fileti, A. M. F.

2010-01-01

This paper focuses on the development of intelligent controllers for use in a process of enzyme recovery from pineapple rind. The proteolytic enzyme bromelain (EC 3.4.22.4) is precipitated with alcohol at low temperature in a fed-batch jacketed tank. Temperature control is crucial to avoid irreversible protein denaturation. Fuzzy or neural controllers offer a way of implementing solutions that cover dynamic and nonlinear processes. The design methodology and a comparative study on the performance of fuzzy-PI, neurofuzzy, and neural network intelligent controllers are presented. To tune the fuzzy PI Mamdani controller, various universes of discourse, rule bases, and membership function support sets were tested. A neurofuzzy inference system (ANFIS), based on Takagi-Sugeno rules, and a model predictive controller, based on neural modeling, were developed and tested as well. Using a Fieldbus network architecture, a coolant variable speed pump was driven by the controllers. The experimental results show the effectiveness of fuzzy controllers in comparison to the neural predictive control. The fuzzy PI controller exhibited a reduced error parameter (ITAE), lower power consumption, and better recovery of enzyme activity. PMID:21234106

A short term quality control tool for biodegradable microspheres.

PubMed

D'Souza, Susan; Faraj, Jabar A; Dorati, Rossella; DeLuca, Patrick P

2014-06-01

Accelerated in vitro release testing methodology has been developed as an indicator of product performance to be used as a discriminatory quality control (QC) technique for the release of clinical and commercial batches of biodegradable microspheres. While product performance of biodegradable microspheres can be verified by in vivo and/or in vitro experiments, such evaluation can be particularly challenging because of slow polymer degradation, resulting in extended study times, labor, and expense. Three batches of Leuprolide poly(lactic-co-glycolic acid) (PLGA) microspheres having varying morphology (process variants having different particle size and specific surface area) were manufactured by the solvent extraction/evaporation technique. Tests involving in vitro release, polymer degradation and hydration of the microspheres were performed on the three batches at 55°C. In vitro peptide release at 55°C was analyzed using a previously derived modification of the Weibull function termed the modified Weibull equation (MWE). Experimental observations and data analysis confirm excellent reproducibility studies within and between batches of the microsphere formulations demonstrating the predictability of the accelerated experiments at 55°C. The accelerated test method was also successfully able to distinguish the in vitro product performance between the three batches having varying morphology (process variants), indicating that it is a suitable QC tool to discriminate product or process variants in clinical or commercial batches of microspheres. Additionally, data analysis utilized the MWE to further quantify the differences obtained from the accelerated in vitro product performance test between process variants, thereby enhancing the discriminatory power of the accelerated methodology at 55°C.

Size characterization of inclusion bodies by sedimentation field-flow fractionation

PubMed Central

Margreiter, Gerd; Messner, Paul; Caldwell, Karin D.; Bayer, Karl

2015-01-01

Sedimentation field-flow fractionation (sedFFF) was evaluated to characterize the size of Δ(4–23)TEM-β-lactamase inclusion bodies (IBs) overexpressed in fed-batch cultivations of Escherichia coli. Heterologous Δ(4–23)TEM-β-lactamase protein formed different sizes of IBs, depending upon the induction conditions. In the early phases of recombinant protein expression, induced with low concentrations of IPTG (isopropyl-β-d-thiogalactoside), IB masses were larger than expected and showed heterogeneous size distributions. During cultivation, IB sizes showed a Gaussian distribution and reached a broad range by the end of the fed-batch cultivations. The obtained result proved the aptitude of sedFFF to rapidly assess the size distribution of IBs in a culture. PMID:18760314

Actual waste demonstration of the nitric-glycolic flowsheet for sludge batch 9 qualification

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

Newell, D.; Pareizs, J.; Martino, C.

For each sludge batch that is processed in the Defense Waste Processing Facility (DWPF), the Savannah River National Laboratory (SRNL) performs qualification testing to demonstrate that the sludge batch is processable. Based on the results of this actual-waste qualification and previous simulant studies, SRNL recommends implementation of the nitric-glycolic acid flowsheet in DWPF. Other recommendations resulting from this demonstration are reported in section 5.0.

Efficiency improvement of an antibody production process by increasing the inoculum density.

PubMed

Hecht, Volker; Duvar, Sevim; Ziehr, Holger; Burg, Josef; Jockwer, Alexander

2014-01-01

Increasing economic pressure is the main driving force to enhance the efficiency of existing processes. We developed a perfusion strategy for a seed train reactor to generate a higher inoculum density for a subsequent fed batch production culture. A higher inoculum density can reduce culture duration without compromising product titers. Hence, a better capacity utilization can be achieved. The perfusion strategy was planned to be implemented in an existing large scale antibody production process. Therefore, facility and process constraints had to be considered. This article describes the initial development steps. Using a proprietary medium and a Chinese hamster ovary cell line expressing an IgG antibody, four different cell retention devices were compared in regard to retention efficiency and reliability. Two devices were selected for further process refinement, a centrifuge and an inclined gravitational settler. A concentrated feed medium was developed to meet facility constraints regarding maximum accumulated perfundate volume. A 2-day batch phase followed by 5 days of perfusion resulted in cell densities of 1.6 × 10(10) cells L(-1) , a 3.5 fold increase compared to batch cultivations. Two reactor volumes of concentrated feed medium were needed to achieve this goal. Eleven cultivations were carried out in bench and 50 L reactors showing acceptable reproducibility and ease of scale up. In addition, it was shown that at least three perfusion phases can be combined within a repeated perfusion strategy. © 2014 American Institute of Chemical Engineers.

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.

Application of byproducts from food processing for production of 2,3-butanediol using Bacillus amyloliquefaciens TUL 308.

PubMed

Sikora, Barbara; Kubik, Celina; Kalinowska, Halina; Gromek, Ewa; Białkowska, Aneta; Jędrzejczak-Krzepkowska, Marzena; Schüett, Fokko; Turkiewicz, Marianna

2016-08-17

A nonpathogenic bacterial strain Bacillus amyloliquefaciens TUL 308 synthesized minor 2,3-butanediol (2,3-BD) amounts from glucose, fructose, sucrose, and glycerol, and efficiently produced the diol from molasses and hydrolysates of food processing residues. Batch fermentations yielded 16.53, 10.72, and 5 g/L 2,3-BD from enzymatic hydrolysates of apple pomace, dried sugar beet pulp, and potato pulp (at initial concentrations equivalent to 45, 20, and 30 g/L glucose, respectively), and 25.3 g/L 2,3-BD from molasses (at its initial concentration equivalent to 60 g/L saccharose). Fed-batch fermentations in the molasses-based medium with four feedings with either glucose or sucrose (in doses increasing their concentration by 25 g/L) resulted in around twice higher maximum 2,3-BD concentration (of about 60 and 50 g/L, respectively). The GRAS Bacillus strain is an efficient 2,3-BD producer from food industry byproducts.

Production of poly(malic acid) from sugarcane juice in fermentation by Aureobasidium pullulans: Kinetics and process economics.

PubMed

Wei, Peilian; Cheng, Chi; Lin, Meng; Zhou, Yipin; Yang, Shang-Tian

2017-01-01

Poly(β-l-malic acid) (PMA) is a biodegradable polymer with many potential biomedical applications. PMA can be readily hydrolyzed to malic acid (MA), which is widely used as an acidulant in foods and pharmaceuticals. PMA production from sucrose and sugarcane juice by Aureobasidium pullulans ZX-10 was studied in shake-flasks and bioreactors, confirming that sugarcane juice can be used as an economical substrate without any pretreatment or nutrients supplementation. A high PMA titer of 116.3g/L and yield of 0.41g/g were achieved in fed-batch fermentation. A high productivity of 0.66g/L·h was achieved in repeated-batch fermentation with cell recycle. These results compared favorably with those obtained from glucose and other biomass feedstocks. A process economic analysis showed that PMA could be produced from sugarcane juice at a cost of $1.33/kg, offering a cost-competitive bio-based PMA for industrial applications. Copyright © 2016 Elsevier Ltd. All rights reserved.

Bioethanol production from fermentable sugar juice.

PubMed

Zabed, Hossain; Faruq, Golam; Sahu, Jaya Narayan; Azirun, Mohd Sofian; Hashim, Rosli; Boyce, Amru Nasrulhaq

2014-01-01

Bioethanol production from renewable sources to be used in transportation is now an increasing demand worldwide due to continuous depletion of fossil fuels, economic and political crises, and growing concern on environmental safety. Mainly, three types of raw materials, that is, sugar juice, starchy crops, and lignocellulosic materials, are being used for this purpose. This paper will investigate ethanol production from free sugar containing juices obtained from some energy crops such as sugarcane, sugar beet, and sweet sorghum that are the most attractive choice because of their cost-effectiveness and feasibility to use. Three types of fermentation process (batch, fed-batch, and continuous) are employed in ethanol production from these sugar juices. The most common microorganism used in fermentation from its history is the yeast, especially, Saccharomyces cerevisiae, though the bacterial species Zymomonas mobilis is also potentially used nowadays for this purpose. A number of factors related to the fermentation greatly influences the process and their optimization is the key point for efficient ethanol production from these feedstocks.

Bioethanol Production from Fermentable Sugar Juice

PubMed Central

Zabed, Hossain; Faruq, Golam; Sahu, Jaya Narayan; Azirun, Mohd Sofian; Hashim, Rosli; Nasrulhaq Boyce, Amru

2014-01-01

Bioethanol production from renewable sources to be used in transportation is now an increasing demand worldwide due to continuous depletion of fossil fuels, economic and political crises, and growing concern on environmental safety. Mainly, three types of raw materials, that is, sugar juice, starchy crops, and lignocellulosic materials, are being used for this purpose. This paper will investigate ethanol production from free sugar containing juices obtained from some energy crops such as sugarcane, sugar beet, and sweet sorghum that are the most attractive choice because of their cost-effectiveness and feasibility to use. Three types of fermentation process (batch, fed-batch, and continuous) are employed in ethanol production from these sugar juices. The most common microorganism used in fermentation from its history is the yeast, especially, Saccharomyces cerevisiae, though the bacterial species Zymomonas mobilis is also potentially used nowadays for this purpose. A number of factors related to the fermentation greatly influences the process and their optimization is the key point for efficient ethanol production from these feedstocks. PMID:24715820

Systematic development of a two-stage fed-batch process for lipid accumulation in Rhodotorula glutinis.

PubMed

Lorenz, Eric; Runge, Dennis; Marbà-Ardébol, Anna-Maria; Schmacht, Maximilian; Stahl, Ulf; Senz, Martin

2017-03-20

The application of oleaginous yeast cells as feed supplement, for instance in aqua culture, can be a meaningful alternative for fish meal and oil additives. Therefore, a two-stage fed-batch process split into growth and lipogenesis phase was systematically developed to enrich the oleaginous yeast Rhodotorula glutinis Rh-00301 with high amounts of lipids at industrial relevant biomasses. Thereby, the different carbon sources glucose, sucrose and glycerol were investigated concerning their abilities to serve as a suited raw material for growth and/or lipid accumulation. With the background of economic efficiency C/N ratios of 40, 50 and 70 were investigated as well. It became apparent that glycerol is an improper carbon source most likely because of the passive diffusion of this compound caused by absence of active transporters. The opposite was observed for sucrose, which is the main carbon source in molasses. Finally, an industrially applicable process was successfully established that ensures biomasses of 106±2gL -1 combined with an attractive lipid content of 63±6% and a high lipid-substrate yield (Y L/S ) of 0.18±0.02gg -1 in a short period of time (84h). Furthermore, during these studies a non-negligible formation of the by-product glycerol was detected. This characteristic of R. glutinis is discussed related to other oleaginous yeasts, where glycerol formation is absent. Nevertheless, due to modifications in the feeding procedure, the formation of glycerol could have been reduced but not avoided. Copyright © 2017 Elsevier B.V. All rights reserved.

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.

Investigation of the performance of fermentation processes using a mathematical model including effects of metabolic bottleneck and toxic product on cells.

PubMed

Sriyudthsak, Kansuporn; Shiraishi, Fumihide

2010-11-01

A number of recent research studies have focused on theoretical and experimental investigation of a bottleneck in a metabolic reaction network. However, there is no study on how the bottleneck affects the performance of a fermentation process when a product is highly toxic and remarkably influences the growth and death of cells. The present work therefore studies the effect of bottleneck on product concentrations under different product toxicity conditions. A generalized bottleneck model in a fed-batch fermentation is constructed including both the bottleneck and the product influences on cell growth and death. The simulation result reveals that when the toxic product strongly influences the cell growth and death, the final product concentration is hardly changed even if the bottleneck is removed, whereas it is markedly changed by the degree of product toxicity. The performance of an ethanol fermentation process is also discussed as a case example to validate this result. In conclusion, when the product is highly toxic, one cannot expect a significant increase in the final product concentration even if removing the bottleneck; rather, it may be more effective to somehow protect the cells so that they can continuously produce the product. Copyright © 2010 Elsevier Inc. All rights reserved.

Understanding the interplay of carbon and nitrogen supply for ectoines production and metabolic overflow in high density cultures of Chromohalobacter salexigens.

PubMed

Salar-García, María J; Bernal, Vicente; Pastor, José M; Salvador, Manuel; Argandoña, Montserrat; Nieto, Joaquín J; Vargas, Carmen; Cánovas, Manuel

2017-02-08

The halophilic bacterium Chromohalobacter salexigens has been proposed as promising cell factory for the production of the compatible solutes ectoine and hydroxyectoine. This bacterium has evolved metabolic adaptations to efficiently grow under high salt concentrations by accumulating ectoines as compatible solutes. However, metabolic overflow, which is a major drawback for the efficient conversion of biological feedstocks, occurs as a result of metabolic unbalances during growth and ectoines production. Optimal production of ectoines is conditioned by the interplay of carbon and nitrogen metabolisms. In this work, we set out to determine how nitrogen supply affects the production of ectoines. Chromohalobacter salexigens was challenged to grow in media with unbalanced carbon/nitrogen ratio. In C. salexigens, overflow metabolism and ectoines production are a function of medium composition. At low ammonium conditions, the growth rate decreased importantly, up to 80%. Shifts in overflow metabolism were observed when changing the C/N ratio in the culture medium. 13 C-NMR analysis of ectoines labelling revealed a high metabolic rigidity, with almost constant flux ratios in all conditions assayed. Unbalanced C/N ratio led to pyruvate accumulation, especially upon N-limitation. Analysis of an ect - mutant demonstrated the link between metabolic overflow and ectoine biosynthesis. Under non ectoine synthesizing conditions, glucose uptake and metabolic overflow decreased importantly. Finally, in fed-batch cultures, biomass yield was affected by the feeding scheme chosen. High growth (up to 42.4 g L -1 ) and volumetric ectoine yields (up to 4.21 g L -1 ) were obtained by minimizing metabolite overflow and nutrient accumulation in high density cultures in a low nitrogen fed-batch culture. Moreover, the yield coefficient calculated for the transformation of glucose into biomass was 30% higher in fed-batch than in the batch culture, demonstrating that the metabolic efficiency of C. salexigens can be improved by careful design of culture feeding schemes. Metabolic shifts observed at low ammonium concentrations were explained by a shift in the energy required for nitrogen assimilation. Carbon-limited fed-batch cultures with reduced ammonium supply were the best conditions for cultivation of C. salexigens, supporting high density growth and maintaining high ectoines production.

Use of soybean oil and ammonium sulfate additions to optimize secondary metabolite production.

PubMed

Junker, B; Mann, Z; Gailliot, P; Byrne, K; Wilson, J

1998-12-05

A valine-overproducing mutant (MA7040, Streptomyces hygroscopicus) was found to produce 1.5 to 2.0 g/L of the immunoregulant, L-683,590, at the 0.6 m3 fermentation scale in a simple batch process using soybean oil and ammonium sulfate-based GYG5 medium. Levels of both lower (L-683,795) and higher (HH1 and HH2) undesirable homolog levels were controlled adequately. This batch process was utilized to produce broth economically at the 19 m3 fermentation scale. Material of acceptable purity was obtained without the multiple pure crystallizations previously required for an earlier culture, MA6678, requiring valine supplementation for impurity control. Investigations at the 0.6 m3 fermentation scale were conducted, varying agitation, pH, initial soybean oil/ammonium sulfate charges, and initial aeration rate to further improve growth and productivity. Mid-cycle ammonia levels and lipase activity appeared to have an important role. Using mid-cycle soybean oil additions, a titer of 2.3 g/L of L-683,590 was obtained, while titers reached 2.7 g/L using mid-cycle soybean oil and ammonium sulfate additions. Both higher and lower homolog levels remained acceptable during this fed-batch process. Optimal timing of mid-cycle oil and ammonium sulfate additions was considered a critical factor to further titer improvements. Copyright 1998 John Wiley & Sons, Inc.

Cultural, Transcriptomic, and Proteomic Analyses of Water-Stressed Cells of Actinobacterial Strains Isolated from Compost: Ecological Implications in the Fed-Batch Composting Process.

PubMed

Narihiro, Takashi; Kanosue, Yuji; Hiraishi, Akira

2016-06-25

This study was undertaken to examine the effects of water activity (aw) on the viability of actinobacterial isolates from a fed-batch composting (FBC) process by comparing culturability and stainability with 5-cyano-2,3-ditoryl tetrazolium chloride (CTC). The FBC reactor as the source of these bacteria was operated with the daily loading of household biowaste for 70 d. During this period of composting, aw in the reactor decreased linearly with time and reached approximately 0.95 at the end of operation. The plate counts of aerobic chemoorganotrophic bacteria were 3.2-fold higher than CTC-positive (CTC+) counts on average at the fully acclimated stage (after 7 weeks of operation), in which Actinobacteria predominated, as shown by lipoquinone profiling and cultivation methods. When the actinobacterial isolates from the FBC process were grown under aw stress, no significant differences were observed in culturability among the cultures, whereas CTC stainability decreased with reductions in aw levels. A cDNA microarray-based transcriptomic analysis of a representative isolate showed that many of the genes involved in cellular metabolism and genetic information processing were down-regulated by aw stress. This result was fully supported by a proteomic analysis. The results of the present study suggest that, in low aw mature compost, the metabolic activity of the community with Actinobacteria predominating is temporarily reduced to a level that hardly reacts with CTC; however, these bacteria are easily recoverable by exposure to a high aw culture medium. This may be a plausible reason why acclimated FBC reactors in which Actinobacteria predominate yields higher plate counts than CTC+ counts.

Successful hydraulic strategies to start up OLAND sequencing batch reactors at lab scale

PubMed Central

Schaubroeck, Thomas; Bagchi, Samik; De Clippeleir, Haydée; Carballa, Marta; Verstraete, Willy; Vlaeminck, Siegfried E.

2012-01-01

Summary Oxygen‐limited autotrophic nitrification/denitrification (OLAND) is a one‐stage combination of partial nitritation and anammox, which can have a challenging process start‐up. In this study, start‐up strategies were tested for sequencing batch reactors (SBR), varying hydraulic parameters, i.e. volumetric exchange ratio (VER) and feeding regime, and salinity. Two sequential tests with two parallel SBR were performed, and stable removal rates > 0.4 g N l−1 day−1 with minimal nitrite and nitrate accumulation were considered a successful start‐up. SBR A and B were operated at 50% VER with 3 g NaCl l−1 in the influent, and the influent was fed over 8% and 82% of the cycle time respectively. SBR B started up in 24 days, but SBR A achieved no start‐up in 39 days. SBR C and D were fed over 65% of the cycle time at 25% VER, and salt was added only to the influent of SBR D (5 g NaCl l−1). Start‐up of both SBR C and D was successful in 9 and 32 days respectively. Reactor D developed a higher proportion of small aggregates (0.10–0.25 mm), with a high nitritation to anammox rate ratio, likely the cause of the observed nitrite accumulation. The latter was overcome by temporarily including an anoxic period at the end of the reaction phase. All systems achieved granulation and similar biomass‐specific nitrogen removal rates (141–220 mg N g−1 VSS day−1). FISH revealed a close juxtapositioning of aerobic and anoxic ammonium‐oxidizing bacteria (AerAOB and AnAOB), also in small aggregates. DGGE showed that AerAOB communities had a lower evenness than Planctomycetes communities. A higher richness of the latter seemed to be correlated with better reactor performance. Overall, the fast start‐up of SBR B, C and D suggests that stable hydraulic conditions are beneficial for OLAND while increased salinity at the tested levels is not needed for good reactor performance. PMID:22236147

Successful hydraulic strategies to start up OLAND sequencing batch reactors at lab scale.

PubMed

Schaubroeck, Thomas; Bagchi, Samik; De Clippeleir, Haydée; Carballa, Marta; Verstraete, Willy; Vlaeminck, Siegfried E

2012-05-01

Oxygen-limited autotrophic nitrification/denitrification (OLAND) is a one-stage combination of partial nitritation and anammox, which can have a challenging process start-up. In this study, start-up strategies were tested for sequencing batch reactors (SBR), varying hydraulic parameters, i.e. volumetric exchange ratio (VER) and feeding regime, and salinity. Two sequential tests with two parallel SBR were performed, and stable removal rates > 0.4 g N l(-1) day(-1) with minimal nitrite and nitrate accumulation were considered a successful start-up. SBR A and B were operated at 50% VER with 3 g NaCl l(-1) in the influent, and the influent was fed over 8% and 82% of the cycle time respectively. SBR B started up in 24 days, but SBR A achieved no start-up in 39 days. SBR C and D were fed over 65% of the cycle time at 25% VER, and salt was added only to the influent of SBR D (5 g NaCl l(-1)). Start-up of both SBR C and D was successful in 9 and 32 days respectively. Reactor D developed a higher proportion of small aggregates (0.10-0.25 mm), with a high nitritation to anammox rate ratio, likely the cause of the observed nitrite accumulation. The latter was overcome by temporarily including an anoxic period at the end of the reaction phase. All systems achieved granulation and similar biomass-specific nitrogen removal rates (141-220 mg N g(-1) VSS day(-1)). FISH revealed a close juxtapositioning of aerobic and anoxic ammonium-oxidizing bacteria (AerAOB and AnAOB), also in small aggregates. DGGE showed that AerAOB communities had a lower evenness than Planctomycetes communities. A higher richness of the latter seemed to be correlated with better reactor performance. Overall, the fast start-up of SBR B, C and D suggests that stable hydraulic conditions are beneficial for OLAND while increased salinity at the tested levels is not needed for good reactor performance. © 2012 The Authors. Microbial Biotechnology © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.

A parametric study ot protease production in batch and fed-batch cultures of Bacillus firmus.

PubMed

Moon, S H; Parulekar, S J

1991-03-05

Proteolytic enzymes produced by Bacillus species find a wide variety of applications in brewing, detergent, food, and leather industries. Owing to significant differences normally observed in culture conditions promoting cell growth and those promoting production of metabolites such as enzymes, for increased efficacy of bioreactor operations it is essential to identify these sets of conditions (including medium formulation). This study is focused on formulation of a semidefined medium that substantially enhances synthesis and secretion of an alkaline protease in batch cultures of Bacillus firmus NRS 783, a known superior producer of this enzyme. The series of experiments conducted to identify culture conditions that lead to improved protease production also enables investigation of the regulatory effects of important culture parameters including pH, dissolved oxygen, and concentrations of nitrogen and phosphorous sources and yeast extract in the medium on cell growth, synthesis and secretion of protease, and production of two major nonbiomass products, viz., acetic acid and ethanol. Cell growth and formation of the three nonbiomass products are hampered significantly under nitrogen, phosphorous, or oxygen limitation, with the cells being unable to grow in an oxygen-free environment. Improvement in protease production is achieved with respect to each culture parameter, leading in the process to 80% enhancement in protease activity over that attained using media reported in the literature. Results of a few fed-batch experiments with constant feed rate, conducted to examine possible enhancement in protease production and to further investigate repression of protease synthesis by excess of the principal carbon and nitrogen sources, are also discussed. The detailed investigation of stimulatory and repressory effects of simple and complex nutrients on protease production and metabolism of Bacillus firmus conducted in this study will provide useful guidelines for design of bioreactors for production of protease and bulk chemicals by this bacterium.

Effect of different fermentation strategies on β-mannanase production in fed-batch bioreactor system.

PubMed

Germec, Mustafa; Yatmaz, Ercan; Karahalil, Ercan; Turhan, İrfan

2017-05-01

Mannanases, one of the important enzyme group for industry, are produced by numerous filamentous fungi, especially Aspergillus species with different fermentation methods. The aim of this study was to show the best fermentation method of β-mannanase production for fungal growth in fermenter. Therefore, different fermentation strategies in fed-batch fermentation (suspended, immobilized cell, biofilm and microparticle-enhanced bioreactor) were applied for β-mannanase production from glucose medium (GM) and carob extract medium (CEM) by using recombinant Aspergillus sojae. The highest β-mannanase activities were obtained from microparticle-enhanced bioreactor strategy. It was found to be 347.47 U/mL by adding 10 g/L of Al 2 O 3 to GM and 439.13 U/mL by adding 1 g/L of talcum into CEM. The maximum β-mannanase activities for suspended, immobilization, and biofilm reactor remained at 72.55 U/mL in GM, 148.81 U/mL in CEM, and 194.09 U/mL in GM, respectively. The reason for that is the excessive, and irregular shaped growth and bulk formation, inadequate oxygen transfer or substrate diffusion in bioreactor. Consequently, the enzyme activity was significantly enhanced by addition of microparticles compared to other fed-batch fermentation strategies. Also, repeatable β-mannanase activities were obtained by controlling of the cell morphology by adding microparticle inside the fermenter.

Stepwise heating in Stille polycondensation toward no batch-to-batch variations in polymer solar cell performance.

PubMed

Lee, Sang Myeon; Park, Kwang Hyun; Jung, Seungon; Park, Hyesung; Yang, Changduk

2018-05-14

For a given π-conjugated polymer, the batch-to-batch variations in molecular weight (M w ) and polydispersity index (Ð) can lead to inconsistent process-dependent material properties and consequent performance variations in the device application. Using a stepwise-heating protocol in the Stille polycondensation in conjunction with optimized processing, we obtained an ultrahigh-quality PTB7 polymer having high M w and very narrow Ð. The resulting ultrahigh-quality polymer-based solar cells demonstrate up to 9.97% power conversion efficiencies (PCEs), which is over 24% enhancement from the control devices fabricated with commercially available PTB7. Moreover, we observe almost negligible batch-to-batch variations in the overall PCE values from ultrahigh-quality polymer-based devices. The proposed stepwise polymerization demonstrates a facile and effective strategy for synthesizing high-quality semiconducting polymers that can significantly improve device yield in polymer-based solar cells, an important factor for the commercialization of organic solar cells, by mitigating device-to-device variations.

Production of the short peptide surfactant DAMP4 from glucose or sucrose in high cell density cultures of Escherichia coli BL21(DE3).

PubMed

Bruschi, Michele; Krömer, Jens O; Steen, Jennifer A; Nielsen, Lars K

2014-08-19

Peptides are increasingly used in industry as highly functional materials. Bacterial production of recombinant peptides has the potential to provide large amounts of renewable and low cost peptides, however, achieving high product titers from Chemically Defined Media (CDM) supplemented with simple sugars remains challenging. In this work, the short peptide surfactant, DAMP4, was used as a model peptide to investigate production in Escherichia coli BL21(DE3), a classical strain used for protein production. Under the same fermentation conditions, switching production of DAMP4 from rich complex media to CDM resulted in a reduction in yield that could be attributed to the reduction in final cell density more so than a significant reduction in specific productivity. To maximize product titer, cell density at induction was maximized using a fed-batch approach. In fed-batch DAMP4 product titer increased 9-fold compared to batch, while maintaining 60% specific productivity. Under the fed-batch conditions, the final product titer of DAMP4 reached more than 7 g/L which is the highest titer of DAMP4 reported to date. To investigate production from sucrose, sucrose metabolism was engineered into BL21(DE3) using a simple plasmid approach. Using this strain, growth and DAMP4 production characteristics obtained from CDM supplemented with sucrose were similar to those obtained when culturing the parent strain on CDM supplemented with glucose. Production of a model peptide was increased to several grams per liter using a CDM medium with either glucose or sucrose feedstock. It is hoped that this work will contribute cost reduction for production of designer peptide surfactants to facilitate their commercial application.

Continuous enzymatic hydrolysis of lignocellulosic biomass with simultaneous detoxification and enzyme recovery.

PubMed

Gurram, Raghu N; Menkhaus, Todd J

2014-07-01

Recovering hydrolysis enzymes and/or alternative enzyme addition strategies are two potential mechanisms for reducing the cost during the biochemical conversion of lignocellulosic materials into renewable biofuels and biochemicals. Here, we show that enzymatic hydrolysis of acid-pretreated pine wood with continuous and/or fed-batch enzyme addition improved sugar conversion efficiencies by over sixfold. In addition, specific activity of the hydrolysis enzymes (cellulases, hemicellulases, etc.) increased as a result of continuously washing the residual solids with removal of glucose (avoiding the end product inhibition) and other enzymatic inhibitory compounds (e.g., furfural, hydroxymethyl furfural, organic acids, and phenolics). As part of the continuous hydrolysis, anion exchange resin was tested for its dual application of simultaneous enzyme recovery and removal of potential enzymatic and fermentation inhibitors. Amberlite IRA-96 showed favorable adsorption profiles of inhibitors, especially furfural, hydroxymethyl furfural, and acetic acid with low affinity toward sugars. Affinity of hydrolysis enzymes to adsorb onto the resin allowed for up to 92 % of the enzymatic activity to be recovered using a relatively low-molar NaCl wash solution. Integration of an ion exchange column with enzyme recovery into the proposed fed-batch hydrolysis process can improve the overall biorefinery efficiency and can greatly reduce the production costs of lignocellulosic biorenewable products.

Engineering of solvent-tolerant Pseudomonas putida S12 for bioproduction of phenol from glucose.

PubMed

Wierckx, Nick J P; Ballerstedt, Hendrik; de Bont, Jan A M; Wery, Jan

2005-12-01

Efficient bioconversion of glucose to phenol via the central metabolite tyrosine was achieved in the solvent-tolerant strain Pseudomonas putida S12. The tpl gene from Pantoea agglomerans, encoding tyrosine phenol lyase, was introduced into P. putida S12 to enable phenol production. Tyrosine availability was a bottleneck for efficient production. The production host was optimized by overexpressing the aroF-1 gene, which codes for the first enzyme in the tyrosine biosynthetic pathway, and by random mutagenesis procedures involving selection with the toxic antimetabolites m-fluoro-dl-phenylalanine and m-fluoro-l-tyrosine. High-throughput screening of analogue-resistant mutants obtained in this way yielded a P. putida S12 derivative capable of producing 1.5 mM phenol in a shake flask culture with a yield of 6.7% (mol/mol). In a fed-batch process, the productivity was limited by accumulation of 5 mM phenol in the medium. This toxicity was overcome by use of octanol as an extractant for phenol in a biphasic medium-octanol system. This approach resulted in accumulation of 58 mM phenol in the octanol phase, and there was a twofold increase in the overall production compared to a single-phase fed batch.

Heat-treated (in single aliquot or batch) colostrum outperforms non-heat-treated colostrum in terms of quality and transfer of immunoglobulin G in neonatal Jersey calves.

PubMed

Kryzer, A A; Godden, S M; Schell, R

2015-03-01

The objective of this randomized clinical trial was to describe the effect on colostrum characteristics and passive transfer of IgG in neonatal calves when using the Perfect Udder colostrum management system (single-aliquot treatment; Dairy Tech Inc., Greeley, CO) compared with a negative control (fresh refrigerated or fresh frozen colostrum) and a positive control (batch heat-treated colostrum). First-milking Jersey colostrum was pooled to achieve 31 unique batches with a minimum of 22.8 L per batch. The batch was then divided into 4 with 3.8 L allocated to each treatment group: (1) heat-treated in Perfect Udder bag at 60°C for 60 min and then stored at -20°C (PU); (2) heat-treated in a batch pasteurizer (Dairy Tech Inc.) at 60°C for 60 min and then stored at -20°C in Perfect Udder bag (DTB; positive control); (3) fresh frozen colostrum stored at -20°C in Perfect Udder bag (FF; negative control); and (4) fresh refrigerated colostrum stored at 4°C in Perfect Udder bag (FR; negative control). Colostrum from all treatments was sampled for analysis of IgG concentration and bacterial culture immediately after batch assembly, after processing, and before feeding. Newborn Jersey calves were randomly assigned to be fed 3.8 L of colostrum from 1 of the 4 treatment groups. A prefeeding, 0-h blood sample was collected, calves were fed by esophageal tube within 2 h of birth, and then a 24-h postfeeding blood sample was collected. Paired serum samples from 0- and 24-h blood samples were analyzed for IgG concentration (mg/mL) using radial immunodiffusion analysis. The overall mean IgG concentration in colostrum was 77.9 g/L and was not affected by treatment. Prefeeding total plate counts (log10 cfu/mL) were significantly different for all 4 treatments and were lower for heat-treated colostrum (PU=4.23, DTB=3.63) compared with fresh colostrum (FF=5.68, FR=6.53). Total coliform counts (log10 cfu/mL) were also significantly different for all 4 treatments and were lower for heat-treated colostrum (PU=0.45, DTB=1.08) compared with fresh colostrum (FF=3.82, FR=4.80). Mean 24-h serum IgG concentrations were significantly higher for calves in the PU (41.0 mg/mL) and DTB (40.6 mg/mL) groups compared with FF (35.1 mg/mL) and FR (35.5 mg/mL) groups. Mean apparent efficiency of absorption of IgG was significantly higher for the PU (37%) and DTB (37%) groups compared with the FF (32%) and FR (32%) groups. Calves fed heat-treated colostrum (PU or DTB) experienced significantly improved AEA and serum IgG concentrations. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Proteomic Analysis of Host Cell Protein Dynamics in the Culture Supernatants of Antibody-Producing CHO Cells

PubMed Central

Park, Jin Hyoung; Jin, Jong Hwa; Lim, Myung Sin; An, Hyun Joo; Kim, Jong Won; Lee, Gyun Min

2017-01-01

Chinese hamster ovary (CHO) cells are the most common cell line used for the production of therapeutic proteins including monoclonal antibodies (mAbs). Host cell proteins (HCPs), secreted and released from lysed cells, accumulate extracellularly during the cultures of recombinant CHO (rCHO) cells, potentially impairing product quality. In an effort to maintain good mAb quality during the cultures, HCPs accumulated extracellularly in batch and fed-batch cultures of a mAb-producing rCHO cell line were identified and quantified by nanoflow liquid chromatography-tandem mass spectrometry, followed by their gene ontology and functional analysis. Due to higher cell concentration and longer culture duration, more HCPs were identified and quantitated in fed-batch culture (2145 proteins identified and 1673 proteins quantified) than in batch culture (1934 proteins identified and 1486 proteins quantified). Clustering analysis of HCPs showed that the concentration profiles of HCPs affecting mAb quality (Lgmn, Ctsd, Gbl1, and B4galt1) correlated with changes in mAb quality attributes such as aggregation, charge variants, and N-glycosylation during the cultures. Taken together, the dataset of HCPs obtained in this study provides insights into determining the appropriate target proteins to be removed during both the cultures and purification steps for ensuring good mAb quality. PMID:28281648

SBR treatment of olive mill wastewaters: dilution or pre-treatment?

PubMed

Farabegoli, G; Chiavola, A; Rolle, E

2012-01-01

The olive-oil extraction industry is an economically important activity for many countries of the Mediterranean Sea area, with Spain, Greece and Italy being the major producers. This activity, however, may represent a serious environmental problem due to the discharge of highly polluted effluents, usually referred to as 'olive mill wastewaters' (OMWs). They are characterized by high values of chemical oxygen demand (COD) (80-300 g/L), lipids, total polyphenols (TPP), tannins and other substances difficult to degrade. An adequate treatment before discharging is therefore required to reduce the pollutant load. The aim of the present paper was to evaluate performances of a biological process in a sequencing batch reactor (SBR) fed with pre-treated OMWs. Pre-treatment consisted of a combined acid cracking (AC) and granular activated carbon (GAC) adsorption process. The efficiency of the system was compared with that of an identical SBR fed with the raw wastewater only diluted. Combined AC and GAC adsorption was chosen to be used prior to the following biological process due to its capability of providing high removal efficiencies of COD and TPP and also appreciable improvement of biodegradability. Comparing results obtained with different influents showed that best performances of the SBR were obtained by feeding it with raw diluted OMWs (dOMWs) and at the lowest dilution ratio (1:25): in this case, the removal efficiencies were 90 and 76%, as average, for COD and TPP, respectively. Feeding the SBR with either the pre-treated or the raw dOMWs at 1:50 gave very similar values of COD reduction (74%); however, an improvement of the TPP removal was observed in the former case.

Lysine production from methanol at 50 degrees C using Bacillus methanolicus: Modeling volume control, lysine concentration, and productivity using a three-phase continuous simulation.

PubMed

Lee, G H; Hur, W; Bremmon, C E; Flickinger, M C

1996-03-20

A simulation was developed based on experimental data obtained in a 14-L reactor to predict the growth and L-lysine accumulation kinetics, and change in volume of a large-scale (250-m(3)) Bacillus methanolicus methanol-based process. Homoserine auxotrophs of B. methanolicus MGA3 are unique methylotrophs because of the ability to secrete lysine during aerobic growth and threonine starvation at 50 degrees C. Dissolved methanol (100 mM), pH, dissolved oxygen tension (0.063 atm), and threonine levels were controlled to obtain threonine-limited conditions and high-cell density (25 g dry cell weight/L) in a 14-L reactor. As a fed-batch process, the additions of neat methanol (fed on demand), threonine, and other nutrients cause the volume of the fermentation to increase and the final lysine concentration to decrease. In addition, water produced as a result of methanol metabolism contributes to the increase in the volume of the reactor. A three-phase approach was used to predict the rate of change of culture volume based on carbon dioxide production and methanol consumption. This model was used for the evaluation of volume control strategies to optimize lysine productivity. A constant volume reactor process with variable feeding and continuous removal of broth and cells (VF(cstr)) resulted in higher lysine productivity than a fed-batch process without volume control. This model predicts the variation in productivity of lysine with changes in growth and in specific lysine productivity. Simple modifications of the model allows one to investigate other high-lysine-secreting strains with different growth and lysine productivity characteristics. Strain NOA2#13A5-2 which secretes lysine and other end-products were modeled using both growth and non-growth-associated lysine productivity. A modified version of this model was used to simulate the change in culture volume of another L-lysine producing mutant (NOA2#13A52-8A66) with reduced secretion of end-products. The modified simulation indicated that growth-associated production dominates in strain NOA2#13A52-8A66. (c) 1996 John Wiley & Sons, Inc.

Effects of chlortetracycline amended feed on anaerobic sequencing batch reactor performance of swine manure digestion.

PubMed

Dreher, Teal M; Mott, Henry V; Lupo, Christopher D; Oswald, Aaron S; Clay, Sharon A; Stone, James J

2012-12-01

The effects of antimicrobial chlortetracycline (CTC) on the anaerobic digestion (AD) of swine manure slurry using anaerobic sequencing batch reactors (ASBRs) was investigated. Reactors were loaded with manure collected from pigs receiving CTC and no-antimicrobial amended diets at 2.5 g/L/d. The slurry was intermittently fed to four 9.5L lab-scale anaerobic sequencing batch reactors, two with no-antimicrobial manure, and two with CTC-amended manure, and four 28 day ASBR cycles were completed. The CTC concentration within the manure was 2 8 mg/L immediately after collection and 1.02 mg/L after dilution and 250 days of storage. CTC did not inhibit ASBR biogas production extent, however the volumetric composition of methane was significantly less (approximately 13% and 15% for cycles 1 and 2, respectively) than the no-antimicrobial through 56 d. CTC decreased soluble chemical oxygen demand and acetic acid utilization through 56 d, after which acclimation to CTC was apparent for the duration of the experiment. Copyright © 2012 Elsevier Ltd. All rights reserved.

HPLC-ESI-MS(n) Analysis, Fed-Batch Cultivation Enhances Bioactive Compound Biosynthesis and Immune-Regulative Effect of Adventitious Roots in Pseudostellaria heterophylla.

PubMed

Wang, Juan; Li, Jing; Li, Hongfa; Wu, Xiaolei; Gao, Wenyuan

2015-09-01

A electrospray ionization tandem mass spectrometry (ESI-MS(n)) analysis was performed in order to identify the active composition in Pseudostellaria heterophylla adventitious roots. Pseudostellarin A, C, D, and G were identified from P. heterophylla adventitious roots on the basis of LC-MS(n) analysis. The culture conditions of adventitious roots were optimized, and datasets were subjected to a partial least squares discriminant analysis (PLS-DA), in which the growth ratio and some compounds showed a positive correlation with an aeration volume of 0.3 vvm and inoculum density of 0.15 %. Fed-batch cultivation enhanced the contents of total saponin, polysaccharides, and specific oxygen uptaker rate (SOUR). The maximum dry root weight (4.728 g l(-1)) was achieved in the 3/4 Murashige and Skoog (MS) medium group. PLS-DA showed that polysaccharides contributed significantly to the clustering of different groups and showed a positive correlation in the MS medium group. The delayed-type hypersensitivity (DTH) reaction on the mice induced by 2,4-dinitrofluorobenzene (DNFB) was applied to compare the immunocompetence effects of adventitious roots (AR) with field native roots (NR) of P. heterophylla. As a result, AR possessed a similar immunoregulation function as NR.

Shifting the balance of fermentation products between hydrogen and volatile fatty acids: microbial community structure and function.

PubMed

Miceli, Joseph F; Torres, César I; Krajmalnik-Brown, Rosa

2016-12-01

Fermentation is a key process in many anaerobic environments. Varying the concentration of electron donor fed to a fermenting community is known to shift the distribution of products between hydrogen, fatty acids and alcohols. Work to date has focused mainly on the fermentation of glucose, and how the microbial community structure is affected has not been explored. We fed ethanol, lactate, glucose, sucrose or molasses at 100 me- eq. L -1 , 200 me- eq. L -1 or 400 me- eq. L -1 to batch-fed cultures with fermenting, methanogenic communities. In communities fed high concentrations of electron donor, the fraction of electrons channeled to methane decreased, from 34% to 6%, while the fraction of electrons channeled to short chain fatty acids increased, from 52% to 82%, averaged across all electron donors. Ethanol-fed cultures did not produce propionate, but did show an increase in electrons directed to acetate as initial ethanol concentration increased. In glucose, sucrose, molasses and lactate-fed cultures, propionate accumulation co-occurred with known propionate producing organisms. Overall, microbial communities were determined by the substrate provided, rather than its initial concentration, indicating that a change in community function, rather than community structure, is responsible for shifts in the fermentation products produced. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

How Many Batches Are Needed for Process Validation under the New FDA Guidance?

PubMed

Yang, Harry

2013-01-01

The newly updated FDA Guidance for Industry on Process Validation: General Principles and Practices ushers in a life cycle approach to process validation. While the guidance no longer considers the use of traditional three-batch validation appropriate, it does not prescribe the number of validation batches for a prospective validation protocol, nor does it provide specific methods to determine it. This potentially could leave manufacturers in a quandary. In this paper, I develop a Bayesian method to address the issue. By combining process knowledge gained from Stage 1 Process Design (PD) with expected outcomes of Stage 2 Process Performance Qualification (PPQ), the number of validation batches for PPQ is determined to provide a high level of assurance that the process will consistently produce future batches meeting quality standards. Several examples based on simulated data are presented to illustrate the use of the Bayesian method in helping manufacturers make risk-based decisions for Stage 2 PPQ, and they highlight the advantages of the method over traditional Frequentist approaches. The discussions in the paper lend support for a life cycle and risk-based approach to process validation recommended in the new FDA guidance. The newly updated FDA Guidance for Industry on Process Validation: General Principles and Practices ushers in a life cycle approach to process validation. While the guidance no longer considers the use of traditional three-batch validation appropriate, it does not prescribe the number of validation batches for a prospective validation protocol, nor does it provide specific methods to determine it. This potentially could leave manufacturers in a quandary. In this paper, I develop a Bayesian method to address the issue. By combining process knowledge gained from Stage 1 Process Design (PD) with expected outcomes of Stage 2 Process Performance Qualification (PPQ), the number of validation batches for PPQ is determined to provide a high level of assurance that the process will consistently produce future batches meeting quality standards. Several examples based on simulated data are presented to illustrate the use of the Bayesian method in helping manufacturers make risk-based decisions for Stage 2 PPQ, and THEY highlight the advantages of the method over traditional Frequentist approaches. The discussions in the paper lend support for a life cycle and risk-based approach to process validation recommended in the new FDA guidance.

Effects of sludge retention time, carbon and initial biomass concentrations on selection process: From activated sludge to polyhydroxyalkanoate accumulating cultures.

PubMed

Chen, Zhiqiang; Huang, Long; Wen, Qinxue; Zhang, Huichao; Guo, Zirui

2017-02-01

Four sequence batch reactors (SBRs) fed by fermented sugar cane wastewater were continuously operated under the aerobic dynamic feeding (ADF) mode with different configurations of sludge retention time (SRT), carbon and initial biomass concentrations to enrich polyhydroxyalkanoate (PHA) accumulating mixed microbial cultures (MMCs) from municipal activated sludge. The stability of SBRs was investigated besides the enrichment performance. The microbial community structures of the enriched MMCs were analyzed using terminal restriction fragment length polymorphism (T-RFLP). The optimum operating conditions for the enrichment process were: SRT of 5days, carbon concentration of 2.52g COD/L and initial biomass concentration of 3.65g/L. The best enrichment performance in terms of both operating stability and PHA storage ability of enriched cultures (with the maximum PHA content and PHA storage yield (Y PHA/S ) of 61.26% and 0.68mg COD/mg COD, respectively) was achieved under this condition. Effects of the SRT, carbon concentration and initial biomass concentration on the PHA accumulating MMCs selection process were discussed respectively. A new model including the segmentation of the enrichment process and the effects of SRT on each phase was proposed. Copyright © 2016. Published by Elsevier B.V.

Novel integration strategy for enhancing chalcopyrite bioleaching by Acidithiobacillus sp. in a 7-L fermenter.

PubMed

Feng, Shoushuai; Yang, Hailin; Zhan, Xiao; Wang, Wu

2014-06-01

An integrated strategy (additional energy substrate-three stage pH control-fed batch) was firstly proposed for efficiently improving chalcopyrite bioleaching by Acidithiobacillus sp. in a 7-L fermenter. The strain adaptive-growing phase was greatly shortened from 8days into 4days with the supplement of additional 2g/L Fe(2+)+2g/L S(0). Jarosite passivation was effectively weakened basing on higher biomass via the three-stage pH-stat control (pH 1.3-1.0-0.7). The mineral substrate inhibition was attenuated by fed-batch fermentation. With the integrated strategy, the biochemical reaction was promoted and achieved a better balance. Meanwhile, the domination course of A. thiooxidans in the microbial community was shortened from 14days to 8days. As the results of integrated strategy, the final copper ion and productivity reached 89.1mg/L and 2.23mg/(Ld), respectively, which was improved by 52.8% compared to the uncontrolled batch bioleaching. The integrated strategy could be further exploited for industrial chalcopyrite bioleaching. Copyright © 2014 Elsevier Ltd. All rights reserved.

Batch Tests To Determine Activity Distribution and Kinetic Parameters for Acetate Utilization in Expanded-Bed Anaerobic Reactors

PubMed Central

Fox, Peter; Suidan, Makram T.

1990-01-01

Batch tests to measure maximum acetate utilization rates were used to determine the distribution of acetate utilizers in expanded-bed sand and expanded-bed granular activated carbon (GAC) reactors. The reactors were fed a mixture of acetate and 3-ethylphenol, and they contained the same predominant aceticlastic methanogen, Methanothrix sp. Batch tests were performed both on the entire reactor contents and with media removed from the reactors. Results indicated that activity was evenly distributed within the GAC reactors, whereas in the sand reactor a sludge blanket on top of the sand bed contained approximately 50% of the activity. The Monod half-velocity constant (Ks) for the acetate-utilizing methanogens in two expanded-bed GAC reactors was searched for by combining steady-state results with batch test data. All parameters necessary to develop a model with Monod kinetics were experimentally determined except for Ks. However, Ks was a function of the effluent 3-ethylphenol concentration, and batch test results demonstrated that maximum acetate utilization rates were not a function of the effluent 3-ethylphenol concentration. Addition of a competitive inhibition term into the Monod expression predicted the dependence of Ks on the effluent 3-ethylphenol concentration. A two-parameter search determined a Ks of 8.99 mg of acetate per liter and a Ki of 2.41 mg of 3-ethylphenol per liter. Model predictions were in agreement with experimental observations for all effluent 3-ethylphenol concentrations. Batch tests measured the activity for a specific substrate and determined the distribution of activity in the reactor. The use of steady-state data in conjunction with batch test results reduced the number of unknown kinetic parameters and thereby reduced the uncertainty in the results and the assumptions made. PMID:16348175

Batch tests to determine activity distribution and kinetic parameters for acetate utilization in expanded-bed anaerobic reactors.

PubMed

Fox, P; Suidan, M T

1990-04-01

Batch tests to measure maximum acetate utilization rates were used to determine the distribution of acetate utilizers in expanded-bed sand and expanded-bed granular activated carbon (GAC) reactors. The reactors were fed a mixture of acetate and 3-ethylphenol, and they contained the same predominant aceticlastic methanogen, Methanothrix sp. Batch tests were performed both on the entire reactor contents and with media removed from the reactors. Results indicated that activity was evenly distributed within the GAC reactors, whereas in the sand reactor a sludge blanket on top of the sand bed contained approximately 50% of the activity. The Monod half-velocity constant (K(s)) for the acetate-utilizing methanogens in two expanded-bed GAC reactors was searched for by combining steady-state results with batch test data. All parameters necessary to develop a model with Monod kinetics were experimentally determined except for K(s). However, K(s) was a function of the effluent 3-ethylphenol concentration, and batch test results demonstrated that maximum acetate utilization rates were not a function of the effluent 3-ethylphenol concentration. Addition of a competitive inhibition term into the Monod expression predicted the dependence of K(s) on the effluent 3-ethylphenol concentration. A two-parameter search determined a K(s) of 8.99 mg of acetate per liter and a K(i) of 2.41 mg of 3-ethylphenol per liter. Model predictions were in agreement with experimental observations for all effluent 3-ethylphenol concentrations. Batch tests measured the activity for a specific substrate and determined the distribution of activity in the reactor. The use of steady-state data in conjunction with batch test results reduced the number of unknown kinetic parameters and thereby reduced the uncertainty in the results and the assumptions made.

Adaptations in bacterial catabolic enzyme activity and community structure in membrane-coupled bioreactors fed simple synthetic wastewater.

PubMed

LaPara, Timothy M; Klatt, Christian G; Chen, Ruoyu

2006-02-10

Membrane-coupled bioreactors (MBRs) offer substantial benefits compared to conventional reactor designs for biological wastewater treatment. MBR treatment efficiency, however, has not been optimized because the effects of the MBR on process microbiology are poorly understood. In this study, the structure and function of the microbial communities growing in MBRs fed simple synthetic wastewater were investigated. In four starch-fed MBRs, the bacterial community substantially increased its alpha-glucosidase affinity (>1000-fold), while the leucine aminopeptidase and heptanoate esterase affinities increased slightly (<40-fold) or remained relatively constant. Concomitant to these physiological adaptations, shifts in the bacterial community structure in two of the starch-fed MBRs were detected by PCR-DGGE. Four of the bacterial populations detected by PCR-DGGE were isolated and exhibited specific growth rates in batch culture ranging from 0.009 to 0.22 h(-1). Our results suggest that bacterial communities growing under increasingly stringent nutrient limitation adapt their enzyme activities primarily for the nutrients provided, but that there is also a more subtle response not linked to the substrates included in the feed medium. Our research also demonstrates that MBRs can support relatively complex bacterial communities even on simple feed media.

Production of citric acid using its extraction wastewater treated by anaerobic digestion and ion exchange in an integrated citric acid-methane fermentation process.

PubMed

Xu, Jian; Chen, Yang-Qiu; Zhang, Hong-Jian; Tang, Lei; Wang, Ke; Zhang, Jian-Hua; Chen, Xu-Sheng; Mao, Zhong-Gui

2014-08-01

In order to solve the problem of extraction wastewater pollution in citric acid industry, an integrated citric acid-methane fermentation process is proposed in this study. Extraction wastewater was treated by mesophilic anaerobic digestion and then used to make mash for the next batch of citric acid fermentation. The recycling process was done for seven batches. Citric acid production (82.4 g/L on average) decreased by 34.1 % in the recycling batches (2nd-7th) compared with the first batch. And the residual reducing sugar exceeded 40 g/L on average in the recycling batches. Pigment substances, acetic acid, ammonium, and metal ions in anaerobic digestion effluent (ADE) were considered to be the inhibitors, and their effects on the fermentation were studied. Results indicated that ammonium, Na(+) and K(+) in the ADE significantly inhibited citric acid fermentation. Therefore, the ADE was treated by acidic cation exchange resin prior to reuse to make mash for citric acid fermentation. The recycling process was performed for ten batches, and citric acid productions in the recycling batches were 126.6 g/L on average, increasing by 1.7 % compared with the first batch. This process could eliminate extraction wastewater discharge and reduce water resource consumption.

A framework supporting the development of a Grid portal for analysis based on ROI.

PubMed

Ichikawa, K; Date, S; Kaishima, T; Shimojo, S

2005-01-01

In our research on brain function analysis, users require two different simultaneous types of processing: interactive processing to a specific part of data and high-performance batch processing to an entire dataset. The difference between these two types of processing is in whether or not the analysis is for data in the region of interest (ROI). In this study, we propose a Grid portal that has a mechanism to freely assign computing resources to the users on a Grid environment according to the users' two different types of processing requirements. We constructed a Grid portal which integrates interactive processing and batch processing by the following two mechanisms. First, a job steering mechanism controls job execution based on user-tagged priority among organizations with heterogeneous computing resources. Interactive jobs are processed in preference to batch jobs by this mechanism. Second, a priority-based result delivery mechanism that administrates a rank of data significance. The portal ensures a turn-around time of interactive processing by the priority-based job controlling mechanism, and provides the users with quality of services (QoS) for interactive processing. The users can access the analysis results of interactive jobs in preference to the analysis results of batch jobs. The Grid portal has also achieved high-performance computation of MEG analysis with batch processing on the Grid environment. The priority-based job controlling mechanism has been realized to freely assign computing resources to the users' requirements. Furthermore the achievement of high-performance computation contributes greatly to the overall progress of brain science. The portal has thus made it possible for the users to flexibly include the large computational power in what they want to analyze.

Risk-based Methodology for Validation of Pharmaceutical Batch Processes.

PubMed

Wiles, Frederick

2013-01-01

In January 2011, the U.S. Food and Drug Administration published new process validation guidance for pharmaceutical processes. The new guidance debunks the long-held industry notion that three consecutive validation batches or runs are all that are required to demonstrate that a process is operating in a validated state. Instead, the new guidance now emphasizes that the level of monitoring and testing performed during process performance qualification (PPQ) studies must be sufficient to demonstrate statistical confidence both within and between batches. In some cases, three qualification runs may not be enough. Nearly two years after the guidance was first published, little has been written defining a statistical methodology for determining the number of samples and qualification runs required to satisfy Stage 2 requirements of the new guidance. This article proposes using a combination of risk assessment, control charting, and capability statistics to define the monitoring and testing scheme required to show that a pharmaceutical batch process is operating in a validated state. In this methodology, an assessment of process risk is performed through application of a process failure mode, effects, and criticality analysis (PFMECA). The output of PFMECA is used to select appropriate levels of statistical confidence and coverage which, in turn, are used in capability calculations to determine when significant Stage 2 (PPQ) milestones have been met. The achievement of Stage 2 milestones signals the release of batches for commercial distribution and the reduction of monitoring and testing to commercial production levels. Individuals, moving range, and range/sigma charts are used in conjunction with capability statistics to demonstrate that the commercial process is operating in a state of statistical control. The new process validation guidance published by the U.S. Food and Drug Administration in January of 2011 indicates that the number of process validation batches or runs required to demonstrate that a pharmaceutical process is operating in a validated state should be based on sound statistical principles. The old rule of "three consecutive batches and you're done" is no longer sufficient. The guidance, however, does not provide any specific methodology for determining the number of runs required, and little has been published to augment this shortcoming. The paper titled "Risk-based Methodology for Validation of Pharmaceutical Batch Processes" describes a statistically sound methodology for determining when a statistically valid number of validation runs has been acquired based on risk assessment and calculation of process capability.

Integrated continuous bioprocessing: Economic, operational, and environmental feasibility for clinical and commercial antibody manufacture.

PubMed

Pollock, James; Coffman, Jon; Ho, Sa V; Farid, Suzanne S

2017-07-01

This paper presents a systems approach to evaluating the potential of integrated continuous bioprocessing for monoclonal antibody (mAb) manufacture across a product's lifecycle from preclinical to commercial manufacture. The economic, operational, and environmental feasibility of alternative continuous manufacturing strategies were evaluated holistically using a prototype UCL decisional tool that integrated process economics, discrete-event simulation, environmental impact analysis, operational risk analysis, and multiattribute decision-making. The case study focused on comparing whole bioprocesses that used either batch, continuous or a hybrid combination of batch and continuous technologies for cell culture, capture chromatography, and polishing chromatography steps. The cost of goods per gram (COG/g), E-factor, and operational risk scores of each strategy were established across a matrix of scenarios with differing combinations of clinical development phase and company portfolio size. The tool outputs predict that the optimal strategy for early phase production and small/medium-sized companies is the integrated continuous strategy (alternating tangential flow filtration (ATF) perfusion, continuous capture, continuous polishing). However, the top ranking strategy changes for commercial production and companies with large portfolios to the hybrid strategy with fed-batch culture, continuous capture and batch polishing from a COG/g perspective. The multiattribute decision-making analysis highlighted that if the operational feasibility was considered more important than the economic benefits, the hybrid strategy would be preferred for all company scales. Further considerations outside the scope of this work include the process development costs required to adopt continuous processing. © 2017 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers Biotechnol. Prog., 33:854-866, 2017. © 2017 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers.

Optimization of medium components and cultural variables for enhanced production of acidic high maltose-forming and Ca2+-independent α-amylase by Bacillus acidicola.

PubMed

Sharma, Archana; Satyanarayana, Tulasi

2011-05-01

The production of acidic α-amylase by a novel acidophilic bacterium Bacillus acidicola TSAS1 was optimized in submerged fermentation using statistical approaches. The process parameters that significantly affected α-amylase production (starch, K(2)HPO(4), inoculum size and temperature) were identified by Plackett and Burman design. The optimum levels of the significant variables as determined using central composite design of response surface methodology are starch (2.75%), K(2)HPO(4) (0.01%), inoculum size [2% (v/v) containing 1.9×10(8) CFU ml(-1)], and temperature (33°C). An overall 2.4 and 2.9-fold increase in enzyme production has been attained in batch and fed-batch fermentations in the laboratory fermentor, respectively. Copyright © 2011 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Impact of different spray-drying conditions on the viability of wine Saccharomyces cerevisiae strains.

PubMed

Aponte, Maria; Troianiello, Gabriele Danilo; Di Capua, Marika; Romano, Raffaele; Blaiotta, Giuseppe

2016-01-01

Spray-drying (SD) is widely considered a suitable method to preserve microorganisms, but data regarding yeasts are still scanty. In this study, the effect of growing media, process variables and carriers over viability of a wild wine Saccharomyces (S.) cerevisiae LM52 was evaluated. For biomass production, the strain was grown (batch and fed-batch fermentation) in a synthetic, as well as in a beet sugar molasses based-medium. Drying of cells resuspended in several combinations of soluble starch and maltose was performed at different inlet and outlet temperatures. Under the best conditions-suspension in soluble starch plus maltose couplet to inlet and outlet temperatures of 110 and 55 °C, respectively-the loss of viability of S. cerevisiae LM52 was 0.8 ± 0.1 and 0.5 ± 0.2 Log c.f.u. g(-1) for synthetic and molasses-based medium, respectively. Similar results were obtained when S. cerevisiae strains Zymoflore F15 and EC1118, isolated from commercial active dry yeast (ADY), were tested. Moreover, powders retained a high vitality and showed good fermentation performances up to 6 month of storage, at both 4 and -20 °C. Finally, fermentation performances of different kinds of dried formulates (SD and ADY) compared with fresh cultures did not show significant differences. The procedure proposed allowed a small-scale production of yeast in continuous operation with relatively simple equipment, and may thus represent a rapid response-on-demand for the production of autochthonous yeasts for local wine-making.

The microbiological safety of duckweed fed chickens: a risk assessment of using duckweed reared on domestic wastewater as a protein source in broiler chickens

NASA Astrophysics Data System (ADS)

Moyo, S.; Dalu, J. M.; Ndamba, J.

The possibility of transmission of pathogens from duckweed supplemented feed to chickens and consequently to the human consumer necessitated the microbiological testing of duckweed fed chickens. This assessment was thus done to determine whether there is transmission of pathogens from the duckweed supplemented feed to the chickens; determine whether such infection would be systemic or be confined to the gastro-intestinal tract of the birds; and to investigate the microbial load and distribution of the microbes with age. The study birds were sacrificed at 3, 6, 8 and 10 weeks of age and examined for the indicator organisms Escherichia coli and Salmonella spp. There was no discernible pattern in the microbial load of both the duckweed fed chickens and control birds with age although the control birds sampled clearly had a lower microbial load than the experimental flock. Some Salmonella and two enteropathogenic E. coli strains were isolated from control and experimental sub-samples at 3 weeks. There were no Salmonellae isolated in the subsequent batches of birds and feed although a number of E. coli were isolated. More isolates were obtained from the three weeks’ sub-samples (collected during wet weather) than from all the other sub-samples. The use of duckweed at this inclusion rate under the processing conditions at Nemanwa was thus concluded to be microbiologically safe as long as due caution is exercised during the processing of the duckweed and handling of the birds. There are indications that the chickens may get contaminated especially during wet weather as evidenced by the isolation of E. coli and Salmonella spp from the first batch sub-samples. This was attributed to poor environmental sanitation at the plant particularly in view of the prevailing wet conditions at the time.

Impact of volatile fatty acids on microbial electrolysis cell performance.

PubMed

Yang, Nan; Hafez, Hisham; Nakhla, George

2015-10-01

This study investigated the performance of microbial electrolysis cells (MECs) fed with three common fermentation products: acetate, butyrate, and propionate. Each substrate was fed to the reactor for three consecutive-batch cycles. The results showed high current densities for acetate, but low current densities for butyrate and propionate (maximum values were 6.0 ± 0.28, 2.5 ± 0.06, 1.6 ± 0.14 A/m(2), respectively). Acetate also showed a higher coulombic efficiency of 87 ± 5.7% compared to 72 ± 2.0 and 51 ± 6.4% for butyrate and propionate, respectively. This paper also revealed that acetate could be easily oxidized by anode respiring bacteria in MEC, while butyrate and propionate could not be oxidized to the same degree. The utilization rate of the substrates in MEC followed the order: acetate > butyrate > propionate. The ratio of suspended biomass to attached biomass was approximately 1:4 for all the three substrates. Copyright © 2015 Elsevier Ltd. All rights reserved.

Influence of iron precipitated condition and light intensity on microalgae activated sludge based wastewater remediation.

PubMed

Anbalagan, Anbarasan; Schwede, Sebastian; Lindberg, Carl-Fredrik; Nehrenheim, Emma

2017-02-01

The indigenous microalgae-activated sludge (MAAS) process during remediation of municipal wastewater was investigated by studying the influence of iron flocculation step and light intensity. In addition, availability of total phosphorous (P) and photosynthetic activity was examined in fed-batch and batch mode under northern climatic conditions and limited lighting. This was followed by a semi-continuous operation with 4 d of hydraulic retention time and mean cell residence time of 6.75 d in a photo-bioreactor (PBR) with varying P availability. The fed-batch condition showed that P concentrations of 3-4 mg L -1 were effective for photosynthetic chl. a development in iron flocculated conditions. In the PBR, the oxygen evolution rate increased with increase in the concentration of MAAS (from 258 to 573 mg TSS L -1 ) at higher surface photosynthetic active radiation (250 and 500 μmol m -2 s -1 ). Additionally, the rate approached a saturation phase at low MAAS (110 mg L -1 ) with higher light intensities. Semi-continuous operation with luxury P uptake and effective P condition showed stable average total nitrogen removal of 88 and 92% respectively, with residual concentrations of 3.77 and 2.21 mg L -1 . The corresponding average P removal was 68 and 59% with residual concentrations of 2.32 and 1.75 mg L -1 . The semi-continuous operation produced a rapidly settleable MAAS under iron flocculated condition with a settling velocity of 92-106 m h -1 and sludge volume index of 31-43 ml g -1 in the studied cases. Copyright © 2016 Elsevier Ltd. All rights reserved.

[Monitoring method for macroporous resin column chromatography process of salvianolic acids based on near infrared spectroscopy].

PubMed

Hou, Xiang-Mei; Zhang, Lei; Yue, Hong-Shui; Ju, Ai-Chun; Ye, Zheng-Liang

2016-07-01

To study and establish a monitoring method for macroporous resin column chromatography process of salvianolic acids by using near infrared spectroscopy (NIR) as a process analytical technology (PAT).The multivariate statistical process control (MSPC) model was developed based on 7 normal operation batches, and 2 test batches (including one normal operation batch and one abnormal operation batch) were used to verify the monitoring performance of this model. The results showed that MSPC model had a good monitoring ability for the column chromatography process. Meanwhile, NIR quantitative calibration model was established for three key quality indexes (rosmarinic acid, lithospermic acid and salvianolic acid B) by using partial least squares (PLS) algorithm. The verification results demonstrated that this model had satisfactory prediction performance. The combined application of the above two models could effectively achieve real-time monitoring for macroporous resin column chromatography process of salvianolic acids, and can be used to conduct on-line analysis of key quality indexes. This established process monitoring method could provide reference for the development of process analytical technology for traditional Chinese medicines manufacturing. Copyright© by the Chinese Pharmaceutical Association.

Gut-Bioreactor and Human Health in Future.

PubMed

Purohit, Hemant J

2018-03-01

Gut-microbiome provides the complementary metabolic potential to the human system. To understand the active participation and the performance of the microbial community in human health, the concept of gut as a plug-flow reactor with the fed-batch mode of operation can provide better insight. The concept suggests the virtual compartmentalized gut with sequential stratification of the microbial community in response to a typical host genotype. It also provides the analysis plan for gut microbiome; and its relevance in developing health management options under the identified clinical conditions.

Animal performance and meat characteristics in steers reared in intensive conditions fed with different vegetable oils.

PubMed

Castro, T; Cabezas, A; De la Fuente, J; Isabel, B; Manso, T; Jimeno, V

2016-03-01

Enhancing the quality of beef meat is an important goal in terms of improving both the nutritional value for the consumer and the commercial value for producers. The aim of this work was to study the effects of different vegetable oil supplements on growth performance, carcass quality and meat quality in beef steers reared under intensive conditions. A total of 240 Blonde D' Aquitaine steers (average BW=293.7±38.88 kg) were grouped into 24 batches (10 steers/batch) and were randomly assigned to one of the three dietary treatments (eight batches per treatment), each supplemented with either 4% hydrogenated palm oil (PALM) or fatty acids (FAs) from olive oil (OLI) or soybean oil (SOY). No differences in growth performance or carcass quality were observed. For the meat quality analysis, a steer was randomly selected from each batch and the 6th rib on the left half of the carcass was dissected. PALM meat had the highest percentage of 16:0 (P<0.05) and the lowest n-6/n-3 polyunsaturated fatty acids (PUFA) ratio (P<0.05), OLI had the highest content of t11-18:1 (P<0.01) and c9,t11-18:2 (P<0.05) and SOY showed the lowest value of monounsaturated fatty acids (MUFA) (P<0.001), the highest percentage of PUFA (P<0.01) and a lower index of atherogenicity (P=0.07) than PALM. No significant differences in the sensory characteristics of the meat were noted. However, the results of the principal component analysis of meat characteristics enabled meat from those steers that consumed fatty acids from olive oil to be differentiated from that of steers that consumed soybean oil.

Enhancement of thermoalkaliphilic xylanase production by Pichia pastoris through novel fed-batch strategy in high cell-density fermentation.

PubMed

Shang, Tingting; Si, Dayong; Zhang, Dongyan; Liu, Xuhui; Zhao, Longmei; Hu, Cong; Fu, Yu; Zhang, Rijun

2017-06-21

Xylanase degrades xylan into monomers of various sizes by catalyzing the endohydrolysis of the 1,4-β-D-xylosidic linkage randomly, possessing potential in wide industrial applications. Most of xylanases are susceptible to be inactive when suffering high temperature and high alkaline process. Therefore, it is necessary to develop a high amount of effective thermoalkaliphilic xylanases. This study aims to enhance thermoalkaliphilic xylanase production in Pichia pastoris through fermentation parameters optimization and novel efficient fed-batch strategy in high cell-density fermentation. Recombinant xylanase activity increased 12.2%, 7.4%, 12.0% and 9.9% by supplementing the Pichia pastoris culture with 20 g/L wheat bran, 5 mg/L L-histidine, 10 mg/L L-tryptophan and 10 mg/L L-methionine in shake flasks, respectively. Investigation of nutritional fermentation parameters, non-nutritional fermentation parameters and feeding strategies in 1 L bioreactor and 1 L shake flask revealed that glycerol and methanol feeding strategies were the critical factors for high cell density and xylanase activity. In 50 L bioreactor, a novel glycerol feeding strategy and a four-stage methanol feeding strategy with a stepwise increase in feeding rate were developed to enhance recombinant xylanase production. In the initial 72 h of methanol induction, the linear dependence of xylanase activity on methanol intake was observed (R 2  = 0.9726). The maximum xylanase activity was predicted to be 591.2 U/mL, while the actual maximum xylanase activity was 560.7 U/mL, which was 7.05 times of that in shake flask. Recombinant xylanase retained 82.5% of its initial activity after pre-incubation at 80 °C for 50 min (pH 8.0), and it exhibited excellent stability in the broad temperature (60-80 °C) and pH (pH 8.0-11.0) ranges. Efficient glycerol and methanol fed-batch strategies resulting in desired cell density and xylanase activity should be applied in other P. pastoris fermentation for other recombinant proteins production. Recombinant xylanases with high pH- and thermal-stability showed potential in various industrial applications.

In silico aided metabolic engineering of Klebsiella oxytoca and fermentation optimization for enhanced 2,3-butanediol production.

PubMed

Park, Jong Myoung; Song, Hyohak; Lee, Hee Jong; Seung, Doyoung

2013-09-01

Klebsiella oxytoca naturally produces a large amount of 2,3-butanediol (2,3-BD), a promising bulk chemical with wide industrial applications, along with various byproducts. In this study, the in silico gene knockout simulation of K. oxytoca was carried out for 2,3-BD overproduction by inhibiting the formation of byproducts. The knockouts of ldhA and pflB genes were targeted with the criteria of maximization of 2,3-BD production and minimization of byproducts formation. The constructed K. oxytoca ΔldhA ΔpflB strain showed higher 2,3-BD yields and higher final concentrations than those obtained from the wild-type and ΔldhA strains. However, the simultaneous deletion of both genes caused about a 50 % reduction in 2,3-BD productivity compared with K. oxytoca ΔldhA strain. Based on previous studies and in silico investigation that the agitation speed during 2,3-BD fermentation strongly affected cell growth and 2,3-BD synthesis, the effect of agitation speed on 2,3-BD production was investigated from 150 to 450 rpm in 5-L bioreactors containing 3-L culture media. The highest 2,3-BD productivity (2.7 g/L/h) was obtained at 450 rpm in batch fermentation. Considering the inhibition of acetoin for 2,3-BD production, fed-batch fermentations were performed using K. oxytoca ΔldhA ΔpflB strain to enhance 2,3-BD production. Altering the agitation speed from 450 to 350 rpm at nearly 10 g/L of acetoin during the fed-batch fermentation allowed for the production of 113 g/L 2,3-BD, with a yield of 0.45 g/g, and for the production of 2.1 g/L/h of 2,3-BD.

Batch Statistical Process Monitoring Approach to a Cocrystallization Process.

PubMed

Sarraguça, Mafalda C; Ribeiro, Paulo R S; Dos Santos, Adenilson O; Lopes, João A

2015-12-01

Cocrystals are defined as crystalline structures composed of two or more compounds that are solid at room temperature held together by noncovalent bonds. Their main advantages are the increase of solubility, bioavailability, permeability, stability, and at the same time retaining active pharmaceutical ingredient bioactivity. The cocrystallization between furosemide and nicotinamide by solvent evaporation was monitored on-line using near-infrared spectroscopy (NIRS) as a process analytical technology tool. The near-infrared spectra were analyzed using principal component analysis. Batch statistical process monitoring was used to create control charts to perceive the process trajectory and define control limits. Normal and non-normal operating condition batches were performed and monitored with NIRS. The use of NIRS associated with batch statistical process models allowed the detection of abnormal variations in critical process parameters, like the amount of solvent or amount of initial components present in the cocrystallization. © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association.

Numerical model of spray combustion in a single cylinder diesel engine

NASA Astrophysics Data System (ADS)

Acampora, Luigi; Sequino, Luigi; Nigro, Giancarlo; Continillo, Gaetano; Vaglieco, Bianca Maria

2017-11-01

A numerical model is developed for predicting the pressure cycle from Intake Valve Closing (IVC) to the Exhaust Valve Opening (EVO) events. The model is based on a modified one-dimensional (1D) Musculus and Kattke spray model, coupled with a zero-dimensional (0D) non-adiabatic transient Fed-Batch reactor model. The 1D spray model provides an estimate of the fuel evaporation rate during the injection phenomenon, as a function of time. The 0D Fed-Batch reactor model describes combustion. The main goal of adopting a 0D (perfectly stirred) model is to use highly detailed reaction mechanisms for Diesel fuel combustion in air, while keeping the computational cost as low as possible. The proposed model is validated by comparing its predictions with experimental data of pressure obtained from an optical single cylinder Diesel engine.

Bioconversion of glycerol to 1,3-propanediol in thin stillage-based media by engineered Lactobacillus panis PM1.

PubMed

Kang, Tae Sun; Korber, Darren R; Tanaka, Takuji

2014-04-01

Thin stillage (TS) is a waste residue that remains after bioethanol production, and its disposal reflects the high costs of bioethanol production. Thus, the development of cost-effective ways to process TS is a pending issue in bioethanol plants. The aim of this study was to evaluate the utilization of TS for the production of the valuable chemical, 1,3-propanediol (1,3-PDO), by Lactobacillus panis PM1. Different fermentation parameters, including temperature, pH and strains [wild-type and a recombinant strain expressing a NADPH-dependent aldehyde reductase (YqhD) gene] were tested in batch and fed-batch cultivations. The highest 1,3-PDO concentration (12.85 g/L) and yield (0.84 g/g) were achieved by batch fermentation at pH-4.5/30 °C by the YqhD recombinant strain. Furthermore, pH-controlled batch fermentation reduced the total fermentation period, resulting in the maximal 1,3-PDO concentration of 16.23 g/L and yield of 0.72 g/g in TS without an expensive nutrient or nitrogen (e.g., yeast extract, beef extract, and peptone) supplementation. The addition of two trace elements, Mg(2+) and Mn(2+), in TS increased 1,3-PDO yield (0.74 g/g) without 3-hydroxypropionaldehyde production, the only intermediate of 1,3-PDO biosynthetic pathway in L. panis PM1. Our results suggest that L. panis PM1 can offer a cost-effective process that utilizes the TS to produce a value-added chemical, 1,3-PDO.

Microbial anodic consortia fed with fermentable substrates in microbial electrolysis cells: Significance of microbial structures.

PubMed

Flayac, Clément; Trably, Eric; Bernet, Nicolas

2018-05-28

Microbial community structure of anodic biofilms plays a key role in bioelectrochemical systems (BESs). When ecosystems are used as inocula, many bacterial species having interconnected ecological interactions are present. The aim of the present study was to identify these interactions for the conversion of single substrates into electrical current. Dual-chamber reactors were inoculated with activated sludge and fed in batch mode with acetate, lactate, butyrate and propionate at 80 mMe - equivalents in quadruplicate. Analyses of biofilms and planktonic microbial communities showed that the anodic biofilms were mainly dominated by the Geobacter genus (62.4% of the total sequences). At the species level, Geobacter sulfurreducens was dominant in presence of lactate and acetate, while Geobacter toluenoxydans and Geobacter pelophilus were dominant with butyrate and propionate as substrates. These results indicate for the first time a specificity within the Geobacter genus towards the electron donor, suggesting a competitive process for electrode colonization and the implementations of syntrophic interactions for complete oxidation of substrates such as propionate and butyrate. All together, these results provide a new insight into the ecological relationships within electroactive biofilms and suggest eco-engineering perspectives to improve the performances of BESs. Copyright © 2018 Elsevier B.V. All rights reserved.

Reductive Dechlorination of Carbon Tetrachloride by Tetrachloroethene and Trichloroethene Respiring Anaerobic Mixed Cultures

NASA Astrophysics Data System (ADS)

Vickstrom, K. E.; Azizian, M.; Semprini, L.

2015-12-01

Carbon tetrachloride (CT) is a toxic and recalcitrant groundwater contaminant with the potential to form a broad range of transformation products. Of the possible biochemical pathways through which CT can be degraded, reductive dehalogenation to less chlorinated compounds and mineralization to carbon dioxide (CO2) appear to be the most frequently utilized pathways by anaerobic organisms. Results will be presented from batch experiments of CT degradation by the Evanite (EV), Victoria Strain (VS) and Point Mugu (PM) anaerobic dechlorinating cultures. The cultures are grown in chemostats and are capable of transforming tetrachloroethene (PCE) or trichloroethene (TCE) to ethene by halorespiration via reductive dehalogenase enzymes. For the batch CT transformation tests, the cells along with supernatant were harvested from chemostats fed PCE or TCE, but never CT. The batch reactors were initially fed 0.0085 mM CT and an excess of formate (EV and VS) or lactate (PM) as electron donor. Transformation of CT was 100% with about 20% converted to chloroform (CF) and undetected products. Multiple additions of CT showed a slowing of pseudo first-order CT transformation rates across all cultures. Batch reactors were then established and fed 0.085 mM CT with an excess of electron donor in order to better quantify the reductive pathway. CT was transformed to CF and dichloromethane (DCM), with trace amounts of chloromethane (CM) detected. Between 60-90% of the mass added to the system was accounted for, showing that the majority of the carbon tetrachloride present is being reductively dehalogenated. Results from batch reactors that were poisoned using sodium azide, and from reactors not provided electron donor will be presented to distinguish between biotic and abiotic reactions. Furthermore, results from reactors prepared with acetylene (a potent, reversible inhibitor of reductive dehalogenases (1)) will be presented as a means of identifying the enzymes involved in the transformation of CT. The results clearly demonstrate that reductive dechlorination of CT can be promoted by anaerobic cultures not previously acclimated to CT. 1. G. Pon, M. R. Hyman, L. Semprini, Environ. Sci. Technol. 37, 3181-3188 (2003).

Effect of crude protein concentration and dietary electrolyte balance on litter quality, foot pad dermatitis, growth performance and processing yields in two medium heavy turkey hybrids.

PubMed

Veldkamp, T; Hocking, P M; Vinco, L J

2017-10-01

1. An experiment was conducted to investigate the effect of crude protein (CP) concentration and dietary electrolyte balance (DEB) on growth performance, processing yields, litter quality and foot pad dermatitis (FPD) in male turkeys from two commercial hybrids. Soya bean meal was replaced by vegetable protein sources selected for lower K concentrations to lower DEB in order to improve litter quality and subsequent quality of foot pads. 2. Effects of CP on litter friability and wetness were not consistent during the production period. FPD in turkeys fed on diets with low CP was significantly lower than FPD in turkeys fed on diets with high CP until 84 d. Growth performance was adversely affected at low CP. Processing yields were not affected by CP. 3. Litter was significantly dryer in pens of turkeys fed on diets with low DEB than in pens of turkeys fed on diets with high DEB. FPD in turkeys fed on diets with low DEB was significantly lower than in turkeys fed on diets with high DEB. Growth performance and processing yields were adversely affected at low DEB. 4. FPD in turkey hybrid A was higher than in turkey hybrid B at 28 d of age. Thereafter, no differences in FPD between turkey hybrids were observed. Growth performance and processing yields were not affected by turkey hybrid. 5. Overall, a significant interaction effect of CP × DEB was observed for FCR: in turkeys fed on the high DEB treatment, FCR of turkeys fed on the high CP diets was lower than FCR of turkeys fed on the low CP (LCP) diets whereas on the low DEB treatment, FCR was not affected by CP treatment. 6. It was concluded that litter quality can be improved and FPD may be decreased in turkeys fed on diets containing lower CP and DEB levels.

Indigenous microbial capability in solid manure residues to start-up solid-phase anaerobic digesters.

PubMed

Yap, S D; Astals, S; Jensen, P D; Batstone, D J; Tait, S

2017-06-01

Batch solid-phase anaerobic digestion is a technology for sustainable on-farm treatment of solid residues, but is an emerging technology that is yet to be optimised with respect to start-up and inoculation. In the present study, spent bedding from two piggeries (site A and B) were batch digested at total solids (TS) concentration of 5, 10 and 20% at mesophilic (37°C) and thermophilic (55°C) temperatures, without adding an external inoculum. The results showed that the indigenous microbial community present in spent bedding was able to recover the full methane potential of the bedding (140±5 and 227±6L CH 4 kgVS fed -1 for site A and B, respectively), but longer treatment times were required than for digestion with an added external inoculum. Nonetheless, at high solid loadings (i.e. TS level>10%), the digestion performance was affected by chemical inhibition due to ammonia and/or humic acid. Thermophilic temperatures did not influence digestion performance but did increase start-up failure risk. Further, inoculation of residues from the batch digestion to subsequent batch enhanced start-up and achieved full methane potential recovery of the bedding. Inoculation with liquid residue (leachate) was preferred over a solid residue, to preserve treatment capacity for fresh substrate. Overall, the study highlighted that indigenous microbial community in the solid manure residue was capable of recovering full methane potential and that solid-phase digestion was ultimately limited by chemical inhibition rather than lack of suitable microbial community. Copyright © 2017 Elsevier Ltd. All rights reserved.

Dry anaerobic digestion of high solids content dairy manure at high organic loading rates in psychrophilic sequence batch reactor.

PubMed

Massé, Daniel I; Saady, Noori M Cata

2015-05-01

Cow manure with bedding is renewable organic biomass available around the year on dairy farms. Developing efficient and cost-effective psychrophilic dry anaerobic digestion (PDAD) processes could contribute to solving farm-related environmental, energy, and manure management problems in cold-climate regions. This study was to increase the organic loading rate (OLR), fed to a novel psychrophilic (20 °C) dry anaerobic digestion of 27% total solid dairy manure (cow feces and wheat straw) in sequence batch reactor (PDAD-SBR), by 133 to 160%. The PDAD-SBR process operated at treatment cycle length of 21 days and OLR of 7.0 and 8.0 g total chemical oxygen demand (TCOD) kg(-1) inoculum day(-1) (5.2 ± 0.1 and 5.8 ± 0.0 g volatile solids (VS) kg(-1) inoculum day(-1)) for four successive cycles (84 days) produced average specific methane yields (SMYs) of 147.1 ± 17.2 and 143.2 ± 11.7 normalized liters (NL) CH4 kg(-1) VS fed, respectively. PDAD of cow feces and wheat straw is possible with VS-based inoculum-to-substrate ratio of 1.45 at OLR of 8.0 g TCOD kg(-1) inoculum day(-1). Hydrolysis was the limiting step reaction. The VS removal averaged around 57.4 ± 0.5 and 60.5 ± 5.7% at OLR 7.0 and 8.0 g TCOD kg(-1) inoculum day(-1), respectively.

40 CFR 63.1325 - Batch process vents-performance test methods and procedures to determine compliance.

Code of Federal Regulations, 2010 CFR

2010-07-01

... either total organic HAP or TOC. For purposes of this paragraph (c), the term “batch emission episode... ensure the measurement of total organic HAP or TOC (minus methane and ethane) concentrations in all batch... organic HAP or TOC, as appropriate. Alternatively, any other method or data that has been validated...

Hybrid modeling as a QbD/PAT tool in process development: an industrial E. coli case study.

PubMed

von Stosch, Moritz; Hamelink, Jan-Martijn; Oliveira, Rui

2016-05-01

Process understanding is emphasized in the process analytical technology initiative and the quality by design paradigm to be essential for manufacturing of biopharmaceutical products with consistent high quality. A typical approach to developing a process understanding is applying a combination of design of experiments with statistical data analysis. Hybrid semi-parametric modeling is investigated as an alternative method to pure statistical data analysis. The hybrid model framework provides flexibility to select model complexity based on available data and knowledge. Here, a parametric dynamic bioreactor model is integrated with a nonparametric artificial neural network that describes biomass and product formation rates as function of varied fed-batch fermentation conditions for high cell density heterologous protein production with E. coli. Our model can accurately describe biomass growth and product formation across variations in induction temperature, pH and feed rates. The model indicates that while product expression rate is a function of early induction phase conditions, it is negatively impacted as productivity increases. This could correspond with physiological changes due to cytoplasmic product accumulation. Due to the dynamic nature of the model, rational process timing decisions can be made and the impact of temporal variations in process parameters on product formation and process performance can be assessed, which is central for process understanding.

40 CFR 63.2460 - What requirements must I meet for batch process vents?

Code of Federal Regulations, 2010 CFR

2010-07-01

.... (ii) When you conduct a performance test or design evaluation for a non-flare control device used to... paragraphs (c)(9)(ii)(A) through (D) of this section. The design evaluation option for small control devices...) of this section. (b) Group status. If a process has batch process vents, as defined in § 63.2550, you...

40 CFR 63.2460 - What requirements must I meet for batch process vents?

Code of Federal Regulations, 2011 CFR

2011-07-01

.... (ii) When you conduct a performance test or design evaluation for a non-flare control device used to... paragraphs (c)(9)(ii)(A) through (D) of this section. The design evaluation option for small control devices...) of this section. (b) Group status. If a process has batch process vents, as defined in § 63.2550, you...

40 CFR 63.2460 - What requirements must I meet for batch process vents?

Code of Federal Regulations, 2014 CFR

2014-07-01

.... (ii) When you conduct a performance test or design evaluation for a non-flare control device used to... paragraphs (c)(9)(ii)(A) through (D) of this section. The design evaluation option for small control devices... (c) of this section. (b) Group status. If a process has batch process vents, as defined in § 63.2550...

40 CFR 63.2460 - What requirements must I meet for batch process vents?

Code of Federal Regulations, 2012 CFR

2012-07-01

.... (ii) When you conduct a performance test or design evaluation for a non-flare control device used to... paragraphs (c)(9)(ii)(A) through (D) of this section. The design evaluation option for small control devices... (c) of this section. (b) Group status. If a process has batch process vents, as defined in § 63.2550...

40 CFR 63.2460 - What requirements must I meet for batch process vents?

Code of Federal Regulations, 2013 CFR

2013-07-01

.... (ii) When you conduct a performance test or design evaluation for a non-flare control device used to... paragraphs (c)(9)(ii)(A) through (D) of this section. The design evaluation option for small control devices... (c) of this section. (b) Group status. If a process has batch process vents, as defined in § 63.2550...

Process performance of high-solids batch anaerobic digestion of sewage sludge.

PubMed

Liao, Xiaocong; Li, Huan; Cheng, Yingchao; Chen, Nan; Li, Chenchen; Yang, Yuning

2014-01-01

The characteristics of high-solids anaerobic digestion (AD) of sewage sludge were investigated by comparison with conventional low-solids processes. A series of batch experiments were conducted under mesophilic condition and the initial solid contents were controlled at four levels of 1.79%, 4.47%, 10.28% and 15.67%. During these experiments, biogas production, organic degradation and intermediate products were monitored. The results verified that high-solids batch AD of sewage sludge was feasible. Compared with the low-solids AD with solid contents of 1.79% or 4.47%, the high-solids processes decreased the specific biogas yield per gram of sludge volatile solids slightly, achieved the same organic degradation rate of about 40% within extended degradation time, but increased the volumetric biogas production rate and the treatment capability of digesters significantly. The blocked mass and energy transfer, the low substrate to inoculum rate and the excessive cumulative free ammonia were the main factors impacting the performance of high-solids batch AD.

Combined strategies for improving expression of Citrobacter amalonaticus phytase in Pichia pastoris.

PubMed

Li, Cheng; Lin, Ying; Zheng, Xueyun; Pang, Nuo; Liao, Xihao; Liu, Xiaoxiao; Huang, Yuanyuan; Liang, Shuli

2015-09-26

Phytase is used as an animal feed additive that degrades phytic acid and reduces feeding costs and pollution caused by fecal excretion of phosphorus. Some phytases have been expressed in Pichia pastoris, among which the phytase from Citrobacter amalonaticus CGMCC 1696 had high specific activity (3548 U/mg). Improvement of the phytase expression level will contribute to facilitate its industrial applications. To improve the phytase expression, we use modification of P AOX1 and the α-factor signal peptide, increasing the gene copy number, and overexpressing HAC1 (i) to enhance folding and secretion of the protein in the endoplasmic reticulum. The genetic stability and fermentation in 10-L scaled-up fed-batch fermenter was performed to prepare for the industrial production. The phytase gene from C. amalonaticus CGMCC 1696 was cloned under the control of the AOX1 promoter (P AOX1 ) and expressed in P. pastoris. The phytase activity achieved was 414 U/mL. Modifications of P AOX1 and the α-factor signal peptide increased the phytase yield by 35 and 12%, respectively. Next, on increasing the copy number of the Phy gene to six, the phytase yield was 141% higher than in the strain containing only a single gene copy. Furthermore, on overexpression of HAC1 (i) (i indicating induced), a gene encoding Hac1p that regulates the unfolded protein response, the phytase yield achieved was 0.75 g/L with an activity of 2119 U/mL, 412% higher than for the original strain. The plasmids in this high-phytase expression strain were stable during incubation at 30 °C in Yeast Extract Peptone Dextrose (YPD) Medium. In a 10-L scaled-up fed-batch fermenter, the phytase yield achieved was 9.58 g/L with an activity of 35,032 U/mL. The production of a secreted protein will reach its limit at a specific gene copy number where further increases in transcription and translation due to the higher abundance of gene copies will not enhance the secretion process any further. Enhancement of protein folding in the ER can alleviate bottlenecks in the folding and secretion pathways during the overexpression of heterologous proteins in P. pastoris. Using modification of P AOX1 and the α-factor signal peptide, increasing the gene copy number, and overexpressing HAC1 (i) to enhance folding and secretion of the protein in the endoplasmic reticulum, we have successfully increased the phytase yield 412% relative to the original strain. In a 10-L fed-batch fermenter, the phytase yield achieved was 9.58 g/L with an activity of 35,032 U/mL. Large-scale production of phytase can be applied towards different biocatalytic and feed additive applications.

Development of a cost-effective production process for Halomonas levan.

PubMed

Erkorkmaz, Burak Adnan; Kırtel, Onur; Ateş Duru, Özlem; Toksoy Öner, Ebru

2018-05-17

Levan polysaccharide is an industrially important natural polymer with unique properties and diverse high-value applications. However, current bottlenecks associated with its large-scale production need to be overcome by innovative approaches leading to economically viable processes. Besides many mesophilic levan producers, halophilic Halomonas smyrnensis cultures hold distinctive industrial potential and, for the first time with this study, the advantage of halophilicity is used and conditions for non-sterile levan production were optimized. Levan productivity of Halomonas cultures in medium containing industrial sucrose from sugar beet and food industry by-product syrup, a total of ten sea, lake and rock salt samples from four natural salterns, as well as three different industrial-grade boron compounds were compared and the most suitable low-cost substitutes for sucrose, salt and boron were specified. Then, the effects of pH control, non-sterile conditions and different bioreactor modes (batch and fed-batch) were investigated. The development of a cost-effective production process was achieved with the highest yield (18.06 g/L) reported so far on this microbial system, as well as the highest theoretical bioconversion efficiency ever reported for levan-producing suspension cultures. Structural integrity and biocompatibility of the final product were also verified in vitro.

Application of gain scheduling to the control of batch bioreactors

NASA Technical Reports Server (NTRS)

Cardello, Ralph; San, Ka-Yiu

1987-01-01

The implementation of control algorithms to batch bioreactors is often complicated by the inherent variations in process dynamics during the course of fermentation. Such a wide operating range may render the performance of fixed gain PID controllers unsatisfactory. In this work, a detailed study on the control of batch fermentation is performed. Furthermore, a simple batch controller design is proposed which incorporates the concept of gain-scheduling, a subclass of adaptive control, with oxygen uptake rate as an auxiliary variable. The control of oxygen tension in the biorector is used as a vehicle to convey the proposed idea, analysis and results. Simulation experiments indicate significant improvement in controller performance can be achieved by the proposed approach even in the presence of measurement noise.

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.

Fed-batch cultivation of Arthrospira (Spirulina) platensis: potassium nitrate and ammonium chloride as simultaneous nitrogen sources.

PubMed

Rodrigues, M S; Ferreira, L S; Converti, A; Sato, S; Carvalho, J C M

2010-06-01

Arthrospiraplatensis was cultivated in minitanks at 13 klux, using a mixture of KNO(3) and NH(4)Cl as nitrogen source. Fed-batch daily supply of NH(4)Cl at exponentially-increasing feeding rate allowed preventing ammonia toxicity and nitrogen deficiency, providing high maximum cell concentration (X(m)) and high-quality biomass (21.85 mg chlorophyll g cells(-1); 20.5% lipids; 49.8% proteins). A central composite design combined to response surface methodology was utilized to determine the relationships between responses (X(m), cell productivity and nitrogen-to-cell conversion factor) and independent variables (KNO(3) and NH(4)Cl concentrations). Under optimum conditions (15.5mM KNO(3); 14.1mM NH(4)Cl), X(m) was 4327 mg L(-1), a value almost coincident with that obtained with only 25.4mM KNO(3), but more than twice that obtained with 21.5mM NH(4)Cl. A 30%-reduction of culture medium cost can be estimated when compared to KNO(3)-batch runs, thus behaving as a cheap alternative for the commercial production of this cyanobacterium. (c) 2010 Elsevier Ltd. All rights reserved.

Deletion of lactate dehydrogenase in Enterobacter aerogenes to enhance 2,3-butanediol production.

PubMed

Jung, Moo-Young; Ng, Chiam Yu; Song, Hyohak; Lee, Jinwon; Oh, Min-Kyu

2012-07-01

2,3-Butanediol is an important bio-based chemical product, because it can be converted into several C4 industrial chemicals. In this study, a lactate dehydrogenase-deleted mutant was constructed to improve 2,3-butanediol productivity in Enterobacter aerogenes. To delete the gene encoding lactate dehydrogenase, λ Red recombination method was successfully adapted for E. aerogenes. The resulting strain produced a very small amount of lactate and 16.7% more 2,3-butanediol than that of the wild-type strain in batch fermentation. The mutant and its parental strain were then cultured with six different carbon sources, and the mutant showed higher carbon source consumption and microbial growth rates in all media. The 2,3-butanediol titer reached 69.5 g/l in 54 h during fed-batch fermentation with the mutant,which was 27.4% higher than that with the parental strain.With further optimization of the medium and aeration conditions,118.05 g/l 2,3-butanediol was produced in 54 h during fed-batch fermentation with the mutant. This is by far the highest titer of 2,3-butanediol with E. aerogenes achieved by metabolic pathway engineering.

Characteristics of aerobic granules grown on glucose a sequential batch shaking reactor.

PubMed

Cai, Chun-guang; Zhu, Nan-wen; Liu, Jun-shen; Wang, Zhen-peng; Cai, Wei-min

2004-01-01

Aerobic heterotrophic granular sludge was cultivated in a sequencing batch shaking reactor (SBSR) in which a synthetic wastewater containing glucose as carbon source was fed. The characteristics of the aerobic granules were investigated. Compared with the conventional activated sludge flocs, the aerobic granules exhibit excellent physical characteristics in terms of settleability, size, shape, biomass density, and physical strength. Scanning electron micrographs revealed that in mature granules little filamentous bacteria could be found, rod-shaped and coccoid bacteria were the dominant microorganisms.

Development of a highly efficient oil degumming process using a novel phosphatidylinositol-specific phospholipase C enzyme.

PubMed

Cerminati, Sebastián; Eberhardt, Florencia; Elena, Claudia E; Peirú, Salvador; Castelli, María E; Menzella, Hugo G

2017-06-01

Enzymatic degumming using phospholipase C (PLC) enzymes may be used in environmentally friendly processes with improved oil recovery yields. In this work, phosphatidylinositol-specific phospholipase C (PIPLC) candidates obtained from an in silico analysis were evaluated for oil degumming. A PIPLC from Lysinibacillus sphaericus was shown to efficiently remove phosphatidylinositol from crude oil, and when combined with a second phosphatidylcholine and phosphatidylethanolamine-specific phospholipase C, the three major phospholipids were completely hydrolyzed, providing an extra yield of oil greater than 2.1%, compared to standard methods. A remarkably efficient fed-batch Escherichia coli fermentation process producing ∼14 g/L of the recombinant PIPLC enzyme was developed, which may facilitate the adoption of this cost-effective oil-refining process.

Poultry offal ensiled with lactobacillus acidophilus for growing and finishing swine diets.

PubMed

Tibbetts, G W; Seerley, R W; McCampbell, H C

1987-01-01

Poultry offal (heads, feet, viscera) from a broiler processing plant was ground and mixed with corn, dried molasses and a Lactobacillus acidophilus culture. Fermentation was shown to be effective in batch sizes of 3.8, 75.7 and 208.1 liters, as well as a 1,360-kg silo. It was generally complete at 72 h. In trial 1, rats were fed silage mixtures of 60:30:5:5, 45:45:5:5 and 30:60:5:5, offal, corn, molasses and inoculant, respectively. Rats did not gain as well when fed the silage diets (P less than .05) as when fed the basal diet; however, the ranking of silages was 45:45, 60:30 and 30:60, offal-to-corn ratio for rat daily gains and feed conversions. In trial 2, growing-finishing pigs were fed the 60:30:5:5 silage at rates of 0, 10, 20 and 30% of the diet. Average daily gains and feed-to-gain ratios were not affected by including offal silage at up to 20% of the diet, but 30% offal silage in diets resulted in depressed gains (P less than .05) and increased feed-to-gain ratios. Carcasses were not different among treatments for dressing percentage, length, average backfat and percentage of ham, loin, shoulder, lean cuts and primal cuts. Carcasses from pigs fed 20 and 30% offal silage had significantly darker and firmer loin-eyes than those from control fed pigs. Marbling was higher (P less than .05) in loin-eyes from pigs fed 30% offal silage as compared with those from pigs fed 0% offal silage.(ABSTRACT TRUNCATED AT 250 WORDS)

ANAEROBIC AND AEROBIC TREATMENT OF CHLORINATED ALIPHATIC COMPOUNDS

EPA Science Inventory

Biological degradation of 12 chlorinated aliphatic compounds (CACs) was assessed in bench-top reactors and in serum bottle tests. Three continuously mixed daily batch-fed reactor systems were evaluated: anaerobic, aerobic, and sequential-anaerobic-aerobic (sequential). Glucose,...

40 CFR Table 4 to Subpart Cccc of... - Continuous Compliance With Emission Limitations

Code of Federal Regulations, 2010 CFR

2010-07-01

... yeast in a fermentation stage (last (Trade), second-to-last (First Generation), or third-to-last (Stock... the applicable maximum concentration. 2. Each fed-batch fermenter producing yeast in a fermentation...

40 CFR Table 3 to Subpart Cccc of... - Initial Compliance With Emission Limitations

Code of Federal Regulations, 2010 CFR

2010-07-01

... demonstrated initial compliance if . . . 1. Each fed-batch fermenter producing yeast in a fermentation stage... yeast in a fermentation stage (last (Trade), second-to-last (First Generation), or third-to-last (Stock...

Enhanced Production of Poly-γ-glutamic Acid by Bacillus licheniformis TISTR 1010 with Environmental Controls.

PubMed

Kongklom, Nuttawut; Shi, Zhongping; Chisti, Yusuf; Sirisansaneeyakul, Sarote

2017-07-01

Bacillus licheniformis TISTR 1010 was used for glutamic acid-independent production of poly-γ-glutamic acid (γ-PGA). A fed-batch production strategy was developed involving feedings of glucose, citric acid, and ammonium chloride at specified stages of the fermentation. With the dissolved oxygen concentration controlled at ≥50% of air saturation and the pH controlled at ~7.4, the fed-batch operation at 37 °C afforded a peak γ-PGA concentration of 39.9 ± 0.3 g L -1 with a productivity of 0.926 ± 0.006 g L -1  h -1 . The observed productivity was nearly threefold greater than previously reported for glutamic acid-independent production using the strain TISTR 1010. The molecular weight of γ-PGA was in the approximate range of 60 to 135 kDa.

Production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by Cupriavidus necator from waste rapeseed oil using propanol as a precursor of 3-hydroxyvalerate.

PubMed

Obruca, Stanislav; Marova, Ivana; Snajdar, Ondrej; Mravcova, Ludmila; Svoboda, Zdenek

2010-12-01

Waste rapeseed oil is a useful substrate for polyhydroxyalkanoates (PHA) production employing Cupriavidus necator H16. In fed-batch mode, we obtained biomass and PHA yields of 138 and 105 g l(-1), respectively. Yield coefficient and volumetric productivity were 0.83 g PHA per g oil and 1.46 g l(-1) h(-1), respectively. Propanol at 1% (v/v) enhanced both PHA and biomass formation significantly and, furthermore, resulted in incorporation of 3-hydroxyvalerate units into PHA structure. Thus, propanol can be used as an effective precursor of 3-hydroxyvalarete for production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer. During the fed-batch cultivation, propanol concentration was maintained at 1% which resulted in 8% content of 3-hydroxyvalerate in copolymer.

Fed-batch cultivation of Arthrospira and Chlorella in ammonia-rich wastewater: Optimization of nutrient removal and biomass production.

PubMed

Markou, Giorgos

2015-10-01

In the present work the cyanobacterium Arthrospira platensis and the microalga Chlorella vulgaris were fed-batch cultivated in ammonia-rich wastewater derived from the anaerobic digestion of poultry litter. Aim of the study was to maximize the biomass production along with the nutrient removal aiming to wastewater treatment. Ammonia and phosphorus removals were very high (>95%) for all cultures investigated. Both microorganisms were able to remove volatile fatty acids to an extent of >90%, indicating that they were capable of mixotrophic growth. Chemical oxygen demand and proteins were also removed in various degrees. In contrast, in all cultures carbohydrate concentration was increased. The biochemical composition of the microorganisms varied greatly and was influenced by the indicate that the nutrient availability. A. platensis accumulated carbohydrates (≈ 40%), while C. vulgaris accumulated lipids (≈ 50%), rendering them interesting for biofuel production. Copyright © 2015 Elsevier Ltd. All rights reserved.

A Different Perspective: How Much Innovation Is Really Needed for Monoclonal Antibody Production Using Mammalian Cell Technology?

PubMed

Kelley, Brian; Kiss, Robert; Laird, Michael

2018-05-03

As biopharmaceutical companies have optimized cell line and production culture process development, titers of recombinant antibodies have risen steadily to 3-8 g/L for fed-batch mammalian cultures at production scales of 10 kL or larger. Most new antibody products are produced from Chinese Hamster Ovary (CHO) cell lines, and there are relatively few alternative production hosts under active evaluation. Many companies have adopted a strategy of using the same production cell line for early clinical phases as well as commercial production, which reduces the risk of product comparability issues during the development lifecycle. Product quality and consistency expectations rest on the platform knowledge of the CHO host cell line and processes used for the production of many licensed antibodies. The lack of impact of low-level product variants common to this platform on product safety and efficacy also builds on the established commercial history of recombinant antibodies, which dates back to 1997.Efforts to increase titers further will likely yield diminishing returns. Very few products would benefit significantly from a titer greater than 8 g/L; in many cases, a downstream processing bottleneck would preclude full recovery from production-scale bioreactors for high titer processes. The benefits of a process platform based on standard fed-batch production culture include predictable scale-up, process transfer, and production within a company's manufacturing network or at a contract manufacturing organization. Furthermore, the confidence in an established platform provides key support towards regulatory flexibility (e.g., design space) for license applications following a quality-by-design strategy.These factors suggest that novel technologies for antibody production may not provide a substantial return on investment. What, then, should be the focus of future process development efforts for companies that choose to launch antibody products using their current platform? This review proposes key focus areas in an effort to continually improve process consistency, assure acceptable product quality, and establish appropriate process parameter limits to enable flexible manufacturing options.

Optimization of the biotechnological production of a novel class of anti-MRSA antibiotics from Chitinophaga sancti.

PubMed

Beckmann, Amelie; Hüttel, Stephan; Schmitt, Viktoria; Müller, Rolf; Stadler, Marc

2017-08-17

Recently, the discovery of the elansolids, a group of macrolides, was reported. The molecules show activity against methicillin-resistant Staphylococcus aureus as well as other gram-positive organisms. This fact renders those substances a promising starting point for future chemical development. The active atropisomers A1/A2 are formed by macrolactonization of the biosynthesis product A3 but are prone to ring opening and subsequent formation of several unwanted side products. Recently it could be shown that addition of different nucleophiles to culture extracts of Chitinophaga sancti enable the formation of new stable elansolid derivatives. Furthermore, addition of such a nucleophile directly into the culture led exclusively to formation of a single active elansolid derivative. Due to low product yields, methods for production of gram amounts of these molecules have to be established to enable further development of this promising compound class. Production of elansolid A2 by C. sancti was enabled using a synthetic medium with sucrose as carbon source to a final concentration of 18.9 mg L -1 . A fed-batch fermentation was ensued that resulted in an elansolid A2 concentration of 55.3 mg L -1 . When using glucose as carbon source in a fed-batch fermentation only 34.4 mg L -1 elansolid A2 but 223.1 mg L -1 elansolid C1 were produced. This finding was not unexpected since elansolids A1/A2 and A3 have been reported to easily react with nucleophiles like anthranilic acid, a precursor of tryptophan biosynthesis. Due to the fact that nucleophiles can be incorporated in vivo, a fed-batch cultivation under identical conditions, with addition of anthranilic acid was carried out and lead to almost exclusive formation of elansolid C1 (257.5 mg L -1 ). Reproducible elansolid A2 and C1 production is feasible in different synthetic media at relatively high concentrations that will allow further investigation and semi-synthetic optimization. The feeding of anthranilic acid enables the exclusive production of the stable elansolid derivative C1, which reduces product loss by unspecific reactions and eases downstream processing. This derivative shows activity in the same range as the elansolids A1/A2. Hence, the method can possibly serve as a model-process for incorporation of other nucleophiles and biotechnological production of specifically designed molecules.

Tracking control of concentration profiles in a fed-batch bioreactor using a linear algebra methodology.

PubMed

Rómoli, Santiago; Serrano, Mario Emanuel; Ortiz, Oscar Alberto; Vega, Jorge Rubén; Eduardo Scaglia, Gustavo Juan

2015-07-01

Based on a linear algebra approach, this paper aims at developing a novel control law able to track reference profiles that were previously-determined in the literature. A main advantage of the proposed strategy is that the control actions are obtained by solving a system of linear equations. The optimal controller parameters are selected through Monte Carlo Randomized Algorithm in order to minimize a proposed cost index. The controller performance is evaluated through several tests, and compared with other controller reported in the literature. Finally, a Monte Carlo Randomized Algorithm is conducted to assess the performance of the proposed controller. Copyright © 2015 ISA. Published by Elsevier Ltd. All rights reserved.

Impact of cultivar selection and process optimization on ethanol yield from different varieties of sugarcane

PubMed Central

2014-01-01

Background The development of ‘energycane’ varieties of sugarcane is underway, targeting the use of both sugar juice and bagasse for ethanol production. The current study evaluated a selection of such ‘energycane’ cultivars for the combined ethanol yields from juice and bagasse, by optimization of dilute acid pretreatment optimization of bagasse for sugar yields. Method A central composite design under response surface methodology was used to investigate the effects of dilute acid pretreatment parameters followed by enzymatic hydrolysis on the combined sugar yield of bagasse samples. The pressed slurry generated from optimum pretreatment conditions (maximum combined sugar yield) was used as the substrate during batch and fed-batch simultaneous saccharification and fermentation (SSF) processes at different solid loadings and enzyme dosages, aiming to reach an ethanol concentration of at least 40 g/L. Results Significant variations were observed in sugar yields (xylose, glucose and combined sugar yield) from pretreatment-hydrolysis of bagasse from different cultivars of sugarcane. Up to 33% difference in combined sugar yield between best performing varieties and industrial bagasse was observed at optimal pretreatment-hydrolysis conditions. Significant improvement in overall ethanol yield after SSF of the pretreated bagasse was also observed from the best performing varieties (84.5 to 85.6%) compared to industrial bagasse (74.5%). The ethanol concentration showed inverse correlation with lignin content and the ratio of xylose to arabinose, but it showed positive correlation with glucose yield from pretreatment-hydrolysis. The overall assessment of the cultivars showed greater improvement in the final ethanol concentration (26.9 to 33.9%) and combined ethanol yields per hectare (83 to 94%) for the best performing varieties with respect to industrial sugarcane. Conclusions These results suggest that the selection of sugarcane variety to optimize ethanol production from bagasse can be achieved without adversely affecting juice ethanol and cane yield, thus maintaining first generation ethanol production levels while maximizing second generation ethanol production. PMID:24725458

Mini review: Recombinant production of tailored bio-pharmaceuticals in different Bacillus strains and future perspectives.

PubMed

Lakowitz, Antonia; Godard, Thibault; Biedendieck, Rebekka; Krull, Rainer

2018-05-01

Bio-pharmaceuticals like antibodies, hormones and growth factors represent about one-fifth of commercial pharmaceuticals. Host candidates of growing interest for recombinant production of these proteins are strains of the genus Bacillus, long being established for biotechnological production of homologous and heterologous proteins. Bacillus strains benefit from development of efficient expression systems in the last decades and emerge as major industrial workhorses for recombinant proteins due to easy cultivation, non-pathogenicity and their ability to secrete recombinant proteins directly into extracellular medium allowing cost-effective downstream processing. Their broad product portfolio of pharmaceutically relevant recombinant proteins described in research include antibody fragments, growth factors, interferons and interleukins, insulin, penicillin G acylase, streptavidin and different kinases produced in various cultivation systems like microtiter plates, shake flasks and bioreactor systems in batch, fed-batch and continuous mode. To further improve production and secretion performance of Bacillus, bottlenecks and limiting factors concerning proteases, chaperones, secretion machinery or feedback mechanisms can be identified on different cell levels from genomics and transcriptomics via proteomics to metabolomics and fluxomics. For systematical identification of recurring patterns characteristic of given regulatory systems and key genetic targets, systems biology and omics-technology provide suitable and promising approaches, pushing Bacillus further towards industrial application for recombinant pharmaceutical protein production. Copyright © 2017. Published by Elsevier B.V.

Modelling of Batch Lactic Acid Fermentation in
the Presence of Anionic Clay

PubMed Central

Jinescu, Cosmin; Aruş, Vasilica Alisa; Nistor, Ileana Denisa

2014-01-01

Summary Batch fermentation of milk inoculated with lactic acid bacteria was conducted in the presence of hydrotalcite-type anionic clay under static and ultrasonic conditions. An experimental study of the effect of fermentation temperature (t=38–43 °C), clay/milk ratio (R=1–7.5 g/L) and ultrasonic field (ν=0 and 35 kHz) on process dynamics was performed. A mathematical model was selected to describe the fermentation process kinetics and its parameters were estimated based on experimental data. A good agreement between the experimental and simulated results was achieved. Consequently, the model can be employed to predict the dynamics of batch lactic acid fermentation with values of process variables in the studied ranges. A statistical analysis of the data based on a 23 factorial experiment was performed in order to express experimental and model-regressed process responses depending on t, R and ν factors. PMID:27904318

Influences of operational parameters on phosphorus removal in batch and continuous electrocoagulation process performance.

PubMed

Nguyen, Dinh Duc; Yoon, Yong Soo; Bui, Xuan Thanh; Kim, Sung Su; Chang, Soon Woong; Guo, Wenshan; Ngo, Huu Hao

2017-11-01

Performance of an electrocoagulation (EC) process in batch and continuous operating modes was thoroughly investigated and evaluated for enhancing wastewater phosphorus removal under various operating conditions, individually or combined with initial phosphorus concentration, wastewater conductivity, current density, and electrolysis times. The results revealed excellent phosphorus removal (72.7-100%) for both processes within 3-6 min of electrolysis, with relatively low energy requirements, i.e., less than 0.5 kWh/m 3 for treated wastewater. However, the removal efficiency of phosphorus in the continuous EC operation mode was better than that in batch mode within the scope of the study. Additionally, the rate and efficiency of phosphorus removal strongly depended on operational parameters, including wastewater conductivity, initial phosphorus concentration, current density, and electrolysis time. Based on experimental data, statistical model verification of the response surface methodology (RSM) (multiple factor optimization) was also established to provide further insights and accurately describe the interactive relationship between the process variables, thus optimizing the EC process performance. The EC process using iron electrodes is promising for improving wastewater phosphorus removal efficiency, and RSM can be a sustainable tool for predicting the performance of the EC process and explaining the influence of the process variables.

Applied in situ product recovery in ABE fermentation.

PubMed

Outram, Victoria; Lalander, Carl-Axel; Lee, Jonathan G M; Davies, E Timothy; Harvey, Adam P

2017-05-01

The production of biobutanol is hindered by the product's toxicity to the bacteria, which limits the productivity of the process. In situ product recovery of butanol can improve the productivity by removing the source of inhibition. This paper reviews in situ product recovery techniques applied to the acetone butanol ethanol fermentation in a stirred tank reactor. Methods of in situ recovery include gas stripping, vacuum fermentation, pervaporation, liquid-liquid extraction, perstraction, and adsorption, all of which have been investigated for the acetone, butanol, and ethanol fermentation. All techniques have shown an improvement in substrate utilization, yield, productivity or both. Different fermentation modes favored different techniques. For batch processing gas stripping and pervaporation were most favorable, but in fed-batch fermentations gas stripping and adsorption were most promising. During continuous processing perstraction appeared to offer the best improvement. The use of hybrid techniques can increase the final product concentration beyond that of single-stage techniques. Therefore, the selection of an in situ product recovery technique would require comparable information on the energy demand and economics of the process. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:563-579, 2017. © 2017 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers.

40 CFR Table 2 to Subpart Sssss of... - Operating Limits

Code of Federal Regulations, 2010 CFR

2010-07-01

... activity level of the catalyst at least every 12 months. 7. Each affected batch process unit For each batch... activity level of the catalyst at least every 12 months. 10. Each new kiln that is used to process clay... across the DLA at or above the minimum levels established during the most recent performance test; and b...

40 CFR Table 2 to Subpart Sssss of... - Operating Limits

Code of Federal Regulations, 2011 CFR

2011-07-01

... activity level of the catalyst at least every 12 months. 7. Each affected batch process unit For each batch... activity level of the catalyst at least every 12 months. 10. Each new kiln that is used to process clay... across the DLA at or above the minimum levels established during the most recent performance test; and b...

Comparison of Reductive Dechlorination of Chlorinated Ethylene in Batch and Continuous-Flow Reactor

NASA Astrophysics Data System (ADS)

Park, S.; Jonghwan, L.; Hong, U.; Kim, N.; Ahn, H.; Lee, S.; Kim, Y.

2010-12-01

A 1.28 L-Batch reactor and continuous-flow stirred tank reactor (CFSTR) fed with formate and trichloriethene (TCE) were operated for 120 days and 72 days, respectively, to study the effect of formate as electron donor on reductive dechlorination of TCE to cis-1,2-dichloroethylene (c-DCE), vinyl chloride (VC), and ethylene (ETH). In batch reactor, injected 60 μmol TCE was completely degraded in presence of 20% hydrogen gas (H2) in less than 8 days by Evanite culture (300 mg-soluble protein) with ability to completely degrade tetrachloroethene (PCE) and TCE to ETH under anaerobic conditions. To determine the effect of formate as electron donor instead of H2, about 3 or 11 mmol of formate injected into batch-reactor every 15 days was enough to support H2 for dechlorination of c-DCE to VC and ETH. Soluble protein concentration of Evanite culture during the batch test increased from 300 mg to 688 mg for 120 days. In CFSTR test, TCE was fed continuously at 9.9 ppm (75.38 μmol/L) and the influent formate feed concentration increased stepwise from 1.3 mmol/L to 14.3 mmol/L. Injected TCE was accumulated at HRT 18 days for 13 days, but TCE was completed degraded at HRT 36 days without accumulation during left of experiment period, getting H2 from fermentative hydrogen production of injected formate. Although c-DCE was also accumulated for 23 days after CFSTR operation, it reached steady-state without accumulation in presence of excessive formate. However, since c-DCE in CFSTR was not completely dechlorinated, we will determine the transcriptional level of enzyme involved in reductive dechlorination of TCE, c-DCE, and VC in our future work.

Dynamic Resource Allocation to Improve Service Performance in Order Fulfillment Systems

DTIC Science & Technology

2009-01-01

efficient system uses economies of scale at two points: orders are batched before processing, which reduces processing costs, and processed or- ders ...the ef- fects of batching on order picking processes is well-researched and well-understood ( van den Berg and Gademann, 1999). Because orders are...a final so- journ time distribution. Our work builds on existing research in matrix-geometric methods by Neuts (1981), Asmussen and M0ller (2001

A strategy for clone selection under different production conditions.

PubMed

Legmann, Rachel; Benoit, Brian; Fedechko, Ronald W; Deppeler, Cynthia L; Srinivasan, Sriram; Robins, Russell H; McCormick, Ellen L; Ferrick, David A; Rodgers, Seth T; Russo, A Peter

2011-01-01

Top performing clones have failed at the manufacturing scale while the true best performer may have been rejected early in the screening process. Therefore, the ability to screen multiple clones in complex fed-batch processes using multiple process variations can be used to assess robustness and to identify critical factors. This dynamic ranking of clones' strategy requires the execution of many parallel experiments than traditional approaches. Therefore, this approach is best suited for micro-bioreactor models which can perform hundreds of experiments quickly and efficiently. In this study, a fully monitored and controlled small scale platform was used to screen eight CHO clones producing a recombinant monoclonal antibody across several process variations, including different feeding strategies, temperature shifts and pH control profiles. The first screen utilized 240 micro-bioreactors were run for two weeks for this assessment of the scale-down model as a high-throughput tool for clone evaluation. The richness of the outcome data enable to clearly identify the best and worst clone as well as process in term of maximum monoclonal antibody titer. The follow-up comparison study utilized 180 micro-bioreactors in a full factorial design and a subset of 12 clone/process combinations was selected to be run parallel in duplicate shake flasks. Good correlation between the micro-bioreactor predictions and those made in shake flasks with a Pearson correlation value of 0.94. The results also demonstrate that this micro-scale system can perform clone screening and process optimization for gaining significant titer improvements simultaneously. This dynamic ranking strategy can support better choices of production clones. Copyright © 2011 American Institute of Chemical Engineers (AIChE).

Tier 3 batch system data locality via managed caches

NASA Astrophysics Data System (ADS)

Fischer, Max; Giffels, Manuel; Jung, Christopher; Kühn, Eileen; Quast, Günter

2015-05-01

Modern data processing increasingly relies on data locality for performance and scalability, whereas the common HEP approaches aim for uniform resource pools with minimal locality, recently even across site boundaries. To combine advantages of both, the High- Performance Data Analysis (HPDA) Tier 3 concept opportunistically establishes data locality via coordinated caches. In accordance with HEP Tier 3 activities, the design incorporates two major assumptions: First, only a fraction of data is accessed regularly and thus the deciding factor for overall throughput. Second, data access may fallback to non-local, making permanent local data availability an inefficient resource usage strategy. Based on this, the HPDA design generically extends available storage hierarchies into the batch system. Using the batch system itself for scheduling file locality, an array of independent caches on the worker nodes is dynamically populated with high-profile data. Cache state information is exposed to the batch system both for managing caches and scheduling jobs. As a result, users directly work with a regular, adequately sized storage system. However, their automated batch processes are presented with local replications of data whenever possible.

Biogas Production from Protein-Rich Biomass: Fed-Batch Anaerobic Fermentation of Casein and of Pig Blood and Associated Changes in Microbial Community Composition

PubMed Central

Kovács, Etelka; Wirth, Roland; Maróti, Gergely; Bagi, Zoltán; Rákhely, Gábor; Kovács, Kornél L.

2013-01-01

It is generally accepted as a fact in the biogas technology that protein-rich biomass substrates should be avoided due to inevitable process inhibition. Substrate compositions with a low C/N ratio are considered difficult to handle and may lead to process failure, though protein-rich industrial waste products have outstanding biogas generation potential. This common belief has been challenged by using protein-rich substrates, i.e. casein and precipitated pig blood protein in laboratory scale continuously stirred mesophilic fed-batch biogas fermenters. Both substrates proved suitable for sustained biogas production (0.447 L CH4/g protein oDM, i.e. organic total solids) in high yield without any additives, following a period of adaptation of the microbial community. The apparent key limiting factors in the anaerobic degradation of these proteinaceous materials were the accumulation of ammonia and hydrogen sulfide. Changes in time in the composition of the microbiological community were determined by next-generation sequencing-based metagenomic analyses. Characteristic rearrangements of the biogas-producing community upon protein feeding and specific differences due to the individual protein substrates were recognized. The results clearly demonstrate that sustained biogas production is readily achievable, provided the system is well-characterized, understood and controlled. Biogas yields (0.45 L CH4/g oDM) significantly exceeding those of the commonly used agricultural substrates (0.25-0.28 L CH4/g oDM) were routinely obtained. The results amply reveal that these high-energy-content waste products can be converted to biogas, a renewable energy carrier with flexible uses that can replace fossil natural gas in its applications. Process control, with appropriate acclimation of the microbial community to the unusual substrate, is necessary. Metagenomic analysis of the microbial community by next-generation sequencing allows a precise determination of the alterations in the community composition in the course of the process. PMID:24146974

Bacterial, viral and turbidity removal by intermittent slow sand filtration for household use in developing countries: experimental investigation and modeling.

PubMed

Jenkins, Marion W; Tiwari, Sangam K; Darby, Jeannie

2011-11-15

A two-factor three-block experimental design was developed to permit rigorous evaluation and modeling of the main effects and interactions of sand size (d(10) of 0.17 and 0.52 mm) and hydraulic head (10, 20, and 30 cm) on removal of fecal coliform (FC) bacteria, MS2 bacteriophage virus, and turbidity, under two batch operating modes ('long' and 'short') in intermittent slow sand filters (ISSFs). Long operation involved an overnight pause time between feeding of two successive 20 L batches (16 h average batch residence time (RT)). Short operation involved no pause between two 20 L batch feeds (5h average batch RT). Conditions tested were representative of those encountered in developing country field settings. Over a ten week period, the 18 experimental filters were fed river water augmented with wastewater (influent turbidity of 5.4-58.6 NTU) and maintained with the wet harrowing method. Linear mixed modeling allowed systematic estimates of the independent marginal effects of each independent variable on each performance outcome of interest while controlling for the effects of variations in a batch's actual residence time, days since maintenance, and influent turbidity. This is the first study in which simultaneous measurement of bacteria, viruses and turbidity removal at the batch level over an extended duration has been undertaken with a large number of replicate units to permit rigorous modeling of ISSF performance variability within and across a range of likely filter design configurations and operating conditions. On average, the experimental filters removed 1.40 log fecal coliform CFU (SD 0.40 log, N=249), 0.54 log MS2 PFU (SD 0.42 log, N=245) and 89.0 percent turbidity (SD 6.9 percent, N=263). Effluent turbidity averaged 1.24 NTU (SD 0.53 NTU, N=263) and always remained below 3 NTU. Under the best performing design configuration and operating mode (fine sand, 10 cm head, long operation, initial HLR of 0.01-0.03 m/h), mean 1.82 log removal of bacteria (98.5%) and mean 0.94 log removal of MS2 viruses (88.5%) were achieved. Results point to new recommendations regarding filter design, manufacture, and operation for implementing ISSFs in local settings in developing countries. Sand size emerged as a critical design factor on performance. A single layer of river sand used in this investigation demonstrated removals comparable to those reported for 2 layers of crushed sand. Pause time and increased residence time each emerged as highly beneficial for improving removal performance on all four outcomes. A relatively large and significant negative effect of influent turbidity on MS2 viral removal in the ISSF was measured in parallel with a much smaller weaker positive effect of influent turbidity on FC bacterial removal. Disturbance of the schmutzdecke by wet harrowing showed no effect on virus removal and a modest reductive effect on the bacterial and turbidity removal as measured 7 days or more after the disturbance. For existing coarse sand ISSFs, this research indicates that a reduction in batch feed volume, effectively reducing the operating head and increasing the pore:batch volume ratio, could improve their removal performance by increasing batch residence time. Copyright © 2011 Elsevier Ltd. All rights reserved.

Bioreactor Performance Parameters for an Industrially-Promising Methanotroph Methylomicrobium buryatense 5GB1

DOE PAGES

Gilman, Alexey; Laurens, Lieve M.; Puri, Aaron W.; ...

2015-11-16

Methane is a feedstock of interest for the future, both from natural gas and from renewable biogas sources. Methanotrophic bacteria have the potential to enable commercial methane bioconversion to value-added products such as fuels and chemicals. A strain of interest for such applications is Methylomicrobium buryatense 5GB1, due to its robust growth characteristics. But, to take advantage of the potential of this methanotroph, it is important to generate comprehensive bioreactor-based datasets for different growth conditions to compare bioprocess parameters. The datasets of growth parameters, gas utilization rates, and products (total biomass, extracted fatty acids, glycogen, excreted acids) were obtained formore » cultures of M. buryatense 5GB1 grown in continuous culture under methane limitation and O2 limitation conditions. Additionally, experiments were performed involving unrestricted batch growth conditions with both methane and methanol as substrate. All four growth conditions show significant differences. The most notable changes are the high glycogen content and high formate excretion for cells grown on methanol (batch), and high O2:CH4 utilization ratio for cells grown under methane limitation. The results presented here represent the most comprehensive published bioreactor datasets for a gamma-proteobacterial methanotroph. This information shows that metabolism by M. buryatense 5GB1 differs significantly for each of the four conditions tested. O2 limitation resulted in the lowest relative O2 demand and fed-batch growth on methane the highest. Future studies are needed to understand the metabolic basis of these differences. However, these results suggest that both batch and continuous culture conditions have specific advantages, depending on the product of interest.« less

Pilot scale biodiesel production from microbial oil of Rhodosporidium toruloides DEBB 5533 using sugarcane juice: Performance in diesel engine and preliminary economic study.

PubMed

Soccol, Carlos Ricardo; Dalmas Neto, Carlos José; Soccol, Vanete Thomaz; Sydney, Eduardo Bittencourt; da Costa, Eduardo Scopel Ferreira; Medeiros, Adriane Bianchi Pedroni; Vandenberghe, Luciana Porto de Souza

2017-01-01

A successful pilot-scale process for biodiesel production from microbial oil (Biooil) produced by Rhodosporidium toruloides DEBB 5533 is presented. Using fed-batch strategy (1000L working volume), a lipid productivity of 0.44g/L.h was obtained using a low-cost medium composed by sugarcane juice and urea. The microbial oil was used for biodiesel production and its performance was evaluated in diesel engine tests, showing very good performance, especially for the blend B20 SCO, when operating at 2500rpm with lower pollutant emissions (CO 2 - 220% less; CO - 7-fold less; NO X 50% less and no detectable HC emissions (<0.11ppm)) when compared with the blends of standard biofuel from soybean oil. A preliminary analysis showed that microbial biodiesel is economically competitive (US$ 0.76/L) when compared to the vegetable biodiesel (US$ 0.81/L). Besides, the yield of biodiesel from microbial oil is higher (4172L/ha of cultivated sugarcane) that represents 6.3-fold the yield of standard biodiesel (661L/ha of cultivated soybean). Copyright © 2016 Elsevier Ltd. All rights reserved.

In-Situ Biocatalytic Production of Trehalose with Autoinduction Expression of Trehalose Synthase.

PubMed

Yan, Xincheng; Zhu, Liying; Yu, Yadong; Xu, Qing; Huang, He; Jiang, Ling

2018-02-14

We developed an in-situ biocatalytic process that couples autoinduction expression of trehalose synthase (TreS) and whole-cell catalysis for trehalose production. With lactose as the autoinducer, the activity of recombinant TreS in recombinant Escherichia coli was optimized through a visualization method, which resulted in a maximum value of 12 033 ± 730 U/mL in pH-stat fed-batch fermentation mode. Meanwhile, the permeability of the autoinduced E. coli increased significantly, which makes it possible to be directly used as a whole-cell biocatalyst for trehalose production, whereby the byproduct glucose can also act as an extra carbon source. In this case, the final yield of trehalose was improved to 90.5 ± 5.7% and remained as high as 83.2 ± 5.0% at the 10th batch, which is the highest value achieved using recombinant TreS. Finally, an integrated strategy for trehalose production was established, and its advantages compared to the traditional mode have been summarized.

Sustainable production of valuable compound 3-succinoyl-pyridine by genetically engineering Pseudomonas putida using the tobacco waste.

PubMed

Wang, Weiwei; Xu, Ping; Tang, Hongzhi

2015-11-17

Treatment of solid and liquid tobacco wastes with high nicotine content remains a longstanding challenge. Here, we explored an environmentally friendly approach to replace tobacco waste disposal with resource recovery by genetically engineering Pseudomonas putida. The biosynthesis of 3-succinoyl-pyridine (SP), a precursor in the production of hypotensive agents, from the tobacco waste was developed using whole cells of the engineered Pseudomonas strain, S16dspm. Under optimal conditions in fed-batch biotransformation, the final concentrations of product SP reached 9.8 g/L and 8.9 g/L from aqueous nicotine solution and crude suspension of the tobacco waste, respectively. In addition, the crystal compound SP produced from aqueous nicotine of the tobacco waste in batch biotransformation was of high purity and its isolation yield on nicotine was 54.2%. This study shows a promising route for processing environmental wastes as raw materials in order to produce valuable compounds.

Modeling of process parameters for enhanced production of coenzyme Q10 from Rhodotorula glutinis.

PubMed

Balakumaran, Palanisamy Athiyaman; Meenakshisundaram, Sankaranarayanan

2015-01-01

Coenzyme Q10 (CoQ10) plays an indispensable role in ATP generation through oxidative phosphorylation and helps in scavenging superoxides generated during electron transfer reactions. It finds extensive applications specifically related to oxidative damage and metabolic dysfunctions. This article reports the use of a statistical approach to optimize the concentration of key variables for the enhanced production of CoQ10 by Rhodotorula glutinis in a lab-scale fermenter. The culture conditions that promote optimum growth and CoQ10 production were optimized and the interaction of significant variables para-hydroxybenzoic acid (PHB, 819.34 mg/L) and soybean oil (7.78% [v/v]) was studied using response surface methodology (RSM). CoQ10 production increased considerably from 10 mg/L (in control) to 39.2 mg/L in batch mode with RSM-optimized precursor concentration. In the fed-batch mode, PHB and soybean oil feeding strategy enhanced CoQ10 production to 78.2 mg/L.

Actual Waste Demonstration of the Nitric-Glycolic Flowsheet for Sludge Batch 9 Qualification

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

J. D. Newell; Pareizs, J. M.; Martino, C. J.

For each sludge batch that is processed in the Defense Waste Processing Facility (DWPF), the Savannah River National Laboratory (SRNL) performs qualification testing to demonstrate that the sludge batch is processable. Testing performed by the Savannah River National Laboratory has shown glycolic acid to be effective in replacing the function of formic acid in the DWPF chemical process. The nitric-glycolic flowsheet reduces mercury, significantly lowers the catalytic generation of hydrogen and ammonia which could allow purge reduction in the Sludge Receipt and Adjustment Tank (SRAT), stabilizes the pH and chemistry in the SRAT and the Slurry Mix Evaporator (SME), allowsmore » for effective rheology adjustment, and is favorable with respect to melter flammability. In order to implement the new flowsheet, SRAT and SME cycles, designated SC-18, were performed using a Sludge Batch (SB) 9 slurry blended from SB8 Tank 40H and Tank 51H samples. The SRAT cycle involved adding nitric and glycolic acids to the sludge, refluxing to steam strip mercury, and dewatering to a targeted solids concentration. Data collected during the SRAT cycle included offgas analyses, process temperatures, heat transfer, and pH measurements. The SME cycle demonstrated the addition of glass frit and the replication of six canister decontamination additions. The demonstration concluded with dewatering to a targeted solids concentration. Data collected during the SME cycle included offgas analyses, process temperatures, heat transfer, and pH measurements. Slurry and condensate samples were collected for subsequent analysis« less

Effect of the inclusion of chestnut in the finishing diet on volatile compounds during the manufacture of dry-cured "Lacón" from Celta pig breed.

PubMed

Lorenzo, José M; Franco, Daniel; Carballo, Javier

2014-01-01

The effect of the finishing diet on the volatile compounds throughout the manufacture of dry-cured "lacón" (a Spanish traditional meat product), from the Celta pig breed was studied. Thirty-six pigs were separated into three groups according to the type of feeding during the finish-fattening period of three months (concentrate, mixed diet and chestnut). From the pigs of each diet, four batches of dry-cured "lacón" were manufactured. From each batch, samples of fresh meat, meat after salting, after post-salting, and after 14, 28, 56 and 84 days of drying-ripening were taken. Volatiles were extracted by a purge-and-trap method and analyzed by gas chromatographic/mass spectrometry (GC/MS). Seventy-six volatile compounds were identified and quantified from dry-cured "lacón" samples in pigs finished with chestnut, eighty-two for concentrate fed pigs and eighty in pigs fed with the mixed diet. The number of identified volatile compounds increased during the manufacturing process; at 84 days of drying-ripening, in the dry-cured "lacón" samples from pigs finished with concentrate, mixed diet and chestnut, 54, 58 and 62 volatile compounds were detected, respectively. The most abundant group of flavour compounds at the end of the manufacturing process was hydrocarbons in the three feeding systems, followed by aldehydes, ketones and alcohols. Discriminant analysis selected six variables (dodecane, butadienol, pentenol, 2-pentenal, decen-3-ona and pyridine-2-methyl) and calculated two discriminating functions which allowed verification of chestnut in the finishing diet. © 2013.

Enhanced nitrogen removal with spent mushroom compost in a sequencing batch reactor.

PubMed

Yang, Yunlong; Tao, Xin; Lin, Ershu; Hu, Kaihui

2017-11-01

In order to remove nitrogen effectively from the wastewater with a low C/N ratio, the feasibility of using spent mushroom compost (SMC) hydrolysates as carbon sources for denitrification was investigated in a sequencing batch reactor (SBR). With SMCs supplement, the SBR performance was improved obviously within the 180days of operation. The total nitrogen removal was promoted from 46.9% to 81-89.4%, and no negative impact induced by different SMCs on the SBR system was observed. The abundance of functional genes including amoA, nirS/K, norB and nosZ in the active sludge was quantified by qPCR, and most of them elevated after SMC was fed. 16S rRNA gene high-throughput sequencing showed that the significant change in microbial community not only promoted pollutants removal but also benefited the stability of the reactor. Therefore, SMC could be an extremely promising carbon source used for nitrogen removal due to its cost-effective and efficient characteristics. Copyright © 2017 Elsevier Ltd. All rights reserved.

Effects of hydrogen peroxide feeding strategies on the photochemical degradation of polyvinyl alcohol.

PubMed

Hamad, Dina; Dhib, Ramdhane; Mehrvar, Mehrab

2016-11-01

The performance of batch and fed-batch photoreactors with that of continuous photoreactor for the treatment of aqueous polyvinyl alcohol (PVA) solutions is compared. Hydrogen peroxide feeding strategies, residence time, and [H2O2]/[PVA] mass ratio are examined for their impacts on the molecular weight distribution (MWD) of PVA and the total organic carbon (TOC) removal. The results prove that a continuous addition of H2O2 during the degradation reaction ensures the utilization of the produced radicals to minimize the oxidant consumption and maximize the TOC removal and the PVA degradation in a short irradiation time. Also, the MWD of PVA is found to be bimodal and shifted towards lower molecular weights with small shoulder peak indicating a progressive disappearance of the higher molecular weight fractions that is in accordance with the random chains scission mechanism. Besides, the hydrogen peroxide feeding strategies are found to have a great effect on the reduction in H2O2 residuals in the effluent.

Robust artificial intelligence tool for automatic start-up of the supplementary medium feeding in recombinant E. coli cultivations.

PubMed

Horta, Antônio Carlos Luperni; da Silva, Adilson José; Sargo, Cíntia Regina; Gonçalves, Viviane Maimoni; Zangirolami, Teresa Cristina; Giordano, Roberto de Campos

2011-09-01

One of the most important events in fed-batch fermentations is the definition of the moment to start the feeding. This paper presents a methodology for a rational selection of the architecture of an artificial intelligence (AI) system, based on a neural network committee (NNC), which identifies the end of the batch phase. The AI system was successfully used during high cell density cultivations of recombinant Escherichia coli. The AI algorithm was validated for different systems, expressing three antigens to be used in human and animal vaccines: fragments of surface proteins of Streptococcus pneumoniae (PspA), clades 1 and 3, and of Erysipelothrix rhusiopathiae (SpaA). Standard feed-forward neural networks (NNs), with a single hidden layer, were the basis for the NNC. The NN architecture with best performance had the following inputs: stirrer speed, inlet air, and oxygen flow rates, carbon dioxide evolution rate, and CO2 molar fraction in the exhaust gas.

On Operating a Nanofiltration Membrane for Olive Mill Wastewater Purification at Sub- and Super-Boundary Conditions.

PubMed

Stoller, Marco; Ochando-Pulido, Javier Miguel; Field, Robert

2017-07-14

In the last decades, membrane processes have gained a significant share of the market for wastewater purification. Although the product (i.e., purified water) is not of high added value, these processes are feasible both technically and from an economic point of view, provided the flux is relatively high and that membrane fouling is strongly inhibited. By controlling membrane fouling, the membrane may work for years without service, thus dramatically reducing operating costs and the need for membrane substitution. There is tension between operating at high permeate fluxes, which enhances fouling but reduces capital costs, and operating at lower fluxes which increases capital costs. Operating batch membrane processes leads to increased difficulties, since the feed fed to the membrane changes as a function of the recovery value. This paper is concerned with the operation of such a process. Membrane process designers should therefore avoid membrane fouling by operating membranes away from the permeate flux point where severe fouling is triggered. The design and operation of membrane purification plants is a difficult task, and the precision to properly describe the evolution of the fouling phenomenon as a function of the operating conditions is a key to success. Many reported works have reported on the control of fouling by operating below the boundary flux. On the other hand, only a few works have successfully sought to exploit super-boundary operating conditions; most super-boundary operations are reported to have led to process failures. In this work, both sub- and super-boundary operating conditions for a batch nanofiltration membrane process used for olive mill wastewater treatment were investigated. A model to identify a priori the point of transition from a sub-boundary to a super-boundary operation during a batch operation was developed, and this will provide membrane designers with a helpful tool to carefully avoid process failures.

Neuro-estimator based GMC control of a batch reactive distillation.

PubMed

Prakash, K J Jithin; Patle, Dipesh S; Jana, Amiya K

2011-07-01

In this paper, an artificial neural network (ANN)-based nonlinear control algorithm is proposed for a simulated batch reactive distillation (RD) column. In the homogeneously catalyzed reactive process, an esterification reaction takes place for the production of ethyl acetate. The fundamental model has been derived incorporating the reaction term in the model structure of the nonreactive distillation process. The process operation is simulated at the startup phase under total reflux conditions. The open-loop process dynamics is also addressed running the batch process at the production phase under partial reflux conditions. In this study, a neuro-estimator based generic model controller (GMC), which consists of an ANN-based state predictor and the GMC law, has been synthesized. Finally, this proposed control law has been tested on the representative batch reactive distillation comparing with a gain-scheduled proportional integral (GSPI) controller and with its ideal performance (ideal GMC). Copyright © 2011 ISA. Published by Elsevier Ltd. All rights reserved.

Effects of a high-fibre diet on ammonia and greenhouse gas emissions from gestating sows and fattening pigs

NASA Astrophysics Data System (ADS)

Philippe, François-Xavier; Laitat, Martine; Wavreille, José; Nicks, Baudouin; Cabaraux, Jean-François

2015-05-01

This study aims to measure under barn conditions the emissions of NH3, N2O, CH4 and CO2 associated with gestating sows (trial 1) and fattening pigs (trial 2) fed either a control diet (CTD) based on cereals or a high-fibre diet (HFD) based on sugar beet pulp (SBP). Three successive batches of 10 Belgian Landrace gestating sows were used for trial 1. Two successive batches of 24 Piétrain × Belgian Landrace fattening pigs were used for trial 2. Animals were kept on slatted floor. The gas emissions were measured by infrared photoacoustic detection and expressed per day and per livestock unit, equals to 500 kg body weight. Similar trends were observed for both animal types. With HFD, the NH3 emissions were reduced (27.2 vs. 36.5 g for the gestating sows, P < 0.001; 23.2 vs. 45.0 g for the fattening pigs, P < 0.001) but the CH4 emissions were increased (41.5 vs. 21.0 g for gestating sows, P < 0.001; 37.9 vs. 27.2 g for fattening pigs, P < 0.001). The fibre content of the diet had not significant impact on N2O emissions (around 1.4 g for gestating sows and 2.1 g for fattening pigs, P > 0.05), and on CO2 emissions (around 6.0 kg for gestating sows and 9.1 kg for fattening pigs, P > 0.05). Most of manure parameters did not statistically differ regarding the treatment. Reproductive performance and body condition of the sows were not affected by the diet. However, growth performance and carcass traits of the HFD-fed fattening pigs were deteriorated compared to CTD.

Sliding mode control of dissolved oxygen in an integrated nitrogen removal process in a sequencing batch reactor (SBR).

PubMed

Muñoz, C; Young, H; Antileo, C; Bornhardt, C

2009-01-01

This paper presents a sliding mode controller (SMC) for dissolved oxygen (DO) in an integrated nitrogen removal process carried out in a suspended biomass sequencing batch reactor (SBR). The SMC performance was compared against an auto-tuning PI controller with parameters adjusted at the beginning of the batch cycle. A method for cancelling the slow DO sensor dynamics was implemented by using a first order model of the sensor. Tests in a lab-scale reactor showed that the SMC offers a better disturbance rejection capability than the auto-tuning PI controller, furthermore providing reasonable performance in a wide range of operation. Thus, SMC becomes an effective robust nonlinear tool to the DO control in this process, being also simple from a computational point of view, allowing its implementation in devices such as industrial programmable logic controllers (PLCs).

Inhibitory effect of carbon dioxide on the fed-batch culture of Ralstonia eutropha: evaluation by CO2 pulse injection and autogenous CO2 methods.

PubMed

Shang, Longan; Jiang, Min; Ryu, Chul Hee; Chang, Ho Nam; Cho, Soon Haeng; Lee, Jong Won

2003-08-05

In order to see the effect of CO(2) inhibition resulting from the use of pure oxygen, we carried out a comparative fed-batch culture study of polyhydroxybutyric acid (PHB) production by Ralstonia eutropha using air and pure oxygen in 5-L, 30-L, and 300-L fermentors. The final PHB concentrations obtained with pure O(2) were 138.7 g/L in the 5-L fermentor and 131.3 g/L in the 30-L fermentor, which increased 2.9 and 6.2 times, respectively, as compared to those obtained with air. In the 300-L fermentor, the fed-batch culture with air yielded only 8.4 g/L PHB. However, the maximal CO(2) concentrations in the 5-L fermentor increased significantly from 4.1% (air) to 15.0% (pure O(2)), while it was only 1.6% in the 30-L fermentor with air, but reached 14.2% in the case of pure O(2). We used two different experimental methods for evaluating CO(2) inhibition: CO(2) pulse injection and autogenous CO(2) methods. A 10 or 22% (v/v) CO(2) pulse with a duration of 3 or 6 h was introduced in a pure-oxygen culture of R. eutropha to investigate how CO(2) affects the synthesis of biomass and PHB. CO(2) inhibited the cell growth and PHB synthesis significantly. The inhibitory effect became stronger with the increase of the CO(2) concentration and pulse duration. The new proposed autogenous CO(2) method makes it possible to place microbial cells under different CO(2) level environments by varying the gas flow rate. Introduction of O(2) gas at a low flow rate of 0.42 vvm resulted in an increase of CO(2) concentration to 30.2% in the exit gas. The final PHB of 97.2 g/L was obtained, which corresponded to 70% of the PHB production at 1.0 vvm O(2) flow rate. This new method measures the inhibitory effect of CO(2) produced autogenously by cells through the entire fermentation process and can avoid the overestimation of CO(2) inhibition without introducing artificial CO(2) into the fermentor. Copyright 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 312-320, 2003.

High-titer and productivity of l-(+)-lactic acid using exponential fed-batch fermentation with Bacillus coagulans arr4, a new thermotolerant bacterial strain.

PubMed

Coelho, Luciana Fontes; Beitel, Susan Michelz; Sass, Daiane Cristina; Neto, Paulo Marcelo Avila; Contiero, Jonas

2018-04-01

Bacillus coagulans arr4 is a thermotolerant microorganism with great biotechnological potential for l-(+)-lactic acid production from granulated sugar and yeast extract. The highest l-(+)-lactic acid production was obtained with Ca(OH) 2 . The maximum production of l-(+)-lactic acid (206.81 g/L) was observed in exponential feeding using granulated sugar solution (900 g/L) and yeast extract (1%) at 50 °C, pH 6.5, and initial granulated sugar concentration of 100 g/L at 39 h. 5.3 g/L h productivity and 97% yield were observed, and no sugar remained. Comparing the simple batch with exponential fed-batch fermentation, the l(+) lactic acid production was improved in 133.22% and dry cell weight was improved in 83.29%, using granulated sugar and yeast extract. This study presents the highest productivity of lactic acid ever observed in the literature, on the fermentation of thermotolerant Bacillus sp. as well as an innovative and high-efficiency purification technology, using low-cost substances as Celite and charcoal. The recovery of lactic acid was 86%, with 100% protein removal, and the fermentation medium (brown color) became a colorless solution.

Enrichment of denitrifying methane-oxidizing microorganisms using up-flow continuous reactors and batch cultures.

PubMed

Hatamoto, Masashi; Kimura, Masafumi; Sato, Takafumi; Koizumi, Masato; Takahashi, Masanobu; Kawakami, Shuji; Araki, Nobuo; Yamaguchi, Takashi

2014-01-01

Denitrifying anaerobic methane oxidizing (DAMO) microorganisms were enriched from paddy field soils using continuous-flow and batch cultures fed with nitrate or nitrite as a sole electron acceptor. After several months of cultivation, the continuous-flow cultures using nitrite showed remarkable simultaneous methane oxidation and nitrite reduction and DAMO bacteria belonging to phylum NC10 were enriched. A maximum volumetric nitrite consumption rate of 70.4±3.4 mg-N·L(-1)·day(-1) was achieved with very short hydraulic retention time of 2.1 hour. In the culture, about 68% of total microbial cells were bacteria and no archaeal cells were detected by fluorescence in situ hybridization. In the nitrate-fed continuous-flow cultures, 58% of total microbial cells were bacteria while archaeal cells accounted for 7% of total cell numbers. Phylogenetic analysis of pmoA gene sequence showed that enriched DAMO bacteria in the continuous-flow cultivation had over 98% sequence similarity to DAMO bacteria in the inoculum. In contrast, for batch culture, the enriched pmoA gene sequences had 89-91% sequence similarity to DAMO bacteria in the inoculum. These results indicate that electron acceptor and cultivation method strongly affect the microbial community structures of DAMO consortia.

Design of neural network model-based controller in a fed-batch microbial electrolysis cell reactor for bio-hydrogen gas production

NASA Astrophysics Data System (ADS)

Azwar; Hussain, M. A.; Abdul-Wahab, A. K.; Zanil, M. F.; Mukhlishien

2018-03-01

One of major challenge in bio-hydrogen production process by using MEC process is nonlinear and highly complex system. This is mainly due to the presence of microbial interactions and highly complex phenomena in the system. Its complexity makes MEC system difficult to operate and control under optimal conditions. Thus, precise control is required for the MEC reactor, so that the amount of current required to produce hydrogen gas can be controlled according to the composition of the substrate in the reactor. In this work, two schemes for controlling the current and voltage of MEC were evaluated. The controllers evaluated are PID and Inverse neural network (NN) controller. The comparative study has been carried out under optimal condition for the production of bio-hydrogen gas wherein the controller output is based on the correlation of optimal current and voltage to the MEC. Various simulation tests involving multiple set-point changes and disturbances rejection have been evaluated and the performances of both controllers are discussed. The neural network-based controller results in fast response time and less overshoots while the offset effects are minimal. In conclusion, the Inverse neural network (NN)-based controllers provide better control performance for the MEC system compared to the PID controller.

Influence of co-substrate on textile wastewater treatment and microbial community changes in the anaerobic biological sulfate reduction process.

PubMed

Rasool, Kashif; Mahmoud, Khaled A; Lee, Dae Sung

2015-12-15

This study investigated the anaerobic treatment of sulfate-rich synthetic textile wastewater in three sulfidogenic sequential batch reactors (SBRs). The experimental protocol was designed to examine the effect of three different co-substrates (lactate, glucose, and ethanol) and their concentrations on wastewater treatment performance. Sulfate reduction and dye degradation were improved when lactate and ethanol were used as electron donors, as compared with glucose. Moreover, under co-substrate limited concentrations, color, sulfate, and chemical oxygen demand (COD) removal efficiencies were declined. By reducing co-substrate COD gradually from 3000 to 500 mg/L, color removal efficiencies were decreased from 98.23% to 78.46%, 63.37%, and 69.10%, whereas, sulfate removal efficiencies were decreased from 98.42%, 82.35%, and 87.0%, to 30.27%, 21.50%, and 10.13%, for lactate, glucose, and ethanol fed reactors, respectively. Fourier transform infrared spectroscopy (FTIR) and total aromatic amine analysis revealed lactate to be a potential co-substrate for further biodegradation of intermediate metabolites formed after dye degradation. Pyrosequencing analysis showed that microbial community structure was significantly affected by the co-substrate. The reactor with lactate as co-substrate showed the highest relative abundance of sulfate reducing bacteria (SRBs), followed by ethanol, whereas the glucose-fed reactor showed the lowest relative abundance of SRB. Copyright © 2015 Elsevier B.V. All rights reserved.

40 CFR Table 1 to Subpart Cccc of... - Emission Limitations

Code of Federal Regulations, 2010 CFR

2010-07-01

... Standards for Hazardous Air Pollutants: Manufacturing of Nutritional Yeast Part 63, Subpt. CCCC, Table 1... emission limitations in the following table: For each fed-batch fermenter producing yeast in the following... limitation does not apply during the production of specialty yeast. ...

40 CFR Table 1 to Subpart Cccc of... - Emission Limitations

Code of Federal Regulations, 2011 CFR

2011-07-01

... Standards for Hazardous Air Pollutants: Manufacturing of Nutritional Yeast Part 63, Subpt. CCCC, Table 1... emission limitations in the following table: For each fed-batch fermenter producing yeast in the following... limitation does not apply during the production of specialty yeast. ...

[Identification of the authentic quality of Longdanxiegan pill by systematic quantified fingerprint method based on three wavelength fusion chromatogram].

PubMed

Sun, Guoxiang; Zhang, Jingxian

2009-05-01

The three wavelength fusion high performance liquid chromatographic fingerprin (TWFFP) of Longdanxiegan pill (LDXGP) was established to identify the quality of LDXGP by the systematic quantified fingerprint method. The chromatographic fingerprints (CFPs) of the 12 batches of LDXGP were determined by reversed-phase high performance liquid chromatography. The technique of multi-wavelength fusion fingerprint was applied during processing the fingerprints. The TWFFPs containing 63 co-possessing peaks were obtained when choosing baicalin peak as the referential peak. The 12 batches of LDXGP were identified with hierarchical clustering analysis by using macro qualitative similarity (S(m)) as the variable. According to the results of classification, the referential fingerprint (RFP) was synthesized from 10 batches of LDXGP. Taking the RFP for the qualified model, all the 12 batches of LDXGP were evaluated by the systematic quantified fingerprint method. Among the 12 batches of LDXGP, 9 batches were completely qualified, the contents of 1 batch were obviously higher while the chemical constituents quantity and distributed proportion in 2 batches were not qualified. The systematic quantified fingerprint method based on the technique of multi-wavelength fusion fingerprint ca effectively identify the authentic quality of traditional Chinese medicine.

Biodegradable poly-ε-caprolactone microcarriers for efficient production of human mesenchymal stromal cells and secreted cytokines in batch and fed-batch bioreactors.

PubMed

Lam, Alan Tin-Lun; Li, Jian; Toh, Jessica Pei-Wen; Sim, Eileen Jia-Hui; Chen, Allen Kuan-Liang; Chan, Jerry Kok-Yen; Choolani, Mahesh; Reuveny, Shaul; Birch, William R; Oh, Steve Kah-Weng

2017-03-01

Large numbers of human mesenchymal stromal cells (MSCs) used for a variety of applications in tissue engineering and cell therapy can be generated by scalable expansion in a bioreactor using microcarriers (MCs) systems. However, the enzymatic digestion process needed to detach cells from the growth surface can affect cell viability and potentially the potency and differentiation efficiency. Thus, the main aim of our study was to develop biocompatible and biodegradable MCs that can support high MSC yields while maintaining their differentiation capability and potency. After cell expansion, the cells that covered MCs can be directly implanted in vivo without the need for cell harvesting or use of scaffold. Poly-ε-caprolactone (PCL) is known as a biocompatible and biodegradable material. However, it cannot be used for generation of MCs because its high density (1.14 g/cm 3 ) would exclude its applicability for suspension MCs in stirred reactors. In this article, we describe expansion and potency of MSCs propagated on low-density (1.06 g/cm 3 ) porous PCL MCs coated with extracellular matrices (LPCLs) in suspended stirred reactors. Using these LPCLs, cell yields of about 4 × 10 4 cells/cm 2 and 7- to 10-fold increases were obtained using four different MSC lines (bone marrow, cord blood, fetal and Wharton's jelly). These yields were comparable with those obtained using non-degradable MCs (Cytodex 3) and higher than two-dimensional monolayer (MNL) cultures. A fed-batch process, which demonstrated faster cell expansion (4.5 × 10 4 cells/cm 2 in 5 days as compared with 7 days in batch culture) and about 70% reduction in growth media usage, was developed and scaled up from 100-mL spinner flask to 1-L controlled bioreactor. Surface marker expression, trilineage differentiation and clonogenic potential of the MSCs expanded on LPCL were not affected. Cytokine secretion kinetics, which occurred mostly during late logarithmic phase, was usually comparable with that obtained in Cytodex 3 cultures and higher than MNL cultures. In conclusion, biodegradable LPCL can be used to efficiently expand a variety of MSC lines in stirred scalable reactors in a cost-effective manner while maintaining surface markers expression, differentiation capability and high levels of cytokine secretion. This study is the first step in testing these cell-biodegradable porous MC aggregates for tissue engineering and cell therapy, such as bone and cartilage regeneration, or wound healing. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

Integrating sequencing batch reactor with bio-electrochemical treatment for augmenting remediation efficiency of complex petrochemical wastewater.

PubMed

Yeruva, Dileep Kumar; Jukuri, Srinivas; Velvizhi, G; Naresh Kumar, A; Swamy, Y V; Venkata Mohan, S

2015-01-01

The present study evaluates the sequential integration of two advanced biological treatment methods viz., sequencing batch reactor (SBR) and bioelectrochemical treatment systems (BET) for the treatment of real-field petrochemical wastewater (PCW). Initially two SBR reactors were operated in aerobic (SBR(Ae)) and anoxic (SBR(Ax)) microenvironments with an organic loading rate (OLR) of 9.68 kg COD/m(3)-day. Relatively, SBR(Ax) showed higher substrate degradation (3.34 kg COD/m(3)-day) compared to SBR(Ae) (2.9 kg COD/m(3)-day). To further improve treatment efficiency, the effluents from SBR process were fed to BET reactors. BET(Ax) depicted higher SDR (1.92 kg COD/m(3)-day) with simultaneous power generation (17.12 mW/m(2)) followed by BET(Ae) (1.80 kg COD/m(3)-day; 14.25 mW/m(2)). Integrating both the processes documented significant improvement in COD removal efficiency due to the flexibility of combining multiple microenvironments sequentially. Results were supported with GC-MS and FTIR, which confirmed the increment in biodegradability of wastewater. Copyright © 2015 Elsevier Ltd. All rights reserved.

Effect of acid detergent fiber in hydrothermally pretreated sewage sludge on anaerobic digestion process

NASA Astrophysics Data System (ADS)

Takasaki, Rikiya; Yuan, Lee Chang; Kamahara, Hirotsugu; Atsuta, Youichi; Daimon, Hiroyuki

2017-10-01

Hydrothermal treatment is one of the pre-treatment method for anaerobic digestion. The application of hydrothermal treatment to sewage sludge of wastewater treatment plant has been succeeded to enhance the biogas production. The purpose of this study is to quantitatively clarify the effect of hydrothermal treatment on anaerobic digestion process focusing on acid detergent fiber (ADF) in sewage sludge, which is low biodegradability. The hydrothermal treatment experiment was carried out for 15 minutes between 160 °C and 200 °C respectively. The ADF content was decreased after hydrothermal treatment compared with untreated sludge. However, ADF content was increased when raising the treatment temperature from 160 °C to 200 °C. During batch anaerobic digestion experiment, untreated and treated sludge were examined for 10 days under 38 °C, and all samples were fed once based on volatile solids of samples. From batch anaerobic digestion experiment, as ADF content in sewage sludge increased, the total biogas production decreased. It was found that ADF content in sewage sludge influence on anaerobic digestion. Therefore, ADF could be one of the indicator to evaluate the effect of hydrothermal treatment to sewage sludge on anaerobic digestion.

Nitrate removal from high strength nitrate-bearing wastes in granular sludge sequencing batch reactors.

PubMed

Krishna Mohan, Tulasi Venkata; Renu, Kadali; Nancharaiah, Yarlagadda Venkata; Satya Sai, Pedapati Murali; Venugopalan, Vayalam Purath

2016-02-01

A 6-L sequencing batch reactor (SBR) was operated for development of granular sludge capable of denitrification of high strength nitrates. Complete and stable denitrification of up to 5420 mg L(-1) nitrate-N (2710 mg L(-1) nitrate-N in reactor) was achieved by feeding simulated nitrate waste at a C/N ratio of 3. Compact and dense denitrifying granular sludge with relatively stable microbial community was developed during reactor operation. Accumulation of large amounts of nitrite due to incomplete denitrification occurred when the SBR was fed with 5420 mg L(-1) NO3-N at a C/N ratio of 2. Complete denitrification could not be achieved at this C/N ratio, even after one week of reactor operation as the nitrite levels continued to accumulate. In order to improve denitrification performance, the reactor was fed with nitrate concentrations of 1354 mg L(-1), while keeping C/N ratio at 2. Subsequently, nitrate concentration in the feed was increased in a step-wise manner to establish complete denitrification of 5420 mg L(-1) NO3-N at a C/N ratio of 2. The results show that substrate concentration plays an important role in denitrification of high strength nitrate by influencing nitrite accumulation. Complete denitrification of high strength nitrates can be achieved at lower substrate concentrations, by an appropriate acclimatization strategy. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

CHO cells knocked out for TSC2 display an improved productivity of antibodies under fed batch conditions.

PubMed

McVey, Duncan; Aronov, Michael; Rizzi, Giovanni; Cowan, Alexis; Scott, Charo; Megill, John; Russell, Reb; Tirosh, Boaz

2016-09-01

The kinase mTOR operates in two cellular complexes, mTORC1 and mTORC2. mTORC1 adjusts metabolic activity according to external growth conditions and nutrients availability. When conditions are prosperous, mTOR facilitates protein and lipid biosyntheses and inhibits autophagy, while under metabolic constraints, however, its attenuation induces a catabolic program, energy preservation and autophagy. CHO is a key cell line for manufacturing of biologics owing to its remarkable ability to grow to high densities and maintain protein production and secretion for extended times. While high mTOR activity has been associated with high productivity in CHO cells, its inhibition by rapamycin has also been documented to augment productivity via promotion of viability. Here using CRISPR/Cas9 editing we engineered CHO cells to enforce high mTORC1 activity by knocking-out TSC2, a major mTOR inhibitory protein, or PTEN, a phosphatase that attenuates the PI3K/AKT/mTOR pathway. Only TSC2-deleted cells exhibited a constitutive activation of mTORC1 under fed batch conditions. Cells grew larger in size, synthesized more proteins and displayed an over twofold elevation in their specific productivity. While peak viable cell density was compromised, overall titers increased to an extent dependent upon the parental clone. Our data underscore manipulation of TSC as a strategy to improve performance of CHO cell in bioreactors. Biotechnol. Bioeng. 2016;113: 1942-1952. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Process integration for biological sulfate reduction in a carbon monoxide fed packed bed reactor.

PubMed

Kumar, Manoj; Sinharoy, Arindam; Pakshirajan, Kannan

2018-08-01

This study examined immobilized anaerobic biomass for sulfate reduction using carbon monoxide (CO) as the sole carbon source under batch and continuous fed conditions. The immobilized bacteria with beads made of 10% polyvinyl alcohol (PVA) showed best results in terms of sulfate reduction (84 ± 3.52%) and CO utilization (98 ± 1.67%). The effect of hydraulic retention time (HRT), sulfate loading rate and CO loading rate on sulfate and CO removal was investigated employing a 1L packed bed bioreactor containing the immobilized biomass. At 48, 24 and 12 h HRT, the sulfate removal was 94.42 ± 0.15%, 89.75 ± 0.47% and 61.08 ± 0.34%, respectively, along with a CO utilization of more than 90%. The analysis of variance (ANOVA) of the results obtained showed that only the initial CO concentration significantly affected the sulfate reduction process. The reactor effluent sulfate concentrations were 27.41 ± 0.44, 59.16 ± 1.08, 315.83 ± 7.33 mg/L for 250, 500 and 1000 mg/L of influent sulfate concentrations respectively, under the optimum operating conditions. The sulfate reduction rates matched well with low inlet sulfate loading rates, indicating stable performance of the bioreactor system. Overall, this study yielded very high sulfate reduction efficiency by the immobilized anaerobic biomass under high CO loading condition using the packed bed reactor system. Copyright © 2018 Elsevier Ltd. All rights reserved.

Comparison of different liquid anaerobic digestion effluents as inocula and nitrogen sources for solid-state batch anaerobic digestion of corn stover.

PubMed

Xu, Fuqing; Shi, Jian; Lv, Wen; Yu, Zhongtang; Li, Yebo

2013-01-01

Effluents from three liquid anaerobic digesters, fed with municipal sewage sludge, food waste, or dairy waste, were evaluated as inocula and nitrogen sources for solid-state batch anaerobic digestion of corn stover in mesophilic reactors. Three feedstock-to-effluent (F/E) ratios (i.e., 2, 4, and 6) were tested for each effluent. At an F/E ratio of 2, the reactor inoculated by dairy waste effluent achieved the highest methane yield of 238.5L/kg VS(feed), while at an F/E ratio of 4, the reactor inoculated by food waste effluent achieved the highest methane yield of 199.6L/kg VS(feed). The microbial population and chemical composition of the three effluents were substantially different. Food waste effluent had the largest population of acetoclastic methanogens, while dairy waste effluent had the largest populations of cellulolytic and xylanolytic bacteria. Dairy waste also had the highest C/N ratio of 8.5 and the highest alkalinity of 19.3g CaCO(3)/kg. The performance of solid-state batch anaerobic digestion reactors was closely related to the microbial status in the liquid anaerobic digestion effluents. Copyright © 2012 Elsevier Ltd. All rights reserved.

Production of ω-3 Polyunsaturated Fatty Acids From Cull Potato Using an Algae Culture Process

NASA Astrophysics Data System (ADS)

Chi, Zhanyou; Hu, Bo; Liu, Yan; Frear, Craig; Wen, Zhiyou; Chen, Shulin

Algal cultivation for converting cull potato to docosahexaenoic acid (DHA) was studied. Schizochytrium limacinum SR21 was selected as the better producing strain, compared with Thraustochytrium aureum because of higher cell density and DHA content. Used as both carbon and nitrogen source, an optimal ratio of hydrolyzed potato broth in the culture medium was determined as 50%, with which the highest production of 21.7 g/L dry algae biomass and 5.35 g/L DHA was obtained, with extra glucose supplemented. Repeat culture further improved the cell density but not fed batch culture, suggesting limited growth was most likely caused by metabolites inhibition.

Optimisation of enzymatic hydrolysis of apple pomace for production of biofuel and biorefinery chemicals using commercial enzymes.

PubMed

Gama, Repson; Van Dyk, J Susan; Pletschke, Brett I

2015-12-01

Apple pomace, a waste product from the apple juice industry is a potential feedstock for biofuel and biorefinery chemical production. Optimisation of hydrolysis conditions for apple pomace hydrolysis using Viscozyme L and Celluclast 1.5L was investigated using 1 L reaction volumes. The effects of temperature, pH, β-glucosidase supplementation and substrate feeding regimes were determined. Hydrolysis at room temperature using an unbuffered system gave optimal performance. Reactors in batch mode resulted in a better performance (4.2 g/L glucose and 16.8 g/L reducing sugar, 75 % yield for both) than fed-batch (3.2 g/L glucose and 14.6 g/L reducing sugar, 65.5 and 73.1 % yield respectively) in 72 h. The addition of Novozyme 188 to the core mixture of Viscozyme L and Celluclast 1.5L resulted in the doubling of glucose released. The main products (yield %) released from apple pomace hydrolysis were galacturonic acid (78 %), glucose (75 %), arabinose (90 %) and galactose (87 %). These products are potential raw materials for biofuel and biorefinery chemical production.

Experimental design of a twin-column countercurrent gradient purification process.

PubMed

Steinebach, Fabian; Ulmer, Nicole; Decker, Lara; Aumann, Lars; Morbidelli, Massimo

2017-04-07

As typical for separation processes, single unit batch chromatography exhibits a trade-off between purity and yield. The twin-column MCSGP (multi-column countercurrent solvent gradient purification) process allows alleviating such trade-offs, particularly in the case of difficult separations. In this work an efficient and reliable procedure for the design of the twin-column MCSGP process is developed. This is based on a single batch chromatogram, which is selected as the design chromatogram. The derived MCSGP operation is not intended to provide optimal performance, but it provides the target product in the selected fraction of the batch chromatogram, but with higher yield. The design procedure is illustrated for the isolation of the main charge isoform of a monoclonal antibody from Protein A eluate with ion-exchange chromatography. The main charge isoform was obtained at a purity and yield larger than 90%. At the same time process related impurities such as HCP and leached Protein A as well as aggregates were at least equally well removed. Additionally, the impact of several design parameters on the process performance in terms of purity, yield, productivity and buffer consumption is discussed. The obtained results can be used for further fine-tuning of the process parameters so as to improve its performance. Copyright © 2017 Elsevier B.V. All rights reserved.

Construction and fed-batch cultivation of Candida famata with enhanced riboflavin production.

PubMed

Dmytruk, Kostyantyn; Lyzak, Oleksy; Yatsyshyn, Valentyna; Kluz, Maciej; Sibirny, Vladimir; Puchalski, Czeslaw; Sibirny, Andriy

2014-02-20

Riboflavin (vitamin B2) is an essential nutrition component serving as a precursor of coenzymes FMN and FAD that are involved mostly in reactions of oxidative metabolism. Riboflavin is produced in commercial scale and is used in feed and food industries, and in medicine. The yeast Candida famata (Candida flareri) belongs to the group of so called "flavinogenic yeasts" which overproduce riboflavin under iron limitation. Three genes SEF1, RIB1 and RIB7 coding for a putative transcription factor, GTP cyclohydrolase II and riboflavin synthase, respectively were simultaneously overexpressed in the background of a non-reverting riboflavin producing mutant AF-4, obtained earlier in our laboratory using methods of classical selection (Dmytruk et al. (2011), Metabolic Engineering 13, 82-88). Cultivation conditions of the constructed strain were optimized for shake-flasks and bioreactor cultivations. The constructed strain accumulated up to 16.4g/L of riboflavin in optimized medium in a 7L laboratory bioreactor during fed-batch fermentation. Copyright © 2013 Elsevier B.V. All rights reserved.

Fed-batch anaerobic valorization of slaughterhouse by-products with mesophilic microbial consortia without methane production.

PubMed

Pessiot, J; Nouaille, R; Jobard, M; Singhania, R R; Bournilhas, A; Christophe, G; Fontanille, P; Peyret, P; Fonty, G; Larroche, C

2012-07-01

This work aimed at setting up a fully instrumented, laboratory-scale bioreactor enabling anaerobic valorization of solid substrates through hydrogen and/or volatile fatty acid (VFA) production using mixed microbial populations (consortia). The substrate used was made of meat-based wastes, especially from slaughterhouses, which are becoming available in large amounts as a consequence of the growing constraints for waste disposal from meat industry. A reconstituted microbial mesophilic consortium without Archaebacteria (methanogens), named PBr, was cultivated in a 5-L anaerobic bioreactor on slaughterhouse wastes. The experiments were carried out with sequential fed-batch operations, including liquid medium removal from the bioreactor and addition of fresh substrate. VFAs and nitrogen were the main metabolites observed, while hydrogen accumulation was very low and no methane production was evidenced. After 1,300 h of culture, yields obtained for VFAs reached 0.38 g/g dry matter. Strain composition of the microbial consortium was also characterized using molecular tools (temporal temperature gradient gel electrophoresis and gene sequencing).

De novo Biosynthesis of Biodiesel by Escherichia coli in Optimized Fed-Batch Cultivation

PubMed Central

Cai, Ke; Tan, Xiaoming; Lu, Xuefeng

2011-01-01

Biodiesel is a renewable alternative to petroleum diesel fuel that can contribute to carbon dioxide emission reduction and energy supply. Biodiesel is composed of fatty acid alkyl esters, including fatty acid methyl esters (FAMEs) and fatty acid ethyl esters (FAEEs), and is currently produced through the transesterification reaction of methanol (or ethanol) and triacylglycerols (TAGs). TAGs are mainly obtained from oilseed plants and microalgae. A sustainable supply of TAGs is a major bottleneck for current biodiesel production. Here we report the de novo biosynthesis of FAEEs from glucose, which can be derived from lignocellulosic biomass, in genetically engineered Escherichia coli by introduction of the ethanol-producing pathway from Zymomonas mobilis, genetic manipulation to increase the pool of fatty acyl-CoA, and heterologous expression of acyl-coenzyme A: diacylglycerol acyltransferase from Acinetobacter baylyi. An optimized fed-batch microbial fermentation of the modified E. coli strain yielded a titer of 922 mg L−1 FAEEs that consisted primarily of ethyl palmitate, -oleate, -myristate and -palmitoleate. PMID:21629774

Simultaneous hydrogen utilization and in situ biogas upgrading in an anaerobic reactor.

PubMed

Luo, Gang; Johansson, Sara; Boe, Kanokwan; Xie, Li; Zhou, Qi; Angelidaki, Irini

2012-04-01

The possibility of converting hydrogen to methane and simultaneous upgrading of biogas was investigated in both batch tests and fully mixed biogas reactor, simultaneously fed with manure and hydrogen. Batch experiments showed that hydrogen could be converted to methane by hydrogenotrophic methanogenesis with conversion of more than 90% of the consumed hydrogen to methane. The hydrogen consumption rates were affected by both P(H₂) (hydrogen partial pressure) and mixing intensity. Inhibition of propionate and butyrate degradation by hydrogen (1 atm) was only observed under high mixing intensity (shaking speed 300 rpm). Continuous addition of hydrogen (flow rate of 28.6 mL/(L/h)) to an anaerobic reactor fed with manure, showed that more than 80% of the hydrogen was utilized. The propionate and butyrate level in the reactor was not significantly affected by the hydrogen addition. The methane production rate of the reactor with H₂ addition was 22% higher, compared to the control reactor only fed with manure. The CO₂ content in the produced biogas was only 15%, while it was 38% in the control reactor. However, the addition of hydrogen resulted in increase of pH (from 8.0 to 8.3) due to the consumption of bicarbonate, which subsequently caused slight inhibition of methanogenesis. Copyright © 2011 Wiley Periodicals, Inc.

21 CFR 211.188 - Batch production and control records.

Code of Federal Regulations, 2012 CFR

2012-04-01

... that each significant step in the manufacture, processing, packing, or holding of the batch was... automated equipment under § 211.68, the identification of the person checking the significant step performed by the automated equipment. (12) Any investigation made according to § 211.192. (13) Results of...

21 CFR 211.188 - Batch production and control records.

Code of Federal Regulations, 2013 CFR

2013-04-01

... that each significant step in the manufacture, processing, packing, or holding of the batch was... automated equipment under § 211.68, the identification of the person checking the significant step performed by the automated equipment. (12) Any investigation made according to § 211.192. (13) Results of...

21 CFR 211.188 - Batch production and control records.

Code of Federal Regulations, 2014 CFR

2014-04-01

... that each significant step in the manufacture, processing, packing, or holding of the batch was... automated equipment under § 211.68, the identification of the person checking the significant step performed by the automated equipment. (12) Any investigation made according to § 211.192. (13) Results of...

Aerobic sludge digestion under low dissolved oxygen concentrations.

PubMed

Arunachalam, RaviSankar; Shah, Hemant K; Ju, Lu-Kwang

2004-01-01

Low dissolved oxygen (DO) concentrations occur commonly in aerobic digesters treating thickened sludge, with benefits of smaller digester size, much reduced aeration cost, and higher digestion temperature (especially important for plants in colder areas). The effects of low DO concentrations on digestion kinetics were studied using the sludge from municipal wastewater treatment plants in Akron, Ohio, and Los Lunas, New Mexico. The experiments were conducted in both batch digestion and a mixed mode of continuous, fed-batch, and batch operations. The low DO condition was clearly advantageous in eliminating the need for pH control because of the simultaneous occurrence of nitrification and denitrification. However, when compared with fully aerobic (high DO) systems under constant pH control (rare in full-scale plants), low DO concentrations and a higher solids loading had a negative effect on the specific volatile solids (VS) digestion kinetics. Nonetheless, the overall (volumetric) digestion performance depends not only on the specific digestion kinetics, but also the solids concentration, pH, and digester temperature. All of the latter factors favor the low DO digestion of thickened sludge. The significant effect of temperature on low DO digestion was confirmed in the mixed-mode study with the Akron sludge. When compared with the well-known empirical correlation between VS reduction and the product (temperature x solids retention time), the experimental data followed the same trend, but were lower than the correlation predictions. The latter was attributed to the lower digestible VS in the Akron sludge, the slower digestion at low DO concentrations, or both. Through model simulation, the first-order decay constant (kd) was estimated as 0.004 h(-1) in the mixed-mode operations, much lower than those (0.011 to 0.029 h(-1)) obtained in batch digestion. The findings suggested that the interactions among sludges with different treatment ages may have a substantially negative effect on digestion kinetics. The use of multistage digesters, especially with small front-end reactors, may be advantageous in both "process" kinetics and "biological reaction" kinetics for sludge digestion.

Network acceleration techniques

NASA Technical Reports Server (NTRS)

Crowley, Patricia (Inventor); Maccabe, Arthur Barney (Inventor); Awrach, James Michael (Inventor)

2012-01-01

Splintered offloading techniques with receive batch processing are described for network acceleration. Such techniques offload specific functionality to a NIC while maintaining the bulk of the protocol processing in the host operating system ("OS"). The resulting protocol implementation allows the application to bypass the protocol processing of the received data. Such can be accomplished this by moving data from the NIC directly to the application through direct memory access ("DMA") and batch processing the receive headers in the host OS when the host OS is interrupted to perform other work. Batch processing receive headers allows the data path to be separated from the control path. Unlike operating system bypass, however, the operating system still fully manages the network resource and has relevant feedback about traffic and flows. Embodiments of the present disclosure can therefore address the challenges of networks with extreme bandwidth delay products (BWDP).

Transfer of a three step mAb chromatography process from batch to continuous: Optimizing productivity to minimize consumable requirements.

PubMed

Gjoka, Xhorxhi; Gantier, Rene; Schofield, Mark

2017-01-20

The goal of this study was to adapt a batch mAb purification chromatography platform for continuous operation. The experiments and rationale used to convert from batch to continuous operation are described. Experimental data was used to design chromatography methods for continuous operation that would exceed the threshold for critical quality attributes and minimize the consumables required as compared to batch mode of operation. Four unit operations comprising of Protein A capture, viral inactivation, flow-through anion exchange (AEX), and mixed-mode cation exchange chromatography (MMCEX) were integrated across two Cadence BioSMB PD multi-column chromatography systems in order to process a 25L volume of harvested cell culture fluid (HCCF) in less than 12h. Transfer from batch to continuous resulted in an increase in productivity of the Protein A step from 13 to 50g/L/h and of the MMCEX step from 10 to 60g/L/h with no impact on the purification process performance in term of contaminant removal (4.5 log reduction of host cell proteins, 50% reduction in soluble product aggregates) and overall chromatography process yield of recovery (75%). The increase in productivity, combined with continuous operation, reduced the resin volume required for Protein A and MMCEX chromatography by more than 95% compared to batch. The volume of AEX membrane required for flow through operation was reduced by 74%. Moreover, the continuous process required 44% less buffer than an equivalent batch process. This significant reduction in consumables enables cost-effective, disposable, single-use manufacturing. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

Near infrared spectroscopy based monitoring of extraction processes of raw material with the help of dynamic predictive modeling

NASA Astrophysics Data System (ADS)

Wang, Haixia; Suo, Tongchuan; Wu, Xiaolin; Zhang, Yue; Wang, Chunhua; Yu, Heshui; Li, Zheng

2018-03-01

The control of batch-to-batch quality variations remains a challenging task for pharmaceutical industries, e.g., traditional Chinese medicine (TCM) manufacturing. One difficult problem is to produce pharmaceutical products with consistent quality from raw material of large quality variations. In this paper, an integrated methodology combining the near infrared spectroscopy (NIRS) and dynamic predictive modeling is developed for the monitoring and control of the batch extraction process of licorice. With the spectra data in hand, the initial state of the process is firstly estimated with a state-space model to construct a process monitoring strategy for the early detection of variations induced by the initial process inputs such as raw materials. Secondly, the quality property of the end product is predicted at the mid-course during the extraction process with a partial least squares (PLS) model. The batch-end-time (BET) is then adjusted accordingly to minimize the quality variations. In conclusion, our study shows that with the help of the dynamic predictive modeling, NIRS can offer the past and future information of the process, which enables more accurate monitoring and control of process performance and product quality.

Expression of a thermotolerant laccase from Pycnoporus sanguineus in Trichoderma reesei and its application in the degradation of bisphenol A.

PubMed

Zhao, Jie; Zeng, Shengquan; Xia, Ying; Xia, Liming

2018-04-01

The laccase gene from Pycnoporus sanguineus was cloned and inserted between the strong Pcbh1 promoter and the Tcbh1 terminator from Trichoderma reesei to form the recombinant plasmid pCH-lac. Using Agrobacterium-mediated technique, the pCH-lac was integrated into the chromosomes of T. reesei. Twenty positive transformants were obtained by employing hygromycin B as a selective agent. PCR was used to confirm that the laccase gene was integrated into the chromosomal DNA of T. reesei. Laccase production by recombinant transformants was performed in shaking flasks, and the activity of laccase reached 8.8 IU/mL after 96-h fermentation under a batch process, and 17.7 IU/mL after 144-h fermentation using a fed-batch process. SDS-PAGE analysis of the fermentation broth showed that the molecular mass of the protein was about 68 kDa, almost the same as that of the laccase produced by P. sanguineus, which indicated that laccase was successfully expressed in T. reesei and secreted out of the cells. The laccase produced by the recombinant T. reesei showed good thermal stability, and could degrade the toxic phenolic material bisphenol A efficiently, after 1-h reaction with 0.06 IU/mL laccase and 0.5 mmol/L ABTS as the mediator at 60 °C and pH 4.5, the degradation rate reached 95%, which demonstrated that it had great potential value in treating the household garbage and wastewater containing the bisphenol A. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Automated batch characterization of inkjet-printed elastomer lenses using a LEGO platform.

PubMed

Sung, Yu-Lung; Garan, Jacob; Nguyen, Hoang; Hu, Zhenyu; Shih, Wei-Chuan

2017-09-10

Small, self-adhesive, inkjet-printed elastomer lenses have enabled smartphone cameras to image and resolve microscopic objects. However, the performance of different lenses within a batch is affected by hard-to-control environmental variables. We present a cost-effective platform to perform automated batch characterization of 300 lens units simultaneously for quality inspection. The system was designed and configured with LEGO bricks, 3D printed parts, and a digital camera. The scheme presented here may become the basis of a high-throughput, in-line inspection tool for quality control purposes and can also be employed for optimization of the manufacturing process.

Production of Bacteriophages by Listeria Cells Entrapped in Organic Polymers.

PubMed

Roy, Brigitte; Philippe, Cécile; Loessner, Martin J; Goulet, Jacques; Moineau, Sylvain

2018-06-13

Applications for bacteriophages as antimicrobial agents are increasing. The industrial use of these bacterial viruses requires the production of large amounts of suitable strictly lytic phages, particularly for food and agricultural applications. This work describes a new approach for phage production. Phages H387 ( Siphoviridae ) and A511 ( Myoviridae ) were propagated separately using Listeria ivanovii host cells immobilised in alginate beads. The same batch of alginate beads could be used for four successive and efficient phage productions. This technique enables the production of large volumes of high-titer phage lysates in continuous or semi-continuous (fed-batch) cultures.

Mixed-mode resins: taking shortcut in downstream processing of raw-starch digesting α-amylases

PubMed Central

Lončar, Nikola; Slavić, Marinela Šokarda; Vujčić, Zoran; Božić, Nataša

2015-01-01

Bacillus licheniformis 9945a α-amylase is known as a potent enzyme for raw starch hydrolysis. In this paper, a mixed mode Nuvia cPrime™ resin is examined with the aim to improve the downstream processing of raw starch digesting amylases and exploit the hydrophobic patches on their surface. This resin combines hydrophobic interactions with cation exchange groups and as such the presence of salt facilitates hydrophobic interactions while the ion-exchange groups enable proper selectivity. α-Amylase was produced using an optimized fed-batch approach in a defined media and significant overexpression of 1.2 g L−1 was achieved. This single step procedure enables simultaneous concentration, pigment removal as well as purification of amylase with yields of 96% directly from the fermentation broth. PMID:26492875

In situ Raman spectroscopy for simultaneous monitoring of multiple process parameters in mammalian cell culture bioreactors.

PubMed

Whelan, Jessica; Craven, Stephen; Glennon, Brian

2012-01-01

In this study, the application of Raman spectroscopy to the simultaneous quantitative determination of glucose, glutamine, lactate, ammonia, glutamate, total cell density (TCD), and viable cell density (VCD) in a CHO fed-batch process was demonstrated in situ in 3 L and 15 L bioreactors. Spectral preprocessing and partial least squares (PLS) regression were used to correlate spectral data with off-line reference data. Separate PLS calibration models were developed for each analyte at the 3 L laboratory bioreactor scale before assessing its transferability to the same bioprocess conducted at the 15 L pilot scale. PLS calibration models were successfully developed for all analytes bar VCD and transferred to the 15 L scale. Copyright © 2012 American Institute of Chemical Engineers (AIChE).

40 CFR Table 3 to Subpart Cccc of... - Initial Compliance With Emission Limitations

Code of Federal Regulations, 2011 CFR

2011-07-01

... CATEGORIES National Emission Standards for Hazardous Air Pollutants: Manufacturing of Nutritional Yeast Part... demonstrated initial compliance if . . . 1. Each fed-batch fermenter producing yeast in a fermentation stage... yeast in a fermentation stage (last (Trade), second-to-last (First Generation), or third-to-last (Stock...

40 CFR Table 4 to Subpart Cccc of... - Continuous Compliance With Emission Limitations

Code of Federal Regulations, 2011 CFR

2011-07-01

... CATEGORIES National Emission Standards for Hazardous Air Pollutants: Manufacturing of Nutritional Yeast Part... yeast in a fermentation stage (last (Trade), second-to-last (First Generation), or third-to-last (Stock... the applicable maximum concentration. 2. Each fed-batch fermenter producing yeast in a fermentation...

Stable acetate production in extreme-thermophilic (70°C) mixed culture fermentation by selective enrichment of hydrogenotrophic methanogens

NASA Astrophysics Data System (ADS)

Zhang, Fang; Zhang, Yan; Ding, Jing; Dai, Kun; van Loosdrecht, Mark C. M.; Zeng, Raymond J.

2014-06-01

The control of metabolite production is difficult in mixed culture fermentation. This is particularly related to hydrogen inhibition. In this work, hydrogenotrophic methanogens were selectively enriched to reduce the hydrogen partial pressure and to realize efficient acetate production in extreme-thermophilic (70°C) mixed culture fermentation. The continuous stirred tank reactor (CSTR) was stable operated during 100 days, in which acetate accounted for more than 90% of metabolites in liquid solutions. The yields of acetate, methane and biomass in CSTR were 1.5 +/- 0.06, 1.0 +/- 0.13 and 0.4 +/- 0.05 mol/mol glucose, respectively, close to the theoretical expected values. The CSTR effluent was stable and no further conversion occurred when incubated for 14 days in a batch reactor. In fed-batch experiments, acetate could be produced up to 34.4 g/L, significantly higher than observed in common hydrogen producing fermentations. Acetate also accounted for more than 90% of soluble products formed in these fed-batch fermentations. The microbial community analysis revealed hydrogenotrophic methanogens (mainly Methanothermobacter thermautotrophicus and Methanobacterium thermoaggregans) as 98% of Archaea, confirming that high temperature will select hydrogenotrophic methanogens over aceticlastic methanogens effectively. This work demonstrated a potential application to effectively produce acetate as a value chemical and methane as an energy gas together via mixed culture fermentation.

Stable acetate production in extreme-thermophilic (70°C) mixed culture fermentation by selective enrichment of hydrogenotrophic methanogens

PubMed Central

Zhang, Fang; Zhang, Yan; Ding, Jing; Dai, Kun; van Loosdrecht, Mark C. M.; Zeng, Raymond J.

2014-01-01

The control of metabolite production is difficult in mixed culture fermentation. This is particularly related to hydrogen inhibition. In this work, hydrogenotrophic methanogens were selectively enriched to reduce the hydrogen partial pressure and to realize efficient acetate production in extreme-thermophilic (70°C) mixed culture fermentation. The continuous stirred tank reactor (CSTR) was stable operated during 100 days, in which acetate accounted for more than 90% of metabolites in liquid solutions. The yields of acetate, methane and biomass in CSTR were 1.5 ± 0.06, 1.0 ± 0.13 and 0.4 ± 0.05 mol/mol glucose, respectively, close to the theoretical expected values. The CSTR effluent was stable and no further conversion occurred when incubated for 14 days in a batch reactor. In fed-batch experiments, acetate could be produced up to 34.4 g/L, significantly higher than observed in common hydrogen producing fermentations. Acetate also accounted for more than 90% of soluble products formed in these fed-batch fermentations. The microbial community analysis revealed hydrogenotrophic methanogens (mainly Methanothermobacter thermautotrophicus and Methanobacterium thermoaggregans) as 98% of Archaea, confirming that high temperature will select hydrogenotrophic methanogens over aceticlastic methanogens effectively. This work demonstrated a potential application to effectively produce acetate as a value chemical and methane as an energy gas together via mixed culture fermentation. PMID:24920064

A comparison of anaerobic 2, 4-dichlorophenoxy acetic acid degradation in single-fed and sequencing batch reactor systems

NASA Astrophysics Data System (ADS)

Elefsiniotis, P.; Wareham, D. G.; Fongsatitukul, P.

2017-08-01

This paper compares the practical limits of 2, 4-dichlorophenoxy acetic acid (2,4-D) degradation that can be obtained in two laboratory-scale anaerobic digestion systems; namely, a sequencing batch reactor (SBR) and a single-fed batch reactor (SFBR) system. The comparison involved synthesizing a decade of research conducted by the lead author and drawing summative conclusions about the ability of each system to accommodate industrial-strength concentrations of 2,4-D. In the main, 2 L liquid volume anaerobic SBRs were used with glucose as a supplemental carbon source for both acid-phase and two-phase conditions. Volatile fatty acids however were used as a supplemental carbon source for the methanogenic SBRs. The anaerobic SBRs were operated at an hydraulic retention time of 48 hours, while being subjected to increasing concentrations of 2,4-D. The SBRs were able to degrade between 130 and 180 mg/L of 2,4-D depending upon whether they were operated in the acid-phase or two-phase regime. The methanogenic-only phase did not achieve 2,4-D degradation however this was primarily attributed to difficulties with obtaining a sufficiently long SRT. For the two-phase SFBR system, 3.5 L liquid-volume digesters were used and no difficulty was experienced with degrading 100 % of the 2,4-D concentration applied (300 mg/L).

Continuous enzymatic hydrolysis of lignocellulosic biomass in a membrane-reactor system: Continuous enzymatic hydrolysis of lignocellulosic biomass in a membrane-reactor system

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

Stickel, Jonathan J.; Adhikari, Birendra; Sievers, David A.

Converting abundant lignocellulosic biomass to sugars as fungible precursors to fuels and chemicals has the potential to diversify the supply chain for those products, but further process improvements are needed to achieve economic viability. In the current work, process intensification of the key enzymatic hydrolysis unit operation is demonstrated by means of a membrane reactor system that was operated continuously. Lignocellulosic biomass (pretreated corn stover) and buffered enzyme solution were fed to a continuously stirred-tank reactor, and clarified sugar solution was withdrawn via a commercial tubular ultrafiltration membrane. The membrane permeance decline and membrane cleaning efficacy were studied and didmore » not vary significantly when increasing fraction insoluble solids (FIS) from 2.5% to 5%. Continuous enzymatic hydrolysis was successfully operated for more than 80 h. A model for the reactor system was able to predict dynamic behavior that was in reasonable agreement with experimental results. The modeled technical performance of anticipated commercial batch and continuous enzymatic hydrolysis processes were compared and showed that continuous operation would provide at least twice the volumetric productivity for the conditions studied. Further improvements are anticipated by better membrane selection and by increasing FIS.« less

Continuous enzymatic hydrolysis of lignocellulosic biomass in a membrane-reactor system: Continuous enzymatic hydrolysis of lignocellulosic biomass in a membrane-reactor system

DOE PAGES

Stickel, Jonathan J.; Adhikari, Birendra; Sievers, David A.; ...

2018-02-21

Converting abundant lignocellulosic biomass to sugars as fungible precursors to fuels and chemicals has the potential to diversify the supply chain for those products, but further process improvements are needed to achieve economic viability. In the current work, process intensification of the key enzymatic hydrolysis unit operation is demonstrated by means of a membrane reactor system that was operated continuously. Lignocellulosic biomass (pretreated corn stover) and buffered enzyme solution were fed to a continuously stirred-tank reactor, and clarified sugar solution was withdrawn via a commercial tubular ultrafiltration membrane. The membrane permeance decline and membrane cleaning efficacy were studied and didmore » not vary significantly when increasing fraction insoluble solids (FIS) from 2.5% to 5%. Continuous enzymatic hydrolysis was successfully operated for more than 80 h. A model for the reactor system was able to predict dynamic behavior that was in reasonable agreement with experimental results. The modeled technical performance of anticipated commercial batch and continuous enzymatic hydrolysis processes were compared and showed that continuous operation would provide at least twice the volumetric productivity for the conditions studied. Further improvements are anticipated by better membrane selection and by increasing FIS.« less

Batch Model for Batched Timestamps Data Analysis with Application to the SSA Disability Program

PubMed Central

Yue, Qingqi; Yuan, Ao; Che, Xuan; Huynh, Minh; Zhou, Chunxiao

2016-01-01

The Office of Disability Adjudication and Review (ODAR) is responsible for holding hearings, issuing decisions, and reviewing appeals as part of the Social Security Administration’s disability determining process. In order to control and process cases, the ODAR has established a Case Processing and Management System (CPMS) to record management information since December 2003. The CPMS provides a detailed case status history for each case. Due to the large number of appeal requests and limited resources, the number of pending claims at ODAR was over one million cases by March 31, 2015. Our National Institutes of Health (NIH) team collaborated with SSA and developed a Case Status Change Model (CSCM) project to meet the ODAR’s urgent need of reducing backlogs and improve hearings and appeals process. One of the key issues in our CSCM project is to estimate the expected service time and its variation for each case status code. The challenge is that the systems recorded job departure times may not be the true job finished times. As the CPMS timestamps data of case status codes showed apparent batch patterns, we proposed a batch model and applied the constrained least squares method to estimate the mean service times and the variances. We also proposed a batch search algorithm to determine the optimal batch partition, as no batch partition was given in the real data. Simulation studies were conducted to evaluate the performance of the proposed methods. Finally, we applied the method to analyze a real CPMS data from ODAR/SSA. PMID:27747132

Dynamic control of nutrient-removal from industrial wastewater in a sequencing batch reactor, using common and low-cost online sensors.

PubMed

Dries, Jan

2016-01-01

On-line control of the biological treatment process is an innovative tool to cope with variable concentrations of chemical oxygen demand and nutrients in industrial wastewater. In the present study we implemented a simple dynamic control strategy for nutrient-removal in a sequencing batch reactor (SBR) treating variable tank truck cleaning wastewater. The control system was based on derived signals from two low-cost and robust sensors that are very common in activated sludge plants, i.e. oxidation reduction potential (ORP) and dissolved oxygen. The amount of wastewater fed during anoxic filling phases, and the number of filling phases in the SBR cycle, were determined by the appearance of the 'nitrate knee' in the profile of the ORP. The phase length of the subsequent aerobic phases was controlled by the oxygen uptake rate measured online in the reactor. As a result, the sludge loading rate (F/M ratio), the volume exchange rate and the SBR cycle length adapted dynamically to the activity of the activated sludge and the actual characteristics of the wastewater, without affecting the final effluent quality.

Hyper-production of butyric acid from delignified rice straw by a novel consolidated bioprocess.

PubMed

Chi, Xue; Li, Jianzheng; Wang, Xin; Zhang, Yafei; Antwi, Philip

2018-04-01

A novel consolidated bioprocess for hyper-production of butyric acid from delignified rice straw without exogenous enzymes involved was developed by co-fermentation of Clostridium thermocellum ATCC 27405 and C. thermobutyricum ATCC 49875. Feasibility of the consolidated bioprocess was approved by batch fermentations, with the optimum pH of 6.5. Fed-batch fermentation with a constant pH of 6.5 at 55 °C could enhance the butyric acid yield to a remarkable 33.9 g/L with a selectivity as high as 78%. Metabolic analysis of the co-culture indicated that sugars liberated by C. thermocellum ATCC 27405 were effectively converted to butyric acid by C. thermobutyricum ATCC 49875. Secondary metabolism of C. thermobutyricum ATCC 49875 also contributed to the hyper-production of butyric acid, resulting in the re-assimilation of by-products such as acetic acid and ethanol. This work provides a more effective fermentation process for butyric acid production from lignocellulosic biomass for future applications. Copyright © 2018 Elsevier Ltd. All rights reserved.

Digester performance and microbial community changes in thermophilic and mesophilic sequencing batch reactors fed with the fine sieved fraction of municipal sewage.

PubMed

Ghasimi, Dara S M; Tao, Yu; de Kreuk, Merle; Abbas, Ben; Zandvoort, Marcel H; van Lier, Jules B

2015-12-15

This study investigates the start-up and operation of bench-scale mesophilic (35 °C) and thermophilic (55 °C) anaerobic sequencing batch reactor (SBR) digesters treating the fine sieved fraction (FSF) from raw municipal sewage. FSF was sequestered from raw municipal wastewater, in the Netherlands, using a rotating belt filter equipped with a 350 micron mesh. For the given wastewater, the major component of FSF was toilet paper, which is estimated to be 10-14 kg per year per average person in the western European countries. A seven months adaptation time was allowed for the thermophilic and mesophilic digesters in order to adapt to FSF as the sole substrate with varying dry solids content of 10-25%. Different SBR cycle durations (14, 9 and 2 days) were applied for both temperature conditions to study methane production rates, volatile fatty acids (VFAs) dynamics, lag phases, as well as changes in microbial communities. The prevailing sludge in the two digesters consisted of very different bacterial and archaeal communities, with OP9 lineage and Methanothermobacter being pre-dominant in the thermophilic digester and Bacteroides and Methanosaeta dominating the mesophilic one. Eventually, decreasing the SBR cycle period, thus increasing the FSF load, resulted in improved digester performances, particularly with regard to the thermophilic digester, i.e. shortened lag phases following the batch feedings, and reduced VFA peaks. Over time, the thermophilic digester outperformed the mesophilic one with 15% increased volatile solids (VS) destruction, irrespective to lower species diversity found at high temperature. Copyright © 2015 Elsevier Ltd. All rights reserved.

Feeding schemes and C/N ratio of a laboratory-scale step-fed sequencing batch reactor for liquid swine manure treatment.

PubMed

Wu, Sarah Xiao; Zhu, Jun; Chen, Lide

2017-07-03

This study was undertaken to investigate the effect of two split feeding schemes (600 mL/200 mL and 400 mL/400 mL, designated as FS1 and FS2, respectively) on the performance of a step-fed sequencing batch reactor (SBR) in treating liquid swine manure for nutrient removal. The SBR was run on an 8-h cycle with a repeated pattern of anaerobic/anoxic/aerobic phases in each cycle and the two feedings always occurred at the beginning of each anaerobic phase. A low-level aeration was used (1.0 L/m 3 .sec) for the anoxic/aerobic phase to facilitate nitrification and phosphorus uptake while reducing the energy consumption. The results showed that FS1 reduced NH 4 + -N by 98.7% and FS2 by 98.3%. FS1 had 12.3 mg/L NO 3 -N left in the effluent, while FS2 had 4.51 mg/L. For soluble phosphorus removal, FS1 achieved 95.2%, while FS2 reached only 68.5%. Both feeding schemes achieved ≥ 95% removal of COD. A good power regression was observed between total nitrogen (sum of all three nitrogen species) and the carbon to nitrogen (C/N) ratio, with the correlation coefficients of 0.9729 and 0.9542 for FS1 and FS2, respectively, based on which it was concluded that higher C/N ratios were required to achieve higher nitrogen removal efficiencies.

Conversion of Aqueous Ammonia-Treated Corn Stover to Lactic Acid by Simultaneous Saccharification and Cofermentation

NASA Astrophysics Data System (ADS)

Zhu, Yongming; Lee, Y. Y.; Elander, Richard T.

Treatment of corn stover with aqueous ammonia removes most of the structural lignin, whereas retaining the majority of the carbohydrates in the solids. After treatment, both the cellulose and hemicellulose in corn stover become highly susceptible to enzymatic digestion. In this study, corn stover treated by aqueous ammonia was investigated as the substrate for lactic acid production by simultaneous saccharification and cofermentation (SSCF). A commercial cellulase (Spezyme-CP) and Lactobacillus pentosus American Type Culture Collection (ATCC) 8041 (Spanish Type Culture Collection [CECT]-4023) were used for hydrolysis and fermentation, respectively. In batch SSCF operation, the carbohydrates in the treated corn stover were converted to lactic acid with high yields, the maximum lactic acid yield reaching 92% of the stoichiometric maximum based on total fermentable carbohydrates (glucose, xylose, and arabinose). A small amount of acetic acid was also produced from pentoses through the phosphoketolase pathway. Among the major process variables for batch SSCF, enzyme loading and the amount of yeast extract were found to be the key factors affecting lactic acid production. Further tests on nutrients indicated that corn steep liquor could be substituted for yeast extract as a nitrogen source to achieve the same lactic acid yield. Fed-batch operation of the SSCF was beneficial in raising the concentration of lactic acid to a maximum value of 75.0 g/L.

Slaughterhouse fatty waste saponification to increase biogas yield.

PubMed

Battimelli, A; Torrijos, M; Moletta, R; Delgenès, J P

2010-05-01

A thermochemical pretreatment, i.e. saponification, was optimised in order to improve anaerobic biodegradation of slaughterhouse wastes such as aeroflotation grease and flesh fats from cattle carcass. Anaerobic digestion of raw wastes, as well as of wastes saponified at different temperatures (60 degrees C, 120 degrees C and 150 degrees C) was conducted in fed-batch reactors under mesophilic condition and the effect of different saponification temperatures on anaerobic biodegradation and on the long-chain fatty acids (LCFAs) relative composition was assessed. Even after increasing loads over a long period of time, raw fatty wastes were biodegraded slowly and the biogas potentials were lower than those of theoretical estimations. In contrast, pretreated wastes exhibited improved batch biodegradation, indicating a better initial bio-availability, particularly obvious for carcass wastes. However, LCFA relative composition was not significantly altered by the pretreatment. Consequently, the enhanced biodegradation should be attributed to an increased initial bio-availability of fatty wastes without any modification of their long chain structure which remained slowly biodegradable. Finally, saponification at 120 degrees C achieved best performances during anaerobic digestion of slaughterhouse wastes. Copyright 2009 Elsevier Ltd. All rights reserved.

Improving municipal wastewater nitrogen and phosphorous removal by feeding sludge fermentation products to sequencing batch reactor (SBR).

PubMed

Yuan, Yue; Liu, Jinjin; Ma, Bin; Liu, Ye; Wang, Bo; Peng, Yongzhen

2016-12-01

This study presents a novel strategy to improve the removal efficiency of nitrogen and phosphorus from municipal wastewater by feeding sequencing batch reactor (SBR) with sludge alkaline fermentation products as carbon sources. The performances of two SBRs treating municipal wastewater (one was fed with sludge fermentation products; F-SBR, and the other without sludge fermentation products; B-SBR) were compared. The removal efficiencies of total nitrogen (TN) and phosphorus (PO 4 3- -P) were found to be 82.9% and 96.0% in F-SBR, while the corresponding values in B-SBR were 55.9% (TN) and -6.1% (PO 4 3- -P). Illumina MiSeq sequencing indicated that ammonium-oxidizing bacteria (Nitrosomonadaceae and Nitrosomonas) and denitrifying polyphosphate accumulating organisms (Dechloromonas) were enriched in F-SBR, which resulted in NO 2 - -N accumulation and denitrifying phosphorus removal via nitrite (DPRN). Moreover, feeding of sludge fermentation products reduced 862.1mg VSS/d of sludge in the F-SBR system (volume: 10L). Copyright © 2016 Elsevier Ltd. All rights reserved.

The use of high pressure CO2 -facilitated pH swings to enhance in situ product recovery of butyric acid in a two-phase partitioning bioreactor.

PubMed

Peterson, Eric C; Daugulis, Andrew J

2014-11-01

Through the use of high partial pressures of CO2 (pCO2 ) to facilitate temporary pH reductions in two-phase partitioning bioreactors (TPPBs), improved pH dependent partitioning of butyric acid was observed which achieved in situ product recovery (ISPR), alleviating end-product inhibition (EPI) during the production of butyric acid by Clostridium tyrobutyricum (ATCC 25755). Through high pressure pCO2 studies, media buffering effects were shown to be substantially overcome at 60 bar pCO2 , resulting in effective extraction of the organic acid by the absorptive polymer Pebax® 2533, yielding a distribution coefficient (D) of 2.4 ± 0.1 after 1 h of contact at this pressure. Importantly, it was also found that C. tyrobutyricum cultures were able to withstand 60 bar pCO2 for 1 h with no decrease in growth ability when returned to atmospheric pressure in batch reactors after several extraction cycles. A fed-batch reactor with cyclic high pCO2 polymer extraction recovered 92 g of butyric acid to produce a total of 213 g compared to 121 g generated in a control reactor. This recovery reduced EPI in the TPPB, resulting in both higher productivity (0.65 vs. 0.33 g L(-1)  h(-1) ) and yield (0.54 vs. 0.40). Fortuitously, it was also found that repeated high pCO2 -facilitated polymer extractions of butyric acid during batch growth of C. tyrobutyricum lessened the need for pH control, and reduced base requirements by approximately 50%. Thus, high pCO2 -mediated absorptive polymer extraction presents a novel method for improving process performance in butyric acid fermentation, and this technique could be applied to the bioproduction of other organic acids as well. © 2014 Wiley Periodicals, Inc.

Comparison of Fenton and Fenton-like oxidation for the treatment of cosmetic wastewater.

PubMed

Bautista, P; Casas, J A; Zazo, J A; Rodriguez, J J; Mohedano, A F

2014-01-01

The treatment of cosmetic wastewaters by Fenton (Fe²⁺/H₂O₂) and Fenton-like (Fe³⁺/H₂O₂) oxidation has been studied. From batch and continuous experiments it has been proved that both versions of the Fenton process lead to quite similar results in terms of chemical oxygen demand (COD) and total organic carbon reduction although the COD shows a slightly higher rate in the early stages of reaction. COD reductions of around 55% after 2 h reaction time and 75-80% with 4 h residence time were reached in batch and continuous experiments, respectively, conducted at pH around 3, ambient temperature (20 °C), with 200 mg/L of Fe dose and an initial H₂O₂/COD weight ratio corresponding to the theoretical stoichiometric value. Achieving the locally allowable limit of COD for industrial wastewater discharge into the municipal sewer system takes no more than 30 min reaction time under those conditions by both Fenton systems. However, the Fenton-like process, where iron is fed as Fe(3+), would be preferable for industrial applications since the ferric sludge resulting upon final neutralization of the effluent can be recycled to the process. A second-order kinetic equation with respect to COD fitted fairly well the experimental results at different temperatures, thus providing a simple practical tool for design purposes.

Safety and reliability analysis in a polyvinyl chloride batch process using dynamic simulator-case study: Loss of containment incident.

PubMed

Rizal, Datu; Tani, Shinichi; Nishiyama, Kimitoshi; Suzuki, Kazuhiko

2006-10-11

In this paper, a novel methodology in batch plant safety and reliability analysis is proposed using a dynamic simulator. A batch process involving several safety objects (e.g. sensors, controller, valves, etc.) is activated during the operational stage. The performance of the safety objects is evaluated by the dynamic simulation and a fault propagation model is generated. By using the fault propagation model, an improved fault tree analysis (FTA) method using switching signal mode (SSM) is developed for estimating the probability of failures. The timely dependent failures can be considered as unavailability of safety objects that can cause the accidents in a plant. Finally, the rank of safety object is formulated as performance index (PI) and can be estimated using the importance measures. PI shows the prioritization of safety objects that should be investigated for safety improvement program in the plants. The output of this method can be used for optimal policy in safety object improvement and maintenance. The dynamic simulator was constructed using Visual Modeler (VM, the plant simulator, developed by Omega Simulation Corp., Japan). A case study is focused on the loss of containment (LOC) incident at polyvinyl chloride (PVC) batch process which is consumed the hazardous material, vinyl chloride monomer (VCM).

Improved cost-effectiveness of the block co-polymer anneal process for DSA

NASA Astrophysics Data System (ADS)

Pathangi, Hari; Stokhof, Maarten; Knaepen, Werner; Vaid, Varun; Mallik, Arindam; Chan, Boon Teik; Vandenbroeck, Nadia; Maes, Jan Willem; Gronheid, Roel

2016-04-01

This manuscript first presents a cost model to compare the cost of ownership of DSA and SAQP for a typical front end of line (FEoL) line patterning exercise. Then, we proceed to a feasibility study of using a vertical furnace to batch anneal the block co-polymer for DSA applications. We show that the defect performance of such a batch anneal process is comparable to the process of record anneal methods. This helps in increasing the cost benefit for DSA compared to the conventional multiple patterning approaches.

Soft sensor for monitoring biomass subpopulations in mammalian cell culture processes.

PubMed

Kroll, Paul; Stelzer, Ines V; Herwig, Christoph

2017-11-01

Biomass subpopulations in mammalian cell culture processes cause impurities and influence productivity, which requires this critical process parameter to be monitored in real-time. For this reason, a novel soft sensor concept for estimating viable, dead and lysed cell concentration was developed, based on the robust and cheap in situ measurements of permittivity and turbidity in combination with a simple model. It could be shown that the turbidity measurements contain information about all investigated biomass subpopulations. The novelty of the developed soft sensor is the real-time estimation of lysed cell concentration, which is directly correlated to process-related impurities such as DNA and host cell protein in the supernatant. Based on data generated by two fed-batch processes the developed soft sensor is described and discussed. The presented soft sensor concept provides a tool for viable, dead and lysed cell concentration estimation in real-time with adequate accuracy and enables further applications with respect to process optimization and control.

Continuous flow operation with appropriately adjusting composites in influent for recovery of Cr(VI), Cu(II) and Cd(II) in self-driven MFC-MEC system.

PubMed

Li, Ming; Pan, Yuzhen; Huang, Liping; Zhang, Yong; Yang, Jinhui

2017-03-01

A self-driven microbial fuel cell (MFC) - microbial electrolysis cell (MEC) system, where electricity generated from MFCs is in situ utilized for powering MECs, has been previously reported for recovering Cr(VI), Cu(II) and Cd(II) with individual metals fed in different units of the system in batch operation. Here it was advanced with treating synthetic mixed metals' solution at appropriately adjusting composites in fed-batch and continuous flow operations for complete separation of Cr(VI), Cu(II) and Cd(II) from each other. Under an optimal condition of hydraulic residence time of 4 h, matching of two serially connected MFCs with one MEC, and fed with a composite of either 5 mg L -1 Cr(VI), 1 mg L -1 Cu(II) and 5 mg L -1 Cd(II), or 1 mg L -1 Cr(VI), 5 mg L -1 Cu(II) and 5 mg L -1 Cd(II), the self-driven MFC-MEC system can completely and sequentially recover Cu(II), Cr(VI) and Cd(II) from mixed metals. This study provides a true sustainable and zero-energy-consumed approach of using bioelectrochemical systems for completely recovering and separating Cr(VI), Cu(II) and Cd(II) from each other or from wastes or contaminated sites.

Exploitation of Trametes versicolor for bioremediation of endocrine disrupting chemicals in bioreactors

PubMed Central

Sannia, Giovanni; Raganati, Francesca; Olivieri, Giuseppe; Marzocchella, Antonio; Schlosser, Dietmar

2017-01-01

Endocrine disrupting chemicals (EDCs) are environmental contaminants causing increasing concerns due to their toxicity, persistence and ubiquity. In the present study, degradative capabilities of Trametes versicolor, Pleurotus ostreatus and Phanerochaete chrysosporium to act on five EDCs, which represent different classes of chemicals (phenols, parabens and phthalate) and were first applied as single compounds, were assessed. T. versicolor was selected due to its efficiency against target EDCs and its potentialities were exploited against a mixture of EDCs in a cost-effective bioremediation process. A fed-batch approach as well as a starvation strategy were applied in order to reduce the need for input of ‘fresh’ biomass, and avoid the requirement for external nutrients. The fungus was successfully operated in two different bioreactors over one week. Semi-batch cultures were carried out by daily adding a mixture of EDCs to the bioreactors in a total of five consecutive degradation cycles. T. versicolor was able to efficiently remove all compounds during each cycle converting up to 21 mg L-1 day-1 of the tested EDCs. The maintained ability of T. versicolor to remove EDCs without any additional nutrients represents the main outcome of this study, which enables to forecast its application in a water treatment process. PMID:28575092

Exploitation of Trametes versicolor for bioremediation of endocrine disrupting chemicals in bioreactors.

PubMed

Pezzella, Cinzia; Macellaro, Gemma; Sannia, Giovanni; Raganati, Francesca; Olivieri, Giuseppe; Marzocchella, Antonio; Schlosser, Dietmar; Piscitelli, Alessandra

2017-01-01

Endocrine disrupting chemicals (EDCs) are environmental contaminants causing increasing concerns due to their toxicity, persistence and ubiquity. In the present study, degradative capabilities of Trametes versicolor, Pleurotus ostreatus and Phanerochaete chrysosporium to act on five EDCs, which represent different classes of chemicals (phenols, parabens and phthalate) and were first applied as single compounds, were assessed. T. versicolor was selected due to its efficiency against target EDCs and its potentialities were exploited against a mixture of EDCs in a cost-effective bioremediation process. A fed-batch approach as well as a starvation strategy were applied in order to reduce the need for input of 'fresh' biomass, and avoid the requirement for external nutrients. The fungus was successfully operated in two different bioreactors over one week. Semi-batch cultures were carried out by daily adding a mixture of EDCs to the bioreactors in a total of five consecutive degradation cycles. T. versicolor was able to efficiently remove all compounds during each cycle converting up to 21 mg L-1 day-1 of the tested EDCs. The maintained ability of T. versicolor to remove EDCs without any additional nutrients represents the main outcome of this study, which enables to forecast its application in a water treatment process.

Defense Waste Processing Facility Simulant Chemical Processing Cell Studies for Sludge Batch 9

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

Smith, Tara E.; Newell, J. David; Woodham, Wesley H.

The Savannah River National Laboratory (SRNL) received a technical task request from Defense Waste Processing Facility (DWPF) and Saltstone Engineering to perform simulant tests to support the qualification of Sludge Batch 9 (SB9) and to develop the flowsheet for SB9 in the DWPF. These efforts pertained to the DWPF Chemical Process Cell (CPC). CPC experiments were performed using SB9 simulant (SB9A) to qualify SB9 for sludge-only and coupled processing using the nitric-formic flowsheet in the DWPF. Two simulant batches were prepared, one representing SB8 Tank 40H and another representing SB9 Tank 51H. The simulant used for SB9 qualification testing wasmore » prepared by blending the SB8 Tank 40H and SB9 Tank 51H simulants. The blended simulant is referred to as SB9A. Eleven CPC experiments were run with an acid stoichiometry ranging between 105% and 145% of the Koopman minimum acid equation (KMA), which is equivalent to 109.7% and 151.5% of the Hsu minimum acid factor. Three runs were performed in the 1L laboratory scale setup, whereas the remainder were in the 4L laboratory scale setup. Sludge Receipt and Adjustment Tank (SRAT) and Slurry Mix Evaporator (SME) cycles were performed on nine of the eleven. The other two were SRAT cycles only. One coupled flowsheet and one extended run were performed for SRAT and SME processing. Samples of the condensate, sludge, and off-gas were taken to monitor the chemistry of the CPC experiments.« less

Anaerobic co-digestion of fruit and vegetable wastes and primary sewage sludge.

PubMed

Velmurugan, B; Arathy, E C; Hemalatha, R; Philip, Jerry Elsa; Alwar Ramanujam, R

2010-01-01

Anaerobic co-digestion of fruit and vegetable wastes (FVW) and primary sewage sludge was carried out in a fed-batch reactor having a volume of 21 under ambient temperature conditions. Three different proportions (25:75, 50:50 and 75:25 in terms ofVS) of fruit and vegetable wastes and primary sewage sludge were studied for an organic loading rate (OLR) of 1.0 g VS/ l.d and with a hydraulic retention time (HRT) of 25 days. The reactor with 75% FVW and 25% sewage sludge (in terms of VS) showed better performance in terms of VS reduction and biogas yield when compared to other two proportions.

40 CFR Table 4 to Subpart Cccc of... - Continuous Compliance With Emission Limitations

Code of Federal Regulations, 2013 CFR

2013-07-01

... Yeast Pt. 63, Subpart CCCC, Table 4 Table 4 to Subpart CCCC of Part 63—Continuous Compliance With... fermenter producing yeast in a fermentation stage (last (Trade), second-to-last (First Generation), or third... the applicable maximum concentration. 2. Each fed-batch fermenter producing yeast in a fermentation...

40 CFR Table 4 to Subpart Cccc of... - Continuous Compliance With Emission Limitations

Code of Federal Regulations, 2014 CFR

2014-07-01

... Yeast Pt. 63, Subpart CCCC, Table 4 Table 4 to Subpart CCCC of Part 63—Continuous Compliance With... fermenter producing yeast in a fermentation stage (last (Trade), second-to-last (First Generation), or third... the applicable maximum concentration. 2. Each fed-batch fermenter producing yeast in a fermentation...

40 CFR Table 4 to Subpart Cccc of... - Continuous Compliance With Emission Limitations

Code of Federal Regulations, 2012 CFR

2012-07-01

... Yeast Pt. 63, Subpart CCCC, Table 4 Table 4 to Subpart CCCC of Part 63—Continuous Compliance With... fermenter producing yeast in a fermentation stage (last (Trade), second-to-last (First Generation), or third... the applicable maximum concentration. 2. Each fed-batch fermenter producing yeast in a fermentation...

Growth of the extremophilic Deinococcus geothermalis DSM 11302 using co-substrate fed-batch culture.

PubMed

Bornot, Julie; Molina-Jouve, Carole; Uribelarrea, Jean-Louis; Gorret, Nathalie

2014-02-01

Deinococcus geothermalis metabolism has been scarcely studied to date, although new developments on its utilization for bioremediation have been carried out. So, large-scale production of this strain and a better understanding of its physiology are required. A fed-batch experiment was conducted to achieve a high cell density non-limiting culture of D. geothermalis DSM 11302. A co-substrate nutritional strategy using glucose and yeast extract was carried out in a 20-L bioreactor in order to maintain a non-limited growth at a maximal growth rate of 1 h(-1) at 45 °C. Substrate supplies were adjusted by monitoring online culture parameters and physiological data (dissolved oxygen, gas analyses, respiratory quotient, biomass concentration). The results showed that yeast extract could serve as both carbon and nitrogen sources, although glucose and ammonia were consumed too. Yeast extract carbon-specific uptake rate reached a value 4.5 times higher than glucose carbon-specific uptake rate. Cell concentration of 9.6 g L(-1) dry cell weight corresponding to 99 g of biomass was obtained using glucose and yeast extract as carbon and nitrogen sources.

Kalman filter based glucose control at small set points during fed-batch cultivation of Saccharomyces cerevisiae.

PubMed

Arndt, Michael; Hitzmann, Bernd

2004-01-01

A glucose control system is presented, which is able to control cultivations of Saccharomyces cerevisiae even at low glucose concentrations. Glucose concentrations are determined using a special flow injection analysis (FIA) system, which does not require a sampling module. An extended Kalman filter is employed for smoothing the glucose measurements as well as for the prediction of glucose and biomass concentration, the maximum specific growth rate, and the volume of the culture broth. The predicted values are utilized for feedforward/feedback control of the glucose concentration at set points of 0.08 and 0.05 g/L. The controller established well-defined conditions over several hours up to biomass concentrations of 13.5 and 20.7 g/L, respectively. The specific glucose uptake rates at both set points were 1.04 and 0.68 g/g/h, respectively. It is demonstrated that during fed-batch cultivation an overall pure oxidative metabolism of glucose is maintained at the lower set point and a specific ethanol production rate of 0.18 g/g/h at the higher set point.

Comparison of different estimation techniques for biomass concentration in large scale yeast fermentation.

PubMed

Hocalar, A; Türker, M; Karakuzu, C; Yüzgeç, U

2011-04-01

In this study, previously developed five different state estimation methods are examined and compared for estimation of biomass concentrations at a production scale fed-batch bioprocess. These methods are i. estimation based on kinetic model of overflow metabolism; ii. estimation based on metabolic black-box model; iii. estimation based on observer; iv. estimation based on artificial neural network; v. estimation based on differential evaluation. Biomass concentrations are estimated from available measurements and compared with experimental data obtained from large scale fermentations. The advantages and disadvantages of the presented techniques are discussed with regard to accuracy, reproducibility, number of primary measurements required and adaptation to different working conditions. Among the various techniques, the metabolic black-box method seems to have advantages although the number of measurements required is more than that for the other methods. However, the required extra measurements are based on commonly employed instruments in an industrial environment. This method is used for developing a model based control of fed-batch yeast fermentations. Copyright © 2010 ISA. Published by Elsevier Ltd. All rights reserved.

Modulation of guanosine nucleotides biosynthetic pathways enhanced GDP-L-fucose production in recombinant Escherichia coli.

PubMed

Lee, Won-Heong; Shin, So-Yeon; Kim, Myoung-Dong; Han, Nam Soo; Seo, Jin-Ho

2012-03-01

Guanosine 5'-triphosphate (GTP) is the key substrate for biosynthesis of guanosine 5'-diphosphate (GDP)-L-fucose. In this study, improvement of GDP-L-fucose production was attempted by manipulating the biosynthetic pathway for guanosine nucleotides in recombinant Escherichia coli-producing GDP-L-fucose. The effects of overexpression of inosine 5'-monophosphate (IMP) dehydrogenase, guanosine 5'-monophosphate (GMP) synthetase (GuaB and GuaA), GMP reductase (GuaC) and guanosine-inosine kinase (Gsk) on GDP-L-fucose production were investigated in a series of fed-batch fermentations. Among the enzymes tested, overexpression of Gsk led to a significant improvement of GDP-L-fucose production. Maximum GDP-L-fucose concentration of 305.5 ± 5.3 mg l(-1) was obtained in the pH-stat fed-batch fermentation of recombinant E. coli-overexpressing Gsk, which corresponds to a 58% enhancement in the GDP-L-fucose production compared with the control strain overexpressing GDP-L-fucose biosynthetic enzymes. Such an enhancement of GDP-L-fucose production could be due to the increase in the intracellular level of GMP.

Comparison of the performance of MBBR and SBR systems for the treatment of anaerobic reactor biowaste effluent.

PubMed

Comett-Ambriz, I; Gonzalez-Martinez, S; Wilderer, P

2003-01-01

Anaerobic reactor biowaste effluent was treated with biofilm and activated sludge sequencing batch reactors to compare the performance of both systems. The treatment targets were organic carbon removal and nitrification. The pilot plant was operated in two phases. During the first phase, it was operated like a Moving Bed Biofilm Reactor (MBBR) with the Natrix media, with a specific surface area of 210 m2/m3. The MBBR was operated under Sequencing Batch Reactor (SBR) modality with three 8-hour cycles per day over 70 days. During the second phase of the experiment, the pilot plant was operated over 79 days as a SBR. In both phases the influent was fed to the reactor at a flow rate corresponding to a Hydraulic Retention Time (HRT) of 4 days. Both systems presented a good carbon removal for this specific wastewater. The Chemical Oxygen Demand (COD) total removal was 53% for MBBR and 55% for SBR. MBBR offered a higher dissolved COD removal (40%) than SBR (30%). The limited COD removal achieved is in agreement with the high COD to BOD5 ratio (1/3) of the influent wastewater. In both systems a complete nitrification was obtained. The different efficiencies in both systems are related to the different biomass concentrations.

A preliminary evaluation of a reusable digital sterilization indicator prototype.

PubMed

Puttaiah, R; Griggs, J; D'Onofrio, M

2014-09-01

Sterilization of critical and semicritical instruments used in patient care must undergo a terminal process of sterilization. Use of chemical and physical indicators are important in providing information on the sterilizer's performance during each cycle. Regular and periodic monitoring of sterilizers using biological indicators is necessary in periodically validating performance of sterilizers. Data loggers or independent digital parametric indicators are innovative devices that provide more information than various classes chemical indicators. In this study we evaluated a prototype of an independent digital parametric indicator's use in autoclaves. The purpose of this study was to evaluate the performance of an independent digital indicator/data logger prototype (DS1922F) that could be used for multiple cycles within an autoclave.MG Materials and methods: Three batches of the DS1922F (150 samples) were used in this study that was conducted in a series. The first batch was challenged with 300 sterilization cycles within an autoclave and the data loggers evaluated to study failures and the reason for failure, make corrections and improve the prototype design. After changes made based on studying the first batch, the second batch of the prototype (150 samples) were challenged once again with 300 sterilization cycles within an autoclave and failure studied again in further improvement of the prototype. The final batch (3rd batch) of the prototype (150 samples) was challenged again but with 600 cycles to see how long they would last. Kaplan-Meier survival analysis analyses of all three batches was conducted (α = 0.05) and failed samples qualitatively studied in understanding the variables involved in the failure of the prototype, and in improving quality. Each tested batch provided crucial information on device failure and helped in improvement of the prototype. Mean lifetime survival of the final batch (Batch 3) of prototype was 498 (480, 516) sterilization cycles in an autoclave. In this study, the final batch of the DS1922F prototype data logger was found to be robust in withstanding the challenge of 600 autoclave cycles, with a mean lifetime of more than 450 cycles, multiple times more than prescribed number of cycles. Instrument reprocessing is among the important aspects of infection control. While stringent procedures are followed in instrument reprocessing within the clinic in assuring patient safety, regular use of sterilization process indicators and periodic biological validation of the sterilizer's performance is necessary. Chemical indicators for use in Autoclaves provide information on whether the particular cycle's parameters were achieved but do not provide at what specific point in time or temperature the failure occurred. Data loggers and associated reader software as the tested prototype in this evaluation (DS1922F), are designed to provide continuous information on time and temperature of the prescribed cycle. Data loggers provide immediate information on the process as opposed to Biological Indicators that take from days to a week in obtaining a confirmatory result. Further, many countries do not have the sterilization monitoring service infrastructure to meet the demands of the end users. In the absence of sterilization monitoring services, use of digital data loggers for each sterilization cycle is more pragmatic.