Acetone-butanol-ethanol competitive sorption simulation from single, binary, and ternary systems in a fixed-bed of KA-I resin.

PubMed

Wu, Jinglan; Zhuang, Wei; Ying, Hanjie; Jiao, Pengfei; Li, Renjie; Wen, Qingshi; Wang, Lili; Zhou, Jingwei; Yang, Pengpeng

2015-01-01

Separation of butanol based on sorption methodology from acetone-butanol-ethanol (ABE) fermentation broth has advantages in terms of biocompatibility and stability, as well as economy, and therefore gains much attention. In this work a chromatographic column model based on the solid film linear driving force approach and the competitive Langmuir isotherm equations was used to predict the competitive sorption behaviors of ABE single, binary, and ternary mixture. It was observed that the outlet concentration of weaker retained components exceeded the inlet concentration, which is an evidence of competitive adsorption. Butanol, the strongest retained component, could replace ethanol almost completely and also most of acetone. In the end of this work, the proposed model was validated by comparison of the experimental and predicted ABE ternary breakthrough curves using the real ABE fermentation broth as a feed solution. © 2014 American Institute of Chemical Engineers.

Acetone-butanol-ethanol production from Kraft paper mill sludge by simultaneous saccharification and fermentation.

PubMed

Guan, Wenjian; Shi, Suan; Tu, Maobing; Lee, Yoon Y

2016-01-01

Paper mill sludge (PS), a solid waste from pulp and paper industry, was investigated as a feedstock for acetone-butanol-ethanol (ABE) production by simultaneous saccharification and fermentation (SSF). ABE fermentation of paper sludge by Clostridium acetobutylicum required partial removal of ash in PS to enhance its enzymatic digestibility. Enzymatic hydrolysis was found to be a rate-limiting step in the SSF. A total of 16.4-18.0g/L of ABE solvents were produced in the SSF of de-ashed PS with solid loading of 6.3-7.4% and enzyme loading of 10-15FPU/g-glucan, and the final solvent yield reached 0.27g/g sugars. No pretreatment and pH control were needed in ABE fermentation of paper sludge, which makes it an attractive feedstock for butanol production. The results suggested utilization of paper sludge should not only consider the benefits of buffering effect of CaCO3 in fermentation, but also take into account its inhibitory effect on enzymatic hydrolysis. Published by Elsevier Ltd.

Two-stage pervaporation process for effective in situ removal acetone-butanol-ethanol from fermentation broth.

PubMed

Cai, Di; Hu, Song; Miao, Qi; Chen, Changjing; Chen, Huidong; Zhang, Changwei; Li, Ping; Qin, Peiyong; Tan, Tianwei

2017-01-01

Two-stage pervaporation for ABE recovery from fermentation broth was studied to reduce the energy cost. The permeate after the first stage in situ pervaporation system was further used as the feedstock in the second stage of pervaporation unit using the same PDMS/PVDF membrane. A total 782.5g/L of ABE (304.56g/L of acetone, 451.98g/L of butanol and 25.97g/L of ethanol) was achieved in the second stage permeate, while the overall acetone, butanol and ethanol separation factors were: 70.7-89.73, 70.48-84.74 and 9.05-13.58, respectively. Furthermore, the theoretical evaporation energy requirement for ABE separation in the consolidate fermentation, which containing two-stage pervaporation and the following distillation process, was estimated less than ∼13.2MJ/kg-butanol. The required evaporation energy was only 36.7% of the energy content of butanol. The novel two-stage pervaporation process was effective in increasing ABE production and reducing energy consumption of the solvents separation system. Copyright © 2016 Elsevier Ltd. All rights reserved.

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.

Recent advances on conversion and co-production of acetone-butanol-ethanol into high value-added bioproducts.

PubMed

Xin, Fengxue; Dong, Weiliang; Jiang, Yujia; Ma, Jiangfeng; Zhang, Wenming; Wu, Hao; Zhang, Min; Jiang, Min

2018-06-01

Butanol is an important bulk chemical and has been regarded as an advanced biofuel. Large-scale production of butanol has been applied for more than 100 years, but its production through acetone-butanol-ethanol (ABE) fermentation process by solventogenic Clostridium species is still not economically viable due to the low butanol titer and yield caused by the toxicity of butanol and a by-product, such as acetone. Renewed interest in biobutanol as a biofuel has spurred technological advances to strain modification and fermentation process design. Especially, with the development of interdisciplinary processes, the sole product or even the mixture of ABE produced through ABE fermentation process can be further used as platform chemicals for high value added product production through enzymatic or chemical catalysis. This review aims to comprehensively summarize the most recent advances on the conversion of acetone, butanol and ABE mixture into various products, such as isopropanol, butyl-butyrate and higher-molecular mass alkanes. Additionally, co-production of other value added products with ABE was also discussed.

Bioproduction of butanol in bioreactors: new insights from simultaneous in situ butanol recovery to eliminate product toxicity

USDA-ARS?s Scientific Manuscript database

Simultaneous acetone butanol ethanol (ABE) fermentation by Clostridium beijerinckii 260 and in situ product recovery was investigated using a vacuum process operated in two modes: continuous and intermittent. Integrated batch fermentations and ABE recovery were conducted at 37 deg C using a 14-L bio...

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.

Integrated in situ gas stripping-salting-out process for high-titer acetone-butanol-ethanol production from sweet sorghum bagasse.

PubMed

Wen, Hao; Chen, Huidong; Cai, Di; Gong, Peiwen; Zhang, Tao; Wu, Zhichao; Gao, Heting; Li, Zhuangzhuang; Qin, Peiyong; Tan, Tianwei

2018-01-01

The production of biobutanol from renewable biomass resources is attractive. The energy-intensive separation process and low-titer solvents production are the key constraints on the economy-feasible acetone-butanol-ethanol (ABE) production by fermentation. To decrease energy consumption and increase the solvents concentration, a novel two-stage gas stripping-salting-out system was established for effective ABE separation from the fermentation broth using sweet sorghum bagasse as feedstock. The ABE condensate (143.6 g/L) after gas stripping, the first-stage separation, was recovered and introduced to salting-out process as the second-stage. K 4 P 2 O 7 and K 2 HPO 4 were used, respectively. The effect of saturated salt solution temperature on final ABE concentration was also investigated. The results showed high ABE recovery (99.32%) and ABE concentration (747.58 g/L) when adding saturated K 4 P 2 O 7 solution at 323.15 K and 3.0 of salting-out factor. On this condition, the energy requirement of the downstream distillation process was 3.72 MJ/kg of ABE. High-titer cellulosic ABE production was separated from the fermentation broth by the novel two-stage gas stripping-salting-out process. The process was effective, which reduced the downstream process energy requirement significantly.

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

PubMed

Li, Jingwen; Wang, Lan; Chen, Hongzhang

2016-11-01

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

Evaluation of asymmetric polydimethylsiloxane-polyvinylidene fluoride composite membrane and incorporated with acetone-butanol-ethanol fermentation for butanol recovery.

PubMed

Xue, Chuang; Du, Guang-Qing; Chen, Li-Jie; Ren, Jian-Gang; Bai, Feng-Wu

2014-10-20

The polydimethylsiloxane-polyvinylidene fluoride (PDMS-PVDF) composite membrane was studied for its pervaporation performance to removal of butanol from butanol/ABE solution, fermentation broth as well as incorporated with acetone-butanol-ethanol (ABE) fermentation. The total flux and butanol titer in permeate through the PDMS-PVDF membrane were up to 769.6 g/m(2)h and 323.5 g/L at 80 °C, respectively. The butanol flux and total flux increased with increasing the feed temperature as well as the feed butanol titer. The butanol separation factor and butanol titer in permeate decreased slightly in the presence of acetone and ethanol in the feed due to their preferential dissolution and competitive permeation through the membrane. In fed-batch fermentation incorporated with pervaporation, butanol titer and flux in permeate maintained at a steady level with the range of 139.9-154.0 g/L and 13.3-16.3 g/m(2)h, respectively, which was attributed to the stable butanol titer in fermentation broth as well as the excellent hydrophobic nature of the PDMS-PVDF matrix. Therefore, the PDMS-PVDF composite membrane had a great potential in the in situ product recovery with ABE fermentation, enabling the economic production of biobutanol. Copyright © 2014 Elsevier B.V. All rights reserved.

Production of Butyric Acid and Butanol from Biomass

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

Ramey, David E.; Yang, Shang-Tian

Butanol replaced gasoline gallon for gallon in a 10,000 miles trip across the United States without the need to highly modify a ’92 Buick (your existing car today). Butanol can now be made for less than ethanol and yields more Btu’s from the same corn, making the plow to tire equation positive – more energy out than it takes to make it and Butanol is much safer. Butanol when substituted for gasoline gives better gas mileage and does not pollute as tested in 10 states. Butanol should now receive the same recognition as ethanol in U.S. legislation “ethanol/butanol”. There ismore » abundant plant biomass present as low-value agricultural commodities or processing wastes requiring proper disposal to avoid pollution problems. One example is in the corn refinery industry, which processes more than 13% of the ~9.5 billion bushels (~240 million metric tons) of corn annually produced in the U.S. to produce high-fructose-corn-syrup, dextrose, starch, and fuel alcohol, and generates more than 10 million metric tons of corn byproducts that are currently of limited use and pose significant environmental problems. The abundant inexpensive renewable resources as feedstock for fermentation, and recent advances in the fields of biotechnology and bioprocessing have resulted in a renewed interest in the fermentation production of chemicals and fuels, including n-butanol. The historic acetone-butanol-ethanol (ABE) fermentation by Clostridium acetobutylicum is one of the oldest known industrial fermentations. It was ranked second only to ethanol fermentation by yeast in its scale of production, and is one of the largest biotechnological processes ever known. However, since the 1950's industrial ABE fermentation has declined continuously, and almost all butanol is now produced via petrochemical routes (Chemical Marketing Reporter, 1993). Butanol is an important industrial solvent and is a better fuel for replacing gasoline – gallon for gallon than ethanol. Current butanol prices as a chemical are at $3.00 per gallon – wholesaling in 55 gallon drums for $6.80, with a worldwide market of 1.4 billion gallon per year. The market demand is expected to increase dramatically since butanol can now be produced economically from low-cost biomass. Butanol’s application as a replacement for gasoline will outpace ethanol, biodiesel and hydrogen when its safety and simplicity of use are seen. Butanol’s application for the Department of Defense as a clean-safe replacement for batteries when used in conjunction with fuel cell technology is seen as an application for the future. Disposable canisters made of PLA that carry butanol to be reformed and used to generate electricity for computers, night vision and stealth equipment can be easily disposed of. In a typical ABE fermentation, butyric, propionic and acetic acids are produced first by C. acetobutylicum; the culture then undergoes a metabolic shift and solvents (butanol, acetone, and ethanol) are formed (Fond et al., 1985). In conventional ABE fermentations, the butanol yield from glucose is low, typically at ~15% (w/w) and rarely exceeds 25% (0.77–1.3 gallons per bushel corn respectfully). The production of butanol is also limited by severe product inhibition. Butanol at a concentration of 10 g/L can significantly inhibit cell growth and the fermentation. Consequently, butanol titers in conventional ABE fermentations are usually lower than 13 g/L. The low butanol yield and butanol concentration made butanol production from glucose by ABE fermentation uneconomical.« less

Improved efficiency of butanol production by absorbed lignocellulose fermentation.

PubMed

He, Qin; Chen, Hongzhang

2013-03-01

Alkali-treated steam-exploded corn stover (SECSAT) was used as solid substrate for acetone-butanol-ethanol (ABE) production by absorbed lignocellulose fermentation (ALF) using Clostridium acetobutylicum ATCC 824. The ABE concentration in ALF culture had increased by 47% compared with that in submerged culture. More surprisingly, the acetone production was promoted and ethanol production was lower in the presence of SECSAT than that in its absence. ALF was also successfully in cofermentation of glucose and xylose, although decreased fermentability with an increase in the proportion of xylose. An invariable chemical composition and dry weight of SECSAT was found in ALF. Partial simultaneous saccharification and fermentation of SECSAT using a certain amount of cellulase could not only enhance the ABE concentration by 71%, but also significantly increase the area proportion of fiber cells in SECSAT from 53% to 90%, which would be an excellent paper making material. Copyright © 2012 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

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.

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.

CaCO3 supplementation alleviates the inhibition of formic acid on acetone/butanol/ethanol fermentation by Clostridium acetobutylicum.

PubMed

Qi, Gaoxiang; Xiong, Lian; Lin, Xiaoqing; Huang, Chao; Li, Hailong; Chen, Xuefang; Chen, Xinde

2017-01-01

To investigate the inhibiting effect of formic acid on acetone/butanol/ethanol (ABE) fermentation and explain the mechanism of the alleviation in the inhibiting effect under CaCO 3 supplementation condition. From the medium containing 50 g sugars l -1 and 0.5 g formic acid l -1 , only 0.75 g ABE l -1 was produced when pH was adjusted by KOH and fermentation ended prematurely before the transformation from acidogenesis to solventogenesis. In contrast, 11.4 g ABE l -1 was produced when pH was adjusted by 4 g CaCO 3 l -1 . The beneficial effect can be ascribed to the buffering capacity of CaCO 3 . Comparative analysis results showed that the undissociated formic acid concentration and acid production coupled with ATP and NADH was affected by the pH buffering capacity of CaCO 3 . Four millimole undissociated formic acid was the threshold at which the transformation to solventogenesis occurred. The inhibiting effect of formic acid on ABE fermentation can be alleviated by CaCO 3 supplementation due to its buffering capacity.

Application of continuous substrate feeding to the ABE fermentation: Relief of product inhibition using extraction, perstraction, stripping, and pervaporation

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

Qureshi, N.; Maddox, I.S.; Friedl, A.

1992-09-01

The technique of continuous substrate feeding has been applied to the batch fermentation process using freely suspended cells, for ABE (acetone-butanol-ethanol) production. To avoid the product inhibition which normally restricts ABE production to less than 20 g/L and sugar utilization to 60 g/L, a product removal technique has been integrated into the fermentation process. The techniques investigated were liquid-liquid extraction, perstraction, gas-stripping, and pervaporation. By using a substrate of whey permeate, the reactor productivity has been improved over that observed in a traditional batch fermentation, while equivalent lactose utilization and ABE production values of 180 g and 69 g, respectively,more » have been achieved in a 1-L culture volume. 17 refs., 14 figs., 5 tabs.« less

Production of Butanol (A Biofuel) from Agricultural Residues: Part I - Use of Barley Straw Hydrolysate

USDA-ARS?s Scientific Manuscript database

Fermentation of dilute sulfuric acid barley straw hydrolyzate (BSH; undiluted/untreated) by Clostridium beijerinckii P260 resulted in the production of 7.09 gL**-1 ABE (acetone butanol ethanol; AB or ABE), an ABE yield of 0.33, and productivity of 0.10 gL**-1h**-1. This level of ABE is much less th...

Butanol production from food waste: a novel process for producing sustainable energy and reducing environmental pollution.

PubMed

Huang, Haibo; Singh, Vijay; Qureshi, Nasib

2015-01-01

Waste is currently a major problem in the world, both in the developing and the developed countries. Efficient utilization of food waste for fuel and chemical production can positively influence both the energy and environmental sustainability. This study investigated using food waste to produce acetone, butanol, and ethanol (ABE) by Clostridium beijerinckii P260. In control fermentation, 40.5 g/L of glucose (initial glucose 56.7 g/L) was used to produce 14.2 g/L of ABE with a fermentation productivity and a yield of 0.22 g/L/h and 0.35 g/g, respectively. In a similar fermentation 81 g/L of food waste (containing equivalent glucose of 60.1 g/L) was used as substrate, and the culture produced 18.9 g/L ABE with a high ABE productivity of 0.46 g/L/h and a yield of 0.38 g/g. Fermentation of food waste at higher concentrations (129, 181 and 228 g/L) did not remarkably increase ABE production but resulted in high residual glucose due to the culture butanol inhibition. An integrated vacuum stripping system was designed and applied to recover butanol from the fermentation broth simultaneously to relieve the culture butanol inhibition, thereby allowing the fermentation of food waste at high concentrations. ABE fermentation integrated with vacuum stripping successfully recovered the ABE from the fermentation broth and controlled the ABE concentrations below 10 g/L during fermentation when 129 g/L food waste was used. The ABE productivity with vacuum fermentation was 0.49 g/L/h, which was 109 % higher than the control fermentation (glucose based). More importantly, ABE vacuum recovery and fermentation allowed near-complete utilization of the sugars (~98 %) in the broth. In these studies it was demonstrated that food waste is a superior feedstock for producing butanol using Clostridium beijerinckii. Compared to costly glucose, ABE fermentation of food waste has several advantages including lower feedstock cost, higher productivity, and less residual sugars.

Mathematical modelling of clostridial acetone-butanol-ethanol fermentation.

PubMed

Millat, Thomas; Winzer, Klaus

2017-03-01

Clostridial acetone-butanol-ethanol (ABE) fermentation features a remarkable shift in the cellular metabolic activity from acid formation, acidogenesis, to the production of industrial-relevant solvents, solventogensis. In recent decades, mathematical models have been employed to elucidate the complex interlinked regulation and conditions that determine these two distinct metabolic states and govern the transition between them. In this review, we discuss these models with a focus on the mechanisms controlling intra- and extracellular changes between acidogenesis and solventogenesis. In particular, we critically evaluate underlying model assumptions and predictions in the light of current experimental knowledge. Towards this end, we briefly introduce key ideas and assumptions applied in the discussed modelling approaches, but waive a comprehensive mathematical presentation. We distinguish between structural and dynamical models, which will be discussed in their chronological order to illustrate how new biological information facilitates the 'evolution' of mathematical models. Mathematical models and their analysis have significantly contributed to our knowledge of ABE fermentation and the underlying regulatory network which spans all levels of biological organization. However, the ties between the different levels of cellular regulation are not well understood. Furthermore, contradictory experimental and theoretical results challenge our current notion of ABE metabolic network structure. Thus, clostridial ABE fermentation still poses theoretical as well as experimental challenges which are best approached in close collaboration between modellers and experimentalists.

Enhanced enzymatic hydrolysis and acetone-butanol-ethanol fermentation of sugarcane bagasse by combined diluted acid with oxidate ammonolysis pretreatment.

PubMed

Li, Hailong; Xiong, Lian; Chen, Xuefang; Wang, Can; Qi, Gaoxiang; Huang, Chao; Luo, Mutan; Chen, Xinde

2017-03-01

This study aims to propose a biorefinery pretreatment technology for the bioconversion of sugarcane bagasse (SB) into biofuels and N-fertilizers. Performance of diluted acid (DA), aqueous ammonia (AA), oxidate ammonolysis (OA) and the combined DA with AA or OA were compared in SB pretreatment by enzymatic hydrolysis, structural characterization and acetone-butanol-ethanol (ABE) fermentation. Results indicated that DA-OA pretreatment improves the digestibility of SB by sufficiently hydrolyzing hemicellulose into fermentable monosaccharides and oxidating lignin into soluble N-fertilizer with high nitrogen content (11.25%) and low C/N ratio (3.39). The enzymatic hydrolysates from DA-OA pretreated SB mainly composed of glucose was more suitable for the production of ABE solvents than the enzymatic hydrolysates from OA pretreated SB containing high ratio of xylose. The fermentation of enzymatic hydrolysates from DA-OA pretreated SB produced 12.12g/L ABE in 120h. These results suggested that SB could be utilized efficient, economic, and environmental by DA-OA pretreatment. Copyright © 2017 Elsevier Ltd. All rights reserved.

Butanol production by fermentation: efficient bioreactors

USDA-ARS?s Scientific Manuscript database

Energy security, environmental concerns, and business opportunities in the emerging bio-economy have generated strong interest in the production of n-butanol by fermentation. Acetone butanol ethanol (ABE or solvent) batch fermentation process is product limiting because butanol even at low concentra...

In situ hydrogen, acetone, butanol, ethanol and microdiesel production by Clostridium acetobutylicum ATCC 824 from oleaginous fungal biomass.

PubMed

Hassan, Elhagag Ahmed; Abd-Alla, Mohamed Hemida; Bagy, Magdy Mohamed Khalil; Morsy, Fatthy Mohamed

2015-08-01

An in situ batch fermentation technique was employed for biohydrogen, acetone, butanol, ethanol and microdiesel production from oleaginous fungal biomass using the anaerobic fermentative bacterium Clostridium acetobutylicum ATCC 824. Oleaginous fungal Cunninghamella echinulata biomass which has ability to accumulate up to 71% cellular lipid was used as the substrate carbon source. The maximum cumulative hydrogen by C. acetobutylicum ATCC 824 from crude C. echinulata biomass was 260 ml H2 l(-1), hydrogen production efficiency was 0.32 mol H2 mole(-1) glucose and the hydrogen production rate was 5.2 ml H2 h(-1). Subsequently, the produced acids (acetic and butyric acids) during acidogenesis phase are re-utilized by ABE-producing clostridia and converted into acetone, butanol, and ethanol. The total ABE produced by C. acetobutylicum ATCC 824 during batch fermentation was 3.6 g l(-1) from crude fungal biomass including acetone (1.05 g l(-1)), butanol (2.19 g l(-1)) and ethanol (0.36 g l(-1)). C. acetobutylicum ATCC 824 has ability to produce lipolytic enzymes with a specific activity 5.59 U/mg protein to hydrolyze ester containing substrates. The lipolytic potential of C. acetobutylicum ATCC 824 was used as a biocatalyst for a lipase transesterification process using the produced ethanol from ABE fermentation for microdiesel production. The fatty acid ethyl esters (microdiesel) generated from the lipase transesterification of crude C. echinulata dry mass was analyzed by GC/MS as 15.4% of total FAEEs. The gross energy content of biohydrogen, acetone, butanol, ethanol and biodiesel generated through C. acetobutylicum fermentation from crude C. echinulata dry mass was 3113.14 kJ mol(-1). These results suggest a possibility of integrating biohydrogen, acetone, butanol and ethanol production technology by C. acetobutylicum with microdiesel production from crude C. echinulata dry mass and therefore improve the feasibility and commercialization of bioenergy production. Copyright © 2015 Elsevier Ltd. All rights reserved.

Mathematical models of ABE fermentation: review and analysis.

PubMed

Mayank, Rahul; Ranjan, Amrita; Moholkar, Vijayanand S

2013-12-01

Among different liquid biofuels that have emerged in the recent past, biobutanol produced via fermentation processes is of special interest due to very similar properties to that of gasoline. For an effective design, scale-up, and optimization of the acetone-butanol-ethanol (ABE) fermentation process, it is necessary to have insight into the micro- and macro-mechanisms of the process. The mathematical models for ABE fermentation are efficient tools for this purpose, which have evolved from simple stoichiometric fermentation equations in the 1980s to the recent sophisticated and elaborate kinetic models based on metabolic pathways. In this article, we have reviewed the literature published in the area of mathematical modeling of the ABE fermentation. We have tried to present an analysis of these models in terms of their potency in describing the overall physiology of the process, design features, mode of operation along with comparison and validation with experimental results. In addition, we have also highlighted important facets of these models such as metabolic pathways, basic kinetics of different metabolites, biomass growth, inhibition modeling and other additional features such as cell retention and immobilized cultures. Our review also covers the mathematical modeling of the downstream processing of ABE fermentation, i.e. recovery and purification of solvents through flash distillation, liquid-liquid extraction, and pervaporation. We believe that this review will be a useful source of information and analysis on mathematical models for ABE fermentation for both the appropriate scientific and engineering communities.

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.

Solvent (acetone-butanol: ab) production

USDA-ARS?s Scientific Manuscript database

This article describes production of butanol [acetone-butanol-ethanol, (also called AB or ABE or solvent)] by fermentation using both traditional and current technologies. AB production from agricultural commodities, such as corn and molasses, was an important historical fermentation. Unfortunately,...

Metabolic engineering of Clostridium acetobutylicum for the enhanced production of isopropanol-butanol-ethanol fuel mixture.

PubMed

Jang, Yu-Sin; Malaviya, Alok; Lee, Joungmin; Im, Jung Ae; Lee, Sang Yup; Lee, Julia; Eom, Moon-Ho; Cho, Jung-Hee; Seung, Do Young

2013-01-01

Butanol is considered as a superior biofuel, which is conventionally produced by clostridial acetone-butanol-ethanol (ABE) fermentation. Among ABE, only butanol and ethanol can be used as fuel alternatives. Coproduction of acetone thus causes lower yield of fuel alcohols. Thus, this study aimed at developing an improved Clostridium acetobutylicum strain possessing enhanced fuel alcohol production capability. For this, we previously developed a hyper ABE producing BKM19 strain was further engineered to convert acetone into isopropanol. The BKM19 strain was transformed with the plasmid pIPA100 containing the sadh (primary/secondary alcohol dehydrogenase) and hydG (putative electron transfer protein) genes from the Clostridium beijerinckii NRRL B593 cloned under the control of the thiolase promoter. The resulting BKM19 (pIPA100) strain produced 27.9 g/l isopropanol-butanol-ethanol (IBE) as a fuel alcohols with negligible amount of acetone (0.4 g/l) from 97.8 g/l glucose in lab-scale (2 l) batch fermentation. Thus, this metabolically engineered strain was able to produce 99% of total solvent produced as fuel alcohols. The scalability and stability of BKM19 (pIPA100) were evaluated at 200 l pilot-scale fermentation, which showed that the fuel alcohol yield could be improved to 0.37 g/g as compared to 0.29 g/g obtained at lab-scale fermentation, while attaining a similar titer. To the best of our knowledge, this is the highest titer of IBE achieved and the first report on the large scale fermentation of C. acetobutylicum for IBE production. © 2013 American Institute of Chemical Engineers.

Butanol production from wood pulping hydrolysate in an integrated fermentation-gas stripping process

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

Lu, CC; Dong, J; Yang, ST

2013-09-01

Wood pulping hydrolysate (WPH) containing mainly xylose and glucose as a potential substrate for acetone-butanol-ethanol (ABE) fermentation was studied. Due to the inhibitors present in the hydrolysate, several dilution levels and detoxification treatments, including overliming, activated charcoal adsorption, and resin adsorption, were evaluated for their effectiveness in relieving the inhibition on fermentation. Detoxification using resin and evaporation was found to be the most effective method in reducing the toxicity of WPH. ABE production in batch fermentation by Clostridium beijerinckii increased 68%, from 6.73 g/L in the non-treated and non-diluted WPH to 11.35 g/L in the resin treated WPH. With gasmore » stripping for in situ product removal, ABE production from WPH increased to 17.73 g/L, demonstrating that gas stripping was effective in alleviating butanol toxicity by selectively separating butanol from the fermentation broth, which greatly improved solvents production and sugar conversion in the fermentation. (C) 2013 Elsevier Ltd. All rights reserved.« less

Butanol production from wood pulping hydrolysate in an integrated fermentation-gas stripping process.

PubMed

Lu, Congcong; Dong, Jie; Yang, Shang-Tian

2013-09-01

Wood pulping hydrolysate (WPH) containing mainly xylose and glucose as a potential substrate for acetone-butanol-ethanol (ABE) fermentation was studied. Due to the inhibitors present in the hydrolysate, several dilution levels and detoxification treatments, including overliming, activated charcoal adsorption, and resin adsorption, were evaluated for their effectiveness in relieving the inhibition on fermentation. Detoxification using resin and evaporation was found to be the most effective method in reducing the toxicity of WPH. ABE production in batch fermentation by Clostridium beijerinckii increased 68%, from 6.73 g/L in the non-treated and non-diluted WPH to 11.35 g/L in the resin treated WPH. With gas stripping for in situ product removal, ABE production from WPH increased to 17.73 g/L, demonstrating that gas stripping was effective in alleviating butanol toxicity by selectively separating butanol from the fermentation broth, which greatly improved solvents production and sugar conversion in the fermentation. Copyright © 2013 Elsevier Ltd. All rights reserved.

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.

Cellulosic Substrates and Challenges Ahead

USDA-ARS?s Scientific Manuscript database

The cost of production of butanol (acetone-butanol-ethanol; or ABE) is determined by feedstock prices, fermentation, recovery, by-product credits and the waste water treatment. Along these lines, we have an intensive research program on the use of various agricultural substrates, fermentation strate...

Comparative shotgun proteomic analysis of Clostridium acetobutylicum from butanol fermentation using glucose and xylose

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

Sivagnanam, Kumaran; Raghavan, Vijaya G. S.; Shah, Manesh B

2011-01-01

Background: Butanol is a second generation biofuel produced by Clostridium acetobutylicum through acetonebutanol- ethanol (ABE) fermentation process. Shotgun proteomics provides a direct approach to study the whole proteome of an organism in depth. This paper focuses on shotgun proteomic profiling of C. acetobutylicum from ABE fermentation using glucose and xylose to understand the functional mechanisms of C. acetobutylicum proteins involved in butanol production. Results: We identified 894 different proteins in C. acetobutylicum from ABE fermentation process by two dimensional - liquid chromatography - tandem mass spectrometry (2D-LC-MS/MS) method. This includes 717 proteins from glucose and 826 proteins from the xylosemore » substrate. A total of 649 proteins were found to be common and 22 significantly differentially expressed proteins were identified between glucose and xylose substrates. Conclusion: Our results demonstrate that flagellar proteins are highly up-regulated with glucose compared to xylose substrate during ABE fermentation. Chemotactic activity was also found to be lost with the xylose substrate due to the absence of CheW and CheV proteins. This is the first report on the shotgun proteomic analysis of C. acetobutylicum ATCC 824 in ABE fermentation between glucose and xylose substrate from a single time data point and the number of proteins identified here is more than any other study performed on this organism up to this report.« less

Combined Detoxification and In-situ Product Removal by a Single Resin During Lignocellulosic Butanol Production

PubMed Central

Gao, Kai; Rehmann, Lars

2016-01-01

Phragmites australis (an invasive plant in North America) was used as feedstock for ABE (acetone-butanol-ethanol) fermentation by Clostridium saccharobutylicum. Sulphuric acid pretreated phragmites hydrolysate (SAEH) without detoxification inhibited butanol production (0.73 g/L butanol from 30 g/L sugars). The treatment of SAEH with resin L-493 prior the fermentation resulted in no inhibitory effects and an ABE titer of 14.44 g/L, including 5.49 g/L butanol was obtained, corresponding to an ABE yield and productivity of 0.49 g/g and 0.60 g/L/h, respectively. Dual functionality of the resin was realized by also using it as an in-situ product removal agent. Integrating in-situ product removal allowed for the use of high substrate concentrations without the typical product inhibition. Resin-detoxified SAEH was supplemented with neat glucose and an effective ABE titer of 33 g/L (including 13.7 g/L acetone, 16.4 g/L butanol and 1.9 g/L ethanol) was achieved with resin-based in-situ product removal, corresponding to an ABE yield and productivity of 0.41 g/g and 0.69 g/L/h, respectively. Both detoxification of the substrate and the products was achieved by the same resin, which was added prior the fermentation. Integrating hydrolysate detoxification and in-situ butanol removal in a batch process through single resin can potentially simplify cellulosic butanol production. PMID:27459906

Combined Detoxification and In-situ Product Removal by a Single Resin During Lignocellulosic Butanol Production

NASA Astrophysics Data System (ADS)

Gao, Kai; Rehmann, Lars

2016-07-01

Phragmites australis (an invasive plant in North America) was used as feedstock for ABE (acetone-butanol-ethanol) fermentation by Clostridium saccharobutylicum. Sulphuric acid pretreated phragmites hydrolysate (SAEH) without detoxification inhibited butanol production (0.73 g/L butanol from 30 g/L sugars). The treatment of SAEH with resin L-493 prior the fermentation resulted in no inhibitory effects and an ABE titer of 14.44 g/L, including 5.49 g/L butanol was obtained, corresponding to an ABE yield and productivity of 0.49 g/g and 0.60 g/L/h, respectively. Dual functionality of the resin was realized by also using it as an in-situ product removal agent. Integrating in-situ product removal allowed for the use of high substrate concentrations without the typical product inhibition. Resin-detoxified SAEH was supplemented with neat glucose and an effective ABE titer of 33 g/L (including 13.7 g/L acetone, 16.4 g/L butanol and 1.9 g/L ethanol) was achieved with resin-based in-situ product removal, corresponding to an ABE yield and productivity of 0.41 g/g and 0.69 g/L/h, respectively. Both detoxification of the substrate and the products was achieved by the same resin, which was added prior the fermentation. Integrating hydrolysate detoxification and in-situ butanol removal in a batch process through single resin can potentially simplify cellulosic butanol production.

Butanol productivity enhancers in wheat straw hydrolyzate: employing potential of enhanced reaction rate

USDA-ARS?s Scientific Manuscript database

Butanol production by fermentation is gaining momentum due to increased prices of fossil fuels. This biofuel is a major product of acetone-butanol-ethanol (ABE) fermentation that can be produced from hydrolyzed agricultural residues and/or corn. A control glucose (60 g/L) based batch fermentation us...

Production of butanol from starch-based waste packing peanuts and agricultural waste.

PubMed

Jesse, T W; Ezeji, T C; Qureshi, N; Blaschek, H P

2002-09-01

We examined the fermentation of starch-based packing peanuts and agricultural wastes as a source of fermentable carbohydrates using Clostridium beijerinckii BA101. Using semidefined P2 medium containing packing peanuts and agricultural wastes, instead of glucose as a carbohydrate source, we measured characteristics of the fermentation including solvent production, productivity, and yield. With starch as substrate (control), the culture produced 24.7 g l(-1) acetone-butanol-ethanol (ABE), while with packing peanuts it produced 21.7 g l(-1) total ABE with a productivity of 0.20 g l(-1) h(-1) and a solvent (ABE) yield of 0.37. Cell growth in starch, packing peanuts, and agricultural wastes medium was different, possibly due to the different nature of these substrates. Using model agricultural waste, 20.3g l(-1) ABE was produced; when using actual waste, 14.8 g l(-1) ABE was produced. The use of inexpensive substrates will increase the economic viability of the conversion of biomass to butanol, and can provide new markets for these waste streams.

Shotgun proteomic monitoring of Clostridium acetobutylicum during stationary phase of butanol fermentation using xylose and comparison with the exponential phase

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

Sivagnanam, Kumaran; Raghavan, Vijaya G. S.; Shah, Manesh B

2012-01-01

Economically viable production of solvents through acetone butanol ethanol (ABE) fermentation requires a detailed understanding of Clostridium acetobutylicum. This study focuses on the proteomic profiling of C. acetobutylicum ATCC 824 from the stationary phase of ABE fermentation using xylose and compares with the exponential growth by shotgun proteomics approach. Comparative proteomic analysis revealed 22.9% of the C. acetobutylicum genome and 18.6% was found to be common in both exponential and stationary phases. The proteomic profile of C. acetobutylicum changed during the ABE fermentation such that 17 proteins were significantly differentially expressed between the two phases. Specifically, the expression of fivemore » proteins namely, CAC2873, CAP0164, CAP0165, CAC3298, and CAC1742 involved in the solvent production pathway were found to be significantly lower in the stationary phase compared to the exponential growth. Similarly, the expression of fucose isomerase (CAC2610), xylulose kinase (CAC2612), and a putative uncharacterized protein (CAC2611) involved in the xylose utilization pathway were also significantly lower in the stationary phase. These findings provide an insight into the metabolic behavior of C. acetobutylicum between different phases of ABE fermentation using xylose.« less

Conversion of food processing wastes to biofuel using clostridia.

PubMed

Abd-Alla, Mohamed Hemida; Zohri, Abdel-Naser Ahmed; El-Enany, Abdel-Wahab Elsadek; Ali, Shimaa Mohamed

2017-12-01

This study aims to demonstrate the recycling of food processing wastes as a low cost-effective substrate for acetone - butanol - ethanol (ABE) production. Potato peels and cheese whey were utilized during fermentation with eight local Clostridium strains in addition to the commercial strain, C. acetobutylicum ATCC 824 for ABE and organic acids production. From potato peels, Clostridium beijerinckii ASU10 produced the highest ABE production (17.91 g/l) representing 61.3% butanol (10.98 g/l), 33.6% acetone (6.02 g/l) and 5.1% ethanol (0.91 g/l). While, C. chauvoei ASU12 showed the highest acid production (8.15 g/l) including 5.50 and 2.61 g/l acetic and butyric acids, respectively. Use of cheese whey as fermentable substrate exhibited a substantial increase in ethanol ratio and decrease in butanol ratio compared to those produced from potato peels. Clostridium beijerinckii ASU5 produced the highest ABE concentration (7.13 g/l) representing 50.91% butanol (3.63 g/l), 35.34% acetone (2.52 g/l) and 13.74% ethanol (0.98 g/l). The highest acid production (8.00 g/l) was obtained by C. beijerinckii ASU5 representing 4.89 and 3.11 g/l for acetic and butyric acid, respectively. Supplementation of potato peels with an organic nitrogen source showed NH 4 NO 3 promoted ABE production more than yeast extract. In conclusion, this study introduced an ecofriendly and economical practice for utilization of food processing wastes (renewable substrates as potato peels and cheese whey) for biofuel production using various Clostridium strains. Copyright © 2017 Elsevier Ltd. All rights reserved.

Integrated, systems metabolic picture of acetone-butanol-ethanol fermentation by Clostridium acetobutylicum.

PubMed

Liao, Chen; Seo, Seung-Oh; Celik, Venhar; Liu, Huaiwei; Kong, Wentao; Wang, Yi; Blaschek, Hans; Jin, Yong-Su; Lu, Ting

2015-07-07

Microbial metabolism involves complex, system-level processes implemented via the orchestration of metabolic reactions, gene regulation, and environmental cues. One canonical example of such processes is acetone-butanol-ethanol (ABE) fermentation by Clostridium acetobutylicum, during which cells convert carbon sources to organic acids that are later reassimilated to produce solvents as a strategy for cellular survival. The complexity and systems nature of the process have been largely underappreciated, rendering challenges in understanding and optimizing solvent production. Here, we present a system-level computational framework for ABE fermentation that combines metabolic reactions, gene regulation, and environmental cues. We developed the framework by decomposing the entire system into three modules, building each module separately, and then assembling them back into an integrated system. During the model construction, a bottom-up approach was used to link molecular events at the single-cell level into the events at the population level. The integrated model was able to successfully reproduce ABE fermentations of the WT C. acetobutylicum (ATCC 824), as well as its mutants, using data obtained from our own experiments and from literature. Furthermore, the model confers successful predictions of the fermentations with various network perturbations across metabolic, genetic, and environmental aspects. From foundation to applications, the framework advances our understanding of complex clostridial metabolism and physiology and also facilitates the development of systems engineering strategies for the production of advanced biofuels.

Integrated, systems metabolic picture of acetone-butanol-ethanol fermentation by Clostridium acetobutylicum

PubMed Central

Liao, Chen; Seo, Seung-Oh; Celik, Venhar; Liu, Huaiwei; Kong, Wentao; Wang, Yi; Blaschek, Hans; Jin, Yong-Su; Lu, Ting

2015-01-01

Microbial metabolism involves complex, system-level processes implemented via the orchestration of metabolic reactions, gene regulation, and environmental cues. One canonical example of such processes is acetone-butanol-ethanol (ABE) fermentation by Clostridium acetobutylicum, during which cells convert carbon sources to organic acids that are later reassimilated to produce solvents as a strategy for cellular survival. The complexity and systems nature of the process have been largely underappreciated, rendering challenges in understanding and optimizing solvent production. Here, we present a system-level computational framework for ABE fermentation that combines metabolic reactions, gene regulation, and environmental cues. We developed the framework by decomposing the entire system into three modules, building each module separately, and then assembling them back into an integrated system. During the model construction, a bottom-up approach was used to link molecular events at the single-cell level into the events at the population level. The integrated model was able to successfully reproduce ABE fermentations of the WT C. acetobutylicum (ATCC 824), as well as its mutants, using data obtained from our own experiments and from literature. Furthermore, the model confers successful predictions of the fermentations with various network perturbations across metabolic, genetic, and environmental aspects. From foundation to applications, the framework advances our understanding of complex clostridial metabolism and physiology and also facilitates the development of systems engineering strategies for the production of advanced biofuels. PMID:26100881

Cellulosic butanol biofuel production from sweet sorghum bagasse (SSB): Impact of hot water pretreatment and solid loadings on fermentation employing Clostridium beijerinckii P260

USDA-ARS?s Scientific Manuscript database

A novel butanol fermentation process was developed in which sweet sorghum bagasse (SSB) was pretreated using liquid hot water (LHW) pretreatment technique followed by enzymatic hydrolysis and butanol (acetone butanol ethanol; ABE) fermentation. A pretreatment temperature of 200 deg C resulted in the...

Butanol production from lignocellulose by simultaneous fermentation, saccharification, and pervaporation or vacuum evaporation.

PubMed

Díaz, Víctor Hugo Grisales; Tost, Gerard Olivar

2016-10-01

Techno-economic study of acetone, butanol and ethanol (ABE) fermentation from lignocellulose was performed. Simultaneous saccharification, fermentation and vacuum evaporation (SFS-V) or pervaporation (SFS-P) were proposed. A kinetic model of metabolic pathways for ABE fermentation with the effect of phenolics and furans in the growth was proposed based on published laboratory results. The processes were optimized in Matlab®. The end ABE purification was carried out by heat-integrated distillation. The objective function of the minimization was the total annualized cost (TAC). Fuel consumption of SFS-P using poly[1-(trimethylsilyl)-1-propyne] membrane was between 13.8 and 19.6% lower than SFS-V. Recovery of furans and phenolics for the hybrid reactors was difficult for its high boiling point. TAC of SFS-P was increased 1.9 times with supplementation of phenolics and furans to 3g/l each one for its high toxicity. Therefore, an additional detoxification method or an efficient pretreatment process will be necessary. Copyright © 2016 Elsevier Ltd. All rights reserved.

Industrial production of acetone and butanol by fermentation-100 years later.

PubMed

Sauer, Michael

2016-07-01

Microbial production of acetone and butanol was one of the first large-scale industrial fermentation processes of global importance. During the first part of the 20th century, it was indeed the second largest fermentation process, superseded in importance only by the ethanol fermentation. After a rapid decline after the 1950s, acetone-butanol-ethanol (ABE) fermentation has recently gained renewed interest in the context of biorefinery approaches for the production of fuels and chemicals from renewable resources. The availability of new methods and knowledge opens many new doors for industrial microbiology, and a comprehensive view on this process is worthwhile due to the new interest. This thematic issue of FEMS Microbiology Letters, dedicated to the 100th anniversary of the first industrial exploitation of Chaim Weizmann's ABE fermentation process, covers the main aspects of old and new developments, thereby outlining a model development in biotechnology. All major aspects of industrial microbiology are exemplified by this single process. This includes new technologies, such as the latest developments in metabolic engineering, the exploitation of biodiversity and discoveries of new regulatory systems such as for microbial stress tolerance, as well as technological aspects, such as bio- and down-stream processing. © FEMS 2016.

Continuous production of butanol from starch-based packing peanuts.

PubMed

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

2003-01-01

Acetone, butanol, ethanol (ABE, or solvents) were produced from starch-based packing peanuts in batch and continuous reactors. In a batch reactor, 18.9 g/L of total ABE was produced from 80 g/L packing peanuts in 110 h of fermentation. The initial and final starch concentrations were 69.6 and 11.1 g/L, respectively. In this fermentation, ABE yield and productivity of 0.32 and 0.17 g/(L h) were obtained, respectively. Compared to the batch fermentation, continuous fermentation of 40 g/L of starchbased packing peanuts in P2 medium resulted in a maximum solvent production of 8.4 g/L at a dilution rate of 0.033 h-1. This resulted in a productivity of 0.27 g/(L h). However, the reactor was not stable and fermentation deteriorated with time. Continuous fermentation of 35 g/L of starch solution resulted in a similar performance. These studies were performed in a vertical column reactor using Clostridium beijerinckii BA101 and P2 medium. It is anticipated that prolonged exposure of culture to acrylamide, which is formed during boiling/autoclaving of starch, affects the fermentation negatively.

Detoxification of Organosolv-Pretreated Pine Prehydrolysates with Anion Resin and Cysteine for Butanol Fermentation.

PubMed

Li, Jing; Shi, Suan; Tu, Maobing; Via, Brain; Sun, Fubao Fuelbio; Adhikari, Sushil

2018-05-02

Bioconversion of lignocellulose to biofuels suffers from the degradation compounds formed during pretreatment and acid hydrolysis. In order to achieve an efficient biomass to biofuel conversion, detoxification is often required before enzymatic hydrolysis and microbial fermentation. Prehydrolysates from ethanol organosolv-pretreated pine wood were used as substrates in butanol fermentation in this study. Six detoxification approaches were studied and compared, including overliming, anion exchange resin, nonionic resin, laccase, activated carbon, and cysteine. It was observed that detoxification by anion exchange resin was the most effective method. The final butanol yield after anion exchange resin treatment was comparable to the control group, but the fermentation was delayed for 72 h. The addition of Ca(OH) 2 was found to alleviate this delay and improve the fermentation efficiency. The combination of Ca(OH) 2 and anion exchange resin resulted in completion of fermentation within 72 h and acetone-butanol-ethanol (ABE) production of 11.11 g/L, corresponding to a yield of 0.21 g/g sugar. The cysteine detoxification also resulted in good detoxification performance, but promoted fermentation towards acid production (8.90 g/L). The effect of salt on ABE fermentation was assessed and the possible role of Ca(OH) 2 was to remove the salts in the prehydrolysates by precipitation.

Enhancing acetone biosynthesis and acetone-butanol-ethanol fermentation performance by co-culturing Clostridium acetobutylicum/Saccharomyces cerevisiae integrated with exogenous acetate addition.

PubMed

Luo, Hongzhen; Ge, Laibing; Zhang, Jingshu; Ding, Jian; Chen, Rui; Shi, Zhongping

2016-01-01

Acetone is the major by-product in ABE fermentations, most researches focused on increasing butanol/acetone ratio by decreasing acetone biosynthesis. However, economics of ABE fermentation industry strongly relies on evaluating acetone as a valuable platform chemical. Therefore, a novel ABE fermentation strategy focusing on bio-acetone production by co-culturing Clostridium acetobutylicum/Saccharomyces cerevisiae with exogenous acetate addition was proposed. Experimental and theoretical analysis revealed the strategy could, enhance C. acetobutylicum survival oriented amino acids assimilation in the cells; control NADH regeneration rate at moderately lower level to enhance acetone synthesis but without sacrificing butanol production; enhance the utilization ability of C. acetobutylicum on glucose and direct most of extra consumed glucose into acetone/butanol synthesis routes. By implementing the strategy using synthetic or acetate fermentative supernatant, acetone concentrations increased to 8.27-8.55g/L from 5.86g/L of the control, while butanol concentrations also elevated to the higher levels of 13.91-14.23g/L from 11.63g/L simultaneously. Copyright © 2015 Elsevier Ltd. All rights reserved.

Glycerol supplementation of the growth medium enhances in situ detoxification of furfural by Clostridium beijerinckii during butanol fermentation.

PubMed

Ujor, Victor; Agu, Chidozie Victor; Gopalan, Venkat; Ezeji, Thaddeus Chukwuemeka

2014-01-01

Lignocellulose-derived microbial inhibitors such as furfural and 5-hydroxymethyl furfural adversely affect fermentation of lignocellulosic biomass hydrolysates to fuels and chemicals due to their toxicity on fermenting microbes. To harness the potential of lignocellulose as a cheap source of fermentable sugars, in situ detoxification of furfural and other lignocellulose-derived microbial inhibitors is essential. To enhance in situ detoxification and tolerance of furfural by Clostridium beijerinckii NCIMB 8052 during acetone-butanol-ethanol (ABE) fermentation, the effect of glycerol on NADH/NADPH generation and ABE production by furfural (4, 5, and 6 g/L)-challenged cultures was investigated in this study. In all instances, beneficial outcomes were observed. For example, the fermentation medium supplemented with glycerol and subjected to 5 g/L furfural elicited up to 1.8- and 3-fold increases, respectively, in NADH and NADPH levels in C. beijerinckii 8052 relative to the control culture. These critical changes are the likely underpinnings for the glycerol-mediated 2.3-fold increase in the rate of detoxification of 5 g/L furfural, substrate consumption, and ABE production compared to the unsupplemented medium. Collectively, these results demonstrate that increased intracellular NADH/NADPH in C. beijerinckii 8052 due to glycerol utilization engenders favorable effects on many aspects of cellular metabolism, including enhanced furfural reduction and increased ABE production.

Novel developments in butanol fermentation: Microbial genetics to agricultural substrates, process technology, and downstream processing

USDA-ARS?s Scientific Manuscript database

Butanol is the major product of acetone-butanol-ethanol (ABE; ratio 3:6:1) fermentation. It can be produced from various carbohydrates such as glucose, corn, molasses, and whey permeate (a by-product of the dairy industry) using microbial strains such as Clostridium beijerinckii and/or C. acetobuty...

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

PubMed

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

2016-07-01

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

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

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

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

2015-02-01

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

Engineering Clostridium acetobutylicum for production of kerosene and diesel blendstock precursors.

PubMed

Bormann, Sebastian; Baer, Zachary C; Sreekumar, Sanil; Kuchenreuther, Jon M; Dean Toste, F; Blanch, Harvey W; Clark, Douglas S

2014-09-01

Processes for the biotechnological production of kerosene and diesel blendstocks are often economically unattractive due to low yields and product titers. Recently, Clostridium acetobutylicum fermentation products acetone, butanol, and ethanol (ABE) were shown to serve as precursors for catalytic upgrading to higher chain-length molecules that can be used as fuel substitutes. To produce suitable kerosene and diesel blendstocks, the butanol:acetone ratio of fermentation products needs to be increased to 2-2.5:1, while ethanol production is minimized. Here we show that the overexpression of selected proteins changes the ratio of ABE products relative to the wild type ATCC 824 strain. Overexpression of the native alcohol/aldehyde dehydrogenase (AAD) has been reported to primarily increase ethanol formation in C. acetobutylicum. We found that overexpression of the AAD(D485G) variant increased ethanol titers by 294%. Catalytic upgrading of the 824(aad(D485G)) ABE products resulted in a blend with nearly 50wt%≤C9 products, which are unsuitable for diesel. To selectively increase butanol production, C. beijerinckii aldehyde dehydrogenase and C. ljungdhalii butanol dehydrogenase were co-expressed (strain designate 824(Cb ald-Cl bdh)), which increased butanol titers by 27% to 16.9gL(-1) while acetone and ethanol titers remained essentially unaffected. The solvent ratio from 824(Cb ald-Cl bdh) resulted in more than 80wt% of catalysis products having a carbon chain length≥C11 which amounts to 9.8gL(-1) of products suitable as kerosene or diesel blendstock based on fermentation volume. To further increase solvent production, we investigated expression of both native and heterologous chaperones in C. acetobutylicum. Expression of a heat shock protein (HSP33) from Bacillus psychrosaccharolyticus increased the total solvent titer by 22%. Co-expression of HSP33 and aldehyde/butanol dehydrogenases further increased ABE formation as well as acetone and butanol yields. HSP33 was identified as the first heterologous chaperone that significantly increases solvent titers above wild type C. acetobutylicum levels, which can be combined with metabolic engineering to further increase solvent production. Copyright © 2014 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

Assessment of potential life-cycle energy and greenhouse gas emission effects from using corn-based butanol as a transportation fuel.

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

Wu, M.; Wang, M.; Liu, J.

2008-01-01

Since advances in the ABE (acetone-butanol-ethanol) fermentation process in recent years have led to significant increases in its productivity and yields, the production of butanol and its use in motor vehicles have become an option worth evaluating. This study estimates the potential life-cycle energy and emission effects associated with using bio-butanol as a transportation fuel. It employs a well-to-wheels (WTW) analysis tool: the Greenhouse Gases, Regulated Emissions, and Energy Use in Transportation (GREET) model. The estimates of life-cycle energy use and greenhouse gas (GHG) emissions are based on an Aspen Plus(reg. sign) simulation for a corn-to-butanol production process, which describesmore » grain processing, fermentation, and product separation. Bio-butanol-related WTW activities include corn farming, corn transportation, butanol production, butanol transportation, and vehicle operation. In this study, we also analyzed the bio-acetone that is coproduced with bio-butanol as an alternative to petroleum-based acetone. We then compared the results for bio-butanol with those of conventional gasoline. Our study shows that driving vehicles fueled with corn-based butanol produced by the current ABE fermentation process could result in substantial fossil energy savings (39%-56%) and avoid large percentage of the GHG emission burden, yielding a 32%-48% reduction relative to using conventional gasoline. On energy basis, a bushel of corn produces less liquid fuel from the ABE process than that from the corn ethanol dry mill process. The coproduction of a significant portion of acetone from the current ABE fermentation presents a challenge. A market analysis of acetone, as well as research and development on robust alternative technologies and processes that minimize acetone while increase the butanol yield, should be conducted.« less

Assessment of potential life-cycle energy and greenhouse gas emission effects from using corn-based butanol as a transportation fuel.

PubMed

Wu, May; Wang, Michael; Liu, Jiahong; Huo, Hong

2008-01-01

Since advances in the ABE (acetone-butanol-ethanol) fermentation process in recent years have led to significant increases in its productivity and yields, the production of butanol and its use in motor vehicles have become an option worth evaluating. This study estimates the potential life-cycle energy and emission effects associated with using bio-butanol as a transportation fuel. It employs a well-to-wheels (WTW) analysis tool: the Greenhouse Gases, Regulated Emissions, and Energy Use in Transportation (GREET) model. The estimates of life-cycle energy use and greenhouse gas (GHG) emissions are based on an Aspen Plus(R) simulation for a corn-to-butanol production process, which describes grain processing, fermentation, and product separation. Bio-butanol-related WTW activities include corn farming, corn transportation, butanol production, butanol transportation, and vehicle operation. In this study, we also analyzed the bio-acetone that is coproduced with bio-butanol as an alternative to petroleum-based acetone. We then compared the results for bio-butanol with those of conventional gasoline. Our study shows that driving vehicles fueled with corn-based butanol produced by the current ABE fermentation process could result in substantial fossil energy savings (39%-56%) and avoid large percentage of the GHG emission burden, yielding a 32%-48% reduction relative to using conventional gasoline. On energy basis, a bushel of corn produces less liquid fuel from the ABE process than that from the corn ethanol dry mill process. The coproduction of a significant portion of acetone from the current ABE fermentation presents a challenge. A market analysis of acetone, as well as research and development on robust alternative technologies and processes that minimize acetone while increase the butanol yield, should be conducted.

Effect of chemical pretreatments on corn stalk bagasse as immobilizing carrier of Clostridium acetobutylicum in the performance of a fermentation-pervaporation coupled system.

PubMed

Cai, Di; Li, Ping; Chen, Changjing; Wang, Yong; Hu, Song; Cui, Caixia; Qin, Peiyong; Tan, Tianwei

2016-11-01

In this study, different pretreatment methods were evaluated for modified the corn stalk bagasse and further used the pretreated bagasse as immobilized carrier in acetone-butanol-ethanol fermentation process. Structural changes of the bagasses pretreated by different methods were analyzed by Fourier transform infrared, crystallinity index and scanning pictures by electron microscope. And the performances of batch fermentation using the corn stalk based carriers were evaluated. Results indicated that the highest ABE concentration of 23.86g/L was achieved using NaOH pretreated carrier in batch fermentation. Immobilized fermentation-pervaporation integration process was further carried out. The integration process showed long-term stability with 225-394g/L of ABE solvents on the permeate side of pervaporation membrane. This novel integration process was found to be an efficient method for biobutanol production. Copyright © 2016 Elsevier Ltd. All rights reserved.

Development of a high-throughput assay for rapid screening of butanologenic strains.

PubMed

Agu, Chidozie Victor; Lai, Stella M; Ujor, Victor; Biswas, Pradip K; Jones, Andy; Gopalan, Venkat; Ezeji, Thaddeus Chukwuemeka

2018-02-21

We report a Thermotoga hypogea (Th) alcohol dehydrogenase (ADH)-dependent spectrophotometric assay for quantifying the amount of butanol in growth media, an advance that will facilitate rapid high-throughput screening of hypo- and hyper-butanol-producing strains of solventogenic Clostridium species. While a colorimetric nitroblue tetrazolium chloride-based assay for quantitating butanol in acetone-butanol-ethanol (ABE) fermentation broth has been described previously, we determined that Saccharomyces cerevisiae (Sc) ADH used in this earlier study exhibits approximately 13-fold lower catalytic efficiency towards butanol than ethanol. Any Sc ADH-dependent assay for primary quantitation of butanol in an ethanol-butanol mixture is therefore subject to "ethanol interference". To circumvent this limitation and better facilitate identification of hyper-butanol-producing Clostridia, we searched the literature for native ADHs that preferentially utilize butanol over ethanol and identified Th ADH as a candidate. Indeed, recombinant Th ADH exhibited a 6-fold higher catalytic efficiency with butanol than ethanol, as measured using the reduction of NADP + to NADPH that accompanies alcohol oxidation. Moreover, the assay sensitivity was not affected by the presence of acetone, acetic acid or butyric acid (typical ABE fermentation products). We broadened the utility of our assay by adapting it to a high-throughput microtiter plate-based format, and piloted it successfully in an ongoing metabolic engineering initiative.

Prospective and development of butanol as an advanced biofuel.

PubMed

Xue, Chuang; Zhao, Xin-Qing; Liu, Chen-Guang; Chen, Li-Jie; Bai, Feng-Wu

2013-12-01

Butanol has been acknowledged as an advanced biofuel, but its production through acetone-butanol-ethanol (ABE) fermentation by clostridia is still not economically competitive, due to low butanol yield and titer. In this article, update progress in butanol production is reviewed. Low price and sustainable feedstocks such as lignocellulosic residues and dedicated energy crops are needed for butanol production at large scale to save feedstock cost, but processes are more complicated, compared to those established for ABE fermentation from sugar- and starch-based feedstocks. While rational designs targeting individual genes, enzymes or pathways are effective for improving butanol yield, global and systems strategies are more reasonable for engineering strains with stress tolerance controlled by multigenes. Compared to solvent-producing clostridia, engineering heterologous species such as Escherichia coli and Saccharomyces cerevisiae with butanol pathway might be a solution for eliminating the formation of major byproducts acetone and ethanol so that butanol yield can be improved significantly. Although batch fermentation has been practiced for butanol production in industry, continuous operation is more productive for large scale production of butanol as a biofuel, but a single chemostat bioreactor cannot achieve this goal for the biphasic ABE fermentation, and tanks-in-series systems should be optimized for alternative feedstocks and new strains. Moreover, energy saving is limited for the distillation system, even total solvents in the fermentation broth are increased significantly, since solvents are distilled to ~40% by the beer stripper, and more than 95% water is removed with the stillage without phase change, even with conventional distillation systems, needless to say that advanced chemical engineering technologies can distil solvents up to ~90% with the beer stripper, and the multistage pressure columns can well balance energy consumption for solvent fraction. Indeed, an increase in butanol titer with ABE fermentation can significantly save energy consumption for medium sterilization and stillage treatment, since concentrated medium can be used, and consequently total mass flow with production systems can be reduced. As for various in situ butanol removal technologies, their energy efficiency, capital investment and contamination risk to the fermentation process need to be evaluated carefully. © 2013 Elsevier Inc. All rights reserved.

Pervaporative stripping of acetone, butanol and ethanol to improve ABE fermentation.

PubMed

Jitesh, K; Pangarkar, V G; Niranjan, K

2000-01-01

Acetone-butanol-ethanol fermentation by anaerobic bacterium C. acetobutylicum is a potential source for feedstock chemicals. The problem of product induced inhibition makes this fermentation economically infeasible. Pervaporation is studied as an effective separation technique to remove the toxic inhibitory products. Various membranes like Styrene Butadiene Rubber (SBR), Ethylene Propylene Diene Rubber (EPDM), plain Poly Dimethyl Siloxane (PDMS) and silicalite filled PDMS were studied for the removal of acetone, butanol and ethanol, from binary aqueous mixtures and from a quaternary mixture. It was found that the overall performance of PDMS filled with 15% w/w of silicalite was the best for removal of butanol in binary mixture study. SBR performance was best for the quaternary mixture studied.

Process integration for simultaneous saccharification, fermentation, and recovery (SSFR): Production of butanol from corn stover using Clostridium beijerinckii P260

USDA-ARS?s Scientific Manuscript database

A simultaneous saccharification, fermentation, and recovery (SSFR) process was developed for production of acetone butanol ethanol (AB or ABE), of which butanol is the main product, from corn stover employing Clostridium beijerinckii P260. Of the 86 gL^-1^ corn stover, over 97% of the sugars were r...

A dynamic metabolic flux analysis of ABE (acetone-butanol-ethanol) fermentation by Clostridium acetobutylicum ATCC 824, with riboflavin as a by-product.

PubMed

Zhao, Xinhe; Kasbi, Mayssa; Chen, Jingkui; Peres, Sabine; Jolicoeur, Mario

2017-12-01

The present study reveals that supplementing sodium acetate (NaAc) strongly stimulates riboflavin production in acetone-butanol-ethanol (ABE) fermentation by Clostridium acetobutylicum ATCC 824 with xylose as carbon source. Riboflavin production increased from undetectable concentrations to ∼0.2 g L -1 (0.53 mM) when supplementing 60 mM NaAc. Of interest, solvents production and biomass yield were also promoted with fivefold acetone, 2.6-fold butanol, and 2.4-fold biomass adding NaAc. A kinetic metabolic model, developed to simulate ABE biosystem, with riboflavin production, revealed from a dynamic metabolic flux analysis (dMFA) simultaneous increase of riboflavin (ribA) and GTP (precursor of riboflavin) (PurM) synthesis flux rates under NaAc supplementation. The model includes 23 fluxes, 24 metabolites, and 72 kinetic parameters. It also suggested that NaAc condition has first stimulated the accumulation of intracellular metabolite intermediates during the acidogenic phase, which have then fed the solventogenic phase leading to increased ABE production. In addition, NaAc resulted in higher intracellular levels of NADH during the whole culture. Moreover, lower GTP-to-adenosine phosphates (ATP, ADP, AMP) ratio under NaAc supplemented condition suggests that GTP may have a minor role in the cell energetic metabolism compared to its contribution to riboflavin synthesis. © 2017 Wiley Periodicals, Inc.

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

PubMed

Boonsombuti, Akarin; Luengnaruemitchai, Apanee; Wongkasemjit, Sujitra

2015-01-01

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

Investigation of gas stripping and pervaporation for improved feasibility of two-stage butanol production process.

PubMed

Setlhaku, Mpho; Heitmann, Sebastian; Górak, Andrzej; Wichmann, Rolf

2013-05-01

Gas stripping and pervaporation are investigated for butanol recovery in a two-stage acetone-butanol-ethanol (ABE) fermentation process. The first stage is operated in a continuous mode and the second stage as a fed-batch. Gas stripping coupled to the second stage and operated intermittently enabled additional glucose feeding in the second stage and up to 59 g/L butanol and 73 g/L total ABE solvents in the condensate. Concentration of 167 g/L butanol and 269 g/L ABE in the permeate was measured in ex situ pervaporation experiments using a PDMS membrane at temperature of 37 °C and pressure of 10mbars. The "operating window" tool is introduced to evaluate the feasibility of the existing ABE fermentations operated as continuous with cell recycle, as two-stages, with biomass immobilization or with integrated product removal. This tool enables the identification of the most favorable process configuration, which is the combination of cell immobilization and integrated product removal. Copyright © 2013 Elsevier Ltd. All rights reserved.

A quantitative metabolomics study of high sodium response in Clostridium acetobutylicum ATCC 824 acetone-butanol-ethanol (ABE) fermentation

PubMed Central

Zhao, Xinhe; Condruz, Stefan; Chen, Jingkui; Jolicoeur, Mario

2016-01-01

Hemicellulose hydrolysates, sugar-rich feedstocks used in biobutanol refinery, are normally obtained by adding sodium hydroxide in the hydrolyze process. However, the resulting high sodium concentration in the hydrolysate inhibits ABE (acetone-butanol-ethanol) fermentation, and thus limits the use of these low-cost feedstocks. We have thus studied the effect of high sodium on the metabolic behavior of Clostridium acetobutyricum ATCC 824, with xylose as the carbon source. At a threshold sodium concentration of 200 mM, a decrease of the maximum cell dry weight (−19.50 ± 0.85%) and of ABE yield (−35.14 ± 3.50% acetone, −33.37 ± 0.74% butanol, −22.95 ± 1.81% ethanol) were observed compared to control culture. However, solvents specific productivities were not affected by supplementing sodium. The main effects of high sodium on cell metabolism were observed in acidogenesis, during which we observed the accumulation of ATP and NADH, and the inhibition of the pentose phosphate (PPP) and the glycolytic pathways with up to 80.73 ± 1.47% and 68.84 ± 3.42% decrease of the associated metabolic intermediates, respectively. However, the NADP+-to-NADPH ratio was constant for the whole culture duration, a phenomenon explaining the robustness of solvents specific productivities. Therefore, high sodium, which inhibited biomass growth through coordinated metabolic effects, interestingly triggered cell robustness on solvents specific productivity. PMID:27321153

Carbon monoxide bioconversion to butanol-ethanol by Clostridium carboxidivorans: kinetics and toxicity of alcohols.

PubMed

Fernández-Naveira, Ánxela; Abubackar, Haris Nalakath; Veiga, María C; Kennes, Christian

2016-05-01

Butanol production from carbon monoxide-rich waste gases or syngas is an attractive novel alternative to the conventional acetone-butanol-ethanol (ABE) fermentation. Solvent toxicity is a key factor reported in ABE fermentation with carbohydrates as substrates. However, in the gas-fermentation process, kinetic aspects and the inhibition effect of solvents have not thoroughly been studied. Therefore, different batch bottle experiments were carried out with the bacterial species Clostridium carboxidivorans using CO as carbon source for butanol-ethanol fermentation. A maximum specific growth rate of 0.086 ± 0.004 h(-1) and a biomass yield of 0.011 gbiomass/gCO were found, which is significantly lower than in other clostridia grown on sugars. Besides, three assays were carried out to check the inhibitory effect of butanol, ethanol, and their mixtures. Butanol had a higher inhibitory effect on the cells than ethanol and showed a lower IC50, reduced growth rate, and slower CO consumption with increasing alcohol concentrations. A concentration of 14-14.50 g/L butanol caused 50 % growth inhibition in C. carboxidivorans, and 20 g/L butanol resulted in complete inhibition, with a growth rate of 0 h(-1). Conversely, 35 g/L ethanol decreased by 50 % the final biomass concentration respect to the control and yielded the lowest growth rate of 0.024 h(-1). The inhibitory effect of mixtures of both alcohols was also checked adding similar, near identical, concentrations of each one. Growth decreased by 50 % in the presence of a total concentration of alcohols of 16.22 g/L, consisting of similar amounts of each alcohol. Occasional differences in initially added concentrations of alcohols were minimal. The lowest growth rate (0.014 h(-1)) was observed at the highest concentration assayed (25 g/L).

Nesterenkonia sp. strain F, a halophilic bacterium producing acetone, butanol, and ethanol under aerobic conditions.

PubMed

Amiri, Hamid; Azarbaijani, Reza; Parsa Yeganeh, Laleh; Shahzadeh Fazeli, Abolhassan; Tabatabaei, Meisam; Salekdeh, Ghasem Hosseini; Karimi, Keikhosro

2016-01-04

The moderately halophilic bacterium Nesterenkonia sp. strain F, which was isolated from Aran-Bidgol Lake (Iran), has the ability to produce acetone, butanol, and ethanol (ABE) as well as acetic and butyric acids under aerobic and anaerobic conditions. This result is the first report of ABE production with a wild microorganism from a family other than Clostridia and also the first halophilic species shown to produce butanol under aerobic cultivation. The cultivation of Nesterenkonia sp. strain F under anaerobic conditions with 50 g/l of glucose for 72 h resulted in the production of 105 mg/l of butanol, 122 mg/l of acetone, 0.2 g/l of acetic acid, and 2.5 g/l of butyric acid. Furthermore, the strain was cultivated on media with different glucose concentrations (20, 50, and 80 g/l) under aerobic and anaerobic conditions. Through fermentation with a 50 g/l initial glucose concentration under aerobic conditions, 66 mg/l of butanol, 125 mg/l of acetone, 291 mg/l of ethanol, 5.9 g/l of acetic acid, and 1.2 g/l of butyric acid were produced. The enzymes pertaining to the fermentation pathway in the strain were compared with the enzymes of Clostridium spp., and the metabolic pathway of fermentation used by Nesterenkonia sp. strain F was investigated.

Enhancement of butanol tolerance and butanol yield in Clostridium acetobutylicum mutant NT642 obtained by nitrogen ion beam implantation.

PubMed

Liu, Xiao-Bo; Gu, Qiu-Ya; Yu, Xiao-Bin; Luo, Wei

2012-12-01

As a promising alternative biofuel, biobutanol can be produced through acetone/butanol/ethanol (ABE) fermentation. Currently, ABE fermentation is still a small-scale industry due to its low production and high input cost. Moreover, butanol toxicity to the Clostridium fermentation host limits the accumulation of butanol in the fermentation broth. The wild-type Clostridium acetobutylicum D64 can only produce about 13 g butanol/L and tolerates less than 2% (v/v) butanol. To improve the tolerance of C. acetobutylicum D64 for enhancing the production of butanol, nitrogen ion beam implantation was employed and finally five mutants with enhanced butanol tolerance were obtained. Among these, the most butanol tolerant mutant C. acetobutylicum NT642 can tolerate above 3% (v/v) butanol while the wide-type strain can only withstand 2% (v/v). In batch fermentation, the production of butanol and ABE yield of C. acetobutylicum NT642 was 15.4 g/L and 22.3 g/L, respectively, which were both higher than those of its parental strain and the other mutants using corn or cassava as substrate. Enhancing butanol tolerance is a great precondition for obtaining a hyper-yield producer. Nitrogen ion beam implantation could be a promising biotechnology to improve butanol tolerance and production of the host strain C. acetobutylicum.

Continuous butanol fermentation and feed starch retrogradation: butanol fermentation sustainability using Clostridium beijerinckii BA101.

PubMed

Ezeji, T C; Qureshi, N; Blaschek, H P

2005-01-26

Use of starch solution as feed for butanol bioconversion processes employing Clostridium beijerinckii BA101 may have added economic advantage over the use of glucose. Acetone butanol ethanol (ABE) was produced from 30 gL(-1) starch solution using a continuous process. The bioreactor was fed at a dilution rate of 0.02 h(-1) and starch solution/feed volume (3 L) was replaced every 72 h. The continuous reactor fed with cornstarch solution (feed temperature 19 degrees C) produced approximately 6.0 gL(-1) total ABE. Increasing the feed storage temperature to 37 degrees C improved ABE production to 7.2 gL(-1) suggesting that retrogradation was occurring more rapidly at 19 degrees C. In both these cases the fermentation drifted toward acid production after approximately 260 h, consistent with the retrogradation of starch overtime. The use of soluble starch, which is less prone to retrogradation, resulted in the production of 9.9 gL(-1) ABE at 37 degrees C feed storage temperature, as compared to 7.2 gL(-1) ABE when cornstarch was used. It should be noted that gelatinized starch retrogradation takes place after sterilization and prior to use of the feed medium, and does not occur during long-term storage of the raw corn material in the months leading up to processing. The degree of hydrolysis of gelatinized starch decreased from 68.8 to 56.2% in 3 days when stored at 37 degrees C. Soluble starch which does not retrograde demonstrated no change in the degree of hydrolysis.

Butanol production from concentrated lactose/whey permeate: Use of pervaporation membrane to recover and concentrate product

USDA-ARS?s Scientific Manuscript database

In these studies butanol (acetone butanol ethanol, or ABE) was produced from concentrated lactose/whey permeate containing 211 gL-1 lactose. Fermentation of such a highly concentrated lactose solution was possible due to simultaneous product removal using a pervaporation membrane. In this system a p...

Agricultural residues and energy crops as potentially economical and novel substrates for microbial production of butanol (a biofuel)

USDA-ARS?s Scientific Manuscript database

This review describes production of acetone butanol ethanol (ABE) from a variety of agricultural residues and energy crops employing biochemical or fermentation processes. A number of organisms are available for this bioconversion including Clostridium beijerinckii P260, C. beijerinckii BA101, C. a...

Efficient acetone-butanol-ethanol production by Clostridium beijerinckii from sugar beet pulp.

PubMed

Bellido, Carolina; Infante, Celia; Coca, Mónica; González-Benito, Gerardo; Lucas, Susana; García-Cubero, María Teresa

2015-08-01

Sugar beet pulp (SBP) has been investigated as a promising feedstock for ABE fermentation by Clostridium beijerinckii. Although lignin content in SBP is low, a pretreatment is needed to enhance enzymatic hydrolysis and fermentation yields. Autohydrolysis at pH 4 has been selected as the best pretreatment for SBP in terms of sugars release and acetone and butanol production. The best overall sugars release yields from raw SBP ranged from 66.2% to 70.6% for this pretreatment. The highest ABE yield achieved was 0.4g/g (5.1g/L of acetone and 6.6g/L butanol) and 143.2g ABE/kg SBP (62.3g acetone and 80.9g butanol) were obtained when pretreated SBP was enzymatically hydrolyzed at 7.5% (w/w) solid loading. Higher solid loadings (10%) offered higher acetone and butanol titers (5.8g/L of acetone and 7.8g/L butanol). All the experiments were carried out under not-controlling pH conditions reaching about 5.3 in the final samples. Copyright © 2015 Elsevier Ltd. All rights reserved.

Allopurinol-mediated lignocellulose-derived microbial inhibitor tolerance by Clostridium beijerinckii during acetone-butanol-ethanol (ABE) fermentation.

PubMed

Ujor, Victor; Agu, Chidozie Victor; Gopalan, Venkat; Ezeji, Thaddeus Chukwuemeka

2015-04-01

In addition to glucans, xylans, and arabinans, lignocellulosic biomass hydrolysates contain significant levels of nonsugar components that are toxic to the microbes that are typically used to convert biomass to biofuels and chemicals. To enhance the tolerance of acetone-butanol-ethanol (ABE)-generating Clostridium beijerinckii NCIMB 8052 to these lignocellulose-derived microbial inhibitory compounds (LDMICs; e.g., furfural), we have been examining different metabolic perturbation strategies to increase the cellular reductant pools and thereby facilitate detoxification of LDMICs. As part of these efforts, we evaluated the effect of allopurinol, an inhibitor of NAD(P)H-generating xanthine dehydrogenase (XDH), on C. beijerinckii grown in furfural-supplemented medium and found that it unexpectedly increased the rate of detoxification of furfural by 1.4-fold and promoted growth, butanol, and ABE production by 1.2-, 2.5-, and 2-fold, respectively. Since NAD(P)H/NAD(P)(+) levels in C. beijerinckii were largely unchanged upon allopurinol treatment, we postulated and validated a possible basis in DNA repair to account for the solventogenic gains with allopurinol. Following the observation that supplementation of allopurinol in the C. beijerinckii growth media mitigates the toxic effects of nalidixic acid, a DNA-damaging antibiotic, we found that allopurinol elicited 2.4- and 6.7-fold increase in the messenger RNA (mRNA) levels of xanthine and hypoxanthine phosphoribosyltransferases, key purine-salvage enzymes. Consistent with this finding, addition of inosine (a precursor of hypoxanthine) and xanthine led to 1.4- and 1.7-fold increase in butanol production in furfural-challenged cultures of C. beijerinckii. Taken together, our results provide a purine salvage-based rationale for the unanticipated effect of allopurinol in improving furfural tolerance of the ABE-fermenting C. beijerinckii.

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

PubMed

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

2016-10-01

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

Enhanced solvent production by metabolic engineering of a twin-clostridial consortium.

PubMed

Wen, Zhiqiang; Minton, Nigel P; Zhang, Ying; Li, Qi; Liu, Jinle; Jiang, Yu; Yang, Sheng

2017-01-01

The efficient fermentative production of solvents (acetone, n-butanol, and ethanol) from a lignocellulosic feedstock using a single process microorganism has yet to be demonstrated. Herein, we developed a consolidated bioprocessing (CBP) based on a twin-clostridial consortium composed of Clostridium cellulovorans and Clostridium beijerinckii capable of producing cellulosic butanol from alkali-extracted, deshelled corn cobs (AECC). To accomplish this a genetic system was developed for C. cellulovorans and used to knock out the genes encoding acetate kinase (Clocel_1892) and lactate dehydrogenase (Clocel_1533), and to overexpress the gene encoding butyrate kinase (Clocel_3674), thereby pulling carbon flux towards butyrate production. In parallel, to enhance ethanol production, the expression of a putative hydrogenase gene (Clocel_2243) was down-regulated using CRISPR interference (CRISPRi). Simultaneously, genes involved in organic acids reassimilation (ctfAB, cbei_3833/3834) and pentose utilization (xylR, cbei_2385 and xylT, cbei_0109) were engineered in C. beijerinckii to enhance solvent production. The engineered twin-clostridia consortium was shown to decompose 83.2g/L of AECC and produce 22.1g/L of solvents (4.25g/L acetone, 11.5g/L butanol and 6.37g/L ethanol). This titer of acetone-butanol-ethanol (ABE) approximates to that achieved from a starchy feedstock. The developed twin-clostridial consortium serves as a promising platform for ABE fermentation from lignocellulose by CBP. Copyright © 2016 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

Sustainable biobutanol production from pineapple waste by using Clostridium acetobutylicum B 527: Drying kinetics study.

PubMed

Khedkar, Manisha A; Nimbalkar, Pranhita R; Gaikwad, Shashank G; Chavan, Prakash V; Bankar, Sandip B

2017-02-01

Present investigation explores the use of pineapple peel, a food industry waste, for acetone-butanol-ethanol (ABE) production using Clostridium acetobutylicum B 527. Proximate analysis of pineapple peel shows that it contains 35% cellulose, 19% hemicellulose, and 16% lignin on dry basis. Drying experiments on pineapple peel waste were carried out in the temperature range of 60-120°C and experimental drying data was modeled using moisture diffusion control model to study its effect on ABE production. The production of ABE was further accomplished via acid hydrolysis, detoxification, and fermentation process. Maximum total sugar release obtained by using acid hydrolysis was 97g/L with 95-97% and 10-50% removal of phenolics and acetic acid, respectively during detoxification process. The maximum ABE titer obtained was 5.23g/L with 55.6% substrate consumption when samples dried at 120°C were used as a substrate (after detoxification). Copyright © 2016 Elsevier Ltd. All rights reserved.

Synergistic dark and photo-fermentation continuous system for hydrogen production from molasses by Clostridium acetobutylicum ATCC 824 and Rhodobacter capsulatus DSM 1710.

PubMed

Morsy, Fatthy Mohamed

2017-04-01

This study investigated synergistic dark and photo-fermentation using continuous fermentation system (CFS). The system relies on connecting several fermenters from bottom of one to top culture level of the next in a manner that allows for delaying movement of the substrate and thus for its full consumption. While H 2 was collected, CFS allowed for moving liquid byproducts toward the outlet and hence continuous productivity. CFS could be efficiently used for: (1) Continuous dark and photo-fermentation H 2 production by Clostridium acetobutylicum and Rhodobacter capsulatus producing 5.65moleH 2 mole -1 hexose; (2) Continuous dark-fermentation synergistic H 2 , acetone, butanol and ethanol (ABE) production by C. acetobutylicum which produced per mole hexose, 2.43mol H 2 along with 73.08g ABE (3) Continuous H 2 and methane production by C. acetobutylicum and bacterial sludge producing, per mole hexose, 1.64mol pure H 2 and 2.56mol CH 4 mixed with 0.37mol H 2 ·The hydraulic retention time (HRT) for whole system was short where organic acids produced in dark-fermentation in first fermenter were synergistically utilized for H 2 production by R. capsulatus in subsequent fermenters. CFS is suitable for fast-digestible sugars but not lignocelluloses or other hard-digestible organics, requiring prolonged HRT, unless such polymeric organics were hydrolyzed prior to fermentation. Copyright © 2017 Elsevier B.V. All rights reserved.

Adsorption of water and butanol in silicalite-1 film studied with in situ attenuated total reflectance-Fourier transform infrared spectroscopy.

PubMed

Farzaneh, Amirfarrokh; Zhou, Ming; Potapova, Elisaveta; Bacsik, Zoltán; Ohlin, Lindsay; Holmgren, Allan; Hedlund, Jonas; Grahn, Mattias

2015-05-05

Biobutanol produced by, e.g., acetone-butanol-ethanol (ABE) fermentation is a promising alternative to petroleum-based chemicals as, e.g., solvent and fuel. Recovery of butanol from dilute fermentation broths by hydrophobic membranes and adsorbents has been identified as a promising route. In this work, the adsorption of water and butanol vapor in a silicalite-1 film was studied using in situ attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy to better understand the adsorption properties of silicalite-1 membranes and adsorbents. Single-component adsorption isotherms were determined in the temperature range of 35-120 °C, and the Langmuir model was successfully fitted to the experimental data. The adsorption of butanol is very favorable compared to that of water. When the silicalite-1 film was exposed to a butanol/water vapor mixture with 15 mol % butanol (which is the vapor composition of an aqueous solution containing 2 wt % butanol, a typical concentration in an ABE fermentation broth, i.e., the composition of the gas obtained from gas stripping of an ABE broth) at 35 °C, the adsorption selectivity toward butanol was as high as 107. These results confirm that silicalite-1 quite selectively adsorbs hydrocarbons from vapor mixtures. To the best of our knowledge, this is the first comprehensive study on the adsorption of water and butanol in silicalite-1 from vapor phase.

Butanol production in a first-generation Brazilian sugarcane biorefinery: technical aspects and economics of greenfield projects.

PubMed

Mariano, Adriano Pinto; Dias, Marina O S; Junqueira, Tassia L; Cunha, Marcelo P; Bonomi, Antonio; Filho, Rubens Maciel

2013-05-01

The techno-economics of greenfield projects of a first-generation sugarcane biorefinery aimed to produce ethanol, sugar, power, and n-butanol was conducted taking into account different butanol fermentation technologies (regular microorganism and mutant strain with improved butanol yield) and market scenarios (chemicals and automotive fuel). The complete sugarcane biorefinery with the batch acetone-butanol-ethanol (ABE) fermentation process was simulated using Aspen Plus®. The biorefinery was designed to process 2 million tonne sugarcane per year and utilize 25%, 50%, and 25% of the available sugarcane juice to produce sugar, ethanol, and butanol, respectively. The investment on a biorefinery with butanol production showed to be more attractive [14.8% IRR, P(IRR>12%)=0.99] than the conventional 50:50 (ethanol:sugar) annexed plant [13.3% IRR, P(IRR>12%)=0.80] only in the case butanol is produced by an improved microorganism and traded as a chemical. Copyright © 2012 Elsevier Ltd. All rights reserved.

Applied in situ product recovery in ABE fermentation

PubMed Central

Lalander, Carl‐Axel; Lee, Jonathan G. M.; Davies, E. Timothy; Harvey, Adam P.

2017-01-01

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 PMID:28188696

New Insight into Sugarcane Industry Waste Utilization (Press Mud) for Cleaner Biobutanol Production by Using C. acetobutylicum NRRL B-527.

PubMed

Nimbalkar, Pranhita R; Khedkar, Manisha A; Gaikwad, Shashank G; Chavan, Prakash V; Bankar, Sandip B

2017-11-01

In the present study, press mud, a sugar industry waste, was explored for biobutanol production to strengthen agricultural economy. The fermentative production of biobutanol was investigated via series of steps, viz. characterization, drying, acid hydrolysis, detoxification, and fermentation. Press mud contains an adequate amount of cellulose (22.3%) and hemicellulose (21.67%) on dry basis, and hence, it can be utilized for further acetone-butanol-ethanol (ABE) production. Drying experiments were conducted in the temperature range of 60-120 °C to circumvent microbial spoilage and enhance storability of press mud. Furthermore, acidic pretreatment variables, viz. sulfuric acid concentration, solid to liquid ratio, and time, were optimized using response surface methodology. The corresponding values were found to be 1.5% (v/v), 1:5 g/mL, and 15 min, respectively. In addition, detoxification studies were also conducted using activated charcoal, which removed almost 93-97% phenolics and around 98% furans, which are toxic to microorganisms during fermentation. Finally, the batch fermentation of detoxified press mud slurry (the sample dried at 100 °C and pretreated) using Clostridium acetobutylicum NRRL B-527 resulted in a higher butanol production of 4.43 g/L with a total ABE of 6.69 g/L.

A novel process for direct production of acetone-butanol-ethanol from native starches using granular starch hydrolyzing enzyme by Clostridium saccharoperbutylacetonicum N1-4.

PubMed

Thang, Vu Hong; Kobayashi, Genta

2014-02-01

In this work, a new approach for acetone-butanol-ethanol (ABE) production has been proposed. Direct fermentation of native starches (uncooked process) was investigated by using granular starch hydrolyzing enzyme (GSHE) and Clostridium saccharoperbutylacetonicum N1-4. Even the process was carried out under suboptimal condition for activity of GSHE, the production of ABE was similar with that observed in conventional process or cooked process in terms of final solvent concentration (21.3 ± 0.4 to 22.4 ± 0.4 g/L), butanol concentration (17.5 ± 0.4 to 17.8 ± 0.3 g/L) and butanol yield (0.33 to 0.37 g/g). The production of solvents was significantly dependent on the source of starches. Among investigated starches, corn starch was more susceptible to GSHE while cassava starch was the most resistant to this enzyme. Fermentation using native corn starch resulted in the solvent productivity of 0.47 g/L h, which was about 15 % higher than that achieved in cooked process. On the contrary, uncooked process using cassava and wheat starch resulted in the solvent productivity of 0.30 and 0.37 g/L h, which were respectively about 30 % lower than those obtained in cooked process. No contamination was observed during all trials even fermentation media were prepared without sterilization. During the fermentation using native starches, no formation of foam is observed. This uncooked process does not require cooking starchy material; therefore, the thermal energy consumption for solvent production would remarkably be reduced in comparison with cooked process.

Acetone-butanol-ethanol (ABE) fermentation in an immobilized cell trickle bed reactor.

PubMed

Park, C H; Okos, M R; Wankat, P C

1989-06-05

Acetone-butanol-ethanol (ABE) fermentation was successfully carried out in an immobilized cell trickle bed reactor. The reactor was composed of two serial columns packed with Clostridium acetobutylicum ATCC 824 entrapped on the surface of natural sponge segments at a cell loading in the range of 2.03-5.56 g dry cells/g sponge. The average cell loading was 3.58 g dry cells/g sponge. Batch experiments indicated that a critical pH above 4.2 is necessary for the initiation of cell growth. One of the media used during continuous experiments consisted of a salt mixture alone and the other a nutrient medium containing a salt mixture with yeast extract and peptone. Effluent pH was controlled by supplying various fractions of the two different types of media. A nutrient medium fraction above 0.6 was crucial for successful fermentation in a trickle bed reactor. The nutrient medium fraction is the ratio of the volume of the nutrient medium to the total volume of nutrient plus salt medium. Supplying nutrient medium to both columns continuously was an effective way to meet both pH and nutrient requirement. A 257-mL reactor could ferment 45 g/L glucose from an initial concentration of 60 g/L glucose at a rate of 70 mL/h. Butanol, acetone, and ethanol concentrations were 8.82, 5.22, and 1.45 g/L, respectively, with a butanol and total solvent yield of 19.4 and 34.1 wt %. Solvent productivity in an immobilized cell trickle bed reactor was 4.2 g/L h, which was 10 times higher than that obtained in a batch fermentation using free cells and 2.76 times higher than that of an immobilized CSTR. If the nutrient medium fraction was below 0.6 and the pH was below 4.2, the system degenerated. Oxygen also contributed to the system degeneration. Upon degeneration, glucose consumption and solvent yield decreased to 30.9 g/L and 23.0 wt %, respectively. The yield of total liquid product (40.0 wt %) and butanol selectivity (60.0 wt %) remained almost constant. Once the cells were degenerated, they could not be recovered.

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.

Improving Butanol Fermentation To Enter the Advanced Biofuel Market

PubMed Central

Tracy, Bryan P.

2012-01-01

ABSTRACT 1-Butanol is a large-volume, intermediate chemical with favorable physical and chemical properties for blending with or directly substituting for gasoline. The per-volume value of butanol, as a chemical, is sufficient for investing into the recommercialization of the classical acetone-butanol-ethanol (ABE) (E. M. Green, Curr. Opin. Biotechnol. 22:337–343, 2011) fermentation process. Furthermore, with modest improvements in three areas of the ABE process, operating costs can be sufficiently decreased to make butanol an economically viable advanced biofuel. The three areas of greatest interest are (i) maximizing yields of butanol on any particular substrate, (ii) expanding substrate utilization capabilities of the host microorganism, and (iii) reducing the energy consumption of the overall production process, in particular the separation and purification operations. In their study in the September/October 2012 issue of mBio, Jang et al. [mBio 3(5):e00314-12, 2012] describe a comprehensive study on driving glucose metabolism in Clostridium acetobutylicum to the production of butanol. Moreover, they execute a metabolic engineering strategy to achieve the highest yet reported yields of butanol on glucose. PMID:23232720

Biobutanol production from brewer's spent grain hydrolysates by Clostridium beijerinckii.

PubMed

Plaza, Pedro E; Gallego-Morales, Luis Javier; Peñuela-Vásquez, Mariana; Lucas, Susana; García-Cubero, M Teresa; Coca, Mónica

2017-11-01

Brewer's spent grain (BSG) is a promising feedstock for ABE fermentation. Sulfuric acid pretreatment of BSG at pH 1, 121°C and different solid loadings (5-15% w/w) was investigated. Enzymatic hydrolysis and ABE fermentation by Clostridium beijerinckii DSM 6422 of non-washed and washed pretreated BSG were performed to compare monosaccharide release and butanol production. Pretreatment at 15% w/w BSG resulted in higher availability of sugars in both the enzymatic hydrolysates and pretreatment liquid, and overall yields of 75gbutanol/kg BSG and 95gABE/kg BSG were obtained. When the enzymatic hydrolysate from the washed pretreated BSG was fermented, butanol (6.0±0.5g/L) and ABE (7.4±1.0g/L) concentrations were lower compared with 7.5±0.6g/L butanol and 10.0±0.8g/L ABE from a control. The fermentation of the liquid released in the pretreatment at 15% w/w resulted in a butanol production of 6.6±0.8g/L with a total ABE of 8.6±1.3g/L after overliming. Copyright © 2017 Elsevier Ltd. All rights reserved.

Biomass, strain engineering, and fermentation processes for butanol production by solventogenic clostridia.

PubMed

Lee, Sang-Hyun; Yun, Eun Ju; Kim, Jungyeon; Lee, Sang Jun; Um, Youngsoon; Kim, Kyoung Heon

2016-10-01

Butanol is considered an attractive biofuel and a commercially important bulk chemical. However, economical production of butanol by solventogenic clostridia, e.g., via fermentative production of acetone-butanol-ethanol (ABE), is hampered by low fermentation performance, mainly as a result of toxicity of butanol to microorganisms and high substrate costs. Recently, sugars from marine macroalgae and syngas were recognized as potent carbon sources in biomass feedstocks that are abundant and do not compete for arable land with edible crops. With the aid of systems metabolic engineering, many researchers have developed clostridial strains with improved performance on fermentation of these substrates. Alternatively, fermentation strategies integrated with butanol recovery processes such as adsorption, gas stripping, liquid-liquid extraction, and pervaporation have been designed to increase the overall titer of butanol and volumetric productivity. Nevertheless, for economically feasible production of butanol, innovative strategies based on recent research should be implemented. This review describes and discusses recent advances in the development of biomass feedstocks, microbial strains, and fermentation processes for butanol production.

Recent advances and state-of-the-art strategies in strain and process engineering for biobutanol production by Clostridium acetobutylicum.

PubMed

Xue, Chuang; Zhao, Jingbo; Chen, Lijie; Yang, Shang-Tian; Bai, Fengwu

Butanol as an advanced biofuel has gained great attention due to its environmental benefits and superior properties compared to ethanol. However, the cost of biobutanol production via conventional acetone-butanol-ethanol (ABE) fermentation by Clostridium acetobutylicum is not economically competitive, which has hampered its industrial application. The strain performance and downstream process greatly impact the economics of biobutanol production. Although various engineered strains with carefully orchestrated metabolic and sporulation-specific pathways have been developed, none of them is ideal for industrial biobutanol production. For further strain improvement, it is necessary to develop advanced genome editing tools and a deep understanding of cellular functioning of genes in metabolic and regulatory pathways. Processes with integrated product recovery can increase fermentation productivity by continuously removing inhibitory products while generating butanol (ABE) in a concentrated solution. In this review, we provide an overview of recent advances in C. acetobutylicum strain engineering and process development focusing on in situ product recovery. With deep understanding of systematic cellular bioinformatics, the exploration of state-of-the-art genome editing tools such as CRISPR-Cas for targeted gene knock-out and knock-in would play a vital role in Clostridium cell engineering for biobutanol production. Developing advanced hybrid separation processes for in situ butanol recovery, which will be discussed with a detailed comparison of advantages and disadvantages of various recovery techniques, is also imperative to the economical development of biobutanol. Copyright © 2017 Elsevier Inc. All rights reserved.

Improvement of the butanol production selectivity and butanol to acetone ratio (B:A) by addition of electron carriers in the batch culture of a new local isolate of Clostridium acetobutylicum YM1.

PubMed

Nasser Al-Shorgani, Najeeb Kaid; Kalil, Mohd Sahaid; Wan Yusoff, Wan Mohtar; Shukor, Hafiza; Hamid, Aidil Abdul

2015-12-01

Improvement in the butanol production selectivity or enhanced butanol:acetone ratio (B:A) is desirable in acetone-butanol-ethanol (ABE) fermentation by Clostridium strains. In this study, artificial electron carriers were added to the fermentation medium of a new isolate of Clostridium acetobutylicum YM1 in order to improve the butanol yield and B:A ratio. The results revealed that medium supplementation with electron carriers changed the metabolism flux of electron and carbon in ABE fermentation by YM1. A decrease in acetone production, which subsequently improved the B:A ratio, was observed. Further improvement in the butanol production and B:A ratios were obtained when the fermentation medium was supplemented with butyric acid. The maximum butanol production (18.20 ± 1.38 g/L) was gained when a combination of methyl red and butyric acid was added. Although the addition of benzyl viologen (0.1 mM) and butyric acid resulted in high a B:A ratio of 16:1 (800% increment compared with the conventional 2:1 ratio), the addition of benzyl viologen to the culture after 4 h resulted in the production of 18.05 g/L butanol. Manipulating the metabolic flux to butanol through the addition of electron carriers could become an alternative strategy to achieve higher butanol productivity and improve the B:A ratio. Copyright © 2015 Elsevier Ltd. All rights reserved.

Production of acetone butanol ethanol (ABE) by a hyper-producing mutant strain of Clostridium beijerinckii BA101 and recovery by pervaporation

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

Qureshi, N.; Blaschek, H.P.

1999-07-01

A silicone membrane was used to study butanol separation from model butanol solutions and fermentation broth. Depending upon the butanol feed concentration in the model solution and pervaporation conditions, butanol selectivities of 20.88--68.32 and flux values of 158.7--215.4 g m{sup {minus}2} h{sup {minus}1} were achieved. Higher flux values were obtained at higher butanol concentrations using air as sweep gas. In an integrated process of butanol fermentation--recovery, solvent productivities were improved to 200% of the control batch fermentation productivities. In a batch reactor the hyper-butanol-producing mutant strain C. beijerinckii BA101 utilized 57.3 g/L glucose and produced 24.2 g/L total solvents, whilemore » in the integrated process it produced 51.5 g/L (culture volume) total solvents. Concentrated glucose medium was also fermented. The C. beijerinckii BA101 mutant strain was not negatively affected by the pervaporative conditions. In the integrated experiment, acids were not produced. With the active fermentation broth, butanol selectivity was reduced by a factor of 2--3. However, the membrane flux was not affected by the active fermentation broth. The butanol permeate concentration ranged from 26.4 to 95.4 g/L, depending upon butanol concentration in the fermentation broth. Since the permeate of most membranes contains acetone, butanol, and ethanol, it is suggested that distillation be used for further purification.« less

Unravelling the influence of carbon dioxide on the adsorptive recovery of butanol from fermentation broth using ITQ-29 and ZIF-8.

PubMed

Martin-Calvo, Ana; Van der Perre, Stijn; Claessens, Benjamin; Calero, Sofia; Denayer, Joeri F M

2018-04-18

The vapor phase adsorption of butanol from ABE fermentation at the head space of the fermenter is an interesting route for the efficient recovery of biobutanol. The presence of gases such as carbon dioxide that are produced during the fermentation process causes a stripping of valuable compounds from the aqueous into the vapor phase. This work studies the effect of the presence of carbon dioxide on the adsorption of butanol at a molecular level. With this aim in mind Monte Carlo simulations were employed to study the adsorption of mixtures containing carbon dioxide, butanol and ethanol. Molecular models for butanol and ethanol that reproduce experimental properties of the molecules such as polarity, vapor-liquid coexistence or liquid density have been developed. Pure component isotherms and heats of adsorption have been computed and compared to experimental data to check the accuracy of the interacting parameters. Adsorption of butanol/ethanol mixtures has been studied in absence and presence of CO2 on two representative materials, a pure silica LTA zeolite and a hydrophobic metal-organic framework ZIF-8. To get a better understanding of the molecular mechanism that governs the adsorption of the targeted mixture in the selected materials, the distribution of the molecules inside the structures was analyzed. The combination of these features allows obtaining a deeper understanding of the process and to identify the role of carbon dioxide in the butanol purification process.

Effect of pretreatment on simultaneous saccharification and fermentation of hardwood into acetone/butanol

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

Shah, M.M.; Song, S.K.; Lee, Y.Y.

1991-12-31

The effectiveness of pretreatments on hardwood substrate was investigated in connection with its subsequent conversion by simultaneous saccharification and fermentation (SSF), using Clostridium acetobutylicum. The main objectives of the pretreatment were to achieve efficient separation of lignin from carbohydrates, and to obtain maximum sugar yield on enzymatic hydrolysis of pretreated wood. Two methods have given promising results: (1) supercritical CO{sub 2}-SO{sub 2} treatment, and (2) monoethanolamine (MEA) treatment. The MEA pretreatment removed above 90% of hardwood lignin while retaining 83% of carbohydrates. With CO{sub 2}-SO{sub 2} pretreatment, the degree of lignin separation was lower. Under the scheme of SSF, themore » pretreated hardwood was converted to acetone, butanol, and ethanol (ABE) via single stage processing by cellulose enzyme system and C. acetobutylicum cells. The product yield in the process was such that 15 g of ABE/100 g of dry aspen wood was produced. In the overall process of SSF, the enzymatic hydrolysis was found to be the rate-limiting step. The ability of C. acetobutylicum to metabolize various 6-carbon and 5-carbon sugars resulted in efficient utilization of all available sugars from hardwood.« less

Enhancing Butanol Production under the Stress Environments of Co-Culturing Clostridium acetobutylicum/Saccharomyces cerevisiae Integrated with Exogenous Butyrate Addition

PubMed Central

Luo, Hongzhen; Ge, Laibing; Zhang, Jingshu; Zhao, Yanli; Ding, Jian; Li, Zhigang; He, Zhenni; Chen, Rui; Shi, Zhongping

2015-01-01

In this study, an efficient acetone-butanol-ethanol (ABE) fermentation strategy integrating Clostridium acetobutylicum/Saccharomyces cerevisiae co-culturing system with exogenous butyrate addition, was proposed and experimentally conducted. In solventogenic phase, by adding 0.2 g-DCW/L-broth viable S. cerevisiae cells and 4.0 g/L-broth concentrated butyrate solution into C. acetobutylicum culture broth, final butanol concentration and butanol/acetone ratio in a 7 L anaerobic fermentor reached the highest levels of 15.74 g/L and 2.83 respectively, with the increments of 35% and 43% as compared with those of control. Theoretical and experimental analysis revealed that, the proposed strategy could, 1) extensively induce secretion of amino acids particularly lysine, which are favorable for both C. acetobutylicum survival and butanol synthesis under high butanol concentration environment; 2) enhance the utilization ability of C. acetobutylicum on glucose and over-produce intracellular NADH for butanol synthesis in C. acetobutylicum metabolism simultaneously; 3) direct most of extra consumed glucose into butanol synthesis route. The synergetic actions of effective amino acids assimilation, high rates of substrate consumption and NADH regeneration yielded highest butanol concentration and butanol ratio in C. acetobutylicum under this stress environment. The proposed method supplies an alternative way to improve ABE fermentation performance by traditional fermentation technology. PMID:26489085

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

PubMed

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

2016-01-14

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

Capturing the response of Clostridium acetobutylicum to chemical stressors using a regulated genome-scale metabolic model

DOE PAGES

Dash, Satyakam; Mueller, Thomas J.; Venkataramanan, Keerthi P.; ...

2014-10-14

Clostridia are anaerobic Gram-positive Firmicutes containing broad and flexible systems for substrate utilization, which have been used successfully to produce a range of industrial compounds. Clostridium acetobutylicum has been used to produce butanol on an industrial scale through acetone-butanol-ethanol (ABE) fermentation. A genome-scale metabolic (GSM) model is a powerful tool for understanding the metabolic capacities of an organism and developing metabolic engineering strategies for strain development. The integration of stress related specific transcriptomics information with the GSM model provides opportunities for elucidating the focal points of regulation.

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

PubMed

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

2016-09-01

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

A carbon nanotube filled polydimethylsiloxane hybrid membrane for enhanced butanol recovery

PubMed Central

Xue, Chuang; Du, Guang-Qing; Chen, Li-Jie; Ren, Jian-Gang; Sun, Jian-Xin; Bai, Feng-Wu; Yang, Shang-Tian

2014-01-01

The carbon nanotubes (CNTs) filled polydimethylsiloxane (PDMS) hybrid membrane was fabricated to evaluate its potential for butanol recovery from acetone-butanol-ethanol (ABE) fermentation broth. Compared with the homogeneous PDMS membrane, the CNTs filled into the PDMS membrane were beneficial for the improvement of butanol recovery in butanol flux and separation factor. The CNTs acting as sorption-active sites with super hydrophobicity could give an alternative route for mass transport through the inner tubes or along the smooth surface. The maximum total flux and butanol separation factor reached up to 244.3 g/m2·h and 32.9, respectively, when the PDMS membrane filled with 10 wt% CNTs was used to separate butanol from the butanol/water solution at 80°C. In addition, the butanol flux and separation factor increased dramatically as temperature increased from 30°C to 80°C in feed solution since the higher temperature produced more free volumes in polymer chains to facilitate butanol permeation. A similar increase was also observed when butanol titer in solution increased from 10 g/L to 25 g/L. Overall, the CNTs/PDMS hybrid membrane with higher butanol flux and selectivity should have good potential for pervaporation separation of butanol from ABE fermentation broth. PMID:25081019

Effects of Averrhoa bilimbi leaf extract on blood glucose and lipids in streptozotocin-diabetic rats.

PubMed

Pushparaj, P; Tan, C H; Tan, B K

2000-09-01

The present study was designed to investigate the hypoglycemic and hypolipidemic activities of an ethanolic extract of Averrhoa bilimbi Linn. leaves (Oxalidaceae, Common name: Bilimbi) in streptozotocin (STZ)-diabetic rats. The optimal hypoglycemic dose (125 mg kg(-1)) was determined by performing the oral glucose tolerance test (OGTT) in both normal and STZ-diabetic rats. To investigate the effect of repeated administration of an ethanolic extract of Averrhoa bilimbi (ABe) leaves, diabetic rats were treated with vehicle (distilled water), ABe (125 mg kg(-1)) or metformin (500 mg kg(-1)) twice a day for 2 weeks. Like metformin, ABe significantly lowered blood glucose by 50% and blood triglyceride by 130% when compared with the vehicle. ABe also significantly increased the HDL-cholesterol concentrations by 60% compared with the vehicle. ABe thus significantly increased the anti-atherogenic index and HDL-cholesterol/total cholesterol ratio. However, like metformin, ABe did not affect total cholesterol and LDL-cholesterol concentrations, but significantly reduced the kidney lipid peroxidation level. These data show that ABe has hypoglycemic, hypotriglyceridemic, anti-lipid peroxidative and anti-atherogenic properties in STZ-diabetic rats.

Fermentation and genomic analysis of acetone-uncoupled butanol production by Clostridium tetanomorphum.

PubMed

Gong, Fuyu; Bao, Guanhui; Zhao, Chunhua; Zhang, Yanping; Li, Yin; Dong, Hongjun

2016-02-01

In typical acetone-butanol-ethanol (ABE) fermentation, acetone is the main by-product (50 % of butanol mass) of butanol production, resulting in a low yield of butanol. It is known that some Clostridium tetanomorphum strains are able to produce butanol without acetone in nature. Here, we described that C. tetanomorphum strain DSM665 can produce 4.16 g/L butanol and 4.98 g/L ethanol at pH 6.0, and 9.81 g/L butanol and 1.01 g/L ethanol when adding 1 mM methyl viologen. Butyrate and acetate could be reassimilated and no acetone was produced. Further analysis indicated that the activity of the acetate/butyrate:acetoacetyl-CoA transferase responsible for acetone production is lost in C. tetanomorphum DSM665. The genome of C. tetanomorphum DSM665 was sequenced and deposited in DDBJ, EMBL, and GenBank under the accession no. APJS00000000. Sequence analysis indicated that there are no typical genes (ctfA/B and adc) that are typically parts of an acetone synthesis pathway in C. tetanomorphum DSM665. This work provides new insights in the mechanism of clostridial butanol production and should prove useful for the design of a high-butanol-producing strain.

Strategies for improved isopropanol-butanol production by a Clostridium strain from glucose and hemicellulose through consolidated bioprocessing.

PubMed

Xin, Fengxue; Chen, Tianpeng; Jiang, Yujiang; Dong, Weiliang; Zhang, Wenming; Zhang, Min; Wu, Hao; Ma, Jiangfeng; Jiang, Min

2017-01-01

High cost of traditional substrates and formation of by-products (such as acetone and ethanol) in acetone-butanol-ethanol (ABE) fermentation hindered the large-scale production of biobutanol. Here, we comprehensively characterized a newly isolated solventogenic and xylanolytic Clostridium species, which could produce butanol at a high ratio with elimination of ethanol and conversion of acetone to more value-added product, isopropanol. Ultimately, direct butanol production from hemicellulose was achieved with efficient expression of indigenous xylanase by the novel strain via consolidated bioprocessing. A novel wild-type Clostridium sp. strain NJP7 was isolated and characterized in this study, which was capable of fermenting monosaccharides, e.g., glucose into butanol via a fermentative acetone-isopropanol-butanol pathway. With enhancement of buffering capacity and alcohol dehydrogenase activities, butanol and isopropanol titer by Clostridium sp. strain NJP7 was improved to 12.21 and 1.92 g/L, respectively, and solvent productivity could be enhanced to 0.44 g/L/h. Furthermore, with in situ extraction with biodiesel, the amount of butanol and isopropanol was finally improved to 25.58 and 5.25 g/L in the fed-batch mode. Meanwhile, Clostridium sp. strain NJP7 shows capability of direct isopropanol-butanol production from hemicelluloses with expression of indigenous xylanase. 2.06 g/L of butanol and 0.54 g/L of isopropanol were finally achieved through the temperature-shift simultaneous saccharification and fermentation, representing the highest butanol production directly from hemicellulose. The co-production of isopropanol with butanol by the newly isolated Clostridium sp. strain NJP7 would add on the economical values for butanol fermentation. Furthermore, the high isopropanol-butanol production with in situ extraction would also greatly enhance the economic feasibility for fermentative production of butanol-isopropanol in large scale. Meanwhile, its direct production of butanol-isopropanol from polysaccharides, hemicellulose through secretion of indigenous thermostable xylanase, shows great potential using lignocellulosic wastes for biofuel production.

Life-cycle assessment of corn-based butanol as a potential transportation fuel.

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

Wu, M.; Wang, M.; Liu, J.

2007-12-31

Butanol produced from bio-sources (such as corn) could have attractive properties as a transportation fuel. Production of butanol through a fermentation process called acetone-butanol-ethanol (ABE) has been the focus of increasing research and development efforts. Advances in ABE process development in recent years have led to drastic increases in ABE productivity and yields, making butanol production worthy of evaluation for use in motor vehicles. Consequently, chemical/fuel industries have announced their intention to produce butanol from bio-based materials. The purpose of this study is to estimate the potential life-cycle energy and emission effects associated with using bio-butanol as a transportation fuel.more » The study employs a well-to-wheels analysis tool--the Greenhouse Gases, Regulated Emissions and Energy Use in Transportation (GREET) model developed at Argonne National Laboratory--and the Aspen Plus{reg_sign} model developed by AspenTech. The study describes the butanol production from corn, including grain processing, fermentation, gas stripping, distillation, and adsorption for products separation. The Aspen{reg_sign} results that we obtained for the corn-to-butanol production process provide the basis for GREET modeling to estimate life-cycle energy use and greenhouse gas emissions. The GREET model was expanded to simulate the bio-butanol life cycle, from agricultural chemical production to butanol use in motor vehicles. We then compared the results for bio-butanol with those of conventional gasoline. We also analyzed the bio-acetone that is coproduced with bio-butanol as an alternative to petroleum-based acetone. Our study shows that, while the use of corn-based butanol achieves energy benefits and reduces greenhouse gas emissions, the results are affected by the methods used to treat the acetone that is co-produced in butanol plants.« less

Direct α-alkylation of ketones with alcohols in water.

PubMed

Xu, Guoqiang; Li, Qiong; Feng, Jiange; Liu, Qiang; Zhang, Zuojun; Wang, Xicheng; Zhang, Xiaoyun; Mu, Xindong

2014-01-01

The direct α-alkylation of ketones with alcohols has emerged as a new green protocol to construct C-C bonds with H2 O as the sole byproduct. In this work, a very simple and convenient Pd/C catalytic system for the direct α-alkylation of ketones with primary alcohols in pure water is developed. Based on this catalytic system, aqueous mixtures of dilute acetone, 1-butanol, and ethanol (mimicking ABE fermentation products) can be directly transformed into C5 -C11 or longer-chain ketones and alcohols, which are precursors to fuels. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Acetone-butanol-ethanol production from substandard and surplus dates by Egyptian native Clostridium strains.

PubMed

Abd-Alla, Mohamed Hemida; Zohri, Abdel-Naser Ahmed; El-Enany, Abdel-Wahab Elsadek; Ali, Shimaa Mohamed

2015-04-01

One hundred and seven mesophilic isolates of Clostridium were isolated from agricultural soils cultivated with different plants in Assuit Governorate, Egypt. Eighty isolates (out of 107) showed the ability to produce ABE (Acetone, butanol and ethanol) on T6 medium ranging from 0.036 to 31.89 g/L. The highest numbers of ABE producing isolates were obtained from soil samples of potato contributing 27 isolates, followed by 18 isolates from wheat and 10 isolates from onion. On the other hand, there were three native isolates that produced ABE more than those produced by the reference isolate Clostridium acetobutylicum ATCC 824 (11.543 g/L). The three isolates were identified based on phenotypic and gene encoding 16S rRNA as Clostridium beijerinckii ASU10 (KF372577), Clostridium chauvoei ASU55 (KF372580) and Clostridium roseum ASU58 (KF372581). The highest ABE level from substandard and surplus dates was produced by C. beijerinckii ASU10 (24.07 g/L) comprising butanol 67.15% (16.16 g/L), acetone 30.73% (7.4 g/L) and ethanol 2.12% (0.51 g/L), while C. roseum ASU58 and C. chauvoei ASU55 produced ABE contributing 20.20 and 13.79 g/L, respectively. ABE production by C. acetobutylicum ATCC 824 was 15.01 g/L. This study proved that the native strains C. beijerinckii ASU10 and C. roseum ASU58 have high competitive efficacy on ABE production from economical substrate as substandard and surplus date fruits. Additionally, using this substrate without any nutritional components is considered to be a commercial substrate for desired ABE production. Copyright © 2014 Elsevier Ltd. All rights reserved.

Development of a High-Efficiency Transformation Method and Implementation of Rational Metabolic Engineering for the Industrial Butanol Hyperproducer Clostridium saccharoperbutylacetonicum Strain N1-4.

PubMed

Herman, Nicolaus A; Li, Jeffrey; Bedi, Ripika; Turchi, Barbara; Liu, Xiaoji; Miller, Michael J; Zhang, Wenjun

2017-01-15

While a majority of academic studies concerning acetone, butanol, and ethanol (ABE) production by Clostridium have focused on Clostridium acetobutylicum, other members of this genus have proven to be effective industrial workhorses despite the inability to perform genetic manipulations on many of these strains. To further improve the industrial performance of these strains in areas such as substrate usage, solvent production, and end product versatility, transformation methods and genetic tools are needed to overcome the genetic intractability displayed by these species. In this study, we present the development of a high-efficiency transformation method for the industrial butanol hyperproducer Clostridium saccharoperbutylacetonicum strain N1-4 (HMT) ATCC 27021. Following initial failures, we found that the key to creating a successful transformation method was the identification of three distinct colony morphologies (types S, R, and I), which displayed significant differences in transformability. Working with the readily transformable type I cells (transformation efficiency, 1.1 × 10 6 CFU/μg DNA), we performed targeted gene deletions in C. saccharoperbutylacetonicum N1-4 using a homologous recombination-mediated allelic exchange method. Using plasmid-based gene overexpression and targeted knockouts of key genes in the native acetone-butanol-ethanol (ABE) metabolic pathway, we successfully implemented rational metabolic engineering strategies, yielding in the best case an engineered strain (Clostridium saccharoperbutylacetonicum strain N1-4/pWIS13) displaying an 18% increase in butanol titers and 30% increase in total ABE titer (0.35 g ABE/g sucrose) in batch fermentations. Additionally, two engineered strains overexpressing aldehyde/alcohol dehydrogenases (encoded by adh11 and adh5) displayed 8.5- and 11.8-fold increases (respectively) in batch ethanol production. This paper presents the first steps toward advanced genetic engineering of the industrial butanol producer Clostridium saccharoperbutylacetonicum strain N1-4 (HMT). In addition to providing an efficient method for introducing foreign DNA into this species, we demonstrate successful rational engineering for increasing solvent production. Examples of future applications of this work include metabolic engineering for improving desirable industrial traits of this species and heterologous gene expression for expanding the end product profile to include high-value fuels and chemicals. Copyright © 2016 American Society for Microbiology.

Development of a High-Efficiency Transformation Method and Implementation of Rational Metabolic Engineering for the Industrial Butanol Hyperproducer Clostridium saccharoperbutylacetonicum Strain N1-4

PubMed Central

Herman, Nicolaus A.; Li, Jeffrey; Bedi, Ripika; Turchi, Barbara; Liu, Xiaoji

2016-01-01

ABSTRACT While a majority of academic studies concerning acetone, butanol, and ethanol (ABE) production by Clostridium have focused on Clostridium acetobutylicum, other members of this genus have proven to be effective industrial workhorses despite the inability to perform genetic manipulations on many of these strains. To further improve the industrial performance of these strains in areas such as substrate usage, solvent production, and end product versatility, transformation methods and genetic tools are needed to overcome the genetic intractability displayed by these species. In this study, we present the development of a high-efficiency transformation method for the industrial butanol hyperproducer Clostridium saccharoperbutylacetonicum strain N1-4 (HMT) ATCC 27021. Following initial failures, we found that the key to creating a successful transformation method was the identification of three distinct colony morphologies (types S, R, and I), which displayed significant differences in transformability. Working with the readily transformable type I cells (transformation efficiency, 1.1 × 106 CFU/μg DNA), we performed targeted gene deletions in C. saccharoperbutylacetonicum N1-4 using a homologous recombination-mediated allelic exchange method. Using plasmid-based gene overexpression and targeted knockouts of key genes in the native acetone-butanol-ethanol (ABE) metabolic pathway, we successfully implemented rational metabolic engineering strategies, yielding in the best case an engineered strain (Clostridium saccharoperbutylacetonicum strain N1-4/pWIS13) displaying an 18% increase in butanol titers and 30% increase in total ABE titer (0.35 g ABE/g sucrose) in batch fermentations. Additionally, two engineered strains overexpressing aldehyde/alcohol dehydrogenases (encoded by adh11 and adh5) displayed 8.5- and 11.8-fold increases (respectively) in batch ethanol production. IMPORTANCE This paper presents the first steps toward advanced genetic engineering of the industrial butanol producer Clostridium saccharoperbutylacetonicum strain N1-4 (HMT). In addition to providing an efficient method for introducing foreign DNA into this species, we demonstrate successful rational engineering for increasing solvent production. Examples of future applications of this work include metabolic engineering for improving desirable industrial traits of this species and heterologous gene expression for expanding the end product profile to include high-value fuels and chemicals. PMID:27836845

Butanol formation from gaseous substrates.

PubMed

Dürre, Peter

2016-03-01

Mostly, butanol is formed as a product by saccharolytic anaerobes, employing the so-called ABE fermentation (for acetone-butanol-ethanol). However, this alcohol can also be produced from gaseous substrates such as syn(thesis) gas (major components are carbon monoxide and hydrogen) by autotrophic acetogens. In view of economic considerations, a biotechnological process based on cheap and abundant gases such as CO and CO2 as a carbon source is preferable to more expensive sugar or starch fermentation. In addition, any conflict for use of substrates that can also serve as human nutrition is avoided. Natural formation of butanol has been found with, e.g. Clostridium carboxidivorans, while metabolic engineering for butanol production was successful using, e.g. C. ljungdahlii. Production of butanol from CO2 under photoautotrophic conditions was also possible by recombinant DNA construction of a respective cyanobacterial Synechococcus sp. PCC 7942 strain. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Artificial symbiosis for acetone-butanol-ethanol (ABE) fermentation from alkali extracted deshelled corn cobs by co-culture of Clostridium beijerinckii and Clostridium cellulovorans.

PubMed

Wen, Zhiqiang; Wu, Mianbin; Lin, Yijun; Yang, Lirong; Lin, Jianping; Cen, Peilin

2014-07-15

Butanol is an industrial commodity and also considered to be a more promising gasoline substitute compared to ethanol. Renewed attention has been paid to solvents (acetone, butanol and ethanol) production from the renewable and inexpensive substrates, for example, lignocellulose, on account of the depletion of oil resources, increasing gasoline prices and deteriorating environment. Limited to current tools for genetic manipulation, it is difficult to develop a genetically engineered microorganism with combined ability of lignocellulose utilization and solvents production. Mixed culture of cellulolytic microorganisms and solventogenic bacteria provides a more convenient and feasible approach for ABE fermentation due to the potential for synergistic utilization of the metabolic pathways of two organisms. But few bacteria pairs succeeded in producing biobutanol of high titer or high productivity without adding butyrate. The aim of this work was to use Clostridium cellulovorans 743B to saccharify lignocellulose and produce butyric acid, instead of adding cellulase and butyric acid to the medium, so that the soluble sugars and butyric acid generated can be subsequently utilized by Clostridium beijerinckii NCIMB 8052 to produce butanol in one pot reaction. A stable artificial symbiotic system was constructed by co-culturing a celluloytic, anaerobic, butyrate-producing mesophile (C. cellulovorans 743B) and a non-celluloytic, solventogenic bacterium (C. beijerinckii NCIMB 8052) to produce solvents by consolidated bioprocessing (CBP) with alkali extracted deshelled corn cobs (AECC), a low-cost renewable feedstock, as the sole carbon source. Under optimized conditions, the co-culture degraded 68.6 g/L AECC and produced 11.8 g/L solvents (2.64 g/L acetone, 8.30 g/L butanol and 0.87 g/L ethanol) in less than 80 h. Besides, a real-time PCR assay based on the 16S rRNA gene sequence was performed to study the dynamics of the abundance of each strain during the co-culturing process, which figured out the roles of each strain at different periods in the symbiosis. Our work illustrated the great potential of artificial symbiosis in biofuel production from lignocellulosic biomass by CBP. The dynamics of the abundance of C. beijerinckii and C. cellulovorans revealed mechanisms of cooperation and competition between the two strains during the co-culture process.

Biobutanol production by Clostridium acetobutylicum using xylose recovered from birch Kraft black liquor.

PubMed

Kudahettige-Nilsson, Rasika L; Helmerius, Jonas; Nilsson, Robert T; Sjöblom, Magnus; Hodge, David B; Rova, Ulrika

2015-01-01

Acetone-butanol-ethanol (ABE) fermentation was studied using acid-hydrolyzed xylan recovered from hardwood Kraft black liquor by CO2 acidification as the only carbon source. Detoxification of hydrolyzate using activated carbon was conducted to evaluate the impact of inhibitor removal and fermentation. Xylose hydrolysis yields as high as 18.4% were demonstrated at the highest severity hydrolysis condition. Detoxification using active carbon was effective for removal of both phenolics (76-81%) and HMF (38-52%). Batch fermentation of the hydrolyzate and semi-defined P2 media resulted in a total solvent yield of 0.12-0.13g/g and 0.34g/g, corresponding to a butanol concentration of 1.8-2.1g/L and 7.3g/L respectively. This work is the first study of a process for the production of a biologically-derived biofuel from hemicelluloses solubilized during Kraft pulping and demonstrates the feasibility of utilizing xylan recovered directly from industrial Kraft pulping liquors as a feedstock for biological production of biofuels such as butanol. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

Development Of Sustainable Biobased Products And Bioenergy In Cooperation With The Midwest Consortium For Sustainable Biobased Products And Energy

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

Michael Ladisch; Randy Woodson

2009-03-18

Collaborative efforts of Midwest Consortium have been put forth to add value to distiller's grains by further processing them into fermentable sugars, ethanol, and a protein rich co-product consistent with a pathway to a biorenewables industry (Schell et al, 2008). These studies were recently published in the enclosed special edition (Volume 99, Issue 12) of Bioresource Technology journal. Part of them have demonstrated the utilization of distillers grains as additional feedstock for increased ethanol production in the current dry grind process (Kim et al., 2008a, b; Dien et al.,2008, Ladisch et al., 2008a, b). Results showed that both liquid hotmore » water (LHW) pretreatment and ammonia fiber expansion (AFEX) were effective for enhancing digestibility of distiller's grains. Enzymatic digestion of distiller's grains resulted in more than 90% glucose yield under standard assay conditions, although the yield tends to drop as the concentration of dry solids increases. Simulated process mass balances estimated that hydrolysis and fermentation of distillers grains can increase the ethanol yield by 14% in the current dry milling process (Kim et al., 2008c). Resulting co-products from the modified process are richer in protein and oil contents than conventional distiller's grains, as determined both experimentally and computationally. Other research topics in the special edition include water solubilization of DDGS by transesterification reaction with phosphite esters (Oshel el al., 2008) to improve reactivity of the DDGS to enzymes, hydrolysis of soluble oligomers derived from DDGS using functionalized mesoporous solid catalysts (Bootsma et al., 2008), and ABE (acetone, butanol, ethanol) production from DDGS by solventogenic Clostridia (Ezeji and Blaschek, 2008). Economic analysis of a modified dry milling process, where the fiber and residual starch is extracted and fermented to produce more ethanol from the distillers grains while producing highly concentrated protein co-product, has shown that the process is economically viable resulting in an increase in net present value (Perkis et al., 2008). According to the study, the revenue is expected to increase further with improved amino acid profile of the protein rich co-products and lower cost of cellulase enzyme mixture. Also, Kim and Dale (2008) discuss using life cycle analysis to enhance the environmental performance of the corn based ethanol. On the second phase of the research, concerted efforts were directed on assessing compositional variability of dry milling co-products collected from 4 different dry grind ethanol plants has been measured and its effect on enzymatic digestibility and fermentability. Fermentation utilized a recombinant glucose/xylose co-fermenting yeast (Saccharomyces cerevisiae 424A (LNH-ST)). No significant compositional variability among the samples was found. Simultaneous saccharification and glucose/xylose co-fermentation of the pretreated distillers grains at solids and cellulase loadings of 150 g dry solids per liter and 6.4 mg protein per g dry substrate, respectively, yielded 74-801% of theoretical maximum ethanol concentration using recombinant Saccharomyces cerevisiae 424A (LNH-ST). The paper summarizing the results from the second phase of the Midwest Consortium is currently submitted to Bioresource Technology journal. The copy of the paper submitted is enclosed.« less

Flow cytometry analysis of Clostridium beijerinckii NRRL B-598 populations exhibiting different phenotypes induced by changes in cultivation conditions.

PubMed

Branska, Barbora; Pechacova, Zora; Kolek, Jan; Vasylkivska, Maryna; Patakova, Petra

2018-01-01

Biobutanol production by clostridia via the acetone-butanol-ethanol (ABE) pathway is a promising future technology in bioenergetics , but identifying key regulatory mechanisms for this pathway is essential in order to construct industrially relevant strains with high tolerance and productivity. We have applied flow cytometric analysis to C. beijerinckii NRRL B-598 and carried out comparative screening of physiological changes in terms of viability under different cultivation conditions to determine its dependence on particular stages of the life cycle and the concentration of butanol. Dual staining by propidium iodide (PI) and carboxyfluorescein diacetate (CFDA) provided separation of cells into four subpopulations with different abilities to take up PI and cleave CFDA, reflecting different physiological states. The development of a staining pattern during ABE fermentation showed an apparent decline in viability, starting at the pH shift and onset of solventogenesis, although an appreciable proportion of cells continued to proliferate. This was observed for sporulating as well as non-sporulating phenotypes at low solvent concentrations, suggesting that the increase in percentage of inactive cells was not a result of solvent toxicity or a transition from vegetative to sporulating stages. Additionally, the sporulating phenotype was challenged with butanol and cultivation with a lower starting pH was performed; in both these experiments similar trends were obtained-viability declined after the pH breakpoint, independent of the actual butanol concentration in the medium. Production characteristics of both sporulating and non-sporulating phenotypes were comparable, showing that in C. beijerinckii NRRL B-598, solventogenesis was not conditional on sporulation. We have shown that the decline in C. beijerinckii NRRL B-598 culture viability during ABE fermentation was not only the result of accumulated toxic metabolites, but might also be associated with a special survival strategy triggered by pH change.

Utilization of acetone-butanol-ethanol-water mixture obtained from biomass fermentation as renewable feedstock for hydrogen production via steam reforming: Thermodynamic and energy analyses.

PubMed

Kumar, Brajesh; Kumar, Shashi; Sinha, Shishir; Kumar, Surendra

2018-08-01

A thermodynamic equilibrium analysis on steam reforming process to utilize acetone-butanol-ethanol-water mixture obtained from biomass fermentation as biorenewable fuel has been performed to produce clean energy carrier H 2 via non-stoichiometric approach namely Gibbs free energy minimization method. The effect of process variables such as temperature (573-1473 K), pressure (1-10 atm), and steam/fuel molar feed ratio (F ABE  = 5.5-12) have been investigated on equilibrium compositions of products, H 2 , CO, CO 2 , CH 4 and solid carbon. The best suitable conditions for maximization of desired product H 2 , suppression of CH 4 , and inhibition of solid carbon are 973 K, 1 atm, steam/fuel molar feed ratio = 12. Under these conditions, the maximum molar production of hydrogen is 8.35 with negligible formation of carbon and methane. Furthermore, the energy requirement per mol of H 2 (48.96 kJ), thermal efficiency (69.13%), exergy efficiency (55.09%), exergy destruction (85.36 kJ/mol), and generated entropy (0.29 kJ/mol.K) have been achieved at same operating conditions. Copyright © 2018 Elsevier Ltd. All rights reserved.

Butanol fermentation of the brown seaweed Laminaria digitata by Clostridium beijerinckii DSM-6422.

PubMed

Hou, Xiaoru; From, Nikolaj; Angelidaki, Irini; Huijgen, Wouter J J; Bjerre, Anne-Belinda

2017-08-01

Seaweed represents an abundant, renewable, and fast-growing biomass resource for 3rd generation biofuel production. This study reports an efficient butanol fermentation process carried out by Clostridium beijerinckii DSM-6422 using enzymatic hydrolysate of the sugar-rich brown seaweed Laminaria digitata harvested from the coast of the Danish North Sea as substrate. The highest butanol yield (0.42g/g-consumed-substrates) compared to literature was achieved, with a significantly higher butanol:acetone-butanol-ethanol (ABE) molar ratio (0.85) than typical (0.6). This demonstrates the possibility of using the seaweed L. digitata as a potential biomass for butanol production. For the first time, consumption of alginate components was observed by C. beijerinckii DSM-6422. The efficient utilization of sugars and lactic acid further highlighted the potential of using this strain for future development of large-scale cost-effective butanol production based on (ensiled) seaweed. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

Technical and economic assessment of processes for the production of butanol and acetone. Phase two: analysis of research advances. Energy Conversion and Utilization Technologies Program

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

None

1984-08-01

The initial objective of this work was to develop a methodology for analyzing the impact of technological advances as a tool to help establish priorities for R and D options in the field of biocatalysis. As an example of a biocatalyzed process, butanol/acetone fermentation (ABE process) was selected as the specific topic of study. A base case model characterizing the technology and economics associated with the ABE process was developed in the previous first phase of study. The project objectives were broadened in this second phase of work to provide parametric estimates of the economic and energy impacts of amore » variety of research advances in the hydrolysis, fermentation and purification sections of the process. The research advances analyzed in this study were based on a comprehensive literature review. The six process options analyzed were: continuous ABE fermentaton; vacuum ABE fermentation; Baelene solvent extraction; HRI's Lignol process; improved prehydrolysis/dual enzyme hydrolysis; and improved microorganism tolerance to butanol toxicity. Of the six options analyzed, only improved microorganism tolerance to butanol toxicity had a significant positive effect on energy efficiency and economics. This particular process option reduced the base case production cost (including 10% DCF return) by 20% and energy consumption by 16%. Figures and tables.« less

The advanced strategy for enhancing biobutanol production and high-efficient product recovery with reduced wastewater generation.

PubMed

Xue, Chuang; Zhang, Xiaotong; Wang, Jufang; Xiao, Min; Chen, Lijie; Bai, Fengwu

2017-01-01

Butanol as an important chemical and potential fuel could be produced via ABE fermentation from lignocellulosic biomass. The use of food-related feedstocks such as maize and sugar cane may not be a sustainable solution to world's energy needs. Recently, Jerusalem artichoke tubers containing inulin have been used as feedstock for butanol production, but this bioprocess is not commercially feasible due to the great value of inulin as functional food. Till now, there is a gap on the utilization of Jerusalem artichoke stalk (JAS) as feedstock for microbial butanol production. Biobutanol production from JAS was investigated in order to improve cellulose digestibility and efficient biobutanol fermentation. Compared with 9.0 g/L butanol (14.7 g/L ABE) production by 2% NaOH pretreatment of JAS, 11.8 g/L butanol (17.6 g/L ABE) was produced in the best scenario conditions of NaOH-H 2 O 2 pretreatment, washing times and citrate buffer strengths etc. Furthermore, more than >64% water in washing pretreated JAS process could be saved, with improving butanol production by >25.0%. To mimic in situ product recovery for ABE fermentation, the vapor stripping-vapor permeation (VSVP) process steadily produced 323.4-348.7 g/L butanol (542.7-594.0 g/L ABE) in condensate, which showed more potentials than pervaporation for butanol recovery. Therefore, the present study demonstrated an effective strategy on efficient biobutanol production using lignocellulosic biomass. The process optimization could contribute to significant reduction of wastewater emission and the improvement of lignocellulosic biomass digestibility and biobutanol production, which makes biobutanol production more efficient using JAS.

Novel clostridial fusants in comparison with co-cultured counterpart species for enhanced production of biobutanol using green renewable and sustainable feedstock.

PubMed

Syed, Kashif; Dahman, Yaser

2015-11-01

In this work, biobutanol was produced through simultaneous saccharification and fermentation (SSF) of wheat straw (WS) that traditionally produces acetone, butanol and ethanol solvents (ABE). Thermal stability was imparted to two mesophilic clostridial wild strains (Clostridium beijerinckii and Clostridium acetobutylicum) through protoplast fusion with that of a corresponding thermophilic clostridial species (Clostridium thermocellum). Production was pursued by the fused strains at 45 °C compared to that of the corresponding co-cultures at 35 °C. Results showed that the fused strains generally achieved higher production at 45 °C than that of the corresponding co-cultures at 35 °C. Highest butanol production of 13.82 g/L was recorded with C. beijerinckii fusant, with ABE solvents production of 23 g/L (yields of 0.17 and 0.57, respectively). Total sugar consumption of this strain was the highest among all strains and was 84%. Fused strains also showed immense level of tolerance towards butanol toxicity compared to the wild strains. Filter paper enzyme assay demonstrated that fused strains were able to produce cellulolytic enzymes in the range of 58.73-68.52 FPU/ml. Cellulosome producing C. thermocellum and its ability to ferment sugars offers a promising future in biofuels through eliminating the need to add external enzymes. Generally, productions reported in the present study were higher than literature where biobutanol stripping systems were employed to eliminate toxicity during production. This demonstrates a clear potential for improving productivity and yield at a larger-scale facility.

Integrative modelling of pH-dependent enzyme activity and transcriptomic regulation of the acetone–butanol–ethanol fermentation of Clostridium acetobutylicum in continuous culture

PubMed Central

Millat, Thomas; Janssen, Holger; Bahl, Hubert; Fischer, Ralf-Jörg; Wolkenhauer, Olaf

2013-01-01

Summary In a continuous culture under phosphate limitation the metabolism of Clostridium acetobutylicum depends on the external pH level. By comparing seven steady-state conditions between pH 5.7 and pH 4.5 we show that the switch from acidogenesis to solventogenesis occurs between pH 5.3 and pH 5.0 with an intermediate state at pH 5.1. Here, an integrative study is presented investigating how a changing external pH level affects the clostridial acetone–butanol–ethanol (ABE) fermentation pathway. This is of particular interest as the biotechnological production of n-butanol as biofuel has recently returned into the focus of industrial applications. One prerequisite is the furthering of the knowledge of the factors determining the solvent production and their integrative regulations. We have mathematically analysed the influence of pH-dependent specific enzyme activities of branch points of the metabolism on the product formation. This kinetic regulation was compared with transcriptomic regulation regarding gene transcription and the proteomic profile. Furthermore, both regulatory mechanisms were combined yielding a detailed projection of their individual and joint effects on the product formation. The resulting model represents an important platform for future developments of industrial butanol production based on C. acetobutylicum. PMID:23332010

High acetone-butanol-ethanol production in pH-stat co-feeding of acetate and glucose.

PubMed

Gao, Ming; Tashiro, Yukihiro; Wang, Qunhui; Sakai, Kenji; Sonomoto, Kenji

2016-08-01

We previously reported the metabolic analysis of butanol and acetone production from exogenous acetate by (13)C tracer experiments (Gao et al., RSC Adv., 5, 8486-8495, 2015). To clarify the influence of acetate on acetone-butanol-ethanol (ABE) production, we first performed an enzyme assay in Clostridium saccharoperbutylacetonicum N1-4. Acetate addition was found to drastically increase the activities of key enzymes involved in the acetate uptake (phosphate acetyltransferase and CoA transferase), acetone formation (acetoacetate decarboxylase), and butanol formation (butanol dehydrogenase) pathways. Subsequently, supplementation of acetate during acidogenesis and early solventogenesis resulted in a significant increase in ABE production. To establish an efficient ABE production system using acetate as a co-substrate, several shot strategies were investigated in batch culture. Batch cultures with two substrate shots without pH control produced 14.20 g/L butanol and 23.27 g/L ABE with a maximum specific butanol production rate of 0.26 g/(g h). Furthermore, pH-controlled (at pH 5.5) batch cultures with two substrate shots resulted in not only improved acetate consumption but also a further increase in ABE production. Finally, we obtained 15.13 g/L butanol and 24.37 g/L ABE at the high specific butanol production rate of 0.34 g/(g h) using pH-stat co-feeding method. Thus, in this study, we established a high ABE production system using glucose and acetate as co-substrates in a pH-stat co-feeding system with C. saccharoperbutylacetonicum N1-4. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Production of Butanol (a Biofuel) from Agricultural Residues: Part II - Use of Corn Stover and Switchgrass Hydrolysates

USDA-ARS?s Scientific Manuscript database

Acetone butanol ethanol (ABE or AB, or solvent) was produced from hydrolyzed corn stover and switchgrass using Clostridium beijerinckii P260. A control experiment using glucose resulted in the production of 21.06 gL**-1 total ABE. In this experiment, an AB yield and productivity of 0.41 and 0.31 g...

Biobutanol production from apple pomace: the importance of pretreatment methods on the fermentability of lignocellulosic agro-food wastes.

PubMed

Hijosa-Valsero, María; Paniagua-García, Ana I; Díez-Antolínez, Rebeca

2017-11-01

Apple pomace was studied as a possible raw material for biobutanol production. Five different soft physicochemical pretreatments (autohydrolysis, acids, alkalis, organic solvents and surfactants) were compared in a high-pressure reactor, whose working parameters (temperature, time and reagent concentration) were optimised to maximise the amount of simple sugars released and to minimise inhibitor generation. The pretreated biomass was subsequently subjected to a conventional enzymatic treatment to complete the hydrolysis. A thermal analysis (DSC) of the solid biomass indicated that lignin was mainly degraded during the enzymatic treatment. The hydrolysate obtained with the surfactant polyethylene glycol 6000 (PEG 6000) (1.96% w/w) contained less inhibitors than any other pretreatment, yet providing 42 g/L sugars at relatively mild conditions (100 °C, 5 min), and was readily fermented by Clostridium beijerinckii CECT 508 in 96 h (3.55 g/L acetone, 9.11 g/L butanol, 0.26 g/L ethanol; 0.276 g B /g S yield; 91% sugar consumption). Therefore, it is possible to optimise pretreatment conditions of lignocellulosic apple pomace to reduce inhibitor concentrations in the final hydrolysate and perform successful ABE fermentations without the need of a detoxification stage.

Predictive modeling in Clostridium acetobutylicum fermentations employing Raman spectroscopy and multivariate data analysis for real-time culture monitoring

NASA Astrophysics Data System (ADS)

Zu, Theresah N. K.; Liu, Sanchao; Germane, Katherine L.; Servinsky, Matthew D.; Gerlach, Elliot S.; Mackie, David M.; Sund, Christian J.

2016-05-01

The coupling of optical fibers with Raman instrumentation has proven to be effective for real-time monitoring of chemical reactions and fermentations when combined with multivariate statistical data analysis. Raman spectroscopy is relatively fast, with little interference from the water peak present in fermentation media. Medical research has explored this technique for analysis of mammalian cultures for potential diagnosis of some cancers. Other organisms studied via this route include Escherichia coli, Saccharomyces cerevisiae, and some Bacillus sp., though very little work has been performed on Clostridium acetobutylicum cultures. C. acetobutylicum is a gram-positive anaerobic bacterium, which is highly sought after due to its ability to use a broad spectrum of substrates and produce useful byproducts through the well-known Acetone-Butanol-Ethanol (ABE) fermentation. In this work, real-time Raman data was acquired from C. acetobutylicum cultures grown on glucose. Samples were collected concurrently for comparative off-line product analysis. Partial-least squares (PLS) models were built both for agitated cultures and for static cultures from both datasets. Media components and metabolites monitored include glucose, butyric acid, acetic acid, and butanol. Models were cross-validated with independent datasets. Experiments with agitation were more favorable for modeling with goodness of fit (QY) values of 0.99 and goodness of prediction (Q2Y) values of 0.98. Static experiments did not model as well as agitated experiments. Raman results showed the static experiments were chaotic, especially during and shortly after manual sampling.

Impact of butyric acid on butanol formation by Clostridium pasteurianum.

PubMed

Regestein, Lars; Doerr, Eric Will; Staaden, Antje; Rehmann, Lars

2015-11-01

The butanol yield of the classic fermentative acetone-butanol-ethanol (ABE) process has been enhanced in the past decades through the development of better strains and advanced process design. Nevertheless, by-product formation and the incomplete conversion of intermediates still decrease the butanol yield. This study demonstrates the potential of increasing the butanol yield from glycerol though the addition of small amounts of butyric acid. The impact of butyric acid was investigated in a 7L stirred tank reactor. The results of this study show the positive impact of butyric acid on butanol yield under pH controlled conditions and the metabolic stages were monitored via online measurement of carbon dioxide formation, pH value and redox potential. Butyric acid could significantly increase the butanol yield at low pH values if sufficient quantities of primary carbon source (glycerol) were present. Copyright © 2015 Elsevier Ltd. All rights reserved.

Performance comparison of ethanol and butanol production in a continuous and closed-circulating fermentation system with membrane bioreactor.

PubMed

Chen, Chunyan; Long, Sihua; Li, Airong; Xiao, Guoqing; Wang, Linyuan; Xiao, Zeyi

2017-03-16

Since both ethanol and butanol fermentations are urgently developed processes with the biofuel-demand increasing, performance comparison of aerobic ethanol fermentation and anerobic butanol fermentation in a continuous and closed-circulating fermentation (CCCF) system was necessary to achieve their fermentation characteristics and further optimize the fermentation process. Fermentation and pervaporation parameters including the average cell concentration, glucose consumption rate, cumulated production concentration, product flux, and separation factor of ethanol fermentation were 11.45 g/L, 3.70 g/L/h, 655.83 g/L, 378.5 g/m 2 /h, and 4.83, respectively, the corresponding parameters of butanol fermentation were 2.19 g/L, 0.61 g/L/h, 28.03 g/L, 58.56 g/m 2 /h, and 10.62, respectively. Profiles of fermentation and pervaporation parameters indicated that the intensity and efficiency of ethanol fermentation was higher than butanol fermentation, but the stability of butanol fermentation was superior to ethanol fermentation. Although the two fermentation processes had different features, the performance indicated the application prospect of both ethanol and butanol production by the CCCF system.

Impact of osmotic stress and ethanol inhibition in yeast cells on process oscillation associated with continuous very-high-gravity ethanol fermentation

PubMed Central

2013-01-01

Background VHG fermentation is a promising process engineering strategy aiming at improving ethanol titer, and thus saving energy consumption for ethanol distillation and distillage treatment. However, sustained process oscillation was observed during continuous VHG ethanol fermentation, which significantly affected ethanol fermentation performance of the system. Results Sustained process oscillation was investigated in continuous VHG ethanol fermentation, and stresses exerted on yeast cells by osmotic pressure from unfermented sugars and ethanol inhibition developed within the fermentation system were postulated to be major factors triggering this phenomenon. In this article, steady state was established for continuous ethanol fermentation with LG medium containing 120 g/L glucose, and then 160 g/L non-fermentable xylose was supplemented into the LG medium to simulate the osmotic stress on yeast cells under the VHG fermentation condition, but the fermentation process was still at steady state, indicating that the impact of osmotic stress on yeast cells was not the main reason for the process oscillation. However, when 30 g/L ethanol was supplemented into the LG medium to simulate the ethanol inhibition in yeast cells under the VHG fermentation condition, process oscillation was triggered, which was augmented with extended oscillation period and exaggerated oscillation amplitude as ethanol supplementation was increased to 50 g/L, but the process oscillation was gradually attenuated when the ethanol supplementations were stopped, and the steady state was restored. Furthermore, gas stripping was incorporated into the continuous VHG fermentation system to in situ remove ethanol produced by Saccharomyces cerevisiae, and the process oscillation was also attenuated, but restored after the gas stripping was interrupted. Conclusions Experimental results indicated that ethanol inhibition rather than osmotic stress on yeast cells is one of the main factors triggering the process oscillation under the VHG fermentation condition, and in the meantime gas stripping was validated to be an effective strategy for attenuating the process oscillation. PMID:24041271

Yeasts in sustainable bioethanol production: A review.

PubMed

Mohd Azhar, Siti Hajar; Abdulla, Rahmath; Jambo, Siti Azmah; Marbawi, Hartinie; Gansau, Jualang Azlan; Mohd Faik, Ainol Azifa; Rodrigues, Kenneth Francis

2017-07-01

Bioethanol has been identified as the mostly used biofuel worldwide since it significantly contributes to the reduction of crude oil consumption and environmental pollution. It can be produced from various types of feedstocks such as sucrose, starch, lignocellulosic and algal biomass through fermentation process by microorganisms. Compared to other types of microoganisms, yeasts especially Saccharomyces cerevisiae is the common microbes employed in ethanol production due to its high ethanol productivity, high ethanol tolerance and ability of fermenting wide range of sugars. However, there are some challenges in yeast fermentation which inhibit ethanol production such as high temperature, high ethanol concentration and the ability to ferment pentose sugars. Various types of yeast strains have been used in fermentation for ethanol production including hybrid, recombinant and wild-type yeasts. Yeasts can directly ferment simple sugars into ethanol while other type of feedstocks must be converted to fermentable sugars before it can be fermented to ethanol. The common processes involves in ethanol production are pretreatment, hydrolysis and fermentation. Production of bioethanol during fermentation depends on several factors such as temperature, sugar concentration, pH, fermentation time, agitation rate, and inoculum size. The efficiency and productivity of ethanol can be enhanced by immobilizing the yeast cells. This review highlights the different types of yeast strains, fermentation process, factors affecting bioethanol production and immobilization of yeasts for better bioethanol production.

Characterization of an immobilized cell, trickle bed reactor during long term butanol (ABE) fermentation.

PubMed

Park, C H; Okos, M R; Wankat, P C

1990-06-20

Acetone-butanol-ethanol (ABE) fermentation was performed continuously in an immobilized cell, trickle bed reactor for 54 days without, degeneration by maintaining the pH above 4.3. Column clogging was minimized by structured packing of immobilization matrix. The reactor contained two serial glass columns packed with Clostridium acetobutylicum adsorbed on 12- and 20-in.-long polyester sponge strips at total flow rates between 38 and 98.7 mL/h. Cells were initially grown at 20 g/L glucose resulting in low butanol (1.15 g/L) production encouraging cell growth. After the initial cell growth phase a higher glucose concentration (38.7 g/L) improved solvent yield from 13.2 to 24.1 wt%, and butanol production rate was the best. Further improvement in solvent yield and butanol production rate was not observed with 60 g/L of glucose. However, when the fresh nutrient supply was limited to only the first column, solvent yield increased to 27.3 wt% and butanol selectivity was improved to 0.592 as compared to 0.541 when fresh feed was fed to both columns. The highest butanol concentration of 5.2 g/L occurred at 55% conversion of the feed with 60 g/L glucose. Liquid product yield of immobilized cells approached the theoretical value reported in the literature. Glucose and product concentration profiles along the column showed that the columns can be divided into production and inhibition regions. The length of each zone was dependent upon the feed glucose concentration and feed pattern. Unlike batch fermentation, there was no clear distinction between acid and solvent production regions. The pH dropped, from 6.18-6.43 to 4.50-4.90 in the first inch of the reactor. The pH dropped further to 4.36-4.65 by the exit of the column. The results indicate that the strategy for long term stable operation with high solvent yield requires a structured packing of biologically stable porous matrix such as polyester sponge, a pH maintenance above 4.3, glucose concentrations up to 60 g/L and nutrient supply only to the inlet of the reactor.

Ethanol production from food waste at high solids content with vacuum recovery technology.

PubMed

Huang, Haibo; Qureshi, Nasib; Chen, Ming-Hsu; Liu, Wei; Singh, Vijay

2015-03-18

Ethanol production from food wastes does not only solve environmental issues but also provides renewable biofuels. This study investigated the feasibility of producing ethanol from food wastes at high solids content (35%, w/w). A vacuum recovery system was developed and applied to remove ethanol from fermentation broth to reduce yeast ethanol inhibition. A high concentration of ethanol (144 g/L) was produced by the conventional fermentation of food waste without a vacuum recovery system. When the vacuum recovery is applied to the fermentation process, the ethanol concentration in the fermentation broth was controlled below 100 g/L, thus reducing yeast ethanol inhibition. At the end of the conventional fermentation, the residual glucose in the fermentation broth was 5.7 g/L, indicating incomplete utilization of glucose, while the vacuum fermentation allowed for complete utilization of glucose. The ethanol yield for the vacuum fermentation was found to be 358 g/kg of food waste (dry basis), higher than that for the conventional fermentation at 327 g/kg of food waste (dry basis).

BIOASPEN: System for technology development

NASA Technical Reports Server (NTRS)

1986-01-01

The public version of ASPEN was installed in the VAX 11/750 computer. To examine the idea of BIOASPEN, a test example (the manufacture of acetone, butanol, and ethanol through a biological route) was chosen for simulation. Previous reports on the BIOASPEN project revealed the limitations of ASPEN in modeling this process. To overcome some of the difficulties, modules were written for the acid and enzyme hydrolyzers, the fermentor, and a sterilizer. Information required for these modules was obtained from the literature whenever possible. Additional support modules necessary for interfacing with ASPEN were also written. Some of ASPEN subroutines were themselves altered in order to ensure the correct running of the simulation program. After testing of these additions and charges was completed, the Acetone-Butanol-Ethanol (ABE) process was simulated. A release of ASPEN (which contained the Economic Subsystem) was obtained and installed. This subsection was tested and numerous charges were made in the FORTRAN code. Capital investment and operating cost studies were performed on the ABE process. Some alternatives in certain steps of the ABE simulation were investigated in order to elucidate their effects on the overall economics of the process.

Bioethanol production: an integrated process of low substrate loading hydrolysis-high sugars liquid fermentation and solid state fermentation of enzymatic hydrolysis residue.

PubMed

Chu, Qiulu; Li, Xin; Ma, Bin; Xu, Yong; Ouyang, Jia; Zhu, Junjun; Yu, Shiyuan; Yong, Qiang

2012-11-01

An integrated process of enzymatic hydrolysis and fermentation was investigated for high ethanol production. The combination of enzymatic hydrolysis at low substrate loading, liquid fermentation of high sugars concentration and solid state fermentation of enzymatic hydrolysis residue was beneficial for conversion of steam explosion pretreated corn stover to ethanol. The results suggested that low substrate loading hydrolysis caused a high enzymatic hydrolysis yield; the liquid fermentation of about 200g/L glucose by Saccharomyces cerevisiae provided a high ethanol concentration which could significantly decrease cost of the subsequent ethanol distillation. A solid state fermentation of enzymatic hydrolysis residue was combined, which was available to enhance ethanol production and cellulose-to-ethanol conversion. The results of solid state fermentation demonstrated that the solid state fermentation process accompanied by simultaneous saccharification and fermentation. Copyright © 2012 Elsevier Ltd. All rights reserved.

Effect of pH on ethanol-type acidogenic fermentation of fruit and vegetable waste.

PubMed

Wu, Yuanyuan; Wang, Cuiping; Zheng, Mingyue; Zuo, Jiane; Wu, Jing; Wang, Kaijun; Yang, Boqiong

2017-02-01

The aim of this study was to investigate the possibility and optimal controlling strategy for ethanol-type acidogenic fermentation of fruit and vegetable waste by mixed microbial cultures. Four continuous stirred tank reactors (CSTR) were operated at various pHs (4.0, 5.0, 5.5, and 6.0) with an organic loading rate of 13gVS/(Ld) and hydraulic retention time of 3d. Butyrate-type fermentation was observed at pH 5.0, 5.5, and 6.0. Conversely, at pH 4.0, ethanol-type fermentation was observed with a high mass concentration and proportion (of total fermentative products) of ethanol, which were 6.7g/L and 88.8%, respectively. However, the total concentration of ethanol-type fermentative products substantially decreased from days 22-25. The optimal pH of ethanol-type fermentative microorganisms was investigated by using batch experiments with pH controlled at 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, and 7.0 and results showed that the maximum ethanol concentration and relatively highest acidogenic rate were found at pH of 5.5. The pH in the long term CSTR was changed from 4.0 to 5.5 to improve ethanol-type fermentation and results showed that ethanol-type fermentation was improved temporarily, however, was followed by the reappearance of butyrate-type fermentation. In addition, ethanol-type fermentation recovered once more when pH was reverted to 4.0. Therefore, the results of this study suggest that a process of dynamic, sequenced pH control with the order pH 4.0, 5.5 and 4.0 might be a feasible controlling strategy for continuous and stable ethanol-type fermentation. Copyright © 2016 Elsevier Ltd. All rights reserved.

Extractive Fermentation of Sugarcane Juice to Produce High Yield and Productivity of Bioethanol

NASA Astrophysics Data System (ADS)

Rofiqah, U.; Widjaja, T.; Altway, A.; Bramantyo, A.

2017-04-01

Ethanol production by batch fermentation requires a simple process and it is widely used. Batch fermentation produces ethanol with low yield and productivity due to the accumulation of ethanol in which poisons microorganisms in the fermenter. Extractive fermentation technique is applied to solve the microorganism inhibition problem by ethanol. Extractive fermentation technique can produce ethanol with high yield and productivity. In this process raffinate still, contains much sugar because conversion in the fermentation process is not perfect. Thus, to enhance ethanol yield and productivity, recycle system is applied by returning the raffinate from the extraction process to the fermentation process. This raffinate also contains ethanol which would inhibit the performance of microorganisms in producing ethanol during the fermentation process. Therefore, this study aims to find the optimum condition for the amount of solvent to broth ratio (S: B) and recycle to fresh feed ratio (R: F) which enter the fermenter to produce high yield and productivity. This research was carried out by experiment. In the experiment, sugarcane juice was fermented using Zymomonasmobilis mutant. The fermentation broth was extracted using amyl alcohol. The process was integrated with the recycle system by varying the recycle ratio. The highest yield and productivity is 22.3901% and 103.115 g / L.h respectively, obtained in a process that uses recycle to fresh feed ratio (R: F) of 50:50 and solvents to both ratio of 1.

Genetic improvement of native xylose-fermenting yeasts for ethanol production.

PubMed

Harner, Nicole K; Wen, Xin; Bajwa, Paramjit K; Austin, Glen D; Ho, Chi-Yip; Habash, Marc B; Trevors, Jack T; Lee, Hung

2015-01-01

Lignocellulosic substrates are the largest source of fermentable sugars for bioconversion to fuel ethanol and other valuable compounds. To improve the economics of biomass conversion, it is essential that all sugars in potential hydrolysates be converted efficiently into the desired product(s). While hexoses are fermented into ethanol and some high-value chemicals, the bioconversion of pentoses in hydrolysates remains inefficient. This remains one of the key challenges in lignocellulosic biomass conversion. Native pentose-fermenting yeasts can ferment both glucose and xylose in lignocellulosic biomass to ethanol. However, they perform poorly in the presence of hydrolysate inhibitors, exhibit low ethanol tolerance and glucose repression, and ferment pentoses less efficiently than the main hexoses glucose and mannose. This paper reviews classical and molecular strain improvement strategies applied to native pentose-fermenting yeasts for improved ethanol production from xylose and lignocellulosic substrates. We focus on Pachysolen tannophilus, Scheffersomyces (Candida) shehatae, Scheffersomyces (Pichia) stipitis, and Spathaspora passalidarum which are good ethanol producers among the native xylose-fermenting yeasts. Strains obtained thus far are not robust enough for efficient ethanol production from lignocellulosic hydrolysates and can benefit from further improvements.

Ethanol Production and Maximum Cell Growth Are Highly Correlated with Membrane Lipid Composition during Fermentation as Determined by Lipidomic Analysis of 22 Saccharomyces cerevisiae Strains

PubMed Central

Henderson, Clark M.; Lozada-Contreras, Michelle; Jiranek, Vladimir; Longo, Marjorie L.

2013-01-01

Optimizing ethanol yield during fermentation is important for efficient production of fuel alcohol, as well as wine and other alcoholic beverages. However, increasing ethanol concentrations during fermentation can create problems that result in arrested or sluggish sugar-to-ethanol conversion. The fundamental cellular basis for these problem fermentations, however, is not well understood. Small-scale fermentations were performed in a synthetic grape must using 22 industrial Saccharomyces cerevisiae strains (primarily wine strains) with various degrees of ethanol tolerance to assess the correlation between lipid composition and fermentation kinetic parameters. Lipids were extracted at several fermentation time points representing different growth phases of the yeast to quantitatively analyze phospholipids and ergosterol utilizing atmospheric pressure ionization-mass spectrometry methods. Lipid profiling of individual fermentations indicated that yeast lipid class profiles do not shift dramatically in composition over the course of fermentation. Multivariate statistical analysis of the data was performed using partial least-squares linear regression modeling to correlate lipid composition data with fermentation kinetic data. The results indicate a strong correlation (R2 = 0.91) between the overall lipid composition and the final ethanol concentration (wt/wt), an indicator of strain ethanol tolerance. One potential component of ethanol tolerance, the maximum yeast cell concentration, was also found to be a strong function of lipid composition (R2 = 0.97). Specifically, strains unable to complete fermentation were associated with high phosphatidylinositol levels early in fermentation. Yeast strains that achieved the highest cell densities and ethanol concentrations were positively correlated with phosphatidylcholine species similar to those known to decrease the perturbing effects of ethanol in model membrane systems. PMID:23064336

Ethanol production and maximum cell growth are highly correlated with membrane lipid composition during fermentation as determined by lipidomic analysis of 22 Saccharomyces cerevisiae strains.

PubMed

Henderson, Clark M; Lozada-Contreras, Michelle; Jiranek, Vladimir; Longo, Marjorie L; Block, David E

2013-01-01

Optimizing ethanol yield during fermentation is important for efficient production of fuel alcohol, as well as wine and other alcoholic beverages. However, increasing ethanol concentrations during fermentation can create problems that result in arrested or sluggish sugar-to-ethanol conversion. The fundamental cellular basis for these problem fermentations, however, is not well understood. Small-scale fermentations were performed in a synthetic grape must using 22 industrial Saccharomyces cerevisiae strains (primarily wine strains) with various degrees of ethanol tolerance to assess the correlation between lipid composition and fermentation kinetic parameters. Lipids were extracted at several fermentation time points representing different growth phases of the yeast to quantitatively analyze phospholipids and ergosterol utilizing atmospheric pressure ionization-mass spectrometry methods. Lipid profiling of individual fermentations indicated that yeast lipid class profiles do not shift dramatically in composition over the course of fermentation. Multivariate statistical analysis of the data was performed using partial least-squares linear regression modeling to correlate lipid composition data with fermentation kinetic data. The results indicate a strong correlation (R(2) = 0.91) between the overall lipid composition and the final ethanol concentration (wt/wt), an indicator of strain ethanol tolerance. One potential component of ethanol tolerance, the maximum yeast cell concentration, was also found to be a strong function of lipid composition (R(2) = 0.97). Specifically, strains unable to complete fermentation were associated with high phosphatidylinositol levels early in fermentation. Yeast strains that achieved the highest cell densities and ethanol concentrations were positively correlated with phosphatidylcholine species similar to those known to decrease the perturbing effects of ethanol in model membrane systems.

Stillage reflux in food waste ethanol fermentation and its by-product accumulation.

PubMed

Ma, Hongzhi; Yang, Jian; Jia, Yan; Wang, Qunhui; Tashiro, Yukihiro; Sonomoto, Kenji

2016-06-01

Raw materials and pollution control are key issues for the ethanol fermentation industry. To address these concerns, food waste was selected as fermentation substrate, and stillage reflux was carried out in this study. Reflux was used seven times during fermentation. Corresponding ethanol and reducing sugar were detected. Accumulation of by-products, such as organic acid, sodium chloride, and glycerol, was investigated. Lactic acid was observed to accumulate up to 120g/L, and sodium chloride reached 0.14mol/L. Other by-products did not accumulate. The first five cycles of reflux increased ethanol concentration, which prolonged fermentation time. Further increases in reflux time negatively influenced ethanol fermentation. Single-factor analysis with lactic acid and sodium chloride demonstrated that both factors affected ethanol fermentation, but lactic acid induced more effects. Copyright © 2016 Elsevier Ltd. All rights reserved.

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

PubMed

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

2010-05-01

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

Impact of pseudo-continuous fermentation on the ethanol tolerance of Scheffersomyces stipitis.

PubMed

Liang, Meng; Kim, Min Hea; He, Qinghua Peter; Wang, Jin

2013-09-01

In this work we conducted the pseudo-continuous fermentation, i.e., continuous fermentation with cell retention, using Scheffersomyces stipitis, and studied its effect on ethanol tolerance of the strain. During the fermentation experiments, S. stipitis was adapted to a mild concentration of ethanol (20-26 g/L) for two weeks. Two substrates (glucose and xylose) were used in different fermentation experiments. After fermentation, various experiments were performed to evaluate the ethanol tolerance of adapted cells and unadapted cells. Compared to the unadapted cells, the viability of adapted cells increased by 8 folds with glucose as the carbon source and 6 folds with xylose as the carbon source following exposure to 60 g/L ethanol for 2 h. Improved ethanol tolerance of the adapted cells was also revealed in the effects of ethanol on plasma membrane permeability, extracellular alkalization and acidification. The mathematical modeling of cell leakage, extracellular alkalization and acidification revealed that cells cultured on glucose show better ethanol tolerance than cells cultured on xylose but the differences become smaller for adapted cells. The results show that pseudo-continuous fermentation can effectively improve cell's ethanol tolerance due to the environmental pressure during the fermentation process. Copyright © 2013 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

The preparation and ethanol fermentation of high-concentration sugars from steam-explosion corn stover.

PubMed

Xie, Hui; Wang, Fengqin; Yin, Shuangyao; Ren, Tianbao; Song, Andong

2015-05-01

In the field of biofuel ethanol, high-concentration- reducing sugars made from cellulosic materials lay the foundation for high-concentration ethanol fermentation. In this study, corn stover was pre-treated in a process combining chemical methods and steam explosion; the cellulosic hydrolyzed sugars obtained by fed-batch saccharification were then used as the carbon source for high-concentration ethanol fermentation. Saccharomyces cerevisiae 1308, Angel yeast, and Issatchenkia orientalis were shake-cultured with Pachysolen tannophilus P-01 for fermentation. Results implied that the ethanol yields from the three types of mixed strains were 4.85 g/100 mL, 4.57 g/100 mL, and 5.02 g/100 mL (separately) at yield rates of 91.6, 89.3, and 92.2%, respectively. Therefore, it was inferred that shock-fermentation using mixed strains achieved a higher ethanol yield at a greater rate in a shorter fermentation period. This study provided a theoretical basis and technical guidance for the fermentation of industrial high-concentrated cellulosic ethanol.

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

PubMed

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

2016-03-01

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

Pre-treatment step with Leuconostoc mesenteroides or L. pseudomesenteroides strains removes furfural from Zymomonas mobilis ethanolic fermentation broth.

PubMed

Hunter, William J; Manter, Daniel K

2014-10-01

Furfural is an inhibitor of growth and ethanol production by Zymomonas mobilis. This study used a naturally occurring (not GMO) biological pre-treatment to reduce that amount of furfural in a model fermentation broth. Pre-treatment involved inoculating and incubating the fermentation broth with strains of Leuconostoc mesenteroides or Leuconostoc pseudomesenteroides. The Leuconostoc strains converted furfural to furfuryl alcohol without consuming large amounts of dextrose in the process. Coupling this pre-treatment to ethanolic fermentation reduced furfural in the broth and improved growth, dextrose uptake and ethanol formation. Pre-treatment permitted ethanol formation in the presence of 5.2 g L(-1) furfural, which was otherwise inhibitive. The pre-treatment and presence of the Leuconostoc strains in the fermentation broth did not interfere with Z. mobilis ethanolic fermentation or the amounts of ethanol produced. The method suggests a possible technique for reducing the effect that furfural has on the production of ethanol for use as a biofuel. Published by Elsevier Ltd.

Transcriptional activator Cat8 is involved in regulation of xylose alcoholic fermentation in the thermotolerant yeast Ogataea (Hansenula) polymorpha.

PubMed

Ruchala, Justyna; Kurylenko, Olena O; Soontorngun, Nitnipa; Dmytruk, Kostyantyn V; Sibirny, Andriy A

2017-02-28

Efficient xylose alcoholic fermentation is one of the key to a successful lignocellulosic ethanol production. However, regulation of this process in the native xylose-fermenting yeasts is poorly understood. In this work, we paid attention to the transcriptional factor Cat8 and its possible role in xylose alcoholic fermentation in Ogataea (Hansenula) polymorpha. In Saccharomyces cerevisiae, organism, which does not metabolize xylose, gene CAT8 encodes a Zn-cluster transcriptional activator necessary for expression of genes involved in gluconeogenesis, respiration, glyoxylic cycle and ethanol utilization. Xylose is a carbon source that could be fermented to ethanol and simultaneously could be used in gluconeogenesis for hexose synthesis. This potentially suggests involvement of CAT8 in xylose metabolism. Here, the role of CAT8 homolog in the natural xylose-fermenting thermotolerant yeast O. polymorpha was characterized. The CAT8 ortholog was identified in O. polymorpha genome and deleted both in the wild-type strain and in advanced ethanol producer from xylose. Constructed cat8Δ strain isolated from wild strain showed diminished growth on glycerol, ethanol and xylose as well as diminished respiration on the last substrate. At the same time, cat8Δ mutant isolated from the best available O. polymorpha ethanol producer showed only visible defect in growth on ethanol. CAT8 deletant was characterized by activated transcription of genes XYL3, DAS1 and RPE1 and slight increase in the activity of several enzymes involved in xylose metabolism and alcoholic fermentation. Ethanol production from xylose in cat8Δ mutants in the background of wild-type strain and the best available ethanol producer from xylose increased for 50 and 30%, respectively. The maximal titer of ethanol during xylose fermentation was 12.5 g ethanol/L at 45 °C. Deletion of CAT8 did not change ethanol production from glucose. Gene CAT8 was also overexpressed under control of the strong constitutive promoter GAP of glyceraldehyde-3-phosphate dehydrogenase. Corresponding strains showed drop in ethanol production in xylose medium whereas glucose alcoholic fermentation remained unchanged. Available data suggest on specific role of Cat8 in xylose alcoholic fermentation. The CAT8 gene is one of the first identified genes specifically involved in regulation of xylose alcoholic fermentation in the natural xylose-fermenting yeast O. polymorpha.

A novel inhibitor of Lactobacillus biofilms prevents stuck fermentations in a shake flask model

USDA-ARS?s Scientific Manuscript database

Yeast ethanol fermentations contain contaminating bacteria and yeast, with Lactobacilli being a frequent contaminant. These bacteria tolerate the low pH and high ethanol concentrations present in the fermentation, and can decrease the ethanol yield. Fermentations are routinely treated with antibioti...

Enhanced ethanol fermentation by engineered Saccharomyces cerevisiae strains with high spermidine contents.

PubMed

Kim, Sun-Ki; Jo, Jung-Hyun; Jin, Yong-Su; Seo, Jin-Ho

2017-05-01

Construction of robust and efficient yeast strains is a prerequisite for commercializing a biofuel production process. We have demonstrated that high intracellular spermidine (SPD) contents in Saccharomyces cerevisiae can lead to improved tolerance against various fermentation inhibitors, including furan derivatives and acetic acid. In this study, we examined the potential applicability of the S. cerevisiae strains with high SPD contents under two cases of ethanol fermentation: glucose fermentation in repeated-batch fermentations and xylose fermentation in the presence of fermentation inhibitors. During the sixteen times of repeated-batch fermentations using glucose as a sole carbon source, the S. cerevisiae strains with high SPD contents maintained higher cell viability and ethanol productivities than a control strain with lower SPD contents. Specifically, at the sixteenth fermentation, the ethanol productivity of a S. cerevisiae strain with twofold higher SPD content was 31% higher than that of the control strain. When the SPD content was elevated in an engineered S. cerevisiae capable of fermenting xylose, the resulting S. cerevisiae strain exhibited much 40-50% higher ethanol productivities than the control strain during the fermentations of synthetic hydrolysate containing high concentrations of fermentation inhibitors. These results suggest that the strain engineering strategy to increase SPD content is broadly applicable for engineering yeast strains for robust and efficient production of ethanol.

Ethanol fermentation with Kluyveromyces marxianus from Jerusalem artichoke grown in salina and irrigated with a mixture of seawater and freshwater.

PubMed

Yuan, W J; Zhao, X Q; Ge, X M; Bai, F W

2008-12-01

To study fuel ethanol fermentation with Kluyveromyces marxianus ATCC8554 from Jerusalem artichoke (Helianthus tuberosus) grown in salina and irrigated with a mixture of seawater and freshwater. The growth and ethanol fermentation of K. marxianus ATCC8554 were studied using inulin as substrate. The activity of inulinase, which attributes to the hydrolysis of inulin, the main carbohydrate in Jerusalem artichoke, was monitored. The optimum temperatures were 38 degrees C for growth and inulinase production, and 35 degrees C for ethanol fermentation. Aeration was not necessary for ethanol fermentation with the K. marxianus from inulin. Then, the fresh Jerusalem artichoke tubers grown in salina and irrigated with 25% and 50% seawater were further examined for ethanol fermentation with the K. marxianus, and a higher ethanol yield was achieved for the Jerusalem artichoke tuber irrigated with 25% seawater. Furthermore, the dry meal of the Jerusalem artichoke tubers irrigated with 25% seawater was examined for ethanol fermentation at three solid concentrations of 200, 225 and 250 g l(-1), and the highest ethanol yield of 0.467, or 91.5% of the theoretical value of 0.511, was achieved for the slurry with a solid concentration of 200 g l(-1). Halophilic Jerusalem artichoke can be used for fuel ethanol production. Halophilic Jerusalem artichoke, not competing with grain crops for arable land, is a sustainable feedstock for fuel ethanol production.

Increased expression of the yeast multidrug resistance ABC transporter Pdr18 leads to increased ethanol tolerance and ethanol production in high gravity alcoholic fermentation

PubMed Central

2012-01-01

Background The understanding of the molecular basis of yeast tolerance to ethanol may guide the design of rational strategies to increase process performance in industrial alcoholic fermentations. A set of 21 genes encoding multidrug transporters from the ATP-Binding Cassette (ABC) Superfamily and Major Facilitator Superfamily (MFS) in S. cerevisiae were scrutinized for a role in ethanol stress resistance. Results A yeast multidrug resistance ABC transporter encoded by the PDR18 gene, proposed to play a role in the incorporation of ergosterol in the yeast plasma membrane, was found to confer resistance to growth inhibitory concentrations of ethanol. PDR18 expression was seen to contribute to decreased 3 H-ethanol intracellular concentrations and decreased plasma membrane permeabilization of yeast cells challenged with inhibitory ethanol concentrations. Given the increased tolerance to ethanol of cells expressing PDR18, the final concentration of ethanol produced during high gravity alcoholic fermentation by yeast cells devoid of PDR18 was lower than the final ethanol concentration produced by the corresponding parental strain. Moreover, an engineered yeast strain in which the PDR18 promoter was replaced in the genome by the stronger PDR5 promoter, leading to increased PDR18 mRNA levels during alcoholic fermentation, was able to attain a 6 % higher ethanol concentration and a 17 % higher ethanol production yield than the parental strain. The improved fermentative performance of yeast cells over-expressing PDR18 was found to correlate with their increased ethanol tolerance and ability to restrain plasma membrane permeabilization induced throughout high gravity fermentation. Conclusions PDR18 gene over-expression increases yeast ethanol tolerance and fermentation performance leading to the production of highly inhibitory concentrations of ethanol. PDR18 overexpression in industrial yeast strains appears to be a promising approach to improve alcoholic fermentation performance for sustainable bio-ethanol production. PMID:22839110

Simultaneous Saccharification and Fermentation and Partial Saccharification and Co-Fermentation of Lignocellulosic Biomass for Ethanol Production

NASA Astrophysics Data System (ADS)

Doran-Peterson, Joy; Jangid, Amruta; Brandon, Sarah K.; Decrescenzo-Henriksen, Emily; Dien, Bruce; Ingram, Lonnie O.

Ethanol production by fermentation of lignocellulosic biomass-derived sugars involves a fairly ancient art and an ever-evolving science. Production of ethanol from lignocellulosic biomass is not avant-garde, and wood ethanol plants have been in existence since at least 1915. Most current ethanol production relies on starch- and sugar-based crops as the substrate; however, limitations of these materials and competing value for human and animal feeds is renewing interest in lignocellulose conversion. Herein, we describe methods for both simultaneous saccharification and fermentation (SSF) and a similar but separate process for partial saccharification and cofermentation (PSCF) of lignocellulosic biomass for ethanol production using yeasts or pentose-fermenting engineered bacteria. These methods are applicable for small-scale preliminary evaluations of ethanol production from a variety of biomass sources.

Presence of glucose, xylose, and glycerol fermenting bacteria in the deep biosphere of the former Homestake gold mine, South Dakota

PubMed Central

Rastogi, Gurdeep; Gurram, Raghu N.; Bhalla, Aditya; Gonzalez, Ramon; Bischoff, Kenneth M.; Hughes, Stephen R.; Kumar, Sudhir; Sani, Rajesh K.

2012-01-01

Eight fermentative bacterial strains were isolated from mixed enrichment cultures of a composite soil sample collected at 1.34 km depth from the former Homestake gold mine in Lead, SD, USA. Phylogenetic analysis of their 16S rRNA gene sequences revealed that these isolates were affiliated with the phylum Firmicutes belonging to genera Bacillus and Clostridium. Batch fermentation studies demonstrated that isolates had the ability to ferment glucose, xylose, or glycerol to industrially valuable products such as ethanol and 1,3-propanediol (PDO). Ethanol was detected as the major fermentation end product in glucose-fermenting cultures at pH 10 with yields of 0.205–0.304 g of ethanol/g of glucose. While a xylose-fermenting strain yielded 0.189 g of ethanol/g of xylose and 0.585 g of acetic acid/g of xylose at the end of fermentation. At pH 7, glycerol-fermenting isolates produced PDO (0.323–0.458 g of PDO/g of glycerol) and ethanol (0.284–0.350 g of ethanol/g of glycerol) as major end products while acetic acid and succinic acid were identified as minor by-products in fermentation broths. These results suggest that the deep biosphere of the former Homestake gold mine harbors bacterial strains which could be used in bio-based production of ethanol and PDO. PMID:23919089

Repeated-batch fermentations of xylose and glucose-xylose mixtures using a respiration-deficient Saccharomyces cerevisiae engineered for xylose metabolism.

PubMed

Kim, Soo Rin; Lee, Ki-Sung; Choi, Jin-Ho; Ha, Suk-Jin; Kweon, Dae-Hyuk; Seo, Jin-Ho; Jin, Yong-Su

2010-11-01

Xylose-fermenting Saccharomyces strains are needed for commercialization of ethanol production from lignocellulosic biomass. Engineered Saccharomyces cerevisiae strains expressing XYL1, XYL2 and XYL3 from Pichia stipitis, however, utilize xylose in an oxidative manner, which results in significantly lower ethanol yields from xylose as compared to glucose. As such, we hypothesized that reconfiguration of xylose metabolism from oxidative into fermentative manner might lead to efficient ethanol production from xylose. To this end, we generated a respiration-deficient (RD) mutant in order to enforce engineered S. cerevisiae to utilize xylose only through fermentative metabolic routes. Three different repeated-batch fermentations were performed to characterize characteristics of the respiration-deficient mutant. When fermenting glucose as a sole carbon source, the RD mutant exhibited near theoretical ethanol yields (0.46 g g(-1)) during repeated-batch fermentations by recycling the cells. As the repeated-batch fermentation progressed, the volumetric ethanol productivity increased (from 7.5 to 8.3 g L(-1)h(-1)) because of the increased biomass from previous cultures. On the contrary, the mutant showed decreasing volumetric ethanol productivities during the repeated-batch fermentations using xylose as sole carbon source (from 0.4 to 0.3 g L(-1)h(-1)). The mutant did not grow on xylose and lost fermenting ability gradually, indicating that the RD mutant cannot maintain a good fermenting ability on xylose as a sole carbon source. However, the RD mutant was capable of fermenting a mixture of glucose and xylose with stable yields (0.35 g g(-1)) and productivities (0.52 g L(-1)h(-1)) during the repeated-batch fermentation. In addition, ethanol yields from xylose during the mixed sugar fermentation (0.30 g g(-1)) were higher than ethanol yields from xylose as a sole carbon source (0.21 g g(-1)). These results suggest that a strategy for increasing ethanol yield through respiration-deficiency can be applied for the fermentation of lignocellulosic hydrolyzates containing glucose and xylose. Copyright © 2010 Elsevier B.V. All rights reserved.

Suitability of anaerobic digestion effluent as process water for corn fuel ethanol fermentation.

PubMed

Wang, Ke; Zhang, Jian-Hua; Liu, Pei; Mao, Zhong-Gui

2014-01-01

A corn fuel ethanol plant integrated with anaerobic digestion treatment of thin stillage increases the net energy balance. Furthermore, the anaerobic digestion effluent (ADE) can be reused as a potential substitute for process water in the ethanol fermentation. In this study, the suitability of ADE as process water for corn ethanol fermentation was investigated by analyzing the potential inhibitory components in the ADE. It was found that ammonium influenced the growth and metabolism of Saccharomyces cerevisiae. Maximum ethanol production was obtained when the concentration of ammonium nitrogen was 200 mg/L, and ammonium could replace urea as the nitrogen source for S. cerevisiae under this concentration. In the ethanol fermentation with a higher concentration of ammonium, more glycerol was produced, thereby resulting in the decrease of ethanol production. In addition, components except ammonium in the ADE caused no inhibition to ethanol production. These results suggest that ADE could be reused as process water for corn ethanol fermentation without negative effect when ammonium concentration is well controlled.

Ethanol production by fermentation using immobilized cells of Saccharomyces cerevisiae in cashew apple bagasse.

PubMed

Pacheco, Alexandre Monteiro; Gondim, Diego Romão; Gonçalves, Luciana Rocha Barros

2010-05-01

In this work, cashew apple bagasse (CAB) was used for Saccharomyces cerevisiae immobilization. The support was prepared through a treatment with a solution of 3% HCl, and delignification with 2% NaOH was also conducted. Optical micrographs showed that high populations of yeast cells adhered to pre-treated CAB surface. Ten consecutive fermentations of cashew apple juice for ethanol production were carried out using immobilized yeasts. High ethanol productivity was observed from the third fermentation assay until the tenth fermentation. Ethanol concentrations (about 19.82-37.83 g L(-1) in average value) and ethanol productivities (about 3.30-6.31 g L(-1) h(-1)) were high and stable, and residual sugar concentrations were low in almost all fermentations (around 3.00 g L(-1)) with conversions ranging from 44.80% to 96.50%, showing efficiency (85.30-98.52%) and operational stability of the biocatalyst for ethanol fermentation. Results showed that cashew apple bagasse is an efficient support for cell immobilization aiming at ethanol production.

Kinetic model of continuous ethanol fermentation in closed-circulating process with pervaporation membrane bioreactor by Saccharomyces cerevisiae.

PubMed

Fan, Senqing; Chen, Shiping; Tang, Xiaoyu; Xiao, Zeyi; Deng, Qing; Yao, Peina; Sun, Zhaopeng; Zhang, Yan; Chen, Chunyan

2015-02-01

Unstructured kinetic models were proposed to describe the principal kinetics involved in ethanol fermentation in a continuous and closed-circulating fermentation (CCCF) process with a pervaporation membrane bioreactor. After ethanol was removed in situ from the broth by the membrane pervaporation, the secondary metabolites accumulated in the broth became the inhibitors to cell growth. The cell death rate related to the deterioration of the culture environment was described as a function of the cell concentration and fermentation time. In CCCF process, 609.8 g L(-1) and 750.1 g L(-1) of ethanol production were obtained in the first run and second run, respectively. The modified Gompertz model, correlating the ethanol production with the fermentation period, could be used to describe the ethanol production during CCCF process. The fitting results by the models showed good agreement with the experimental data. These models could be employed for the CCCF process technology development for ethanol fermentation. Copyright © 2014 Elsevier Ltd. All rights reserved.

Development and application of co-culture for ethanol production by co-fermentation of glucose and xylose: a systematic review.

PubMed

Chen, Yanli

2011-05-01

This article reviews current co-culture systems for fermenting mixtures of glucose and xylose to ethanol. Thirty-five co-culture systems that ferment either synthetic glucose and xylose mixture or various biomass hydrolysates are examined. Strain combinations, fermentation modes and conditions, and fermentation performance for these co-culture systems are compared and discussed. It is noted that the combination of Pichia stipitis with Saccharomyces cerevisiae or its respiratory-deficient mutant is most commonly used. One of the best results for fermentation of glucose and xylose mixture is achieved by using co-culture of immobilized Zymomonas mobilis and free cells of P. stipitis, giving volumetric ethanol production of 1.277 g/l/h and ethanol yield of 0.49-0.50 g/g. The review discloses that, as a strategy for efficient conversion of glucose and xylose, co-culture fermentation for ethanol production from lignocellulosic biomass can increase ethanol yield and production rate, shorten fermentation time, and reduce process costs, and it is a promising technology although immature.

High ethanol producing derivatives of Thermoanaerobacter ethanolicus

DOEpatents

Ljungdahl, L.G.; Carriera, L.H.

1983-05-24

Derivatives of the newly discovered microorganism Thermoanaerobacter ethanolicus which under anaerobic and thermophilic conditions continuously ferment substrates such as starch, cellobiose, glucose, xylose and other sugars to produce recoverable amounts of ethanol solving the problem of fermentations yielding low concentrations of ethanol using the parent strain of the microorganism Thermoanaerobacter ethanolicus are disclosed. These new derivatives are ethanol tolerant up to 10% (v/v) ethanol during fermentation. The process includes the use of an aqueous fermentation medium, containing the substrate at a substrate concentration greater than 1% (w/v).

High ethanol producing derivatives of Thermoanaerobacter ethanolicus

DOEpatents

Ljungdahl, Lars G.; Carriera, Laura H.

1983-01-01

Derivatives of the newly discovered microorganism Thermoanaerobacter ethanolicus which under anaerobic and thermophilic conditions continuously ferment substrates such as starch, cellobiose, glucose, xylose and other sugars to produce recoverable amounts of ethanol solving the problem of fermentations yielding low concentrations of ethanol using the parent strain of the microorganism Thermoanaerobacter ethanolicus are disclosed. These new derivatives are ethanol tolerant up to 10% (v/v) ethanol during fermentation. The process includes the use of an aqueous fermentation medium, containing the substrate at a substrate concentration greater than 1% (w/v).

Microbial contamination of fuel ethanol fermentations.

PubMed

Beckner, M; Ivey, M L; Phister, T G

2011-10-01

Microbial contamination is a pervasive problem in any ethanol fermentation system. These infections can at minimum affect the efficiency of the fermentation and at their worse lead to stuck fermentations causing plants to shut down for cleaning before beginning anew. These delays can result in costly loss of time as well as lead to an increased cost of the final product. Lactic acid bacteria (LAB) are the most common bacterial contaminants found in ethanol production facilities and have been linked to decreased ethanol production during fermentation. Lactobacillus sp. generally predominant as these bacteria are well adapted for survival under high ethanol, low pH and low oxygen conditions found during fermentation. It has been generally accepted that lactobacilli cause inhibition of Saccharomyces sp. and limit ethanol production through two basic methods; either production of lactic and acetic acids or through competition for nutrients. However, a number of researchers have demonstrated that these mechanisms may not completely account for the amount of loss observed and have suggested other means by which bacteria can inhibit yeast growth and ethanol production. While LAB are the primary contaminates of concern in industrial ethanol fermentations, wild yeast may also affect the productivity of these fermentations. Though many yeast species have the ability to thrive in a fermentation environment, Dekkera bruxellensis has been repeatedly targeted and cited as one of the main contaminant yeasts in ethanol production. Though widely studied for its detrimental effects on wine, the specific species-species interactions between D. bruxellensis and S. cerevisiae are still poorly understood. © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

Ethanol production by Saccharomyces cerevisiae using lignocellulosic hydrolysate from Chrysanthemum waste degradation.

PubMed

Quevedo-Hidalgo, Balkys; Monsalve-Marín, Felipe; Narváez-Rincón, Paulo César; Pedroza-Rodríguez, Aura Marina; Velásquez-Lozano, Mario Enrique

2013-03-01

Ethanol production derived from Saccharomyces cerevisiae fermentation of a hydrolysate from floriculture waste degradation was studied. The hydrolysate was produced from Chrysanthemum (Dendranthema grandiflora) waste degradation by Pleurotus ostreatus and characterized to determine the presence of compounds that may inhibit fermentation. The products of hydrolysis confirmed by HPLC were cellobiose, glucose, xylose and mannose. The hydrolysate was fermented by S. cerevisiae, and concentrations of biomass, ethanol, and glucose were determined as a function of time. Results were compared to YGC modified medium (yeast extract, glucose and chloramphenicol) fermentation. Ethanol yield was 0.45 g g(-1), 88 % of the maximal theoretical value. Crysanthemum waste hydrolysate was suitable for ethanol production, containing glucose and mannose with adequate nutrients for S. cerevisiae fermentation and low fermentation inhibitor levels.

Fuel ethanol from raw corn by Aspergilli hydrolysis with concurrent yeast fermentation

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

Weller, C.L.; Steinberg, M.P.; Rodda, E.D.

Crude amylase preparations were produced by growing Aspergillus awamori and A. niger on raw ground whole corn. These Koji preparations were used to hydrolyze the starch of raw ground whole corn to sugars during simultaneous fermentation of the sugars to ethanol by distillers active dry yeast. Ethanol concentrations of the fermentation beers were determined with gas chromatography. These fermentations yielded an average of 89.6% theoretical ethanol compared to control, conventional, fermentations that had an average of 89.9%. Carbon dioxide evolutions were determined with use of Alwood valves. Both the Koji and conventional fermentations produced an average of 0.48 g ofmore » carbon dioxide per gram of dry substrate starch within 72 h. However, initially the conventional fermentation rate was greater. Koji dehydrated at 41/sup 0/C had no apparent detrimental effects on theoretical ethanol yield. 41 references, 1 figure, 2 tables.« less

Combined enzyme mediated fermentation of cellulous and xylose to ethanol by Schizosaccharoyces pombe, cellulase, .beta.-glucosidase, and xylose isomerase

DOEpatents

Lastick, Stanley M.; Mohagheghi, Ali; Tucker, Melvin P.; Grohmann, Karel

1994-01-01

A process for producing ethanol from mixed sugar streams from pretreated biomass comprising xylose and cellulose using enzymes to convert these substrates to fermentable sugars; selecting and isolating a yeast Schizosaccharomyces pombe ATCC No. 2476, having the ability to ferment these sugars as they are being formed to produce ethanol; loading the substrates with the fermentation mix composed of yeast, enzymes and substrates; fermenting the loaded substrates and enzymes under anaerobic conditions at a pH range of between about 5.0 to about 6.0 and at a temperature range of between about 35.degree. C. to about 40.degree. C. until the fermentation is completed, the xylose being isomerized to xylulose, the cellulose being converted to glucose, and these sugars being concurrently converted to ethanol by yeast through means of the anaerobic fermentation; and recovering the ethanol.

Combined enzyme mediated fermentation of cellulose and xylose to ethanol by Schizosaccharomyces pombe, cellulase, [beta]-glucosidase, and xylose isomerase

DOEpatents

Lastick, S.M.; Mohagheghi, A.; Tucker, M.P.; Grohmann, K.

1994-12-13

A process for producing ethanol from mixed sugar streams from pretreated biomass comprising xylose and cellulose using enzymes to convert these substrates to fermentable sugars; selecting and isolating a yeast Schizosaccharomyces pombe ATCC No. 2476, having the ability to ferment these sugars as they are being formed to produce ethanol; loading the substrates with the fermentation mix composed of yeast, enzymes and substrates; fermenting the loaded substrates and enzymes under anaerobic conditions at a pH range of between about 5.0 to about 6.0 and at a temperature range of between about 35 C to about 40 C until the fermentation is completed, the xylose being isomerized to xylulose, the cellulose being converted to glucose, and these sugars being concurrently converted to ethanol by yeast through means of the anaerobic fermentation; and recovering the ethanol. 2 figures.

Evaluation of fermentation kinetics of acid-treated corn cob hydrolysate for xylose fermentation in the presence of acetic acid by Pichia stipitis.

PubMed

Kashid, Mohan; Ghosalkar, Anand

2017-08-01

The efficient utilization of lignocellulosic biomass for ethanol production depends on the fermentability of the biomass hydrolysate obtained after pretreatment. In this work we evaluated the kinetics of ethanol production from xylose using Pichia stipitis in acid-treated corn cob hydrolysate. Acetic acid is one of the main inhibitors in corn cob hydrolysate that negatively impacts kinetics of xylose fermentation by P. stipitis. Unstructured kinetic model has been formulated that describes cell mass growth and ethanol production as a function of xylose, oxygen, ethanol, and acetic acid concentration. Kinetic parameters were estimated under different operating conditions affecting xylose fermentation. This is the first report on kinetics of xylose fermentation by P. stipitis which includes inhibition of acetic acid on growth and product formation. In the presence of acetic acid in the hydrolysate, the model accurately predicted reduction in maximum specific growth rate (from 0.23 to 0.15 h -1 ) and increase in ethanol yield per unit biomass (from 3 to 6.2 gg -1 ), which was also observed during experimental trials. Presence of acetic acid in the fermentation led to significant reduction in the cell growth rate, reduction in xylose consumption and ethanol production rate. The developed model accurately described physiological state of P. stipitis during corn cob hydrolysate fermentation. Proposed model can be used to predict the influence of xylose, ethanol, oxygen, and acetic acid concentration on cell growth and ethanol productivity in industrial fermentation.

Sequential Mixed Cultures: From Syngas to Malic Acid

PubMed Central

Oswald, Florian; Dörsam, Stefan; Veith, Nicolas; Zwick, Michaela; Neumann, Anke; Ochsenreither, Katrin; Syldatk, Christoph

2016-01-01

Synthesis gas (syngas) fermentation using acetogenic bacteria is an approach for production of bulk chemicals like acetate, ethanol, butanol, or 2,3-butandiol avoiding the fuel vs. food debate by using carbon monoxide, carbon dioxide, and hydrogen from gasification of biomass or industrial waste gases. Suffering from energetic limitations, yields of C4-molecules produced by syngas fermentation are quite low compared with ABE fermentation using sugars as a substrate. On the other hand, fungal production of malic acid has high yields of product per gram metabolized substrate but is currently limited to sugar containing substrates. In this study, it was possible to show that Aspergilus oryzae is able to produce malic acid using acetate as sole carbon source which is a main product of acetogenic syngas fermentation. Bioreactor cultivations were conducted in 2.5 L stirred tank reactors. During the syngas fermentation part of the sequential mixed culture, Clostridium ljungdahlii was grown in modified Tanner medium and sparged with 20 mL/min of artificial syngas mimicking a composition of clean syngas from entrained bed gasification of straw (32.5 vol-% CO, 32.5 vol-% H2, 16 vol-% CO2, and 19 vol-% N2) using a microsparger. Syngas consumption was monitored via automated gas chromatographic measurement of the off-gas. For the fungal fermentation part gas sparging was switched to 0.6 L/min of air and a standard sparger. Ammonia content of medium for syngas fermentation was reduced to 0.33 g/L NH4Cl to meet the requirements for fungal production of dicarboxylic acids. Malic acid production performance of A. oryzae in organic acid production medium and syngas medium with acetate as sole carbon source was verified and gave YP∕S values of 0.28 g/g and 0.37 g/g respectively. Growth and acetate formation of C. ljungdahlii during syngas fermentation were not affected by the reduced ammonia content and 66 % of the consumed syngas was converted to acetate. The overall conversion of CO and H2 into malic acid was calculated to be 3.5 g malic acid per mol of consumed syngas or 0.22 g malic acid per gram of syngas. PMID:27445993

Effects of fermentation substrate conditions on corn-soy co-fermentation for fuel ethanol production.

PubMed

Yao, Linxing; Lee, Show-Ling; Wang, Tong; de Moura, Juliana M L N; Johnson, Lawrence A

2012-09-01

Soy skim, a protein-rich liquid co-product from the aqueous extraction of soybeans, was co-fermented with corn to produce ethanol. Effects of soy skim addition level, type of skim, corn particle size, water-to-solids ratio, and urea on co-fermentation were determined. The addition of 20-100% skim increased the fermentation rate by 18-27% and shortened the fermentation time by 5-7h without affecting ethanol yield. Finely ground corn or high water-to-solids ratio (≥ 3.0) in the mash gave higher fermentation rates, but did not increase the ethanol yield. When the water was completely replaced with soy skim, the addition of urea became unnecessary. Soy skim retentate that was concentrated by nanofiltration increased fermentation rate by 25%. The highest level of skim addition resulted in a finished beer with 16% solids, 47% protein (dwb) containing 3.6% lysine, and an ethanol yield of 39 g/100g dry corn. Copyright © 2012 Elsevier Ltd. All rights reserved.

Separate hydrolysis and co-fermentation for improved xylose utilization in integrated ethanol production from wheat meal and wheat straw

PubMed Central

2012-01-01

Background The commercialization of second-generation bioethanol has not been realized due to several factors, including poor biomass utilization and high production cost. It is generally accepted that the most important parameters in reducing the production cost are the ethanol yield and the ethanol concentration in the fermentation broth. Agricultural residues contain large amounts of hemicellulose, and the utilization of xylose is thus a plausible way to improve the concentration and yield of ethanol during fermentation. Most naturally occurring ethanol-fermenting microorganisms do not utilize xylose, but a genetically modified yeast strain, TMB3400, has the ability to co-ferment glucose and xylose. However, the xylose uptake rate is only enhanced when the glucose concentration is low. Results Separate hydrolysis and co-fermentation of steam-pretreated wheat straw (SPWS) combined with wheat-starch hydrolysate feed was performed in two separate processes. The average yield of ethanol and the xylose consumption reached 86% and 69%, respectively, when the hydrolysate of the enzymatically hydrolyzed (18.5% WIS) unwashed SPWS solid fraction and wheat-starch hydrolysate were fed to the fermentor after 1 h of fermentation of the SPWS liquid fraction. In the other configuration, fermentation of the SPWS hydrolysate (7.0% WIS), resulted in an average ethanol yield of 93% from fermentation based on glucose and xylose and complete xylose consumption when wheat-starch hydrolysate was included in the feed. Increased initial cell density in the fermentation (from 5 to 20 g/L) did not increase the ethanol yield, but improved and accelerated xylose consumption in both cases. Conclusions Higher ethanol yield has been achieved in co-fermentation of xylose and glucose in SPWS hydrolysate when wheat-starch hydrolysate was used as feed, then in co-fermentation of the liquid fraction of SPWS fed with the mixed hydrolysates. Integration of first-generation and second-generation processes also increases the ethanol concentration, resulting in a reduction in the cost of the distillation step, thus improving the process economics. PMID:22410131

Efficient production of ethanol from waste paper and the biochemical methane potential of stillage eluted from ethanol fermentation.

PubMed

Nishimura, Hiroto; Tan, Li; Sun, Zhao-Yong; Tang, Yue-Qin; Kida, Kenji; Morimura, Shigeru

2016-02-01

Waste paper can serve as a feedstock for ethanol production due to being rich in cellulose and not requiring energy-intensive thermophysical pretreatment. In this study, an efficient process was developed to convert waste paper to ethanol. To accelerate enzymatic saccharification, pH of waste paper slurry was adjusted to 4.5-5.0 with H2SO4. Presaccharification and simultaneous saccharification and fermentation (PSSF) with enzyme loading of 40 FPU/g waste paper achieved an ethanol yield of 91.8% and productivity of 0.53g/(Lh) with an ethanol concentration of 32g/L. Fed-batch PSSF was used to decrease enzyme loading to 13 FPU/g waste paper by feeding two separate batches of waste paper slurry. Feeding with 20% w/w waste paper slurry increased ethanol concentration to 41.8g/L while ethanol yield decreased to 83.8%. To improve the ethanol yield, presaccharification was done prior to feeding and resulted in a higher ethanol concentration of 45.3g/L, a yield of 90.8%, and productivity of 0.54g/(Lh). Ethanol fermentation recovered 33.2% of the energy in waste paper as ethanol. The biochemical methane potential of the stillage eluted from ethanol fermentation was 270.5mL/g VTS and 73.0% of the energy in the stillage was recovered as methane. Integrating ethanol fermentation with methane fermentation, recovered a total of 80.4% of the energy in waste paper as ethanol and methane. Copyright © 2015 Elsevier Ltd. All rights reserved.

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.

An economic comparison of different fermentation configurations to convert corn stover to ethanol using Z. mobilis and Saccharomyces.

PubMed

Dutta, Abhijit; Dowe, Nancy; Ibsen, Kelly N; Schell, Daniel J; Aden, Andy

2010-01-01

Numerous routes are being explored to lower the cost of cellulosic ethanol production and enable large-scale production. One critical area is the development of robust cofermentative organisms to convert the multiple, mixed sugars found in biomass feedstocks to ethanol at high yields and titers without the need for processing to remove inhibitors. Until such microorganisms are commercialized, the challenge is to design processes that exploit the current microorganisms' strengths. This study explored various process configurations tailored to take advantage of the specific capabilities of three microorganisms, Z. mobilis 8b, S. cerevisiae, and S. pastorianus. A technoeconomic study, based on bench-scale experimental data generated by integrated process testing, was completed to understand the resulting costs of the different process configurations. The configurations included whole slurry fermentation with a coculture, and separate cellulose simultaneous saccharification and fermentation (SSF) and xylose fermentations with none, some or all of the water to the SSF replaced with the fermented liquor from the xylose fermentation. The difference between the highest and lowest ethanol cost for the different experimental process configurations studied was $0.27 per gallon ethanol. Separate fermentation of solid and liquor streams with recycle of fermented liquor to dilute the solids gave the lowest ethanol cost, primarily because this option achieved the highest concentrations of ethanol after fermentation. Further studies, using methods similar to ones employed here, can help understand and improve the performance and hence the economics of integrated processes involving enzymes and fermentative microorganisms.

Relationship of trehalose accumulation with ethanol fermentation in industrial Saccharomyces cerevisiae yeast strains.

PubMed

Wang, Pin-Mei; Zheng, Dao-Qiong; Chi, Xiao-Qin; Li, Ou; Qian, Chao-Dong; Liu, Tian-Zhe; Zhang, Xiao-Yang; Du, Feng-Guang; Sun, Pei-Yong; Qu, Ai-Min; Wu, Xue-Chang

2014-01-01

The protective effect and the mechanisms of trehalose accumulation in industrial Saccharomyces cerevisiae strains were investigated during ethanol fermentation. The engineered strains with more intercellular trehalose achieved significantly higher fermentation rates and ethanol yields than their wild strain ZS during very high gravity (VHG) fermentation, while their performances were not different during regular fermentation. The VHG fermentation performances of these strains were consistent with their growth capacity under osmotic stress and ethanol stress, the key stress factors during VHG fermentation. These results suggest that trehalose accumulation is more important for VHG fermentation of industrial yeast strains than regular one. The differences in membrane integrity and antioxidative capacity of these strains indicated the possible mechanisms of trehalose as a protectant under VHG condition. Therefore, trehalose metabolic engineering may be a useful strategy for improving the VHG fermentation performance of industrial yeast strains. Copyright © 2013 Elsevier Ltd. All rights reserved.

Fuel ethanol from raw corn

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

Weller, C.L.; Rodda, E.D.; Steinberg, M.P.

Crude amylase preparations were produced by growing Aspergillus awamori and A. niger on raw ground whole corn. These Koji preparations were used to hydrolyze the starch of raw ground whole corn to sugars during simultaneous fermentation of the sugars to ethanol by distillers active dry yeast. Ethanol concentrations of the fermentation beers were determined with gas-chromatography. These fermentations yielded an average of 89.6% theoretical ethanol compared to control, conventional, fermentations that had an average of 89.8%. Carbon dioxide evolutions were determined with use of Alwood valves. Both the Koji and conventional fermentations produced an average of 0.48 gram of carbonmore » dioxide per gram of dry substrate starch within 72 hours. However, initially the conventional fermentation rate was greater. Koji dehydrated at 41 degrees C had no apparent detrimental effects on theoretical ethanol yield.« less

Fuel ethanol from raw corn

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

Weller, C.L.; Rodda, E.D.; Steinberg, M.P.

Crude amylase preparations were produced by growing Aspergillus awamori and A. niger on raw ground whole corn. These Koji preparations were used to hydrolyze the starch of raw ground whole corn to sugars during simultaneous fermentation of the sugars to ethanol by distillers active dry yeast. Ethanol concentrations of the fermentation beers were determined with gas-chromatography. These fermentations yielded an average of 89.6% theoretical ethanol compared to control, conventional, fermentations that had an average of 89.8%. Carbon dioxide evolutions were determined with use of Alwood valves. Both the Koji and conventional fermentations produced an average of 0.48 gram of carbonmore » dioxide per gram of dry substrate starch within 72 hours. However, initially the conventional fermentation rate was greater. Koji dehydrated at 41/sup 0/C had no apparent detrimental effects on theoretical ethanol yield.« less

Characteristics of an immobilized yeast cell system using very high gravity for the fermentation of ethanol.

PubMed

Ji, Hairui; Yu, Jianliang; Zhang, Xu; Tan, Tianwei

2012-09-01

The characteristics of ethanol production by immobilized yeast cells were investigated for both repeated batch fermentation and continuous fermentation. With an initial sugar concentration of 280 g/L during the repeated batch fermentation, more than 98% of total sugar was consumed in 65 h with an average ethanol concentration and ethanol yield of 130.12 g/L and 0.477 g ethanol/g consumed sugar, respectively. The immobilized yeast cell system was reliable for at least 10 batches and for a period of 28 days without accompanying the regeneration of Saccharomyces cerevisiae inside the carriers. The multistage continuous fermentation was carried out in a five-stage column bioreactor with a total working volume of 3.75 L. The bioreactor was operated for 26 days at a dilution rate of 0.015 h(-1). The ethanol concentration of the effluent reached 130.77 g/L ethanol while an average 8.18 g/L residual sugar remained. Due to the high osmotic pressure and toxic ethanol, considerable yeast cells died without regeneration, especially in the last two stages, which led to the breakdown of the whole system of multistage continuous fermentation.

75 FR 16388 - Approval and Promulgation of Implementation Plans; Commonwealth of Kentucky: Prevention of...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-04-01

... ``chemical process plants'' that produce ethanol through a natural fermentation process (hereafter referred... for excluding ``chemical process plants'' that produce ethanol through a natural fermentation process... facilities that produce ethanol by natural fermentation processes. Kentucky's February 5, 2010, SIP...

Ethanol fermentation characteristics of recycled water by Saccharomyces cerevisiae in an integrated ethanol-methane fermentation process.

PubMed

Yang, Xinchao; Wang, Ke; Wang, Huijun; Zhang, Jianhua; Mao, Zhonggui

2016-11-01

An process of integrated ethanol-methane fermentation with improved economics has been studied extensively in recent years, where the process water used for a subsequent fermentation of carbohydrate biomass is recycled. This paper presents a systematic study of the ethanol fermentation characteristics of recycled process water. Compared with tap water, fermentation time was shortened by 40% when mixed water was employed. However, while the maximal ethanol production rate increased from 1.07g/L/h to 2.01g/L/h, ethanol production was not enhanced. Cell number rose from 0.6×10(8) per mL in tap water to 1.6×10(8) per mL in mixed water but although biomass increased, cell morphology was not affected. Furthermore, the use of mixed water increased the glycerol yield but decreased that of acetic acid, and the final pH with mixed water was higher than when using tap water. Copyright © 2016 Elsevier Ltd. All rights reserved.

Effect of Different Silage Storing Conditions on the Oxygen Concentration in the Silo and Fermentation Quality of Rice.

PubMed

Uegaki, Ryuichi; Kawano, Kazuo; Ohsawa, Ryo; Kimura, Toshiyuki; Yamamura, Kohji

2017-06-21

We investigated the effects of different silage storing conditions on the oxygen concentration in the silo and fermentation quality of rice (Oryza sativa L.). Forage rice was ensiled in bottles (with or without space at the bottlemouth, with solid or pinhole cap, and with oxygen scavenger, ethanol transpiration agent, oxygen scavenger and ethanol transpiration agent, or no adjuvant) and stored for 57 days. The oxygen concentration decreased with the addition of the oxygen scavenger and increased with that of the ethanol transpiration agent. The oxygen scavenger facilitated silage fermentation and fungus generation, whereas the ethanol transpiration agent suppressed silage fermentation and fungus generation. However, the combined use of the oxygen scavenger and ethanol transpiration agent facilitated silage fermentation and also suppressed fungus generation. Overall, this study revealed the negative effects of oxygen on the internal silo and the positive effects of the combined use of the oxygen scavenger and ethanol transpiration agent on silage fermentation quality.

Influence of fiber degradation and concentration of fermentable sugars on simultaneous saccharification and fermentation of high-solids spruce slurry to ethanol.

PubMed

Hoyer, Kerstin; Galbe, Mats; Zacchi, Guido

2013-10-08

Saccharification and fermentation of pretreated lignocellulosic materials, such as spruce, should be performed at high solids contents in order to reduce the cost of the produced bioethanol. However, this has been shown to result in reduced ethanol yields or a complete lack of ethanol production. Previous studies have shown inconsistent results when prehydrolysis is performed at a higher temperature prior to the simultaneous saccharification and fermentation (SSF) of steam-pretreated lignocellulosic materials. In some cases, a significant increase in overall ethanol yield was reported, while in others, a slight decrease in ethanol yield was observed. In order to investigate the influence of prehydrolysis on high-solids SSF of steam-pretreated spruce slurry, in the present study, the presence of fibers and inhibitors, degree of fiber degradation and initial fermentable sugar concentration has been studied. SSF of whole steam-pretreated spruce slurry at a solids content of 13.7% water-insoluble solids (WIS) resulted in a very low overall ethanol yield, mostly due to poor fermentation. The yeast was, however, able to ferment the washed slurry and the liquid fraction of the pretreated slurry. Performing prehydrolysis at 48°C for 22 hours prior to SSF of the whole pretreated slurry increased the overall ethanol yield from 3.9 to 62.1%. The initial concentration of fermentable sugars in SSF could not explain the increase in ethanol yield in SSF with prehydrolysis. Although the viscosity of the material did not appear to decrease significantly during prehydrolysis, the degradation of the fibers prior to the addition of the yeast had a positive effect on ethanol yield when using whole steam-pretreated spruce slurry. The results of the present study suggest that the increase in ethanol yield from SSF when performing prehydrolysis is a result of fiber degradation rather than a decrease in viscosity. The increased concentration of fermentable sugars at the beginning of the fermentation phase in SSF following prehydrolysis did not affect the overall ethanol yield in the present study.

76 FR 36875 - Approval and Promulgation of Implementation Plans; South Carolina: Prevention of Significant...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-06-23

... ethanol through a natural fermentation process (hereafter referred to as the ``Ethanol Rule'') from the... exception of the phrase ``except ethanol production facilities producing ethanol by natural fermentation...

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

PubMed

Xu, Lei; Tschirner, Ulrike

2014-08-01

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

Xylose-fermenting Pichia stipitis by genome shuffling for improved ethanol production.

PubMed

Shi, Jun; Zhang, Min; Zhang, Libin; Wang, Pin; Jiang, Li; Deng, Huiping

2014-03-01

Xylose fermentation is necessary for the bioconversion of lignocellulose to ethanol as fuel, but wild-type Saccharomyces cerevisiae strains cannot fully metabolize xylose. Several efforts have been made to obtain microbial strains with enhanced xylose fermentation. However, xylose fermentation remains a serious challenge because of the complexity of lignocellulosic biomass hydrolysates. Genome shuffling has been widely used for the rapid improvement of industrially important microbial strains. After two rounds of genome shuffling, a genetically stable, high-ethanol-producing strain was obtained. Designated as TJ2-3, this strain could ferment xylose and produce 1.5 times more ethanol than wild-type Pichia stipitis after fermentation for 96 h. The acridine orange and propidium iodide uptake assays showed that the maintenance of yeast cell membrane integrity is important for ethanol fermentation. This study highlights the importance of genome shuffling in P. stipitis as an effective method for enhancing the productivity of industrial strains. © 2013 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

Solid-phase fermentation and juice expression systems for sweet sorghum

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

Bryan, W.L.; Monroe, G.E.; Caussariel, P.M.

1985-01-01

Two systems to recover fermented juice from variety M 81E sweet sorghum stalks that contained about 11% fermentable sugar were compared. (a) Stalks with leaves and tops removed were chopped and inoculated with 0.2% yeast in a forage harvester, stored under anaerobic conditions for 75 hours in insulated fermentors and pressed in a screw press to recover fermented juice (5-6% ethanol). (b) Mechanically harvested sweet sorghum billets (30 cm length) without leaves or seed heads were shredded and milled in a 3-roll mill; and bagasse was inoculated with 0.2% yeast, fermented for 100 h and pressed to recover fermented juicemore » (4 to 5% ethanol). Potential ethanol yields were 75% of theoretical for the forage harvest system and 78% for the shredder mill system, based on 95% of theoretical ethanol yield from juice expressed during milling and no loss of ethanol during fermentation, handling and pressing in the screw press. 20 references.« less

Diffusion-driven proton exchange membrane fuel cell for converting fermenting biomass to electricity.

PubMed

Malati, P; Mehrotra, P; Minoofar, P; Mackie, D M; Sumner, J J; Ganguli, R

2015-10-01

A membrane-integrated proton exchange membrane fuel cell that enables in situ fermentation of sugar to ethanol, diffusion-driven separation of ethanol, and its catalytic oxidation in a single continuous process is reported. The fuel cell consists of a fermentation chamber coupled to a direct ethanol fuel cell. The anode and fermentation chambers are separated by a reverse osmosis (RO) membrane. Ethanol generated from fermented biomass in the fermentation chamber diffuses through the RO membrane into a glucose solution contained in the DEFC anode chamber. The glucose solution is osmotically neutral to the biomass solution in the fermentation chamber preventing the anode chamber from drying out. The fuel cell sustains >1.3 mW cm(-2) at 47°C with high discharge capacity. No separate purification or dilution is necessary, resulting in an efficient and portable system for direct conversion of fermenting biomass to electricity. Copyright © 2015 Elsevier Ltd. All rights reserved.

Simultaneous hydrolysis and co-fermentation of whey lactose with wheat for ethanol production.

PubMed

Jin, Yiqiong; Parashar, Archana; Mason, Beth; Bressler, David C

2016-12-01

Whey permeate was used as a co-substrate to replace part of the wheat for ethanol production by Saccharomyces cerevisiae. The simultaneous saccharification and fermentation was achieved with β-galactosidase added at the onset of the fermentation to promote whey lactose hydrolysis. Aspergillus oryzae and Kluyveromyces lactis β-galactosidases were two enzymes selected and used in the co-fermentation of wheat and whey permeate for the comparison of their effectiveness on lactose hydrolysis. The possibility of co-fermentations in both STARGEN and jet cooking systems was investigated in 5L bioreactors. Ethanol yields from the co-fermentations of wheat and whey permeate were evaluated. It was found that A. oryzae β-galactosidase was more efficient for lactose hydrolysis during the co-fermentation and that whey permeate supplementation can contribute to ethanol yield in co-fermentations with wheat. Copyright © 2016 Elsevier Ltd. All rights reserved.

Study of Sugarcane Pieces as Yeast Supports for Ethanol Production from Sugarcane Juice and Molasses Using Newly Isolated Yeast from Toddy Sap

PubMed Central

Satyanarayana, Botcha; Balakrishnan, Kesavapillai; Raghava Rao, Tamanam; Seshagiri Rao, Gudapaty

2012-01-01

A repeated batch fermentation system was used to produce ethanol using Saccharomyces cerevisiae strain (NCIM 3640) immobilized on sugarcane (Saccharum officinarum L.) pieces. For comparison free cells were also used to produce ethanol by repeated batch fermentation. Scanning electron microscopy evidently showed that cell immobilization resulted in firm adsorption of the yeast cells within subsurface cavities, capillary flow through the vessels of the vascular bundle structure, and attachment of the yeast to the surface of the sugarcane pieces. Repeated batch fermentations using sugarcane supported biocatalyst were successfully carried out for at least ten times without any significant loss in ethanol production from sugarcane juice and molasses. The number of cells attached to the support increased during the fermentation process, and fewer yeast cells leaked into fermentation broth. Ethanol concentrations (about 72.65~76.28 g/L in an average value) and ethanol productivities (about 2.27~2.36 g/L/hr in an average value) were high and stable, and residual sugar concentrations were low in all fermentations (0.9~3.25 g/L) with conversions ranging from 98.03~99.43%, showing efficiency 91.57~95.43 and operational stability of biocatalyst for ethanol fermentation. The results of the work pertaining to the use of sugarcane as immobilized yeast support could be promising for industrial fermentations. PMID:22783132

Fermentation of Soybean Meal Hydrolyzates with Saccharomyces cerevisiae and Zymomonas mobilis for Ethanol Production.

PubMed

Luján-Rhenals, Deivis E; Morawicki, Rubén O; Gbur, Edward E; Ricke, Steven C

2015-07-01

Most of the ethanol currently produced by fermentation is derived from sugar cane, corn, or beets. However, it makes good ecological and economic sense to use the carbohydrates contained in by-products and coproducts of the food processing industry for ethanol production. Soybean meal, a co-product of the production of soybean oil, has a relatively high carbohydrate content that could be a reasonable substrate for ethanol production after fermentable sugars are released via hydrolysis. In this research, the capability of Saccharomyces cerevisiae NRRL Y-2233 and Zymomonas mobilis subsp. mobilis NRRL B-4286 to produce ethanol was evaluated using soybean meal hydrolyzates as substrates for the fermentation. These substrates were produced from the dilute-acid hydrolysis of soybean meal at 135 °C for 45 min with 0, 0.5%, 1.25%, and 2% H2 SO4 and at 120 °C for 30 min with 1.25% H2 SO4 . Kinetic parameters of the fermentation were estimated using the logistic model. Ethanol production using S. cerevisiae was highest with the substrates obtained at 135 °C, 45 min, and 0.5% H2 SO4 and fermented for 8 h, 8 g/L (4 g ethanol/100 g fresh SBM), while Z. mobilis reached its maximum ethanol production, 9.2 g/L (4.6 g ethanol/100 g fresh SBM) in the first 20 h of fermentation with the same hydrolyzate. © 2015 Institute of Food Technologists®

Homo- and heterofermentative lactobacilli differently affect sugarcane-based fuel ethanol fermentation.

PubMed

Basso, Thiago Olitta; Gomes, Fernanda Sgarbosa; Lopes, Mario Lucio; de Amorim, Henrique Vianna; Eggleston, Gillian; Basso, Luiz Carlos

2014-01-01

Bacterial contamination during industrial yeast fermentation has serious economic consequences for fuel ethanol producers. In addition to deviating carbon away from ethanol formation, bacterial cells and their metabolites often have a detrimental effect on yeast fermentative performance. The bacterial contaminants are commonly lactic acid bacteria (LAB), comprising both homo- and heterofermentative strains. We have studied the effects of these two different types of bacteria upon yeast fermentative performance, particularly in connection with sugarcane-based fuel ethanol fermentation process. Homofermentative Lactobacillus plantarum was found to be more detrimental to an industrial yeast strain (Saccharomyces cerevisiae CAT-1), when compared with heterofermentative Lactobacillus fermentum, in terms of reduced yeast viability and ethanol formation, presumably due to the higher titres of lactic acid in the growth medium. These effects were only noticed when bacteria and yeast were inoculated in equal cell numbers. However, when simulating industrial fuel ethanol conditions, as conducted in Brazil where high yeast cell densities and short fermentation time prevail, the heterofermentative strain was more deleterious than the homofermentative type, causing lower ethanol yield and out competing yeast cells during cell recycle. Yeast overproduction of glycerol was noticed only in the presence of the heterofermentative bacterium. Since the heterofermentative bacterium was shown to be more deleterious to yeast cells than the homofermentative strain, we believe our findings could stimulate the search for more strain-specific antimicrobial agents to treat bacterial contaminations during industrial ethanol fermentation.

Enzymatic hydrolysis and fermentation of corn for fuel alcohol

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

Mullins, J.T.

1985-01-01

The integration of enzyme saccharification with fermentation reduces the total time required to produce acceptable levels of ethanol. The use of a more concentrated mash (84.8 L total mash/bu corn) results in a 26.6% increase in ethanol productivity and a 21.4% increase in beer ethanol concentration compared to standard corn mash (96.6 L total mash/bu corn). Thus, the energy requirement and cost of distillation can be reduced. The addition of waste cola syrup at 30 g invert sugar/L total mash gave a 19% increase in ethanol concentration in the final beer and required only a small increase in period ofmore » fermentation. Surplus laundry starch can replace 30-50% of the weight of corn normally used in fermentation without influencing ethanol production or the time required for fermentation. Both of these waste materials reduce the unit cost of ethanol and demonstrate the value of such substances in ethanol systems.« less

Inhibitory effects of phenolic compounds of rice straw formed by saccharification during ethanol fermentation by Pichia stipitis.

PubMed

Wang, Xiahui; Tsang, Yiu Fai; Li, Yuhao; Ma, Xiubing; Cui, Shouqing; Zhang, Tian-Ao; Hu, Jiajun; Gao, Min-Tian

2017-11-01

In this study, it was found that the type of phenolic acids derived from rice straw was the major factor affecting ethanol fermentation by Pichia stipitis. The aim of this study was to investigate the inhibitory effect of phenolic acids on ethanol fermentation with rice straw. Different cellulases produced different ratios of free phenolic acids to soluble conjugated phenolic acids, resulting in different fermentation efficiencies. Free phenolic acids exhibited much higher inhibitory effect than conjugated phenolic acids. The flow cytometry results indicated that the damage to cell membranes was the primary mechanism of inhibition of ethanol fermentation by phenolic acids. The removal of free phenolic acids from the hydrolysates increased ethanol productivity by 2.0-fold, indicating that the free phenolic acids would be the major inhibitors formed during saccharification. The integrated process for ethanol and phenolic acids may constitute a new strategy for the production of low-cost ethanol. Copyright © 2017 Elsevier Ltd. All rights reserved.

75 FR 55988 - Approval and Promulgation of Implementation Plans; Commonwealth of Kentucky; Prevention of...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-09-15

... exclude facilities that produce ethanol through a natural fermentation process from the definition of... action on Kentucky's provisions to exclude facilities that produce ethanol through a natural fermentation... facilities that produce ethanol through a natural fermentation process from the definition of ``chemical...

Incorporation of whey permeate, a dairy effluent, in ethanol fermentation to provide a zero waste solution for the dairy industry.

PubMed

Parashar, Archana; Jin, Yiqiong; Mason, Beth; Chae, Michael; Bressler, David C

2016-03-01

This study proposes a novel alternative for utilization of whey permeate, a by-product stream from the dairy industry, in wheat fermentation for ethanol production using Saccharomyces cerevisiae. Whey permeates were hydrolyzed using enzymes to release fermentable sugars. Hydrolyzed whey permeates were integrated into wheat fermentation as a co-substrate or to partially replace process water. Cold starch hydrolysis-based simultaneous saccharification and fermentation was done as per the current industrial protocol for commercial wheat-to-ethanol production. Ethanol production was not affected; ethanol yield efficiency did not change when up to 10% of process water was replaced. Lactic acid bacteria in whey permeate did not negatively affect the co-fermentation or reduce ethanol yield. Whey permeate could be effectively stored for up to 4 wk at 4 °C with little change in lactose and lactic acid content. Considering the global abundance and nutrient value of whey permeate, the proposed strategy could improve economics of the dairy and biofuel sectors, and reduce environmental pollution. Furthermore, our research may be applied to fermentation strategies designed to produce value-added products other than ethanol. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Comparison of ethanol production performance in 10 varieties of sweet potato at different growth stages

NASA Astrophysics Data System (ADS)

Jin, Yanling; Fang, Yang; Zhang, Guohua; Zhou, Lingling; Zhao, Hai

2012-10-01

The performance in the ethanol production of 10 varieties of sweet potato was evaluated, and the consumption in raw materials, land occupation and fermentation waste residue in producing 1 ton of anhydrous ethanol were investigated. The comparative results between 10 varieties of sweet potato at 3 growth stages indicated that NS 007 and SS 19 were better feedstocks for ethanol production, exhibiting less feedstock consumption (6.19 and 7.59 tons/ton ethanol, respectively), the least land occupation (0.24 and 0.24 ha/ton ethanol, respectively), less fermentation waste residue (0.56 and 0.55 tons/ton ethanol, respectively), the highest level of ethanol output per unit area (4.17 and 4.17 ton/ha, respectively), and a lower viscosity of the fermentation culture (591 and 612 mPa S, respectively). The data above are average data. In most varieties, the ethanol output speed at day 130 was the highest. Therefore, NS 007 and SS 19 could be used for ethanol production and harvested after 130 days of growth from an economic point of view. In addition, the high content of fermentable sugars and low content of fiber in sweet potatoes are criteria for achieving low viscosity in ethanol fermentation cultures.

Ethanol addition enhances acid treatment to eliminate Lactobacillus fermentum from the fermentation process for fuel ethanol production.

PubMed

Costa, M A S; Cerri, B C; Ceccato-Antonini, S R

2018-01-01

Fermentation is one of the most critical steps of the fuel ethanol production and it is directly influenced by the fermentation system, selected yeast, and bacterial contamination, especially from the genus Lactobacillus. To control the contamination, the industry applies antibiotics and biocides; however, these substances can result in an increased cost and environmental problems. The use of the acid treatment of cells (water-diluted sulphuric acid, adjusted to pH 2·0-2·5) between the fermentation cycles is not always effective to combat the bacterial contamination. In this context, this study aimed to evaluate the effect of ethanol addition to the acid treatment to control the bacterial growth in a fed-batch system with cell recycling, using the industrial yeast strain Saccharomyces cerevisiae PE-2. When only the acid treatment was used, the population of Lactobacillus fermentum had a 3-log reduction at the end of the sixth fermentation cycle; however, when 5% of ethanol was added to the acid solution, the viability of the bacterium was completely lost even after the first round of cell treatment. The acid treatment +5% ethanol was able to kill L. fermentum cells without affecting the ethanol yield and with a low residual sugar concentration in the fermented must. In Brazilian ethanol-producing industry, water-diluted sulphuric acid is used to treat the cell mass at low pH (2·0) between the fermentative cycles. This procedure reduces the number of Lactobacillus fermentum from 10 7 to 10 4  CFU per ml. However, the addition of 5% ethanol to the acid treatment causes the complete loss of bacterial cell viability in fed-batch fermentation with six cell recycles. The ethanol yield and yeast cell viability are not affected. These data indicate the feasibility of adding ethanol to the acid solution replacing the antibiotic use, offering a low cost and a low amount of residue in the biomass. © 2017 The Society for Applied Microbiology.

Cellular mechanisms contributing to multiple stress tolerance in Saccharomyces cerevisiae strains with potential use in high-temperature ethanol fermentation.

PubMed

Kitichantaropas, Yasin; Boonchird, Chuenchit; Sugiyama, Minetaka; Kaneko, Yoshinobu; Harashima, Satoshi; Auesukaree, Choowong

2016-12-01

High-temperature ethanol fermentation has several benefits including a reduction in cooling cost, minimizing risk of bacterial contamination, and enabling simultaneous saccharification and fermentation. To achieve the efficient ethanol fermentation at high temperature, yeast strain that tolerates to not only high temperature but also the other stresses present during fermentation, e.g., ethanol, osmotic, and oxidative stresses, is indispensable. The C3253, C3751, and C4377 Saccharomyces cerevisiae strains, which have been previously isolated as thermotolerant yeasts, were found to be multiple stress-tolerant. In these strains, continuous expression of heat shock protein genes and intracellular trehalose accumulation were induced in response to stresses causing protein denaturation. Compared to the control strains, these multiple stress-tolerant strains displayed low intracellular reactive oxygen species levels and effective cell wall remodeling upon exposures to almost all stresses tested. In response to simultaneous multi-stress mimicking fermentation stress, cell wall remodeling and redox homeostasis seem to be the primary mechanisms required for protection against cell damage. Moreover, these strains showed better performances of ethanol production than the control strains at both optimal and high temperatures, suggesting their potential use in high-temperature ethanol fermentation.

Fuel ethanol production from sweet sorghum using repeated-batch fermentation.

PubMed

Chohnan, Shigeru; Nakane, Megumi; Rahman, M Habibur; Nitta, Youji; Yoshiura, Takanori; Ohta, Hiroyuki; Kurusu, Yasurou

2011-04-01

Ethanol was efficiently produced from three varieties of sweet sorghum using repeated-batch fermentation without pasteurization or acidification. Saccharomyces cerevisiae cells could be recycled in 16 cycles of the fermentation process with good ethanol yields. This technique would make it possible to use a broader range of sweet sorghum varieties for ethanol production. Copyright © 2010 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Ethanol fermentation of raw cassava starch with Rhizopus koji in a gas circulation type fermentor

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

Fujio, Y.; Ogato, M.; Ueda, S.

Studies have been conducted in a gas circulation type fermentor in order to characterize the ethanol fermentation of uncooked cassava starch with Rhizopus koji. Results showed that ethanol concentration reached 13-14% (v/v) in 4-day broth, and the maximum productivity of ethanol was 2.3 g ethanol/l broth h. This productivity was about 50% compared to the productivity of a glucose-yeast system. Ethanol yield reached 83.5-72.3% of the theoretical yield for the cassava starch used. The fermentor used in the present work has been proven by experiment to be suitable for ethanol fermentation of the broth with solid substrate. 10 references.

Fermentation of xylose into ethanol by a new fungus strain Pestalotiopsis sp. XE-1.

PubMed

Pang, Zong-wen; Liang, Jing-juan; Huang, Ri-bo

2011-08-01

A new fungus, Pestalotiopsis sp. XE-1, which produced ethanol from xylose with yield of 0.47 g ethanol/g of consumed xylose was isolated. It also produced ethanol from arabinose, glucose, fructose, mannose, galactose, cellobiose, maltose, and sucrose with yields of 0.38, 0.47, 0.45, 0.46, 0.31, 0.25, 0.31, and 0.34 g ethanol/g of sugar consumed, respectively. It produced maximum ethanol from xylose at pH 6.5, 30°C under a semi-aerobic condition. Acetic acid produced in xylose fermenting process inhibited ethanol production of XE-1. The ethanol yield in the pH-uncontrolled batch fermentation was about 27% lower than that in the pH-controlled one. The ethanol tolerance of XE-1 was higher than most xylose-fermenting, ethanol-producing microbes, but lower than Saccharomyces cerevisiae and Hansenula polymorpha. XE-1 showed tolerance to high concentration of xylose, and was able to grow and produce ethanol even when it was cultivated in 97.71 g/l xylose.

Ethanol fermentation of cassava starch pretreated with alkali

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

Shin, Y.C.; Lee, S.Y.; Choe, Y.K.

1986-04-01

In view of the current industrial process for the conventional ethanol fermentation, in which raw starch materials are heated at 120 degrees C for 2 h, conditions for an alternative process were set: an overall time from saccharification to ethanol fermentation of within 3-4 days, an operation temperature of below 60 degrees C, an ethanol yield of over 93%, and a ratio of raw material to fermentation volume of within 1:4. To meet these conditions, previously a steeping method of starch materials in 0.5N HCl solution at 60 degrees C for 12 h were used, followed by combined actions ofmore » ..cap alpha..-amylase and glucoamylase. The ethanol yield from uncooked cassava starch treated under the conditions described was 95% after fermentation for 3 days with Saccharomyces cerevisiae. However, the use of a relatively higher concentration of acid for steeping is still a problem and gelatinization of starch materials is insufficient. This communication, therefore, describes effects of alkali steeping and structural change of starch granules on the ethanol fermentation. 8 references.« less

Thermophilic ethanol fermentation from lignocellulose hydrolysate by genetically engineered Moorella thermoacetica.

PubMed

Rahayu, Farida; Kawai, Yuto; Iwasaki, Yuki; Yoshida, Koichiro; Kita, Akihisa; Tajima, Takahisa; Kato, Junichi; Murakami, Katsuji; Hoshino, Tamotsu; Nakashimada, Yutaka

2017-12-01

A transformant of Moorella thermoacetica was constructed for thermophilic ethanol production from lignocellulosic biomass by deleting two phosphotransacetylase genes, pdul1 and pdul2, and introducing the native aldehyde dehydrogenase gene (aldh) controlled by the promoter from glyceraldehyde-3-phosphate dehydrogenase. The transformant showed tolerance to 540mM and fermented sugars including fructose, glucose, galactose and xylose to mainly ethanol. In a mixed-sugar medium of glucose and xylose, all of the sugars were consumed to produce ethanol at the yield of 1.9mol/mol-sugar. The transformant successfully fermented sugars in hydrolysate prepared through the acid hydrolysis of lignocellulose to ethanol, suggesting that this transformant can be used to ferment the sugars in lignocellulosic biomass for ethanol production. Copyright © 2017 Elsevier Ltd. All rights reserved.

Recent trends in bioethanol production from food processing byproducts.

PubMed

Akbas, Meltem Yesilcimen; Stark, Benjamin C

2016-11-01

The widespread use of corn starch and sugarcane as sources of sugar for the production of ethanol via fermentation may negatively impact the use of farmland for production of food. Thus, alternative sources of fermentable sugars, particularly from lignocellulosic sources, have been extensively investigated. Another source of fermentable sugars with substantial potential for ethanol production is the waste from the food growing and processing industry. Reviewed here is the use of waste from potato processing, molasses from processing of sugar beets into sugar, whey from cheese production, byproducts of rice and coffee bean processing, and other food processing wastes as sugar sources for fermentation to ethanol. Specific topics discussed include the organisms used for fermentation, strategies, such as co-culturing and cell immobilization, used to improve the fermentation process, and the use of genetic engineering to improve the performance of ethanol producing fermenters.

Improvement in fermentation characteristics of degermed ground corn by lipid supplementation.

PubMed

Murthy, Ganti S; Singh, Vijay; Johnston, David B; Rausch, Kent D; Tumbleson, M E

2006-08-01

With rapid growth of fuel ethanol industry, and concomitant increase in distillers dried grains with solubles (DDGS), new corn fractionation technologies that reduce DDGS volume and produce higher value coproducts in dry grind ethanol process have been developed. One of the technologies, a dry degerm, defiber (3D) process (similar to conventional corn dry milling) was used to separate germ and pericarp fiber prior to the endosperm fraction fermentation. Recovery of germ and pericarp fiber in the 3D process results in removal of lipids from the fermentation medium. Biosynthesis of lipids, which is important for cell growth and viability, cannot proceed in strictly anaerobic fermentations. The effects of ten different lipid supplements on improving fermentation rates and ethanol yields were studied and compared to the conventional dry grind process. Endosperm fraction (from the 3D process) was mixed with water and liquefied by enzymatic hydrolysis and was fermented using simultaneous saccharification and fermentation. The highest ethanol concentration (13.7% v/v) was achieved with conventional dry grind process. Control treatment (endosperm fraction from 3D process without lipid supplementation) produced the lowest ethanol concentration (11.2% v/v). Three lipid treatments (fatty acid ester, alkylphenol, and ethoxylated sorbitan ester 1836) were most effective in improving final ethanol concentrations. Fatty acid ester treatment produced the highest final ethanol concentration (12.3% v/v) among all lipid supplementation treatments. Mean final ethanol concentrations of alkylphenol and ethoxylated sorbitan ester 1836 supplemented samples were 12.3 and 12.0% v/v, respectively.

Production of ethanol from a mixture of waste paper and kitchen waste via a process of successive liquefaction, presaccharification, and simultaneous saccharification and fermentation.

PubMed

Nishimura, Hiroto; Tan, Li; Kira, Noriko; Tomiyama, Shigeo; Yamada, Kazuo; Sun, Zhao-Yong; Tang, Yue-Qin; Morimura, Shigeru; Kida, Kenji

2017-09-01

Efficient ethanol production from waste paper requires the addition of expensive nutrients. To reduce the production cost of ethanol from waste paper, a study on how to produce ethanol efficiently by adding kitchen waste (potentially as a carbon source, nutrient source, and acidity regulator) to waste paper was performed and a process of successive liquefaction, presaccharification, and simultaneous saccharification and fermentation (L+PSSF) was developed. The individual saccharification performances of waste paper and kitchen waste were not influenced by their mixture. Liquefaction of kitchen waste at 90°C prior to presaccharification and simultaneous saccharification and fermentation (PSSF) was essential for efficient ethanol fermentation. Ethanol at concentrations of 46.6 or 43.6g/l was obtained at the laboratory scale after fermentation for 96h, even without pH adjustment and/or the addition of extra nutrients. Similarly, ethanol at a concentration of 45.5g/l was obtained at the pilot scale after fermentation for 48h. The ethanol concentration of L+PSSF of the mixture of waste paper and kitchen waste was comparable to that of PSSF of waste paper with added nutrients (yeast extract and peptone) and pH adjustment using H 2 SO 4 , indicating that kitchen waste is not only a carbon source but also an excellent nutrient source and acidity regulator for fermentation of the mixture of waste paper and kitchen waste. Copyright © 2017. Published by Elsevier Ltd.

Process engineering and scale-up of autotrophic Clostridium strain P11 syngas fermentation

NASA Astrophysics Data System (ADS)

Kundiyana, Dimple Kumar Aiyanna

Scope and Method of Study. Biomass gasification followed by fermentation of syngas to ethanol is a potential process to produce bioenergy. The process is currently being researched under laboratory- and pilot-scale in an effort to optimize the process conditions and make the process feasible for commercial production of ethanol and other biofuels such as butanol and propanol. The broad research objectives for the research were to improve ethanol yields during syngas fermentation and to design a economical fermentation process. The research included four statistically designed experimental studies in serum bottles, bench-scale and pilot-scale fermentors to screen alternate fermentation media components, to determine the effect of process parameters such as pH, temperature and buffer on syngas fermentation, to determine the effect of key limiting nutrients of the acetyl-CoA pathway in a continuous series reactor design, and to scale-up the syngas fermentation in a 100-L pilot scale fermentor. Findings and Conclusions. The first experimental study identified cotton seed extract (CSE) as a feasible medium for Clostridium strain P11 fermentation. The study showed that CSE at 0.5 g L-1 can potentially replace all the standard Clostridium strain P11 fermentation media components while using a media buffer did not significantly improve the ethanol production when used in fermentation with CSE. Scale-up of the CSE fermentation in 2-L and 5-L stirred tank fermentors showed 25% increase in ethanol yield. The second experimental study showed that syngas fermentation at 32°C without buffer was associated with higher ethanol concentration and reduced lag time in switching to solventogenesis. Conducting fermentation at 40°C or by lowering incubation pH to 5.0 resulted in reduced cell growth and no production of ethanol or acetic acid. The third experiment studied the effect of three limiting nutrients, calcium pantothenate, vitamin B12 and CoCl2 on syngas fermentation. Results indicated that it is possible to modulate the product formation by limiting key nutrients of acetyl-CoA pathway and using a continuous fermentation in two-stage fermentor design to improve ethanol yields. The last experimental study was conducted to commission a pilot scale fermentor, and subsequently scale-up the Clostridium strain P11 fermentation from a bench-scale to a pilot scale 100-L fermentor. Results indicated a six-fold improvement in ethanol concentration (25.3 g L-1 at the end of 59 d) compared to previous Clostridium strain P11 and Clostridium carboxidivorans fermentations plus the formation of other compounds such as isopropyl alcohol, acetic acid and butanol, which are of commercial importance.

Ethanol effect on metabolic activity of the ethalogenic fungus Fusarium oxysporum.

PubMed

Paschos, Thomas; Xiros, Charilaos; Christakopoulos, Paul

2015-03-12

Fusarium oxysporum is a filamentous fungus which has attracted a lot of scientific interest not only due to its ability to produce a variety of lignocellulolytic enzymes, but also because it is able to ferment both hexoses and pentoses to ethanol. Although this fungus has been studied a lot as a cell factory, regarding applications for the production of bioethanol and other high added value products, no systematic study has been performed concerning its ethanol tolerance levels. In aerobic conditions it was shown that both the biomass production and the specific growth rate were affected by the presence of ethanol. The maximum allowable ethanol concentration, above which cells could not grow, was predicted to be 72 g/L. Under limited aeration conditions the ethanol-producing capability of the cells was completely inhibited at 50 g/L ethanol. The lignocellulolytic enzymatic activities were affected to a lesser extent by the presence of ethanol, while the ethanol inhibitory effect appears to be more severe at elevated temperatures. Moreover, when the produced ethanol was partially removed from the broth, it led to an increase in fermenting ability of the fungus up to 22.5%. The addition of F. oxysporum's system was shown to increase the fermentation of pretreated wheat straw by 11%, in co-fermentation with Saccharomyces cerevisiae. The assessment of ethanol tolerance levels of F. oxysporum on aerobic growth, on lignocellulolytic activities and on fermentative performance confirmed its biotechnological potential for the production of bioethanol. The cellulolytic and xylanolytic enzymes of this fungus could be exploited within the biorefinery concept as their ethanol resistance is similar to that of the commercial enzymes broadly used in large scale fermentations and therefore, may substantially contribute to a rational design of a bioconversion process involving F. oxysporum. The SSCF experiments on liquefied wheat straw rich in hemicellulose indicated that the contribution of the metabolic system of F. oxysporum in a co-fermentation with S. cerevisiae may play a secondary role.

Dry-grind processing using amylase corn and superior yeast to reduce the exogenous enzyme requirements in bioethanol production.

PubMed

Kumar, Deepak; Singh, Vijay

2016-01-01

Conventional corn dry-grind ethanol production process requires exogenous alpha and glucoamylases enzymes to breakdown starch into glucose, which is fermented to ethanol by yeast. This study evaluates the potential use of new genetically engineered corn and yeast, which can eliminate or minimize the use of these external enzymes, improve the economics and process efficiencies, and simplify the process. An approach of in situ ethanol removal during fermentation was also investigated for its potential to improve the efficiency of high-solid fermentation, which can significantly reduce the downstream ethanol and co-product recovery cost. The fermentation of amylase corn (producing endogenous α-amylase) using conventional yeast and no addition of exogenous α-amylase resulted in ethanol concentration of 4.1 % higher compared to control treatment (conventional corn using exogenous α-amylase). Conventional corn processed with exogenous α-amylase and superior yeast (producing glucoamylase or GA) with no exogenous glucoamylase addition resulted in ethanol concentration similar to control treatment (conventional yeast with exogenous glucoamylase addition). Combination of amylase corn and superior yeast required only 25 % of recommended glucoamylase dose to complete fermentation and achieve ethanol concentration and yield similar to control treatment (conventional corn with exogenous α-amylase, conventional yeast with exogenous glucoamylase). Use of superior yeast with 50 % GA addition resulted in similar increases in yield for conventional or amylase corn of approximately 7 % compared to that of control treatment. Combination of amylase corn, superior yeast, and in situ ethanol removal resulted in a process that allowed complete fermentation of 40 % slurry solids with only 50 % of exogenous GA enzyme requirements and 64.6 % higher ethanol yield compared to that of conventional process. Use of amylase corn and superior yeast in the dry-grind processing industry can reduce the total external enzyme usage by more than 80 %, and combining their use with in situ removal of ethanol during fermentation allows efficient high-solid fermentation.

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

PubMed

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

2007-12-01

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

Process for producing ethanol from plant biomass using the fungus paecilomyces sp.

DOEpatents

Wu, Jung Fu

1989-01-01

A process for producing ethanol from plant biomass is disclosed. The process in cludes forming a substrate from the biomass with the substrate including hydrolysates of cellulose and hemicellulose. A species of the fungus Paecilomyces, which has the ability to ferment both cellobiose and xylose to ethanol, is then selected and isolated. The substrate is inoculated with this fungus, and the inoculated substrate is then fermented under conditions favorable for cell viability and conversion of hydrolysates to ethanol. Finally, ethanol is recovered from the fermented substrate.

Process for producing ethanol from plant biomass using the fungus Paecilomyces sp

DOEpatents

Wu, J.F.

1985-08-08

A process for producing ethanol from plant biomass is disclosed. The process includes forming a substrate from the biomass with the substrate including hydrolysates of cellulose and hemicellulose. A species of the fungus Paecilomyces which has the ability to ferment both cellobiose and xylose to ethanol is then selected and isolated. The substrate is inoculated with this fungus, and the inoculated substrate is then fermented under conditions favorable for cell viability and conversion of hydrolysates to ethanol. Finally, ethanol is recovered from the fermented substrate. 5 figs., 3 tabs.

Bioethanol production from the dry powder of Jerusalem artichoke tubers by recombinant Saccharomyces cerevisiae in simultaneous saccharification and fermentation.

PubMed

Wang, Yi-Zhou; Zou, Shan-Mei; He, Mei-Lin; Wang, Chang-Hai

2015-04-01

It has been found that recombinant Saccharomyces cerevisiae 6525 can produce high concentration of ethanol in one-step fermentation from the extract of Jerusalem artichoke tubers or inulin. However, the utilization rate of raw materials was low and the fermentation process was costly and complicated. Therefore, in this study, after the optimum processing conditions for ethanol production in fed-batch fermentation were determined in flask, the recombinant S. cerevisiae 6525 was first used to produce ethanol from the dry powder of Jerusalem artichoke tubers in 5-L agitating fermentor. After 72 h of fermentation, around 84.3 g/L ethanol was produced in the fermentation liquids, and the conversion efficiency of inulin-type sugars to ethanol was 0.453, or 88.6 % of the theoretical value of 0.511. This study showed high feasibility of bioethanol industrial production from the Jerusalem artichoke tubers and provided a basis for it in the future.

Improved Ethanol Production from Xylose by Candida shehatae Induced by Dielectric Barrier Discharge Air Plasma

NASA Astrophysics Data System (ADS)

Chen, Huixia; Xiu, Zhilong; Bai, Fengwu

2014-06-01

Xylose fermentation is essential for ethanol production from lignocellulosic biomass. Exposure of the xylose-fermenting yeast Candida shehatae (C. shehatae) CICC1766 to atmospheric pressure dielectric barrier discharge (DBD) air plasma yields a clone (designated as C81015) with stability, which exhibits a higher ethanol fermentation rate from xylose, giving a maximal enhancement in ethanol production of 36.2% compared to the control (untreated). However, the biomass production of C81015 is lower than that of the control. Analysis of the NADH (nicotinamide adenine dinucleotide)- and NADPH (nicotinamide adenine dinucleotide phosphate)-linked xylose reductases and NAD+-linked xylitol dehydrogenase indicates that their activities are enhanced by 34.1%, 61.5% and 66.3%, respectively, suggesting that the activities of these three enzymes are responsible for improving ethanol fermentation in C81015 with xylose as a substrate. The results of this study show that DBD air plasma could serve as a novel and effective means of generating microbial strains that can better use xylose for ethanol fermentation.

Production of ethanol 3G from Kappaphycus alvarezii: evaluation of different process strategies.

PubMed

Hargreaves, Paulo Iiboshi; Barcelos, Carolina Araújo; da Costa, Antonio Carlos Augusto; Pereira, Nei

2013-04-01

This study evaluated the potential of Kappaphycus alvarezii as feedstock for ethanol production, i.e. ethanol 3G. First, aquatic biomass was subjected to a diluted acid pretreatment. This acid pretreatment generated two streams--a galactose-containing liquid fraction and a cellulose-containing solid fraction, which were investigated to determine their fermentability with the following strategies: a single-stream process (simultaneous saccharification and co-fermentation (SSCF) of both fractions altogether), which achieved 64.3 g L(-1) of ethanol, and a two-stream process (fractions were fermented separately), which resulted in 38 g L(-1) of ethanol from the liquid fraction and 53.0 g L(-1) from the simultaneous saccharification and fermentation (SSF) of the solid fraction. Based on the average fermentable carbohydrate concentration, it was possible to obtain 105 L of ethanol per ton of dry seaweed. These preliminaries results indicate that the use of the macro-algae K. alvarezii has a good potential feedstock for bioethanol production. Copyright © 2013. Published by Elsevier Ltd.

Manufacturing Ethyl Acetate From Fermentation Ethanol

NASA Technical Reports Server (NTRS)

Rohatgi, Naresh K.; Ingham, John D.

1991-01-01

Conceptual process uses dilute product of fermentation instead of concentrated ethanol. Low-concentration ethanol, extracted by vacuum from fermentation tank, and acetic acid constitutes feedstock for catalytic reaction. Product of reaction goes through steps that increases ethyl acetate content to 93 percent by weight. To conserve energy, heat exchangers recycle waste heat to preheat process streams at various points.

Simultaneous co-fermentation of mixed sugars: a promising strategy for producing cellulosic ethanol.

PubMed

Kim, Soo Rin; Ha, Suk-Jin; Wei, Na; Oh, Eun Joong; Jin, Yong-Su

2012-05-01

The lack of microbial strains capable of fermenting all sugars prevalent in plant cell wall hydrolyzates to ethanol is a major challenge. Although naturally existing or engineered microorganisms can ferment mixed sugars (glucose, xylose and galactose) in these hydrolyzates sequentially, the preferential utilization of glucose to non-glucose sugars often results in lower overall yield and productivity of ethanol. Therefore, numerous metabolic engineering approaches have been attempted to construct optimal microorganisms capable of co-fermenting mixed sugars simultaneously. Here, we present recent findings and breakthroughs in engineering yeast for improved ethanol production from mixed sugars. In particular, this review discusses new sugar transporters, various strategies for simultaneous co-fermentation of mixed sugars, and potential applications of co-fermentation for producing fuels and chemicals. Copyright © 2012 Elsevier Ltd. All rights reserved.

Simultaneous saccharification and fermentation (SSF) using cellobiose fermenting yeast Brettanomyces custersii

DOEpatents

Spindler, Diane D.; Grohmann, Karel; Wyman, Charles E.

1992-01-01

A process for producing ethanol from plant biomass includes forming a substrate from the biomass with the substrate including hydrolysates of cellulose and hemicellulose. A species of the yeast Brettanomyces custersii (CBS 5512), which has the ability to ferment both cellobiose and glucose to ethanol, is then selected and isolated. The substrate is inoculated with this yeast, and the inoculated substrate is then fermented under conditions favorable for cell viability and conversion of hydrolysates to ethanol.

Performance of several Saccharomyces strains for the alcoholic fermentation of sugar-sweetened high-strength wastewaters: Comparative analysis and kinetic modelling.

PubMed

Comelli, Raúl N; Seluy, Lisandro G; Isla, Miguel A

2016-12-25

This work focuses on the performance of ten commercial Saccharomyces yeast strains in the batch alcoholic fermentation of sugars contained in selected industrial wastewaters from the soft drink industry. Fermentation has been applied successfully to treat these effluents prior to their disposal. Although many strains were investigated, similar behaviour was observed between all of the Saccharomyces strains tested. When media were inoculated with 2gL -1 of yeast, all strains were able to completely consume the available sugars in less than 14h. Thus, any of the strains studied in this work could be used in non-conventional wastewater treatment processes based on alcoholic fermentation. However, ethanol production varied between strains, and these differences could be significant from a production point of view. Saccharomyces bayanus produced the most ethanol, with a mean yield of 0.44g ethanol g sugarconsumed -1 and an ethanol specific production rate of 5.96g ethanol (Lh) -1 . As the assayed soft drinks wastewaters contain about 105g sugar /L of fermentable sugars, the concentration of ethanol achieved after the fermentations process was 46.2g ethanol /L. A rigorous kinetic modelling methodology was used to model the Saccharomyces bayanus fermentation process. The kinetic model included coupled mass balances and a minimal number of parameters. A simple unstructured model based on the Andrews equation (substrate inhibition) was developed. This model satisfactorily described biomass growth, sugar consumption and bioethanol production. In addition to providing insights into the fermentative performance of potentially relevant strains, this work can facilitate the design of large-scale ethanol production processes that use wastewaters from the sugar-sweetened beverage industry as feedstock. Copyright © 2016 Elsevier B.V. All rights reserved.

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

Hang, Y.D.; Lee, C.Y.; Woodams, E.E.

A solid state fermentation system for the production of ethanol from apple pomace with a Montrachet strain of Saccharomyces cerevisiae is described. The yields of ethanol varied from about 29 g to more than 40 g/kg of apple pomace, depending on the samples fermented. Separation of up to 99% of the ethanol from spent qpple pomace was achieved with a rotary vacuum evaporator. Alcohol fermentation of apple pomace might be an efficient method of alleviating waste disposal problems with the concomitant production of ethanol.

Evaluation of UV-C mutagenized Scheffersomyces stipitis strains for ethanol production.

PubMed

Geiger, Melanie; Gibbons, Jaimie; West, Thomas; Hughes, Stephen R; Gibbons, William

2012-12-01

We evaluated fermentation capabilities of five strains of Scheffersomyces stipitis (WT-2-1, WT-1-11, 14-2-6, 22-1-1, and 22-1-12) that had been produced by UV-C mutagenesis and selection for improved xylose fermentation to ethanol using an integrated automated robotic work cell. They were incubated under both facultative and anaerobic conditions to evaluate ethanol production on glucose, xylose, cellobiose, and a combination of all three sugars. The medium contained 50 g/L total sugar and 5 g/L yeast extract. The strains performed significantly better under facultative compared with anaerobic conditions. As expected, glucose was the most readily fermented sugar with ~100% fermentation efficiency (FE) under facultative conditions but only 5% to 16% FE anaerobically. Xylose utilization was 20% to 40% FE under facultative conditions but 9% to 25% FE anaerobically. Cellobiose was the least fermented sugar, at 18% to 27% FE facultatively and 8% to 11% anaerobically. Similar trends occurred in the sugar mixture. Under facultative conditions, strain 22-1-12 produced 19.6 g/L ethanol on glucose, but strain 14-2-6 performed best on xylose (4.5 g/L ethanol) and the sugar combination (8.0 g/L ethanol). Ethanol titers from glucose under anaerobic conditions were again highest with strain 22-1-12, but none of the strains produced ethanol from xylose. Future trials will evaluate nutrient addition to boost microaerophilic xylose fermentation.

76 FR 49313 - Approval and Promulgation of Implementation Plans North Carolina: Prevention of Significant...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-08-10

... exception of the phrase ``except ethanol production facilities producing ethanol by natural fermentation... ethanol by natural fermentation under the North American Industry Classification System (NAICS) codes...

Ethanol fermentation from molasses at high temperature by thermotolerant yeast Kluyveromyces sp. IIPE453 and energy assessment for recovery.

PubMed

Dasgupta, Diptarka; Ghosh, Prasenjit; Ghosh, Debashish; Suman, Sunil Kumar; Khan, Rashmi; Agrawal, Deepti; Adhikari, Dilip K

2014-10-01

High temperature ethanol fermentation from sugarcane molasses B using thermophilic Crabtree-positive yeast Kluyveromyces sp. IIPE453 was carried out in batch bioreactor system. Strain was found to have a maximum specific ethanol productivity of 0.688 g/g/h with 92 % theoretical ethanol yield. Aeration and initial sugar concentration were tuning parameters to regulate metabolic pathways of the strain for either cell mass or higher ethanol production during growth with an optimum sugar to cell ratio 33:1 requisite for fermentation. An assessment of ethanol recovery from fermentation broth via simulation study illustrated that distillation-based conventional recovery was significantly better in terms of energy efficiency and overall mass recovery in comparison to coupled solvent extraction-azeotropic distillation technique for the same.

Methods for increasing the production of ethanol from microbial fermentation

DOEpatents

Gaddy, James L [Fayetteville, AR; Arora, Dinesh K [Fayetteville, AR; Ko, Ching-Whan [Fayetteville, AR; Phillips, John Randall [Fayetteville, AR; Basu, Rahul [Bethlehem, PA; Wikstrom, Carl V [Fayetteville, AR; Clausen, Edgar C [Fayetteville, AR

2007-10-23

A stable continuous method for producing ethanol from the anaerobic bacterial fermentation of a gaseous substrate containing at least one reducing gas involves culturing a fermentation bioreactor anaerobic, acetogenic bacteria in a liquid nutrient medium; supplying the gaseous substrate to the bioreactor; and manipulating the bacteria in the bioreactor by reducing the redox potential, or increasing the NAD(P)H TO NAD(P) ratio, in the fermentation broth after the bacteria achieves a steady state and stable cell concentration in the bioreactor. The free acetic acid concentration in the bioreactor is maintained at less than 5 g/L free acid. This method allows ethanol to be produced in the fermentation broth in the bioreactor at a productivity greater than 10 g/L per day. Both ethanol and acetate are produced in a ratio of ethanol to acetate ranging from 1:1 to 20:1.

Production of ethanol and feed by high dry matter hydrolysis and fermentation of palm kernel press cake.

PubMed

Jørgensen, Henning; Sanadi, Anand R; Felby, Claus; Lange, Niels Erik Krebs; Fischer, Morten; Ernst, Steffen

2010-05-01

Palm kernel press cake (PKC) is a residue from palm oil extraction presently only used as a low protein feed supplement. PKC contains 50% fermentable hexose sugars present in the form of glucan and mainly galactomannan. This makes PKC an interesting feedstock for processing into bioethanol or in other biorefinery processes. Using a combination of mannanase, beta-mannosidase, and cellulases, it was possible without any pretreatment to hydrolyze PKC at solid concentrations of 35% dry matter with mannose yields up to 88% of theoretical. Fermentation was tested using Saccharomyces cerevisiae in both a separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) setup. The hydrolysates could readily be fermented without addition of nutrients and with average fermentation yields of 0.43 +/- 0.02 g/g based on consumed mannose and glucose. Employing SSF, final ethanol concentrations of 70 g/kg was achieved in 216 h, corresponding to an ethanol yield of 70% of theoretical or 200 g ethanol/kg PKC. Testing various enzyme mixtures revealed that including cellulases in combination with mannanases significantly improved ethanol yields. Processing PKC to ethanol resulted in a solid residue enriched in protein from 17% to 28%, a 70% increase, thereby potentially making a high-protein containing feed supplement.

Improved bioethanol production using fusants of Saccharomyces cerevisiae and xylose-fermenting yeasts.

PubMed

Kumari, Rajni; Pramanik, K

2012-06-01

The present research deals with the development of a hybrid yeast strain with the aim of converting pentose and hexose sugar components of lignocellulosic substrate to bioethanol by fermentation. Different fusant strains were obtained by fusing protoplasts of Saccharomyces cerevisiae and xylose-fermenting yeasts such as Pachysolen tannophilus, Candida shehatae and Pichia stipitis. The fusants were sorted by fluorescent-activated cell sorter and further confirmed by molecular characterization. The fusants were evaluated by fermentation of glucose-xylose mixture and the highest ethanol producing fusant was used for further study to ferment hydrolysates produced by acid pretreatment and enzymatic hydrolysis of cotton gin waste. Among the various fusant and parental strains used under present study, RPR39 was found to be stable and most efficient strain giving maximum ethanol concentration (76.8 ± 0.31 g L(-1)), ethanol productivity (1.06 g L(-1) h(-1)) and ethanol yield (0.458 g g(-1)) by fermentation of glucose-xylose mixture under test conditions. The fusant has also shown encouraging result in fermenting hydrolysates of cotton gin waste with ethanol concentration of 7.08 ± 0.142 g L(-1), ethanol yield of 0.44 g g(-1), productivity of 0.45 g L(-1) h(-1) and biomass yield of 0.40 g g(-1).

Identification of candidate genes for yeast engineering to improve bioethanol production in very high gravity and lignocellulosic biomass industrial fermentations.

PubMed

Pereira, Francisco B; Guimarães, Pedro Mr; Gomes, Daniel G; Mira, Nuno P; Teixeira, Miguel C; Sá-Correia, Isabel; Domingues, Lucília

2011-12-09

The optimization of industrial bioethanol production will depend on the rational design and manipulation of industrial strains to improve their robustness against the many stress factors affecting their performance during very high gravity (VHG) or lignocellulosic fermentations. In this study, a set of Saccharomyces cerevisiae genes found, through genome-wide screenings, to confer resistance to the simultaneous presence of different relevant stresses were identified as required for maximal fermentation performance under industrial conditions. Chemogenomics data were used to identify eight genes whose expression confers simultaneous resistance to high concentrations of glucose, acetic acid and ethanol, chemical stresses relevant for VHG fermentations; and eleven genes conferring simultaneous resistance to stresses relevant during lignocellulosic fermentations. These eleven genes were identified based on two different sets: one with five genes granting simultaneous resistance to ethanol, acetic acid and furfural, and the other with six genes providing simultaneous resistance to ethanol, acetic acid and vanillin. The expression of Bud31 and Hpr1 was found to lead to the increase of both ethanol yield and fermentation rate, while Pho85, Vrp1 and Ygl024w expression is required for maximal ethanol production in VHG fermentations. Five genes, Erg2, Prs3, Rav1, Rpb4 and Vma8, were found to contribute to the maintenance of cell viability in wheat straw hydrolysate and/or the maximal fermentation rate of this substrate. The identified genes stand as preferential targets for genetic engineering manipulation in order to generate more robust industrial strains, able to cope with the most significant fermentation stresses and, thus, to increase ethanol production rate and final ethanol titers.

Improving ethanol productivity through self-cycling fermentation of yeast: a proof of concept.

PubMed

Wang, Jie; Chae, Michael; Sauvageau, Dominic; Bressler, David C

2017-01-01

The cellulosic ethanol industry has developed efficient strategies for converting sugars obtained from various cellulosic feedstocks to bioethanol. However, any further major improvements in ethanol productivity will require development of novel and innovative fermentation strategies that enhance incumbent technologies in a cost-effective manner. The present study investigates the feasibility of applying self-cycling fermentation (SCF) to cellulosic ethanol production to elevate productivity. SCF is a semi-continuous cycling process that employs the following strategy: once the onset of stationary phase is detected, half of the broth volume is automatically harvested and replaced with fresh medium to initiate the next cycle. SCF has been shown to increase product yield and/or productivity in many types of microbial cultivation. To test whether this cycling process could increase productivity during ethanol fermentations, we mimicked the process by manually cycling the fermentation for five cycles in shake flasks, and then compared the results to batch operation. Mimicking SCF for five cycles resulted in regular patterns with regards to glucose consumption, ethanol titer, pH, and biomass production. Compared to batch fermentation, our cycling strategy displayed improved ethanol volumetric productivity (the titer of ethanol produced in a given cycle per corresponding cycle time) and specific productivity (the amount of ethanol produced per cellular biomass) by 43.1 ± 11.6 and 42.7 ± 9.8%, respectively. Five successive cycles contributed to an improvement of overall productivity (the aggregate amount of ethanol produced at the end of a given cycle per total processing time) and the estimated annual ethanol productivity (the amount of ethanol produced per year) by 64.4 ± 3.3 and 33.1 ± 7.2%, respectively. This study provides proof of concept that applying SCF to ethanol production could significantly increase productivities, which will help strengthen the cellulosic ethanol industry.

A dynamic flux balance model and bottleneck identification of glucose, xylose, xylulose co-fermentation in Saccharomyces cerevisiae

USDA-ARS?s Scientific Manuscript database

Economically viable production of lignocellulosic ethanol requires efficient conversion of feedstock sugars to ethanol. Saccharomyces cerevisiae cannot ferment xylose, the main five-carbon sugars in biomass, but can ferment xylulose, an enzymatically derived isomer. Xylulose fermentation is slow rel...

Simultaneous saccharification and fermentation (SSF) using cellobiose fermenting yeast Brettanomyces custersii (CBS 5512)

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

Spindler, D.D.; Grohmann, K.; Wyman, C.E.

1991-01-16

A process for producing ethanol from plant biomass includes forming a substrate from the biomass with the substrate including hydrolysates of cellulose and hemicellulose. A species of the yeast Brettanomyces custersii (CBS 5512), which has the ability to ferment both cellobiose and glucose to ethanol, is then selected and isolated. The substrate is inoculated with this yeast, and the inoculated substrate is then fermented under conditions favorable for cell viability and conversion of hydrolysates to ethanol.

Simultaneous saccharification and fermentation (SSF) using cellobiose fermenting yeast Brettanomyces custersii

DOEpatents

Spindler, D.D.; Grohmann, K.; Wyman, C.E.

1992-03-31

A process for producing ethanol from plant biomass includes forming a substrate from the biomass with the substrate including hydrolysates of cellulose and hemicellulose. A species of the yeast Brettanomyces custersii (CBS 5512), which has the ability to ferment both cellobiose and glucose to ethanol, is then selected and isolated. The substrate is inoculated with this yeast, and the inoculated substrate is then fermented under conditions favorable for cell viability and conversion of hydrolysates to ethanol. 2 figs.

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

PubMed

Cysewski, G R; Wilke, C R

1976-09-01

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

Fungal invertase as an aid for fermentation of cane molasses into ethanol

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

Park, Y.K.; Sato, H.H.

1982-10-01

Comparative studies of the fermentation of cane molasses into ethanol by Saccharomyces cerevisiae in the presence or absence of fungal invertase were performed. When cane molasses was fermented by the yeast at 30 degrees Centigrade and pH 5.0, the presence of the enzyme had no effect on ethanol production. At pH 3.4, ethanol production was increased by the addition of invertase. At 40 degrees C, the addition of invertase increased ethanol production by 5.5% at pH 5.0 and by 20.9% at pH 3.5. (Refs. 8).

Alleviation of harmful effect in stillage reflux in food waste ethanol fermentation based on metabolic and side-product accumulation regulation.

PubMed

Ma, Hongzhi; Yang, Jian; Jia, Yan; Wang, Qunhui; Ma, Xiaoyu; Sonomoto, Kenji

2016-10-01

Stillage reflux fermentation in food waste ethanol fermentation could reduce sewage discharge but exert a harmful effect because of side-product accumulation. In this study, regulation methods based on metabolic regulation and side-product alleviation were conducted. Result demonstrated that controlling the proper oxidation-reduction potential value (-150mV to -250mV) could reduce the harmful effect, improve ethanol yield by 21%, and reduce fermentation time by 20%. The methods of adding calcium carbonate to adjust the accumulated lactic acid showed that ethanol yield increased by 17.3%, and fermentation time decreased by 20%. The accumulated glyceal also shows that these two methods can reduce the harmful effect. Fermentation time lasted for seven times without effect, and metabolic regulation had a better effect than side-product regulation. Copyright © 2016 Elsevier Ltd. All rights reserved.

A novel fermentation pathway in an Escherichia coli mutant producing succinic acid, acetic acid, and ethanol.

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

Donnelly, M. I.; Millard, C. S.; Clark, D. P.

1998-04-01

Escherichia coli strain NZN111, which is unable to grow fermentatively because of insertional inactivation of the genes encoding pyruvate: formate lyase and the fermentative lactate dehydrogenase, gave rise spontaneously to a chromosomal mutation that restored its ability to ferment glucose. The mutant strain, named AFP111, fermented glucose more slowly than did its wild-type ancestor, strain W1485, and generated a very different spectrum of products. AFP111 produced succinic acid, acetic acid, and ethanol in proportions of approx 2:1:1. Calculations of carbon and electron balances accounted fully for the observed products; 1 mol of glucose was converted to 1 mol of succinicmore » acid and 0.5 mol each of acetic acid and ethanol. The data support the emergence in E.coli of a novel succinic acid:acetic acid:ethanol fermentation pathway.« less

The transcription factor Ace2 and its paralog Swi5 regulate ethanol production during static fermentation through their targets Cts1 and Rps4a in Saccharomyces cerevisiae.

PubMed

Wu, Yao; Du, Jie; Xu, Guoqiang; Jiang, Linghuo

2016-05-01

Saccharomyces cerevisiae is the most widely used fermentation organism for ethanol production. However, the gene expression regulatory networks behind the ethanol fermentation are still not fully understood. Using a static fermentation model, we examined the ethanol yields on biomass of deletion mutants for 77 yeast genes encoding nonessential transcription factors, and found that deletion mutants for ACE2 and SWI5 showed dramatically increased ethanol yields. Overexpression of ACE2 or SWI5 in wild type cells reduced their ethanol yields. Furthermore, among the 34 target genes regulated by Ace2 and Swi5, deletion of CTS1,RPS4a,SIC1,EGT2,DSE2, or SCP160 led to increased ethanol yields, with the former two showing higher effects. Overexpression of CTS1 or RPS4a in both ace2/ace2 and swi5/swi5 mutants reduced their ethanol yields. In contrast, deletion of MCR1 or HO significantly decreased ethanol yields, with the former one showing the highest effect. Therefore, Ace2 and Swi5 are two negative regulators of ethanol yield during static fermentation of yeast cells, and both CTS1 and RPS4a are major effectors mediating these two transcription factors in regulating ethanol production. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Ethanol production from xylose with the yeast Pichia stipitis and simultaneous product recovery by gas stripping using a gas-lift loop fermentor with attached side-arm (GLSA).

PubMed

Domínguez, J M; Cao, N; Gong, C S; Tsao, G T

2000-02-05

The bioconversion of xylose into ethanol with the yeast Pichia stipitis CBS 5773 is inhibited when 20 g/L of ethanol are present in the fermentation broth. In order to avoid this limitation, the fermentation was carried out with simultaneous recovery of product by CO(2) stripping. The fermentation was also improved by attaching a side-arm to the main body of a classical gas-lift loop fermentor. This side-arm increases the liquid circulation, mass transfer, and gas distribution, reducing the amount of oxygen in the inlet gas necessary to perform the fermentation of xylose under microaerobic conditions (K(L)a approximately 16 h(-1)). The continuous stripping of ethanol from the fermentation broth in this new bioreactor system allowed the consumption of higher xylose concentrations than using Erlenmeyer shaker flasks, improved significantly the process productivity and provided a clean ethanol solution by using an ice-cooled condenser system. Finally, a fed-batch fermentation was carried out with a K(L)a = 15.8 h(-1). Starting with 248.2 g of xylose, 237.6 g of xylose was consumed to produce 88.1 g of ethanol which represents 72.6% of the theoretical yield (47.2 g/L of ethanol was recovered in the condenser, while 9.6 g/L remained in the fermentation broth). Copyright 2000 John Wiley & Sons, Inc.

Impact of recycling stillage on conversion of dilute sulfuric acid pretreated corn stover to ethanol.

PubMed

Mohagheghi, Ali; Schell, Daniel J

2010-04-01

Both the current corn starch to ethanol industry and the emerging lignocellulosic biofuels industry view recycling of spent fermentation broth or stillage as a method to reduce fresh water use. The objective of this study was to understand the impact of recycling stillage on conversion of corn stover to ethanol. Sugars in a dilute-acid pretreated corn stover hydrolysate were fermented to ethanol by the glucose-xylose fermenting bacteria Zymomonas mobilis 8b. Three serial fermentations were performed at two different initial sugar concentrations using either 10% or 25% of the stillage as makeup water for the next fermentation in the series. Serial fermentations were performed to achieve near steady state concentration of inhibitors and other compounds in the corn stover hydrolysate. Little impact on ethanol yields was seen at sugar concentrations equivalent to pretreated corn stover slurry at 15% (w/w) with 10% recycle of the stillage. However, ethanol yields became progressively poorer as the sugar concentration increased and fraction of the stillage recycled increased. At an equivalent corn stover slurry concentration of 20% with 25% recycled stillage the ethanol yield was only 5%. For this microorganism with dilute-acid pretreated corn stover, recycling a large fraction of the stillage had a significant negative impact on fermentation performance. Although this finding is of concern for biochemical-based lignocellulose conversion processes, other microorganism/pretreatment technology combinations will likely perform differently. (c) 2009 Wiley Periodicals, Inc.

Engineering industrial Saccharomyces cerevisiae strains for xylose fermentation and comparison for switchgrass conversion.

PubMed

Hector, Ronald E; Dien, Bruce S; Cotta, Michael A; Qureshi, Nasib

2011-09-01

Saccharomyces' physiology and fermentation-related properties vary broadly among industrial strains used to ferment glucose. How genetic background affects xylose metabolism in recombinant Saccharomyces strains has not been adequately explored. In this study, six industrial strains of varied genetic background were engineered to ferment xylose by stable integration of the xylose reductase, xylitol dehydrogenase, and xylulokinase genes. Aerobic growth rates on xylose were 0.04-0.17 h(-1). Fermentation of xylose and glucose/xylose mixtures also showed a wide range of performance between strains. During xylose fermentation, xylose consumption rates were 0.17-0.31 g/l/h, with ethanol yields 0.18-0.27 g/g. Yields of ethanol and the metabolite xylitol were positively correlated, indicating that all of the strains had downstream limitations to xylose metabolism. The better-performing engineered and parental strains were compared for conversion of alkaline pretreated switchgrass to ethanol. The engineered strains produced 13-17% more ethanol than the parental control strains because of their ability to ferment xylose.

Effect of lignocellulosic degradation compounds from steam explosion pretreatment on ethanol fermentation by thermotolerant yeast Kluyveromyces marxianus.

PubMed

Oliva, Jose Miguel; Sáez, Felicia; Ballesteros, Ignacio; González, Alberto; Negro, Maria José; Manzanares, Paloma; Ballesteros, Mercedes

2003-01-01

The filtrate from steam-pretreated poplar was analyzed to identify degradation compounds. The effect of selected compounds on growth and ethanolic fermentation of the thermotolerant yeast strain Kluyveromyces marxianus CECT 10875 was tested. Several fermentations on glucose medium, containing individual inhibitory compounds found in the hydrolysate, were carried out. The degree of inhibition on yeast strain growth and ethanolic fermentation was determined. At concentrations found in the prehy-drolysate, none of the individual compounds significantly affected the fermentation. For all tested compounds, growth was inhibited to a lesser extent than ethanol production. Lower concentrations of catechol (0.96 g/L) and 4-hydroxybenzaldehyde (1.02 g/L) were required to produce the 50% reduction in cell mass in comparison to other tested compounds.

Comparing cell viability and ethanol fermentation of the thermotolerant yeast Kluyveromyces marxianus and Saccharomyces cerevisiae on steam-exploded biomass treated with laccase.

PubMed

Moreno, Antonio D; Ibarra, David; Ballesteros, Ignacio; González, Alberto; Ballesteros, Mercedes

2013-05-01

In this study, the thermotolerant yeast Kluyveromyces marxianus CECT 10875 was compared to the industrial strain Saccharomyces cerevisiae Ethanol Red for lignocellulosic ethanol production. For it, whole slurry from steam-exploded wheat straw was used as raw material, and two process configurations, simultaneous saccharification and fermentation (SSF) and presaccharification and simultaneous saccharification and fermentation (PSSF), were evaluated. Compared to S. cerevisiae, which was able to produce ethanol in both process configurations, K. marxianus was inhibited, and neither growth nor ethanol production occurred during the processes. However, laccase treatment of the whole slurry removed specifically lignin phenols from the overall inhibitory compounds present in the slurry and triggered the fermentation by K. marxianus, attaining final ethanol concentrations and yields comparable to those obtained by S. cerevisiae. Copyright © 2012 Elsevier Ltd. All rights reserved.

Accounting for all sugars produced during integrated production of ethanol from lignocellulosic biomass

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

Schell, Daniel J.; Dowe, Nancy; Chapeaux, Alexandre

This study explored integrated conversion of corn stover to ethanol and highlights techniques for accurate yield calculations. Acid pretreated corn stover (PCS) produced in a pilot-scale reactor was enzymatically hydrolyzed and the resulting sugars were fermented to ethanol by the glucose–xylose fermenting bacteria, Zymomonas mobilis 8b. The calculations account for high solids operation and oligomeric sugars produced during pretreatment, enzymatic hydrolysis, and fermentation, which, if not accounted for, leads to overestimating ethanol yields. The calculations are illustrated for enzymatic hydrolysis and fermentation of PCS at 17.5% and 20.0% total solids achieving 80.1% and 77.9% conversion of cellulose and xylan tomore » ethanol and ethanol titers of 63 g/L and 69 g/L, respectively. In the future, these techniques, including the TEA results, will be applied to fully integrated pilot-scale runs.« less

Accounting for all sugars produced during integrated production of ethanol from lignocellulosic biomass

DOE PAGES

Schell, Daniel J.; Dowe, Nancy; Chapeaux, Alexandre; ...

2016-01-19

This study explored integrated conversion of corn stover to ethanol and highlights techniques for accurate yield calculations. Acid pretreated corn stover (PCS) produced in a pilot-scale reactor was enzymatically hydrolyzed and the resulting sugars were fermented to ethanol by the glucose–xylose fermenting bacteria, Zymomonas mobilis 8b. The calculations account for high solids operation and oligomeric sugars produced during pretreatment, enzymatic hydrolysis, and fermentation, which, if not accounted for, leads to overestimating ethanol yields. The calculations are illustrated for enzymatic hydrolysis and fermentation of PCS at 17.5% and 20.0% total solids achieving 80.1% and 77.9% conversion of cellulose and xylan tomore » ethanol and ethanol titers of 63 g/L and 69 g/L, respectively. In the future, these techniques, including the TEA results, will be applied to fully integrated pilot-scale runs.« less

Alcoholic Fermentation of d-Xylose by Yeasts

PubMed Central

Toivola, Ansa; Yarrow, David; van den Bosch, Eduard; van Dijken, Johannes P.; Scheffers, W. Alexander

1984-01-01

Type strains of 200 species of yeasts able to ferment glucose and grow on xylose were screened for fermentation of d-xylose. In most of the strains tested, ethanol production was negligible. Nineteen were found to produce between 0.1 and 1.0 g of ethanol per liter. Strains of the following species produce more than 1 g of ethanol per liter in the fermentation test with 2% xylose: Brettanomyces naardenensis, Candida shehatae, Candida tenuis, Pachysolen tannophilus, Pichia segobiensis, and Pichia stipitis. Subsequent screening of these yeasts for their capacity to ferment d-cellobiose revealed that only Candida tenuis CBS 4435 was a good fermenter of both xylose and cellobiose under the test conditions used. PMID:16346558

Enhanced Ethanol Production from De-Ashed Paper Sludge by Simultaneous Saccharification and Fermentation and Simultaneous Saccharification and Co-Fermentation

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

Kang, L.; Wang, W.; Pallapolu, V. R.

2011-11-01

A previous study demonstrated that paper sludges with high ash contents can be converted to ethanol by simultaneous saccharification and fermentation (SSF) or simultaneous saccharification and co-fermentation (SSCF). High ash content in the sludge, however, limited solid loading in the bioreactor, causing low product concentration. To overcome this problem, sludges were de-ashed before SSF and SSCF. Low ash content in sludges also increased the ethanol yield to the extent that the enzyme dosage required to achieve 70% yield in the fermentation process was reduced by 30%. High solid loading in SSF and SSCF decreased the ethanol yield. High agitation andmore » de-ashing of the sludges were able to restore the part of the yield loss caused by high solid loading. Substitution of the laboratory fermentation medium (peptone and yeast extract) with corn steep liquor did not bring about any adverse effects in the fermentation. Fed-batch operation of the SSCF and SSF using low-ash content sludges was effective in raising the ethanol concentration, achieving 47.8 g/L and 60.0 g/L, respectively.« less

Simultaneous Saccharification and Fermentation of Sugar Beet Pulp for Efficient Bioethanol Production.

PubMed

Berłowska, Joanna; Pielech-Przybylska, Katarzyna; Balcerek, Maria; Dziekońska-Kubczak, Urszula; Patelski, Piotr; Dziugan, Piotr; Kręgiel, Dorota

2016-01-01

Sugar beet pulp, a byproduct of sugar beet processing, can be used as a feedstock in second-generation ethanol production. The objective of this study was to investigate the effects of pretreatment, of the dosage of cellulase and hemicellulase enzyme preparations used, and of aeration on the release of fermentable sugars and ethanol yield during simultaneous saccharification and fermentation (SSF) of sugar beet pulp-based worts. Pressure-thermal pretreatment was applied to sugar beet pulp suspended in 2% w/w sulphuric acid solution at a ratio providing 12% dry matter. Enzymatic hydrolysis was conducted using Viscozyme and Ultraflo Max (Novozymes) enzyme preparations (0.015-0.02 mL/g dry matter). Two yeast strains were used for fermentation: Ethanol Red ( S. cerevisiae ) (1 g/L) and Pichia stipitis (0.5 g/L), applied sequentially. The results show that efficient simultaneous saccharification and fermentation of sugar beet pulp was achieved. A 6 h interval for enzymatic activation between the application of enzyme preparations and inoculation with Ethanol Red further improved the fermentation performance, with the highest ethanol concentration reaching 26.9 ± 1.2 g/L and 86.5 ± 2.1% fermentation efficiency relative to the theoretical yield.

Starch saccharification and fermentation of uncooked sweet potato roots for fuel ethanol production.

PubMed

Zhang, Peng; Chen, Caifa; Shen, Yanhu; Ding, Tielin; Ma, Daifu; Hua, Zichun; Sun, Dongxu

2013-01-01

An energy-saving ethanol fermentation technology was developed using uncooked fresh sweet potato as raw material. A mutant strain of Aspergillus niger isolated from mildewed sweet potato was used to produce abundant raw starch saccharification enzymes for treating uncooked sweet potato storage roots. The viscosity of the fermentation paste of uncooked sweet potato roots was lower than that of the cooked roots. The ethanol fermentation was carried out by Zymomonas mobilis, and 14.4 g of ethanol (87.2% of the theoretical yield) was produced from 100g of fresh sweet potato storage roots. Based on this method, an energy-saving, high efficient and environment-friendly technology can be developed for large-scale production of fuel ethanol from sweet potato roots. Copyright © 2012 Elsevier Ltd. All rights reserved.

Fate of Fumonisin B1 in Naturally Contaminated Corn during Ethanol Fermentation

PubMed Central

Bothast, R. J.; Bennett, G. A.; Vancauwenberge, J. E.; Richard, J. L.

1992-01-01

Two lots of corn naturally contaminated with fumonisin B1 (15 and 36 ppm) and a control lot (no fumonisin B1 detected) were used as substrates for ethanol production in replicate 8.5-liter yeast fermentations. Ethanol yields were 8.8% for both the control and low-fumonisin corn, while the high-fumonisin corn contained less starch and produced 7.2% ethanol. Little degradation of fumonisin occurred during fermentation, and most was recovered in the distillers' grains, thin stillage, and distillers' solubles fractions. No toxin was detected in the distilled alcohol or centrifuge solids. Ethanol fermentation of fumonisin-contaminated corn coupled with effective detoxification of distillers' grains and aqueous stillage is suggested as a practical process strategy for salvaging contaminated corn. PMID:16348623

Effects of culture conditions on the fermentation of xylose to ethanol by Candida shehatae

Treesearch

T. W. Jeffries

1985-01-01

This research examined four factors on the fermentation of xylose by Candida shehatae, and the following conclusions were reached: (1) A minimal medium is effective for producing ethanol. (2) Peptone and casamino acids stimulate ethanol production. (3) Aeration is important in obtaining good ethanol production rates and yields. (4) The maximal rate of ethanol...

Mutant strain of C. acetobutylicum and process for making butanol

DOEpatents

Jain, Mahendra K.; Beacom, Daniel; Datta, Rathin

1993-01-01

A biologically pure asporogenic mutant of Clostridium acetobutylicum is produced by growing sporogenic C. acetobutylicum ATCC 4259 and treating the parent strain with ethane methane sulfonate. The mutant which as been designated C. acetobutylicum ATCC 55025 is useful in an improved ABE fermentation process, and produces high concentrations of butanol and total solvents.

Fermentation of Agave tequilana juice by Kloeckera africana: influence of amino-acid supplementations.

PubMed

Valle-Rodríguez, Juan Octavio; Hernández-Cortés, Guillermo; Córdova, Jesús; Estarrón-Espinosa, Mirna; Díaz-Montaño, Dulce María

2012-02-01

This study aimed to improve the fermentation efficiency of Kloeckera africana K1, in tequila fermentations. We investigated organic and inorganic nitrogen source requirements in continuous K. africana fermentations fed with Agave tequilana juice. The addition of a mixture of 20 amino-acids greatly improved the fermentation efficiency of this yeast, increasing the consumption of reducing sugars and production of ethanol, compared with fermentations supplemented with ammonium sulfate. The preference of K. africana for each of the 20 amino-acids was further determined in batch fermentations and we found that asparagine supplementation increased K. africana biomass production, reducing sugar consumption and ethanol production (by 30, 36.7 and 45%, respectively) over fermentations supplemented with ammonium sulfate. Therefore, asparagine appears to overcome K. africana nutritional limitation in Agave juice. Surprisingly, K. africana produced a high concentration of ethanol. This contrasts to poor ethanol productivities reported for other non-Saccharomyces yeasts indicating a relatively high ethanol tolerance for the K. africana K1 strain. Kloeckera spp. strains are known to synthesize a wide variety of volatile compounds and we have shown that amino-acid supplements influenced the synthesis by K. africana of important metabolites involved in the bouquet of tequila. The findings of this study have revealed important nutritional limitations of non-Saccharomyces yeasts fermenting Agave tequilana juice, and have highlighted the potential of K. africana in tequila production processes.

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.

Effects of soya fatty acids on cassava ethanol fermentation.

PubMed

Xiao, Dongguang; Wu, Shuai; Zhu, Xudong; Chen, Yefu; Guo, Xuewu

2010-01-01

Ethanol tolerance is a key trait of microbes in bioethanol production. Previous studies have shown that soya flour contributed to the increase of ethanol tolerance of yeast cells. In this paper, the mechanism of this ethanol tolerance improvement was investigated in cassava ethanol fermentation supplemented with soya flour or defatted soya flour, respectively. Experiment results showed that ethanol tolerance of cells from soya flour supplemented medium increased by 4-6% (v/v) than the control with defatted soya flour. Microscopic observation found that soya flour can retain the cell shape while dramatic elongations of cells were observed with the defatted soya flour supplemented medium. Unsaturated fatty acids (UFAs) compositions of cell membrane were analyzed and the UFAs amounts increased significantly in all tested strains grown in soya flour supplemented medium. Growth study also showed that soya flour stimulated the cell growth rate by approximately tenfolds at 72-h fermentation. All these results suggested that soya fatty acids play an important role to protect yeast cells from ethanol stress during fermentation process.

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

PubMed

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

2011-07-01

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

A novel wild-type Saccharomyces cerevisiae strain TSH1 in scaling-up of solid-state fermentation of ethanol from sweet sorghum stalks.

PubMed

Du, Ran; Yan, Jianbin; Feng, Quanzhou; Li, Peipei; Zhang, Lei; Chang, Sandra; Li, Shizhong

2014-01-01

The rising demand for bioethanol, the most common alternative to petroleum-derived fuel used worldwide, has encouraged a feedstock shift to non-food crops to reduce the competition for resources between food and energy production. Sweet sorghum has become one of the most promising non-food energy crops because of its high output and strong adaptive ability. However, the means by which sweet sorghum stalks can be cost-effectively utilized for ethanol fermentation in large-scale industrial production and commercialization remains unclear. In this study, we identified a novel Saccharomyces cerevisiae strain, TSH1, from the soil in which sweet sorghum stalks were stored. This strain exhibited excellent ethanol fermentative capacity and ability to withstand stressful solid-state fermentation conditions. Furthermore, we gradually scaled up from a 500-mL flask to a 127-m3 rotary-drum fermenter and eventually constructed a 550-m3 rotary-drum fermentation system to establish an efficient industrial fermentation platform based on TSH1. The batch fermentations were completed in less than 20 hours, with up to 96 tons of crushed sweet sorghum stalks in the 550-m3 fermenter reaching 88% of relative theoretical ethanol yield (RTEY). These results collectively demonstrate that ethanol solid-state fermentation technology can be a highly efficient and low-cost solution for utilizing sweet sorghum, providing a feasible and economical means of developing non-food bioethanol.

A Novel Wild-Type Saccharomyces cerevisiae Strain TSH1 in Scaling-Up of Solid-State Fermentation of Ethanol from Sweet Sorghum Stalks

PubMed Central

Feng, Quanzhou; Li, Peipei; Zhang, Lei; Chang, Sandra; Li, Shizhong

2014-01-01

The rising demand for bioethanol, the most common alternative to petroleum-derived fuel used worldwide, has encouraged a feedstock shift to non-food crops to reduce the competition for resources between food and energy production. Sweet sorghum has become one of the most promising non-food energy crops because of its high output and strong adaptive ability. However, the means by which sweet sorghum stalks can be cost-effectively utilized for ethanol fermentation in large-scale industrial production and commercialization remains unclear. In this study, we identified a novel Saccharomyces cerevisiae strain, TSH1, from the soil in which sweet sorghum stalks were stored. This strain exhibited excellent ethanol fermentative capacity and ability to withstand stressful solid-state fermentation conditions. Furthermore, we gradually scaled up from a 500-mL flask to a 127-m3 rotary-drum fermenter and eventually constructed a 550-m3 rotary-drum fermentation system to establish an efficient industrial fermentation platform based on TSH1. The batch fermentations were completed in less than 20 hours, with up to 96 tons of crushed sweet sorghum stalks in the 550-m3 fermenter reaching 88% of relative theoretical ethanol yield (RTEY). These results collectively demonstrate that ethanol solid-state fermentation technology can be a highly efficient and low-cost solution for utilizing sweet sorghum, providing a feasible and economical means of developing non-food bioethanol. PMID:24736641

Production of cellulosic ethanol from sugarcane bagasse by steam explosion: Effect of extractives content, acid catalysis and different fermentation technologies.

PubMed

Neves, P V; Pitarelo, A P; Ramos, L P

2016-05-01

The production of cellulosic ethanol was carried out using samples of native (NCB) and ethanol-extracted (EECB) sugarcane bagasse. Autohydrolysis (AH) exhibited the best glucose recovery from both samples, compared to the use of both H3PO4 and H2SO4 catalysis at the same pretreatment time and temperature. All water-insoluble steam-exploded materials (SEB-WI) resulted in high glucose yields by enzymatic hydrolysis. SHF (separate hydrolysis and fermentation) gave ethanol yields higher than those obtained by SSF (simultaneous hydrolysis and fermentation) and pSSF (pre-hydrolysis followed by SSF). For instance, AH gave 25, 18 and 16 g L(-1) of ethanol by SHF, SSF and pSSF, respectively. However, when the total processing time was taken into account, pSSF provided the best overall ethanol volumetric productivity of 0.58 g L(-1) h(-1). Also, the removal of ethanol-extractable materials from cane bagasse had no influence on the cellulosic ethanol production of SEB-WI, regardless of the fermentation strategy used for conversion. Copyright © 2016 Elsevier Ltd. All rights reserved.

Production of fuel ethanol from bamboo by concentrated sulfuric acid hydrolysis followed by continuous ethanol fermentation.

PubMed

Sun, Zhao-Yong; Tang, Yue-Qin; Iwanaga, Tomohiro; Sho, Tomohiro; Kida, Kenji

2011-12-01

An efficient process for the production of fuel ethanol from bamboo that consisted of hydrolysis with concentrated sulfuric acid, removal of color compounds, separation of acid and sugar, hydrolysis of oligosaccharides and subsequent continuous ethanol fermentation was developed. The highest sugar recovery efficiency was 81.6% when concentrated sulfuric acid hydrolysis was carried out under the optimum conditions. Continuous separation of acid from the saccharified liquid after removal of color compounds with activated carbon was conducted using an improved simulated moving bed (ISMB) system, and 98.4% of sugar and 90.5% of acid were recovered. After oligosaccharide hydrolysis and pH adjustment, the unsterilized saccharified liquid was subjected to continuous ethanol fermentation using Saccharomycescerevisiae strain KF-7. The ethanol concentration, the fermentation yield based on glucose and the ethanol productivity were approximately 27.2 g/l, 92.0% and 8.2 g/l/h, respectively. These results suggest that the process is effective for production of fuel ethanol from bamboo. Copyright © 2011 Elsevier Ltd. All rights reserved.

Utilization of concentrate after membrane filtration of sugar beet thin juice for ethanol production.

PubMed

Kawa-Rygielska, Joanna; Pietrzak, Witold; Regiec, Piotr; Stencel, Piotr

2013-04-01

The subject of this study was to investigate the feasibility of the concentrate obtained after membrane ultrafiltration of sugar beet thin juice for ethanol production and selection of fermentation conditions (yeast strain and media supplementation). Resulting concentrate was subjected to batch ethanol fermentation using two strains of Saccharomyces cerevisiae (Ethanol Red and Safdistill C-70). The effect of different forms of media supplementation (mineral salts: (NH4)2SO4, K2HPO4, MgCl2; urea+Mg3(PO4)2 and yeast extract) on the fermentation course was also studied. It was stated that sugar beet juice concentrate is suitable for ethanol production yielding, depending on the yeast strain, ca. 85-87 g L(-1) ethanol with ca. 82% practical yield and more than 95% of sugars consumption after 72 h of fermentation. Nutrients enrichment further increased ethanol yield. The best results were obtained for media supplemented with urea+Mg3(PO4)2 yielding 91.16-92.06 g L(-1) ethanol with practical yield ranging 84.78-85.62% and full sugars consumption. Copyright © 2013. Published by Elsevier Ltd.

Effect of Specific Growth Rate on Fermentative Capacity of Baker’s Yeast

PubMed Central

Van Hoek, Pim; Van Dijken, Johannes P.; Pronk, Jack T.

1998-01-01

The specific growth rate is a key control parameter in the industrial production of baker’s yeast. Nevertheless, quantitative data describing its effect on fermentative capacity are not available from the literature. In this study, the effect of the specific growth rate on the physiology and fermentative capacity of an industrial Saccharomyces cerevisiae strain in aerobic, glucose-limited chemostat cultures was investigated. At specific growth rates (dilution rates, D) below 0.28 h−1, glucose metabolism was fully respiratory. Above this dilution rate, respirofermentative metabolism set in, with ethanol production rates of up to 14 mmol of ethanol · g of biomass−1 · h−1 at D = 0.40 h−1. A substantial fermentative capacity (assayed offline as ethanol production rate under anaerobic conditions) was found in cultures in which no ethanol was detectable (D < 0.28 h−1). This fermentative capacity increased with increasing dilution rates, from 10.0 mmol of ethanol · g of dry yeast biomass−1 · h−1 at D = 0.025 h−1 to 20.5 mmol of ethanol · g of dry yeast biomass−1 · h−1 at D = 0.28 h−1. At even higher dilution rates, the fermentative capacity showed only a small further increase, up to 22.0 mmol of ethanol · g of dry yeast biomass−1 · h−1 at D = 0.40 h−1. The activities of all glycolytic enzymes, pyruvate decarboxylase, and alcohol dehydrogenase were determined in cell extracts. Only the in vitro activities of pyruvate decarboxylase and phosphofructokinase showed a clear positive correlation with fermentative capacity. These enzymes are interesting targets for overexpression in attempts to improve the fermentative capacity of aerobic cultures grown at low specific growth rates. PMID:9797269

Impact of an acid fungal protease in high gravity fermentation for ethanol production using Indian sorghum as a feedstock.

PubMed

Gohel, V; Duan, G; Maisuria, V B

2013-01-01

This study evaluated the conventional jet cooking liquefaction process followed by simultaneous saccharification and fermentation (SSF) at 30% and 35% dry solids (DS) concentration of Indian sorghum feedstock for ethanol production, with addition of acid fungal protease or urea. To evaluate the efficacy of thermostable α-amylase in liquefaction at 30% and 35% DS concentration of Indian sorghum, liquefact solubility, higher dextrins, and fermentable sugars were analyzed at the end of the process. The liquefact was further subjected to SSF using yeast. In comparison with urea, addition of an acid fungal protease during SSF process was observed to accelerate yeast growth (μ), substrate consumption (Q(s)), ultimately ethanol yield based on substrate (Y(p/s)) and ethanol productivity based on fermentation time (Q(p)). The fermentation efficiency and ethanol recovery were determined for both concentrations of Indian sorghum and found to be increased with use of acid fungal protease in SSF process. Copyright © 2013 American Institute of Chemical Engineers.

Contamination issues in continuous fermentation for ethanol production

USDA-ARS?s Scientific Manuscript database

Continuous fermentation processes are employed by corn wet milling plants all over world to convert starch to ethanol. Contaminations by bacterial microorganisms like Lactobacillus and wild yeasts like Brettanomyces are common and result in lower ethanol yields. Contaminants compete with inoculate...

System for extracting protein from a fermentation product

DOEpatents

Lawton, Jr., John Warren; Bootsma, Jason Alan; Lewis, Stephen Michael

2016-04-26

A method of producing bioproducts from a feedstock in a system configured to produce ethanol and distillers grains from a fermentation product is disclosed. A system configured to process feedstock into a fermentation product and bioproducts including ethanol and meal is disclosed. A bioproduct produced from a fermentation product produced from a feedstock in a biorefining system is disclosed.

Method for extracting protein from a fermentation product

DOEpatents

Lawton, Jr., John Warren; Bootsma, Jason Alan; Lewis, Stephen Michael

2014-02-18

A method of producing bioproducts from a feedstock in a system configured to produce ethanol and distillers grains from a fermentation product is disclosed. A system configured to process feedstock into a fermentation product and bioproducts including ethanol and meal is disclosed. A bioproduct produced from a fermentation product produced from a feedstock in a biorefining system is disclosed.

Techno-economic analysis of extraction-based separation systems for acetone, butanol, and ethanol recovery and purification.

PubMed

Grisales Díaz, Víctor Hugo; Olivar Tost, Gerard

2017-01-01

Dual extraction, high-temperature extraction, mixture extraction, and oleyl alcohol extraction have been proposed in the literature for acetone, butanol, and ethanol (ABE) production. However, energy and economic evaluation under similar assumptions of extraction-based separation systems are necessary. Hence, the new process proposed in this work, direct steam distillation (DSD), for regeneration of high-boiling extractants was compared with several extraction-based separation systems. The evaluation was performed under similar assumptions through simulation in Aspen Plus V7.3 ® software. Two end distillation systems (number of non-ideal stages between 70 and 80) were studied. Heat integration and vacuum operation of some units were proposed reducing the energy requirements. Energy requirement of hybrid processes, substrate concentration of 200 g/l, was between 6.4 and 8.3 MJ-fuel/kg-ABE. The minimum energy requirements of extraction-based separation systems, feeding a water concentration in the substrate equivalent to extractant selectivity, and ideal assumptions were between 2.6 and 3.5 MJ-fuel/kg-ABE, respectively. The efficiencies of recovery systems for baseline case and ideal evaluation were 0.53-0.57 and 0.81-0.84, respectively. The main advantages of DSD were the operation of the regeneration column at atmospheric pressure, the utilization of low-pressure steam, and the low energy requirements of preheating. The in situ recovery processes, DSD, and mixture extraction with conventional regeneration were the approaches with the lowest energy requirements and total annualized costs.

Simultaneous saccharification and fermentation of ground corn stover for the production of fuel ethanol using Phanerochaete chrysosporium, Gloeophyllum trabeum, Saccharomyces cerevisiae, and Escherichia coli K011.

PubMed

Vincent, Micky; Pometto, Anthony L; van Leeuwen, J Hans

2011-07-01

Enzymatic saccharification of corn stover using Phanerochaete chrysosporium and Gloeophyllum trabeum and subsequent fermentation of the saccharification products to ethanol by Saccharomyces cerevisiae and Escherichia coli K011 were achieved. Prior to simultaneous saccharification and fermentation (SSF) for ethanol production, solid-state fermentation was performed for four days on ground corn stover using either P. chrysosporium or G. trabeum to induce in situ cellulase production. During SSF with S. cerevisiae or E. coli, ethanol production was the highest on day 4 for all samples. For corn stover treated with P. chrysosporium, the conversion to ethanol was 2.29 g/100 g corn stover with S. cerevisiae as the fermenting organism, whereas for the sample inoculated with E. coli K011, the ethanol production was 4.14 g/100 g corn stover. Corn stover treated with G. trabeum showed a conversion 1.90 and 4.79 g/100 g corn stover with S. cerevisiae and E. coli K011 as the fermenting organisms, respectively. Other fermentation co-products, such as acetic acid and lactic acid, were also monitored. Acetic acid production ranged between 0.45 and 0.78 g/100 g corn stover, while no lactic acid production was detected throughout the 5 days of SSF. The results of our experiment suggest that it is possible to perform SSF of corn stover using P. chrysosporium, G. trabeum, S. cerevisiae and E. coli K011 for the production of fuel ethanol.

Real-time monitoring of high-gravity corn mash fermentation using in situ raman spectroscopy.

PubMed

Gray, Steven R; Peretti, Steven W; Lamb, H Henry

2013-06-01

In situ Raman spectroscopy was employed for real-time monitoring of simultaneous saccharification and fermentation (SSF) of corn mash by an industrial strain of Saccharomyces cerevisiae. An accurate univariate calibration model for ethanol was developed based on the very strong 883 cm(-1) C-C stretching band. Multivariate partial least squares (PLS) calibration models for total starch, dextrins, maltotriose, maltose, glucose, and ethanol were developed using data from eight batch fermentations and validated using predictions for a separate batch. The starch, ethanol, and dextrins models showed significant prediction improvement when the calibration data were divided into separate high- and low-concentration sets. Collinearity between the ethanol and starch models was avoided by excluding regions containing strong ethanol peaks from the starch model and, conversely, excluding regions containing strong saccharide peaks from the ethanol model. The two-set calibration models for starch (R(2)  = 0.998, percent error = 2.5%) and ethanol (R(2)  = 0.999, percent error = 2.1%) provide more accurate predictions than any previously published spectroscopic models. Glucose, maltose, and maltotriose are modeled to accuracy comparable to previous work on less complex fermentation processes. Our results demonstrate that Raman spectroscopy is capable of real time in situ monitoring of a complex industrial biomass fermentation. To our knowledge, this is the first PLS-based chemometric modeling of corn mash fermentation under typical industrial conditions, and the first Raman-based monitoring of a fermentation process with glucose, oligosaccharides and polysaccharides present. Copyright © 2013 Wiley Periodicals, Inc.

Enhancing ethanol yields through d-xylose and l-arabinose co-fermentation after construction of a novel high efficient l-arabinose-fermenting Saccharomyces cerevisiae strain.

PubMed

Caballero, Antonio; Ramos, Juan Luis

2017-04-01

Lignocellulose contains two pentose sugars, l-arabinose and d-xylose, neither of which is naturally fermented by first generation (1G) ethanol-producing Saccharomyces cerevisiae yeast. Since these sugars are inaccessible to 1G yeast, a significant percentage of the total carbon in bioethanol production from plant residues, which are used in second generation (2G) ethanol production, remains unused. Recombinant Saccharomyces cerevisiae strains capable of fermenting d-xylose are available on the market; however, there are few examples of l-arabinose-fermenting yeasts, and commercially, there are no strains capable of fermenting both d-xylose and l-arabinose because of metabolic incompatibilities when both metabolic pathways are expressed in the same cell. To attempt to solve this problem we have tested d-xylose and l-arabinose co-fermentation. To find efficient alternative l-arabinose utilization pathways to the few existing ones, we have used stringent methodology to screen for new genes (metabolic and transporter functions) to facilitate l-arabinose fermentation in recombinant yeast. We demonstrate the feasibility of this approach in a successfully constructed yeast strain capable of using l-arabinose as the sole carbon source and capable of fully transforming it to ethanol, reaching the maximum theoretical fermentation yield (0.43 g g-1). We demonstrate that efficient co-fermentation of d-xylose and l-arabinose is feasible using two different co-cultured strains, and observed no fermentation delays, yield drops or accumulation of undesired byproducts. In this study we have identified a technically efficient strategy to enhance ethanol yields by 10 % in 2G plants in a process based on C5 sugar co-fermentation.

Enhanced bioprocessing of lignocellulose: Wood-rot fungal saccharification and fermentation of corn fiber to ethanol

NASA Astrophysics Data System (ADS)

Shrestha, Prachand

This research aims at developing a biorefinery platform to convert corn-ethanol coproduct, corn fiber, into fermentable sugars at a lower temperature with minimal use of chemicals. White-rot (Phanerochaete chrysosporium), brown-rot (Gloeophyllum trabeum) and soft-rot (Trichoderma reesei) fungi were used in this research to biologically break down cellulosic and hemicellulosic components of corn fiber into fermentable sugars. Laboratory-scale simultaneous saccharification and fermentation (SSF) process proceeded by in-situ cellulolytic enzyme induction enhanced overall enzymatic hydrolysis of hemi/cellulose from corn fiber into simple sugars (mono-, di-, tri-saccharides). The yeast fermentation of hydrolyzate yielded 7.1, 8.6 and 4.1 g ethanol per 100 g corn fiber when saccharified with the white-, brown-, and soft-rot fungi, respectively. The highest corn-to-ethanol yield (8.6 g ethanol/100 g corn fiber) was equivalent to 42 % of the theoretical ethanol yield from starch and cellulose in corn fiber. Cellulase, xylanase and amylase activities of these fungi were also investigated over a week long solid-substrate fermentation of corn fiber. G. trabeum had the highest activities for starch (160 mg glucose/mg protein.min) and on day three of solid-substrate fermentation. P. chrysosporium had the highest activity for xylan (119 mg xylose/mg protein.min) on day five and carboxymethyl cellulose (35 mg glucose/mg protein.min) on day three of solid-substrate fermentation. T. reesei showed the highest activity for Sigma cell 20 (54.8 mg glucose/mg protein.min) on day 5 of solid-substrate fermentation. The effect of different pretreatments on SSF of corn fiber by fungal processes was examined. Corn fiber was treated at 30 °C for 2 h with alkali [2% NaOH (w/w)], alkaline peroxide [2% NaOH (w/w) and 1% H2O 2 (w/w)], and by steaming at 100 °C for 2 h. Mild pretreatment resulted in improved ethanol yields for brown- and soft-rot SSF, while white-rot and Spezyme CP SSFs showed no improvement in ethanol yields. We showed that saccharification of lignocellulosic material with a wood-rot fungal process is quite feasible. Corn fiber from wet milling was best degraded to sugars using aerobic solid state fermentation with the soft-rot fungus T. reesei. However, it was shown that both the white-rot fungus P. chrysosporium and brown-rot fungus G. trabeum had the ability to produce additional consortia of hemi/cellulose degrading enzymes. It is likely that a consortium of enzymes from these fungi would be the best approach in saccharification of lignocellulose. In all cases, a subsequent anaerobic yeast process under submerged conditions is required to ferment the released sugars to ethanol. To our knowledge, this is the first time report on production of cellulolytic enzymes from wet-milled corn fiber using white- and brown-rot fungi for sequential fermentation of corn fiber hydrolyzate to ethanol. Keywords: lignocellulose, ethanol, biofuel, bioeconomy, biomass, renewable resources, corn fiber, pretreatment, solid-substrate fermentation, simultaneous saccharification and fermentation (SSF), white-rot fungus, brown-rot fungus, soft-rot fungus, fermentable sugars, enzyme activities, cellulytic enzymes Phanerochaete chrysosporium, Gloleophyllum trabeum, Trichoderma reesei, Saccharomyces cerevisiae.

Saccharomyces cerevisiae expressing bacteriophage endolysins reduce Lactobacillus contamination during fermentation

USDA-ARS?s Scientific Manuscript database

One of the challenges facing the fuel ethanol industry is the management of bacterial contamination during fermentation. Lactobacillus species are the predominant contaminants that decrease the profitability of biofuel production by reducing ethanol yields and causing “stuck” fermentations, which i...

Biofilm formation and ethanol inhibition by bacterial contaminants of biofuel fermentation

USDA-ARS?s Scientific Manuscript database

Bacterial contaminants can inhibit ethanol production in biofuel fermentations, and even result in stuck fermentations. Contaminants may persist in production facilities by forming recalcitrant biofilms. A two-year longitudinal study was conducted of bacterial contaminants from a Midwestern dry grin...

Vapor-fed bio-hybrid fuel cell.

PubMed

Benyamin, Marcus S; Jahnke, Justin P; Mackie, David M

2017-01-01

Concentration and purification of ethanol and other biofuels from fermentations are energy-intensive processes, with amplified costs at smaller scales. To circumvent the need for these processes, and to potentially reduce transportation costs as well, we have previously investigated bio-hybrid fuel cells (FCs), in which a fermentation and FC are closely coupled. However, long-term operation requires strictly preventing the fermentation and FC from harming each other. We introduce here the concept of the vapor-fed bio-hybrid FC as a means of continuously extracting power from ongoing fermentations at ambient conditions. By bubbling a carrier gas (N 2 ) through a yeast fermentation and then through a direct ethanol FC, we protect the FC anode from the catalyst poisons in the fermentation (which are non-volatile), and also protect the yeast from harmful FC products (notably acetic acid) and from build-up of ethanol. Since vapor-fed direct ethanol FCs at ambient conditions have never been systematically characterized (in contrast to vapor-fed direct methanol FCs), we first assess the effects on output power and conversion efficiency of ethanol concentration, vapor flow rate, and FC voltage. The results fit a continuous stirred-tank reactor model. Over a wide range of ethanol partial pressures (2-8 mmHg), power densities are comparable to those for liquid-fed direct ethanol FCs at the same temperature, with power densities >2 mW/cm 2 obtained. We then demonstrate the continuous operation of a vapor-fed bio-hybrid FC with fermentation for 5 months, with no indication of performance degradation due to poisoning (of either the FC or the fermentation). It is further shown that the system is stable, recovering quickly from disturbances or from interruptions in maintenance. The vapor-fed bio-hybrid FC enables extraction of power from dilute bio-ethanol streams without costly concentration and purification steps. The concept should be scalable to both large and small operations and should be generalizable to other biofuels and waste-to-energy systems.

Physiological and fermentation properties of Bacillus coagulans and a mutant lacking fermentative lactate dehydrogenase activity.

PubMed

Su, Yue; Rhee, Mun Su; Ingram, Lonnie O; Shanmugam, K T

2011-03-01

Bacillus coagulans, a sporogenic lactic acid bacterium, grows optimally at 50-55 °C and produces lactic acid as the primary fermentation product from both hexoses and pentoses. The amount of fungal cellulases required for simultaneous saccharification and fermentation (SSF) at 55 °C was previously reported to be three to four times lower than for SSF at the optimum growth temperature for Saccharomyces cerevisiae of 35 °C. An ethanologenic B. coagulans is expected to lower the cellulase loading and production cost of cellulosic ethanol due to SSF at 55 °C. As a first step towards developing B. coagulans as an ethanologenic microbial biocatalyst, activity of the primary fermentation enzyme L-lactate dehydrogenase was removed by mutation (strain Suy27). Strain Suy27 produced ethanol as the main fermentation product from glucose during growth at pH 7.0 (0.33 g ethanol per g glucose fermented). Pyruvate dehydrogenase (PDH) and alcohol dehydrogenase (ADH) acting in series contributed to about 55% of the ethanol produced by this mutant while pyruvate formate lyase and ADH were responsible for the remainder. Due to the absence of PDH activity in B. coagulans during fermentative growth at pH 5.0, the l-ldh mutant failed to grow anaerobically at pH 5.0. Strain Suy27-13, a derivative of the l-ldh mutant strain Suy27, that produced PDH activity during anaerobic growth at pH 5.0 grew at this pH and also produced ethanol as the fermentation product (0.39 g per g glucose). These results show that construction of an ethanologenic B. coagulans requires optimal expression of PDH activity in addition to the removal of the LDH activity to support growth and ethanol production.

Biofuel production from Jerusalem artichoke tuber inulins: a review

DOE PAGES

Bhagia, Samarthya; Akinosho, Hannah; Ferreira, Jorge F. S.; ...

2017-06-01

Jerusalem artichoke (JA) has a high productivity of tubers that are rich in inulins, a fructan polymer. These inulins can be easily broken down into fructose and glucose for conversion into ethanol by fermentation. This paper discusses tuber and inulin yields, effect of cultivar and environment on tuber productivity, and approaches to fermentation for ethanol production. Consolidated bioprocessing with Kluyveromyces marxianus has been the most popular approach for fermentation into ethanol. Apart from ethanol, fructose can be dehydrated into into 5-hydrolxymethylfurfural followed by catalytic conversion into hydrocarbons. Finally, findings from several studies indicate that this plant from tubers alone canmore » produce ethanol at yields that rival corn and sugarcane ethanol. JA has tremendous potential for use as a bioenergy feedstock.« less

Biofuel production from Jerusalem artichoke tuber inulins: a review

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

Bhagia, Samarthya; Akinosho, Hannah; Ferreira, Jorge F. S.

Jerusalem artichoke (JA) has a high productivity of tubers that are rich in inulins, a fructan polymer. These inulins can be easily broken down into fructose and glucose for conversion into ethanol by fermentation. This paper discusses tuber and inulin yields, effect of cultivar and environment on tuber productivity, and approaches to fermentation for ethanol production. Consolidated bioprocessing with Kluyveromyces marxianus has been the most popular approach for fermentation into ethanol. Apart from ethanol, fructose can be dehydrated into into 5-hydrolxymethylfurfural followed by catalytic conversion into hydrocarbons. Finally, findings from several studies indicate that this plant from tubers alone canmore » produce ethanol at yields that rival corn and sugarcane ethanol. JA has tremendous potential for use as a bioenergy feedstock.« less

Troubleshooting fermentation in corn wet milling ethanol production

USDA-ARS?s Scientific Manuscript database

To convert starch to ethanol, continuous fermentation processes are employed by corn wet milling plants all over world. Contaminations by bacterial microorganisms like Lactobacillus and wild yeasts like Brettanomyces are common and result in lower ethanol yields (Abbott and Ingledew 2005, Skinner an...

SEPARATION AND CONCENTRATION OF ETHANOL BY PERVAPORATION

EPA Science Inventory

A significant issue affecting widespread acceptance of bioethanol as a sustainable fuel is the energy used to grow the feedstock, ferment the feedstock to ethanol, and separate dry ethanol from the fermentation broth. For the latter, the best current technology is two-step disti...

Direct fermentation of raw starch using a Kluyveromyces marxianus strain that expresses glucoamylase and alpha-amylase to produce ethanol.

PubMed

Wang, Rongliang; Wang, Dongmei; Gao, Xiaolian; Hong, Jiong

2014-01-01

Raw starch and raw cassava tuber powder were directly and efficiently fermented at elevated temperatures to produce ethanol using the thermotolerant yeast Kluyveromyces marxianus that expresses α-amylase from Aspergillus oryzae as well as α-amylase and glucoamylase from Debaryomyces occidentalis. Among the constructed K. marxianus strains, YRL 009 had the highest efficiency in direct starch fermentation. Raw starch from corn, potato, cassava, or wheat can be fermented at temperatures higher than 40°C. At the optimal fermentation temperature 42°C, YRL 009 produced 66.52 g/L ethanol from 200 g/L cassava starch, which was the highest production among the selected raw starches. This production increased to 79.75 g/L ethanol with a 78.3% theoretical yield (with all cassava starch were consumed) from raw cassava starch at higher initial cell densities. Fermentation was also carried out at 45 and 48°C. By using 200 g/L raw cassava starch, 137.11 and 87.71 g/L sugar were consumed with 55.36 and 32.16 g/L ethanol produced, respectively. Furthermore, this strain could directly ferment 200 g/L nonsterile raw cassava tuber powder (containing 178.52 g/L cassava starch) without additional nutritional supplements to produce 69.73 g/L ethanol by consuming 166.07 g/L sugar at 42°C. YRL 009, which has consolidated bioprocessing ability, is the best strain for fermenting starches at elevated temperatures that has been reported to date. © 2014 American Institute of Chemical Engineers.

Fermentative and growth performances of Dekkera bruxellensis in different batch systems and the effect of initial low cell counts in co-cultures with Saccharomyces cerevisiae.

PubMed

Meneghin, Maria Cristina; Bassi, Ana Paula Guarnieri; Codato, Carolina Brito; Reis, Vanda Renata; Ceccato-Antonini, Sandra Regina

2013-08-01

Dekkera bruxellensis is a multifaceted yeast present in the fermentative processes used for alcoholic beverage and fuel alcohol production - in the latter, normally regarded as a contaminant. We evaluated the fermentation and growth performance of a strain isolated from water in an alcohol-producing unit, in batch systems with/without cell recycling in pure and co-cultures with Saccharomyces cerevisiae. The ethanol resistance and aeration dependence for ethanol/acid production were verified. Ethanol had an effect on the growth of D. bruxellensis in that it lowered or inhibited growth depending on the concentration. Acid production was verified in agitated cultures either with glucose or sucrose, but more ethanol was produced with glucose in agitated cultures. Regardless of the batch system, low sugar consumption and alcohol production and expressive growth were found with D. bruxellensis. Despite a similar ethanol yield compared to S. cerevisiae in the batch system without cell recycling, ethanol productivity was approximately four times lower. However, with cell recycling, ethanol yield was almost half that of S. cerevisiae. At initial low cell counts of D. bruxellensis (10 and 1000 cells/ml) in co-cultures with S. cerevisiae, a decrease in fermentative efficiency and a substantial growth throughout the fermentative cycles were displayed by D. bruxellensis. Due to the peculiarity of cell repitching in Brazilian fermentation processes, D. bruxellensis is able to establish itself in the process, even when present in low numbers initially, substantially impairing bioethanol production due to the low ethanol productivity, in spite of comparable ethanol yields. Copyright © 2013 John Wiley & Sons, Ltd.

Mathematical modeling of the fermentation of acid-hydrolyzed pyrolytic sugars to ethanol by the engineered strain Escherichia coli ACCC 11177.

PubMed

Chang, Dongdong; Yu, Zhisheng; Islam, Zia Ul; Zhang, Hongxun

2015-05-01

Pyrolysate from waste cotton was acid hydrolyzed and detoxified to yield pyrolytic sugars, which were fermented to ethanol by the strain Escherichia coli ACCC 11177. Mathematical models based on the fermentation data were also constructed. Pyrolysate containing an initial levoglucosan concentration of 146.34 g/L gave a glucose yield of 150 % after hydrolysis, suggesting that other compounds were hydrolyzed to glucose as well. Ethyl acetate-based extraction of bacterial growth inhibitors with an ethyl acetate/hydrolysate ratio of 1:0.5 enabled hydrolysate fermentation by E. coli ACCC 11177, without a standard absorption treatment. Batch processing in a fermenter exhibited a maximum ethanol yield and productivity of 0.41 g/g and 0.93 g/L·h(-1), respectively. The cell growth rate (r x ) was consistent with a logistic equation [Formula: see text], which was determined as a function of cell growth (X). Glucose consumption rate (r s ) and ethanol formation rate (r p ) were accurately validated by the equations [Formula: see text] and [Formula: see text], respectively. Together, our results suggest that combining mathematical models with fermenter fermentation processes can enable optimized ethanol production from cellulosic pyrolysate with E. coli. Similar approaches may facilitate the production of other commercially important organic substances.

Simultaneous Saccharification and Fermentation of Sugar Beet Pulp for Efficient Bioethanol Production

PubMed Central

Berłowska, Joanna; Balcerek, Maria; Dziekońska-Kubczak, Urszula; Patelski, Piotr; Dziugan, Piotr

2016-01-01

Sugar beet pulp, a byproduct of sugar beet processing, can be used as a feedstock in second-generation ethanol production. The objective of this study was to investigate the effects of pretreatment, of the dosage of cellulase and hemicellulase enzyme preparations used, and of aeration on the release of fermentable sugars and ethanol yield during simultaneous saccharification and fermentation (SSF) of sugar beet pulp-based worts. Pressure-thermal pretreatment was applied to sugar beet pulp suspended in 2% w/w sulphuric acid solution at a ratio providing 12% dry matter. Enzymatic hydrolysis was conducted using Viscozyme and Ultraflo Max (Novozymes) enzyme preparations (0.015–0.02 mL/g dry matter). Two yeast strains were used for fermentation: Ethanol Red (S. cerevisiae) (1 g/L) and Pichia stipitis (0.5 g/L), applied sequentially. The results show that efficient simultaneous saccharification and fermentation of sugar beet pulp was achieved. A 6 h interval for enzymatic activation between the application of enzyme preparations and inoculation with Ethanol Red further improved the fermentation performance, with the highest ethanol concentration reaching 26.9 ± 1.2 g/L and 86.5 ± 2.1% fermentation efficiency relative to the theoretical yield. PMID:27722169

Effect of agitation rate on ethanol production from sugar maple hemicellulosic hydrolysate by Pichia stipitis.

PubMed

Shupe, Alan M; Liu, Shijie

2012-09-01

Concentrated dilute acid hydrolysate was obtained from hot water extracts of Acer saccharum (sugar maple) and was fermented to ethanol by Pichia stipitis in a 1.3-L-benchtop bioreactor. The conditions under which the highest ethanol yield was achieved were when the air flow rate was set to 100 cm(3) and the agitation rate was set to 150 rpm resulting in an overall mass transfer coefficient (K(L)a) of 0.108 min(-1). A maximum ethanol concentration of 29.7 g/L was achieved after 120 h of fermentation; however, after 90 h of fermentation, the ethanol concentration was only slightly lower at 29.1 g/L with a yield of 0.39 g ethanol per gram of sugar consumed. Using the same air flow rate and adjusting the agitation rate resulted in lower ethanol yields of 0.25 g/g at 50 rpm and 0.30 g/g at 300 rpm. The time it takes to reach the maximum ethanol concentration was also affected by the agitation rate. The ethanol concentration continued to increase even after 130 h of fermentation when the agitation rate was set at 50 rpm, whereas the maximum ethanol concentration was reached after only 68.5 h at 300 rpm.

Bacteriophage endolysins expressed in yeast kill strains of Lactobacillus that contaminate fermentations

USDA-ARS?s Scientific Manuscript database

One of the challenges facing the fuel ethanol industry is the management of bacterial contamination during fermentation. Species of Lactobacillus are the predominant contaminants that reduce ethanol yields and cause “stuck” fermentations, decreasing the profitability of biofuel production with expen...

Overcoming bacterial contamination of fuel ethanol fermentations -- alterntives to antibiotics

USDA-ARS?s Scientific Manuscript database

Fuel ethanol fermentations are not performed under aseptic conditions and microbial contamination reduces yields and can lead to costly "stuck fermentations". Antibiotics are commonly used to combat contaminants, but these may persist in the distillers grains co-product. Among contaminants, it is kn...

Xylose fermentation to ethanol. A review

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

McMillan, J D

1993-01-01

The past several years have seen tremendous progress in the understanding of xylose metabolism and in the identification, characterization, and development of strains with improved xylose fermentation characteristics. A survey of the numerous microorganisms capable of directly fermenting xylose to ethanol indicates that wild-type yeast and recombinant bacteria offer the best overall performance in terms of high yield, final ethanol concentration, and volumetric productivity. The best performing bacteria, yeast, and fungi can achieve yields greater than 0.4 g/g and final ethanol concentrations approaching 5%. Productivities remain low for most yeast and particularly for fungi, but volumetric productivities exceeding 1.0 g/L-hmore » have been reported for xylose-fermenting bacteria. In terms of wild-type microorganisms, strains of the yeast Pichia stipitis show the most promise in the short term for direct high-yield fermentation of xylose without byproduct formation. Of the recombinant xylose-fermenting microorganisms developed, recombinant E. coli ATTC 11303 (pLOI297) exhibits the most favorable performance characteristics reported to date.« less

Xylose fermentation to ethanol

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

McMillan, J.D.

1993-01-01

The past several years have seen tremendous progress in the understanding of xylose metabolism and in the identification, characterization, and development of strains with improved xylose fermentation characteristics. A survey of the numerous microorganisms capable of directly fermenting xylose to ethanol indicates that wild-type yeast and recombinant bacteria offer the best overall performance in terms of high yield, final ethanol concentration, and volumetric productivity. The best performing bacteria, yeast, and fungi can achieve yields greater than 0.4 g/g and final ethanol concentrations approaching 5%. Productivities remain low for most yeast and particularly for fungi, but volumetric productivities exceeding 1.0 g/L-hmore » have been reported for xylose-fermenting bacteria. In terms of wild-type microorganisms, strains of the yeast Pichia stipitis show the most promise in the short term for direct high-yield fermentation of xylose without byproduct formation. Of the recombinant xylose-fermenting microorganisms developed, recombinant E. coli ATTC 11303 (pLOI297) exhibits the most favorable performance characteristics reported to date.« less

Immobilized Kluyveromyces marxianus cells in carboxymethyl cellulose for production of ethanol from cheese whey: experimental and kinetic studies.

PubMed

Roohina, Fatemeh; Mohammadi, Maedeh; Najafpour, Ghasem D

2016-09-01

Cheese whey fermentation to ethanol using immobilized Kluyveromyces marxianus cells was investigated in batch and continuous operation. In batch fermentation, the yeast cells were immobilized in carboxymethyl cellulose (CMC) polymer and also synthesized graft copolymer of CMC with N-vinyl-2-pyrrolidone, denoted as CMC-g-PVP, and the efficiency of the two developed cell entrapped beads for lactose fermentation to ethanol was examined. The yeast cells immobilized in CMC-g-PVP performed slightly better than CMC with ethanol production yields of 0.52 and 0.49 g ethanol/g lactose, respectively. The effect of supplementation of cheese whey with lactose (42, 70, 100 and 150 g/l) on fermentative performance of K. marxianus immobilized in CMC beads was considered and the results were used for kinetic studies. The first order reaction model was suitable to describe the kinetics of substrate utilization and modified Gompertz model was quite successful to predict the ethanol production. For continuous ethanol fermentation, a packed-bed immobilized cell reactor (ICR) was operated at several hydraulic retention times; HRTs of 11, 15 and 30 h. At the HRT of 30 h, the ethanol production yield using CMC beads was 0.49 g/g which implies that 91.07 % of the theoretical yield was achieved.

Evaluation of hardboard manufacturing process wastewater as a feedstream for ethanol production.

PubMed

Groves, Stephanie; Liu, Jifei; Shonnard, David; Bagley, Susan

2013-07-01

Waste streams from the wood processing industry can serve as feedstream for ethanol production from biomass residues. Hardboard manufacturing process wastewater (HPW) was evaluated on the basis of monomeric sugar recovery and fermentability as a novel feedstream for ethanol production. Dilute acid hydrolysis, coupled with concentration of the wastewater resulted in a hydrolysate with 66 g/l total fermentable sugars. As xylose accounted for 53 % of the total sugars, native xylose-fermenting yeasts were evaluated for their ability to produce ethanol from the hydrolysate. The strains selected were, in decreasing order by ethanol yields from xylose (Y p/s, based on consumed sugars), Scheffersomyces stipitis ATCC 58785 (CBS 6054), Pachysolen tannophilus ATCC 60393, and Kluyveromyces marxianus ATCC 46537. The yeasts were compared on the basis of substrate utilization and ethanol yield during fermentations of the hydrolysate, measured using an HPLC. S. stipitis, P. tannophilus, and K. marxianus produced 0.34, 0.31, and 0.36 g/g, respectively. The yeasts were able to utilize between 58 and 75 % of the available substrate. S. stipitis outperformed the other yeast during the fermentation of the hydrolysate; consuming the highest concentration of available substrate and producing the highest ethanol concentration in 72 h. Due to its high sugar content and low inhibitor levels after hydrolysis, it was concluded that HPW is a suitable feedstream for ethanol production by S. stipitis.

Ethanol production in Brazil: a bridge between science and industry.

PubMed

Lopes, Mario Lucio; Paulillo, Silene Cristina de Lima; Godoy, Alexandre; Cherubin, Rudimar Antonio; Lorenzi, Marcel Salmeron; Giometti, Fernando Henrique Carvalho; Bernardino, Claudemir Domingues; Amorim Neto, Henrique Berbert de; Amorim, Henrique Vianna de

2016-12-01

In the last 40 years, several scientific and technological advances in microbiology of the fermentation have greatly contributed to evolution of the ethanol industry in Brazil. These contributions have increased our view and comprehension about fermentations in the first and, more recently, second-generation ethanol. Nowadays, new technologies are available to produce ethanol from sugarcane, corn and other feedstocks, reducing the off-season period. Better control of fermentation conditions can reduce the stress conditions for yeast cells and contamination by bacteria and wild yeasts. There are great research opportunities in production processes of the first-generation ethanol regarding high-value added products, cost reduction and selection of new industrial yeast strains that are more robust and customized for each distillery. New technologies have also focused on the reduction of vinasse volumes by increasing the ethanol concentrations in wine during fermentation. Moreover, conversion of sugarcane biomass into fermentable sugars for second-generation ethanol production is a promising alternative to meet future demands of biofuel production in the country. However, building a bridge between science and industry requires investments in research, development and transfer of new technologies to the industry as well as specialized personnel to deal with new technological challenges. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Matt Wecker | NREL

Science.gov Websites

acetaldehyde from bacteria. The idea was to short-sheet the ethanol fermentation pathway to produce ; Biochem. (1995) "Fermentation strategies: Acetaldehyde or ethanol?," Process Biochem. (1987

Amylolysis of raw corn by Aspergillus niger for simultaneous ethanol fermentation

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

Han, I.Y.; Steinberg, M.P.

The novelty of this approach was hydrolysis of the raw starch in ground corn to fermentable sugars that are simultaneously fermented to ethanol by yeast in a nonsterile environment. Thus, the conventional cooking step can be eliminated for energy conservation. A koji of Aspergillus niger grown on whole corn for 3 days was the crude enzyme source. A ratio of 0.2 g dry koji/g total solids was found sufficient. Optimum pH was 4.2. Ethanol concentration was 7.7% (w/w) in the aqueous phase with 92% raw starch conversion. Agitation increased rate. Sacharification was the rate-limiting step. The initial ethanol concentration preventingmore » fermentation was estimated to be 8.3% by weight. (Refs. 96).« less

Kinetics and thermodynamics of ethanol production by Saccharomyces cerevisiae MLD10 using molasses.

PubMed

Arshad, Muhammad; Ahmed, Sibtain; Zia, Muhammad Anjum; Rajoka, Muhammad Ibrahim

2014-03-01

In this study, we have used ultraviolet (UV) and γ-ray induction to get a catabolite repression resistant and thermotolerant mutant with enhanced ethanol production along with optimization of sugar concentration and temperature of fermentation. Classical mutagenesis in two consecutive cycles of UV- and γ-ray-induced mutations evolved one best catabolite-resistant and thermotolerant mutant Saccharomyces cerevisiae MLD10 which showed improved ethanol yield (0.48 ± 0.02 g g(-1)), theoretical yield (93 ± 3%), and extracellular invertase productivity (1,430 ± 50 IU l(-1) h(-1)), respectively, when fermenting 180 g sugars l(-1) in molasses medium at 43 °C in 300 m(3) working volume fermenter. Ethanol production was highly dependent on invertase production. Enthalpy (ΔH*) (32.27 kJ M(-1)) and entropy (ΔS*) (-202.88 J M(-1) K(-1)) values at 43 °C by the mutant MLD10 were significantly lower than those of β-glucosidase production by a thermophilic mutant derivative of Thermomyces lanuginosus. These results confirmed the enhanced production of ethanol and invertase by this mutant derivative. These studies proved that mutant was significantly improved for ethanol production and was thermostable in nature. Lower fermentation time for ethanol production and maintenance of ethanol production rates (3.1 g l(-1) h(-1)) at higher temperature (43 °C) by this mutant could decrease the overall cost of fermentation process and increase the quality of ethanol production.

Cellulosic ethanol production via consolidated bioprocessing by a novel thermophilic anaerobic bacterium isolated from a Himalayan hot spring.

PubMed

Singh, Nisha; Mathur, Anshu S; Tuli, Deepak K; Gupta, Ravi P; Barrow, Colin J; Puri, Munish

2017-01-01

Cellulose-degrading thermophilic anaerobic bacterium as a suitable host for consolidated bioprocessing (CBP) has been proposed as an economically suited platform for the production of second-generation biofuels. To recognize the overall objective of CBP, fermentation using co-culture of different cellulolytic and sugar-fermenting thermophilic anaerobic bacteria has been widely studied as an approach to achieving improved ethanol production. We assessed monoculture and co-culture fermentation of novel thermophilic anaerobic bacterium for ethanol production from real substrates under controlled conditions. In this study, Clostridium sp. DBT-IOC-C19, a cellulose-degrading thermophilic anaerobic bacterium, was isolated from the cellulolytic enrichment cultures obtained from a Himalayan hot spring. Strain DBT-IOC-C19 exhibited a broad substrate spectrum and presented single-step conversion of various cellulosic and hemicellulosic substrates to ethanol, acetate, and lactate with ethanol being the major fermentation product. Additionally, the effect of varying cellulose concentrations on the fermentation performance of the strain was studied, indicating a maximum cellulose utilization ability of 10 g L -1 cellulose. Avicel degradation kinetics of the strain DBT-IOC-C19 displayed 94.6% degradation at 5 g L -1 and 82.74% degradation at 10 g L -1 avicel concentration within 96 h of fermentation. In a comparative study with Clostridium thermocellum DSM 1313, the ethanol and total product concentrations were higher by the newly isolated strain on pretreated rice straw at an equivalent substrate loading. Three different co-culture combinations were used on various substrates that presented two-fold yield improvement than the monoculture during batch fermentation. This study demonstrated the direct fermentation ability of the novel thermophilic anaerobic bacteria on various cellulosic and hemicellulosic substrates into ethanol without the aid of any exogenous enzymes, representing CBP-based fermentation approach. Here, the broad substrate utilization spectrum of isolated cellulolytic thermophilic anaerobic bacterium was shown to be of potential utility. We demonstrated that the co-culture strategy involving novel strains is efficient in improving ethanol production from real substrate.

Poisoning of mixed matrix membranes by fermentation components in pervaporation of ethanol

USDA-ARS?s Scientific Manuscript database

Pervaporation is an alternative to distillation for recovering ethanol produced by fermentation of grains and biomass. Ethanol-selective mixed matrix membranes of the hydrophobic zeolite ZSM-5 in polydimethylsiloxane (PDMS) have superior performance compared to pure PDMS membranes in pervaporation o...

CARBOXYLIC ACID EFFECTS ON ETHANOL RECOVERY FROM AQUEOUS MIXTURES USING PERVAPORATION THROUGH MFI ZEOLITE-FILLED POLYDIMETHYLSILOXANE MEMBRANES

EPA Science Inventory

Most bioethanol is produced by fermenting sugars released from biomass and using distillation to recover the ethanol. Recovering ethanol from the fermentation broths using pervaporation through hydrophobic membranes is potentially economically competitive with distillation for s...

Construction and Analysis of High-Ethanol-Producing Fusants with Co-Fermentation Ability through Protoplast Fusion and Double Labeling Technology

PubMed Central

Ge, Jingping; Zhao, Jingwen; Zhang, Luyan; Zhang, Mengyun; Ping, Wenxiang

2014-01-01

Double labeling of resistance markers and report genes can be used to breed engineered Saccharomyces cerevisiae strains that can assimilate xylose and glucose as a mixed carbon source for ethanol fermentation and increased ethanol production. In this study Saccharomyces cerevisiae W5 and Candida shehatae 20335 were used as parent strains to conduct protoplast fusion and the resulting fusants were screened by double labeling. High performance liquid chromatography (HPLC) was used to assess the ethanol yield following the fermentation of xylose and glucose, as both single and mixed carbon sources, by the fusants. Interestingly, one fusant (ZLYRHZ7) was demonstrated to have an excellent fermentation performance, with an ethanol yield using the mixed carbon source of 0.424 g g−1, which compares with 0.240 g g−1 (W5) and 0.353 g g−1 (20335) for the parent strains. This indicates an improvement in the ethanol yield of 43.4% and 16.7%, respectively. PMID:25268957

Mini review: hydrogen and ethanol co-production from waste materials via microbial fermentation.

PubMed

Soo, Chiu-Shyan; Yap, Wai-Sum; Hon, Wei-Min; Phang, Lai-Yee

2015-10-01

The simultaneous production of hydrogen and ethanol by microorganisms from waste materials in a bioreactor system would establish cost-effective and time-saving biofuel production. This review aims to present the current status of fermentation processes producing hydrogen accompanied by ethanol as a co-product. We outlined the microbes used and their fundamental pathways for hydrogen and ethanol fermentation. Moreover, we discussed the exploitation of renewable and sustainable waste materials as promising feedstock and the limitations encountered. The low substrate bioconversion rate in hydrogen and ethanol co-production is regarded as the primary constraint towards the development of large scale applications. Thus, microbes with an enhanced capability have been generated via genetic manipulation to diminish the inefficiency of substrate consumption. In this review, other potential approaches to improve the performance of co-production through fermentation were also elaborated. This review will be a useful guide for the future development of hydrogen and ethanol co-production using waste materials.

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

PubMed

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

2014-09-01

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

Construction and analysis of high-ethanol-producing fusants with co-fermentation ability through protoplast fusion and double labeling technology.

PubMed

Ge, Jingping; Zhao, Jingwen; Zhang, Luyan; Zhang, Mengyun; Ping, Wenxiang

2014-01-01

Double labeling of resistance markers and report genes can be used to breed engineered Saccharomyces cerevisiae strains that can assimilate xylose and glucose as a mixed carbon source for ethanol fermentation and increased ethanol production. In this study Saccharomyces cerevisiae W5 and Candida shehatae 20335 were used as parent strains to conduct protoplast fusion and the resulting fusants were screened by double labeling. High performance liquid chromatography (HPLC) was used to assess the ethanol yield following the fermentation of xylose and glucose, as both single and mixed carbon sources, by the fusants. Interestingly, one fusant (ZLYRHZ7) was demonstrated to have an excellent fermentation performance, with an ethanol yield using the mixed carbon source of 0.424 g g-1, which compares with 0.240 g g-1 (W5) and 0.353 g g-1 (20335) for the parent strains. This indicates an improvement in the ethanol yield of 43.4% and 16.7%, respectively.

Accounting for all sugars produced during integrated production of ethanol from lignocellulosic biomass.

PubMed

Schell, Daniel J; Dowe, Nancy; Chapeaux, Alexandre; Nelson, Robert S; Jennings, Edward W

2016-04-01

Accurate mass balance and conversion data from integrated operation is needed to fully elucidate the economics of biofuel production processes. This study explored integrated conversion of corn stover to ethanol and highlights techniques for accurate yield calculations. Acid pretreated corn stover (PCS) produced in a pilot-scale reactor was enzymatically hydrolyzed and the resulting sugars were fermented to ethanol by the glucose-xylose fermenting bacteria, Zymomonas mobilis 8b. The calculations presented here account for high solids operation and oligomeric sugars produced during pretreatment, enzymatic hydrolysis, and fermentation, which, if not accounted for, leads to overestimating ethanol yields. The calculations are illustrated for enzymatic hydrolysis and fermentation of PCS at 17.5% and 20.0% total solids achieving 80.1% and 77.9% conversion of cellulose and xylan to ethanol and ethanol titers of 63g/L and 69g/L, respectively. These procedures will be employed in the future and the resulting information used for techno-economic analysis. Copyright © 2016 Elsevier Ltd. All rights reserved.

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

PubMed

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

2012-01-01

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

Comparative fermentation behaviour and chemical characteristics of Saccharomyces and Zymomonas fermented culled apple juice.

PubMed

Sandhu, D K; Joshi, V K

1994-12-01

Ethanol production from culled apple juice showed that fermentability of the juice could be enhanced by addition of DAHP or ammonium sulphate in Saccharomyces and DAHP in Zymomonas fermentation. Addition of trace elements inhibited both the fermentations and ethanol, consequently. With respect to by-products of fermentation, no clear advantage of Zymomnas fermentation of culled apple juice could be observed. Differences in physico-chemical characteristics of the fermented apple juice were also noted. Saccharomyces cerevisiae proved to be better than Zymomonas in most of the parameters and is preferrable from handling and spoilage point of view.

Ethanol at levels produced by Saccharomyces cerevisiae during wheat dough fermentation has a strong impact on dough properties.

PubMed

Jayaram, Vinay B; Rezaei, Mohammad N; Cuyvers, Sven; Verstrepen, Kevin J; Delcour, Jan A; Courtin, Christophe M

2014-09-24

Yeast's role in bread making is primarily the fermentative production of carbon dioxide to leaven the dough. Fermentation also impacts dough matrix rheology, thereby affecting the quality of the end product. Surprisingly, the role of ethanol, the other yeast primary metabolite, has been ill studied in this context. Therefore, this study aims to assess the potential impact of ethanol on yeastless dough extensibility and spread and gluten agglomeration at concentrations at which it is produced in fermenting dough, i.e., up to 60 mmol per 100 g of flour. Reduced dough extensibility and dough spread were observed upon incorporation of ethanol in the dough formula, and were more pronounced for a weak than for a strong flour. Uniaxial and biaxial extension tests showed up to 50% decrease in dough extensibility and a dough strength increase of up to 18% for 60 mmol of ethanol/100 g of flour. Ethanol enhanced gluten agglomeration of a weak flour. Sequential extraction of flour in increasing ethanol concentrations showed that better gluten-solvent interaction is a possible explanation for the changed dough behavior.

Effect of Temperature on Chinese Rice Wine Brewing with High Concentration Presteamed Whole Sticky Rice

PubMed Central

Zhang, Hong-Tao; Xiong, Weili; Hu, Jianhua; Xu, Baoguo; Lin, Chi-Chung; Xu, Ling; Jiang, Lihua

2014-01-01

Production of high quality Chinese rice wine largely depends on fermentation temperature. However, there is no report on the ethanol, sugars, and acids kinetics in the fermentation mash of Chinese rice wine treated at various temperatures. The effects of fermentation temperatures on Chinese rice wine quality were investigated. The compositions and concentrations of ethanol, sugars, glycerol, and organic acids in the mash of Chinese rice wine samples were determined by HPLC method. The highest ethanol concentration and the highest glycerol concentration both were attained at the fermentation mash treated at 23°C. The highest peak value of maltose (90 g/L) was obtained at 18°C. Lactic acid and acetic acid both achieved maximum values at 33°C. The experimental results indicated that temperature contributed significantly to the ethanol production, acid flavor contents, and sugar contents in the fermentation broth of the Chinese rice wines. PMID:24672788

Resolving bacterial contamination of fuel ethanol fermentations with beneficial bacteria - An alternative to antibiotic treatment.

PubMed

Rich, Joseph O; Bischoff, Kenneth M; Leathers, Timothy D; Anderson, Amber M; Liu, Siqing; Skory, Christopher D

2018-01-01

Fuel ethanol fermentations are not performed under aseptic conditions and microbial contamination reduces yields and can lead to costly "stuck fermentations". Antibiotics are commonly used to combat contaminants, but these may persist in the distillers grains co-product. Among contaminants, it is known that certain strains of lactic acid bacteria are capable of causing stuck fermentations, while other strains appear to be harmless. However, it was not previously known whether or how these strains interact one with another. In this study, more than 500 harmless strains of lactic acid bacteria were tested in a model system in combination with strains that cause stuck fermentations. Among these harmless strains, a group of beneficial strains was identified that restored ethanol production to near normal levels. Such beneficial strains may serve as an alternative approach to the use of antibiotics in fuel ethanol production. Published by Elsevier Ltd.

Effect of temperature on Chinese rice wine brewing with high concentration presteamed whole sticky rice.

PubMed

Liu, Dengfeng; Zhang, Hong-Tao; Xiong, Weili; Hu, Jianhua; Xu, Baoguo; Lin, Chi-Chung; Xu, Ling; Jiang, Lihua

2014-01-01

Production of high quality Chinese rice wine largely depends on fermentation temperature. However, there is no report on the ethanol, sugars, and acids kinetics in the fermentation mash of Chinese rice wine treated at various temperatures. The effects of fermentation temperatures on Chinese rice wine quality were investigated. The compositions and concentrations of ethanol, sugars, glycerol, and organic acids in the mash of Chinese rice wine samples were determined by HPLC method. The highest ethanol concentration and the highest glycerol concentration both were attained at the fermentation mash treated at 23 °C. The highest peak value of maltose (90 g/L) was obtained at 18 °C. Lactic acid and acetic acid both achieved maximum values at 33 °C. The experimental results indicated that temperature contributed significantly to the ethanol production, acid flavor contents, and sugar contents in the fermentation broth of the Chinese rice wines.

Process simulation of ethanol production from biomass gasification and syngas fermentation.

PubMed

Pardo-Planas, Oscar; Atiyeh, Hasan K; Phillips, John R; Aichele, Clint P; Mohammad, Sayeed

2017-12-01

The hybrid gasification-syngas fermentation platform can produce more bioethanol utilizing all biomass components compared to the biochemical conversion technology. Syngas fermentation operates at mild temperatures and pressures and avoids using expensive pretreatment processes and enzymes. This study presents a new process simulation model developed with Aspen Plus® of a biorefinery based on a hybrid conversion technology for the production of anhydrous ethanol using 1200tons per day (wb) of switchgrass. The simulation model consists of three modules: gasification, fermentation, and product recovery. The results revealed a potential production of about 36.5million gallons of anhydrous ethanol per year. Sensitivity analyses were also performed to investigate the effects of gasification and fermentation parameters that are keys for the development of an efficient process in terms of energy conservation and ethanol production. Copyright © 2017 Elsevier Ltd. All rights reserved.

Fermentation of biomass sugars to ethanol using native industrial yeast strains.

PubMed

Yuan, Dawei; Rao, Kripa; Relue, Patricia; Varanasi, Sasidhar

2011-02-01

In this paper, the feasibility of a technology for fermenting sugar mixtures representative of cellulosic biomass hydrolyzates with native industrial yeast strains is demonstrated. This paper explores the isomerization of xylose to xylulose using a bi-layered enzyme pellet system capable of sustaining a micro-environmental pH gradient. This ability allows for considerable flexibility in conducting the isomerization and fermentation steps. With this method, the isomerization and fermentation could be conducted sequentially, in fed-batch, or simultaneously to maximize utilization of both C5 and C6 sugars and ethanol yield. This system takes advantage of a pH-dependent complexation of xylulose with a supplemented additive to achieve up to 86% isomerization of xylose at fermentation conditions. Commercially-proven Saccharomyces cerevisiae strains from the corn-ethanol industry were used and shown to be very effective in implementation of the technology for ethanol production. Copyright © 2010 Elsevier Ltd. All rights reserved.

Simultaneous glucose and xylose uptake by an acetone/butanol/ethanol producing laboratory Clostridium beijerinckii strain SE-2.

PubMed

Zhang, Jie; Zhu, Wen; Xu, Haipeng; Li, Yan; Hua, Dongliang; Jin, Fuqiang; Gao, Mintian; Zhang, Xiaodong

2016-04-01

Most butanol-producing strains of Clostridium prefer glucose over xylose, leading to a slower butanol production from lignocellulose hydrolysates. It is therefore beneficial to find and use a strain that can simultaneously use both glucose and xylose. Clostridium beijerinckii SE-2 strain assimilated glucose and xylose simultaneously and produced ABE (acetone/butanol/ethanol). The classic diauxic growth behavior was not seen. Similar rates of sugar consumption (4.44 mM glucose h(-1) and 6.66 mM xylose h(-1)) were observed suggesting this strain could use either glucose or xylose as the substrate and it has a similar capability to degrade these two sugars. With different initial glucose:xylose ratios, glucose and xylose were consumed simultaneously at rates roughly proportional to their individual concentrations in the medium, leading to complete utilization of both sugars at the same time. ABE production profiles were similar on different substrates. Transcriptional studies on the effect of glucose and xylose supplementation, however, suggests a clear glucose inhibition on xylose metabolism-related genes is still present.

[Process development for continuous ethanol fermentation by the flocculating yeast under stillage backset conditions].

PubMed

Zi, Lihan; Liu, Chenguang; Bai, Fengwu

2014-02-01

Propionic acid, a major inhibitor to yeast cells, was accumulated during continuous ethanol fermentation from corn meal hydrolysate by the flocculating yeast under stillage backset conditions. Based on its inhibition mechanism in yeast cells, strategies were developed for alleviating this effect. Firstly, high temperature processes such as medium sterilization generated more propionic acid, which should be avoided. Propionic acid was reduced significantly during ethanol fermentation without medium sterilization, and concentrations of biomass and ethanol increased by 59.3% and 7.4%, respectively. Secondly, the running time of stillage backset should be controlled so that propionic acid accumulated would be lower than its half inhibition concentration IC50 (40 mmol/L). Finally, because low pH augmented propionic acid inhibition in yeast cells, a higher pH of 5.5 was validated to be suitable for ethanol fermentation under the stillage backset condition.

Biofilm formation and ethanol inhibition by bacterial contaminants of biofuel fermentation.

PubMed

Rich, Joseph O; Leathers, Timothy D; Bischoff, Kenneth M; Anderson, Amber M; Nunnally, Melinda S

2015-11-01

Bacterial contaminants can inhibit ethanol production in biofuel fermentations, and even result in stuck fermentations. Contaminants may persist in production facilities by forming recalcitrant biofilms. A two-year longitudinal study was conducted of bacterial contaminants from a Midwestern dry grind corn fuel ethanol facility. Among eight sites sampled in the facility, the combined liquefaction stream and yeast propagation tank were consistently contaminated, leading to contamination of early fermentation tanks. Among 768 contaminants isolated, 92% were identified as Lactobacillus sp., with the most abundant species being Lactobacillus plantarum, Lactobacillus casei, Lactobacillus mucosae, and Lactobacillus fermentum. Seven percent of total isolates showed the ability to form biofilms in pure cultures, and 22% showed the capacity to significantly inhibit ethanol production. However, these traits were not correlated. Ethanol inhibition appeared to be related to acetic acid production by contaminants, particularly by obligately heterofermentative species such as L. fermentum and L. mucosae. Published by Elsevier Ltd.

Effects of lactic acid bacteria contamination on lignocellulosic ethanol fermentation

USDA-ARS?s Scientific Manuscript database

Slower fermentation rates, mixed sugar compositions, and lower sugar concentrations may make lignocellulosic fermentations more susceptible to contamination by lactic acid bacteria (LAB), which is a common and costly problem to the corn-based fuel ethanol industry. To examine the effects of LAB con...

Improvement of dry fractionation ethanol fermentation by partial germ supplementation

USDA-ARS?s Scientific Manuscript database

Ethanol fermentation of dry fractionated grits (corn endosperm pieces) containing different levels of germ was studied using the dry grind process. Partial removal of germ fraction allows for marketing the germ fraction and potentially more efficient fermentation. Grits obtained from a dry milling p...

Alcoholic fermentation of d-xylose by yeasts. [Brettanomyces naardenensis; Candida shehatae; Candida tenuis; Pachysolen tannaphilus, Pichia segobiensis; Pichia stipitis

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

Toivola, A.; Yarrow, D.; van den Bosch, E.

1984-06-01

Type strains of 200 species of yeasts able to ferment glucose and grow on xylose were screened for fermentation of D-xylose. In most of the strains tested, ethanol production was negligible. Nineteen were found to produce between 0.1 and 1.0 g of ethanol per liter. Strains of the following species produce more than 1 g of ethanol per liter in the fermentation test with 2% xylose: Brettanomyces naardenensis, Candida shehatae, Candida tenuis, Pachysolen tannophilus, Pichia segobiensis, and Pichia stipitis. Subsequent screening of these yeasts for their capacity to ferment D-cellobiose revealed that only Candida tenuis CBS 4435 was a goodmore » fermenter of both xylose and cellobiose under the test conditions used.« less

Very high gravity ethanol and fatty acid production of Zymomonas mobilis without amino acid and vitamin.

PubMed

Wang, Haoyong; Cao, Shangzhi; Wang, William Tianshuo; Wang, Kaven Tianyv; Jia, Xianhui

2016-06-01

Very high gravity (VHG) fermentation is the mainstream technology in ethanol industry, which requires the strains be resistant to multiple stresses such as high glucose concentration, high ethanol concentration, high temperature and harsh acidic conditions. To our knowledge, it was not reported previously that any ethanol-producing microbe showed a high performance in VHG fermentations without amino acid and vitamin. Here we demonstrate the engineering of a xylose utilizing recombinant Zymomonas mobilis for VHG ethanol fermentations. The recombinant strain can produce ethanol up to 136 g/L without amino acid and vitamin with a theoretical yield of 90 %, which is significantly superior to that produced by all the reported ethanol-producing strains. The intracellular fatty acids of the bacterial were about 16 % of the bacterial dry biomass, with the ratio of ethanol:fatty acids was about 273:1 (g/g). The recombinant strain was achieved by a multivariate-modular strategy tackles with the multiple stresses which are closely linked to the ethanol productivity of Z. mobilis. The over-expression of metB/yfdZ operon enabled the growth of the recombinant Z. mobilis in a chemically defined medium without amino acid and vitamin; and the fatty acids overproduction significantly increased ethanol tolerance and ethanol production. The coupled production of ethanol with fatty acids of the Z. mobilis without amino acid and vitamin under VHG fermentation conditions may permit a significant reduction of the production cost of ethanol and microbial fatty acids.

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.

Lower-cost cellulosic ethanol production using cellobiose fermenting yeast Clavispora NRRL Y-50464

USDA-ARS?s Scientific Manuscript database

For ethanol production from cellulosic materials, there are generally two major steps needed including enzymatic hydrolysis to break down biomass sugars and microbial fermentation to convert available simple sugars into ethanol. It often requires two different kinds of microorganisms since ethanolog...

Bacteriophage-encoded lytic enzymes control growth of contaminating Lactobacillus found in fuel ethanol fermentations

USDA-ARS?s Scientific Manuscript database

Background: Reduced yields of ethanol due to bacterial contamination in fermentation cultures weakens the economics of biofuel production. Lactic acid bacteria are considered the most problematic, and surveys of commercial fuel ethanol facilities have found that species of Lactobacillus are predomin...

Bacteriophage application restores ethanol fermentation characteristics disrupted by Lactobacillus fermentum

USDA-ARS?s Scientific Manuscript database

Background: Contamination of corn mash by lactic acid bacteria (LAB) reduces ethanol yields and the overall efficiency of the ethanol fermentation process, and the industry relies heavily on antibiotics for contamination control. There is a need to develop alternative methods for the control of cont...

Biocatalyzed processes for production of commodity chemicals: Assessment of future research advances for N-butanol production

NASA Technical Reports Server (NTRS)

Ingham, J. D.

1984-01-01

This report is a summary of assessments by Chem Systems Inc. and a further evaluation of the impacts of research advances on energy efficiency and the potential for future industrial production of acetone-butanol-ethanol (ABE) solvents and other products by biocatalyzed processes. Brief discussions of each of the assessments made by CSI, followed by estimates of minimum projected energy consumption and costs for production of solvents by ABE biocatalyzed processes are included. These assessments and further advances discussed in this report show that substantial decreases in energy consumption and costs are possible on the basis of specific research advances; therefore, it appears that a biocatalyzed process for ABE can be developed that will be competitive with conventional petrochemical processes for production of n-butanol and acetone. (In this work, the ABE process was selected and utilized only as an example for methodology development; other possible bioprocesses for production of commodity chemicals are not intended to be excluded.) It has been estimated that process energy consumption can be decreased by 50%, with a corresponding cost reduction of 15-30% (in comparison with a conventional petrochemical process) by increasing microorganism tolerance to n-butanol and efficient recovery of product solvents from the vapor phase.

Yeast Derived LysA2 Can Control Bacterial Contamination in Ethanol Fermentation.

PubMed

Kim, Jun-Seob; Daum, M Angela; Jin, Yong-Su; Miller, Michael J

2018-05-24

Contamination of fuel-ethanol fermentations continues to be a significant problem for the corn and sugarcane-based ethanol industries. In particular, members of the Lactobacillaceae family are the primary bacteria of concern. Currently, antibiotics and acid washing are two major means of controlling contaminants. However, antibiotic use could lead to increased antibiotic resistance, and the acid wash step stresses the fermenting yeast and has limited effectiveness. Bacteriophage endolysins such as LysA2 are lytic enzymes with the potential to contribute as antimicrobials to the fuel ethanol industries. Our goal was to evaluate the potential of yeast-derived LysA2 as a means of controlling Lactobacillaceae contamination. LysA2 intracellularly produced by Pichia pastoris showed activity comparable to Escherichia coli produced LysA2. Lactic Acid Bacteria (LAB) with the A4α peptidoglycan chemotype (L-Lys-D-Asp crosslinkage) were the most sensitive to LysA2, though a few from that chemotype were insensitive. Pichia -expressed LysA2, both secreted and intracellularly produced, successfully improved ethanol productivity and yields in glucose (YPD60) and sucrose-based (sugarcane juice) ethanol fermentations in the presence of a LysA2 susceptible LAB contaminant. LysA2 secreting Sacharomyces cerevisiae did not notably improve production in sugarcane juice, but it did control bacterial contamination during fermentation in YPD60. Secretion of LysA2 by the fermenting yeast, or adding it in purified form, are promising alternative tools to control LAB contamination during ethanol fermentation. Endolysins with much broader lytic spectrums than LysA2 could supplement or replace the currently used antibiotics or the acidic wash.

Bioethanol production from ball milled bagasse using an on-site produced fungal enzyme cocktail and xylose-fermenting Pichia stipitis.

PubMed

Buaban, Benchaporn; Inoue, Hiroyuki; Yano, Shinichi; Tanapongpipat, Sutipa; Ruanglek, Vasimon; Champreda, Verawat; Pichyangkura, Rath; Rengpipat, Sirirat; Eurwilaichitr, Lily

2010-07-01

Sugarcane bagasse is one of the most promising agricultural by-products for conversion to biofuels. Here, ethanol fermentation from bagasse has been achieved using an integrated process combining mechanical pretreatment by ball milling, with enzymatic hydrolysis and fermentation. Ball milling for 2 h was sufficient for nearly complete cellulose structural transformation to an accessible amorphous form. The pretreated cellulosic residues were hydrolyzed by a crude enzyme preparation from Penicillium chrysogenum BCC4504 containing cellulase activity combined with Aspergillus flavus BCC7179 preparation containing complementary beta-glucosidase activity. Saccharification yields of 84.0% and 70.4% for glucose and xylose, respectively, were obtained after hydrolysis at 45 degrees C, pH 5 for 72 h, which were slightly higher than those obtained with a commercial enzyme mixture containing Acremonium cellulase and Optimash BG. A high conversion yield of undetoxified pretreated bagasse (5%, w/v) hydrolysate to ethanol was attained by separate hydrolysis and fermentation processes using Pichia stipitis BCC15191, at pH 5.5, 30 degrees C for 24 h resulting in an ethanol concentration of 8.4 g/l, corresponding to a conversion yield of 0.29 g ethanol/g available fermentable sugars. Comparable ethanol conversion efficiency was obtained by a simultaneous saccharification and fermentation process which led to production of 8.0 g/l ethanol after 72 h fermentation under the same conditions. This study thus demonstrated the potential use of a simple integrated process with minimal environmental impact with the use of promising alternative on-site enzymes and yeast for the production of ethanol from this potent lignocellulosic biomass. 2009. Published by Elsevier B.V.

Lignocellulosic ethanol: Technology design and its impact on process efficiency.

PubMed

Paulova, Leona; Patakova, Petra; Branska, Barbora; Rychtera, Mojmir; Melzoch, Karel

2015-11-01

This review provides current information on the production of ethanol from lignocellulosic biomass, with the main focus on relationships between process design and efficiency, expressed as ethanol concentration, yield and productivity. In spite of unquestionable advantages of lignocellulosic biomass as a feedstock for ethanol production (availability, price, non-competitiveness with food, waste material), many technological bottlenecks hinder its wide industrial application and competitiveness with 1st generation ethanol production. Among the main technological challenges are the recalcitrant structure of the material, and thus the need for extensive pretreatment (usually physico-chemical followed by enzymatic hydrolysis) to yield fermentable sugars, and a relatively low concentration of monosaccharides in the medium that hinder the achievement of ethanol concentrations comparable with those obtained using 1st generation feedstocks (e.g. corn or molasses). The presence of both pentose and hexose sugars in the fermentation broth, the price of cellulolytic enzymes, and the presence of toxic compounds that can inhibit cellulolytic enzymes and microbial producers of ethanol are major issues. In this review, different process configurations of the main technological steps (enzymatic hydrolysis, fermentation of hexose/and or pentose sugars) are discussed and their efficiencies are compared. The main features, benefits and drawbacks of simultaneous saccharification and fermentation (SSF), simultaneous saccharification and fermentation with delayed inoculation (dSSF), consolidated bioprocesses (CBP) combining production of cellulolytic enzymes, hydrolysis of biomass and fermentation into one step, together with an approach combining utilization of both pentose and hexose sugars are discussed and compared with separate hydrolysis and fermentation (SHF) processes. The impact of individual technological steps on final process efficiency is emphasized and the potential for use of immobilized biocatalysts is considered. Copyright © 2014 Elsevier Inc. All rights reserved.

KINETICS OF GROWTH AND ETHANOL PRODUCTION ON DIFFERENT CARBON SUBSTRATES USING GENETICALLY ENGINEERED XYLOSE-FERMENTING YEAST

EPA Science Inventory

Saccharomyces cerevisiae 424A (LNH-ST) strain was used for fermentation of glucose and xylose. Growth kinetics and ethanol productivity were calculated for batch fermentation on media containing different combinations of glucose and xylose to give a final sugar concentra...

75 FR 81868 - Approval and Promulgation of Implementation Plans; Kentucky: Prevention of Significant...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-12-29

... ethanol through a natural fermentation process from the definition of ``chemical process plants'' in the... through a natural fermentation process from the definition of ``chemical process plants'' in the major NSR... facilities producing ethanol by natural fermentation under the North American Industry Classification System...

Multi-stage Continuous Culture Fermentation of Glucose-Xylose Mixtures to Fuel Ethanol using Genetically Engineered Saccharomyces cerevisiae 424A

EPA Science Inventory

Multi-stage continuous (chemostat) culture fermentation (MCCF) with variable fermentor volumes was carried out to study utilizing glucose and xylose for ethanol production by means of mixed sugar fermentation (MSF). Variable fermentor volumes were used to enable enhanced sugar u...

Cloning and Expression of Laccase from Trametes versicolor in Saccharomyces cerevisiae using a Novel Vector System

USDA-ARS?s Scientific Manuscript database

The long-term goal of this research is to increase efficiency and decrease cost of ethanol fermentation of lignocellulosic feedstocks by combining pre-treatment using laccase enzyme and subsequent fermentation to ethanol through simultaneous saccharification and fermentation paradigms. The first st...

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

PubMed

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

2015-09-01

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

The consequences of Lactobacillus vini and Dekkera bruxellensis as contaminants of the sugarcane-based ethanol fermentation.

PubMed

de Souza, Rafael Barros; dos Santos, Billy Manoel; de Fátima Rodrigues de Souza, Raquel; da Silva, Paula Katharina Nogueira; Lucena, Brígida Thais Luckwu; de Morais, Marcos Antonio

2012-11-01

This work describes the effects of the presence of the yeast Dekkera bruxellensis and the bacterium Lactobacillus vini on the industrial production of ethanol from sugarcane fermentation. Both contaminants were quantified in industrial samples, and their presence was correlated to a decrease in ethanol concentration and accumulation of sugar. Then, laboratory mixed-cell fermentations were carried out to evaluate the effects of these presumed contaminants on the viability of Saccharomyces cerevisiae and the overall ethanol yield. The results showed that high residual sugar seemed the most significant factor arising from the presence of D. bruxellensis in the industrial process when compared to pure S. cerevisiae cultures. Moreover, when L. vini was added to S. cerevisiae cultures it did not appear to affect the yeast cells by any kind of antagonistic effect under stable fermentations. In addition, when L. vini was added to D. bruxellensis cultures, it showed signs of being able to stimulate the fermentative activity of the yeast cells in a way that led to an increase in the ethanol yield.

Process Design and Economics of On-Site Cellulase Production on Various Carbon Sources in a Softwood-Based Ethanol Plant

PubMed Central

Barta, Zsolt; Kovacs, Krisztina; Reczey, Kati; Zacchi, Guido

2010-01-01

On-site cellulase enzyme fermentation in a softwood-to-ethanol process, based on SO2-catalysed steam pretreatment followed by simultaneous saccharification and fermentation, was investigated from a techno-economic aspect using Aspen Plus© and Aspen Icarus Process Evaluator© softwares. The effect of varying the carbon source of enzyme fermentation, at constant protein and mycelium yields, was monitored through the whole process. Enzyme production step decreased the overall ethanol yield (270 L/dry tonne of raw material in the case of purchased enzymes) by 5–16 L/tonne. Capital cost was found to be the main cost contributor to enzyme fermentation, constituting to 60–78% of the enzyme production cost, which was in the range of 0.42–0.53 SEK/L ethanol. The lowest minimum ethanol selling prices (4.71 and 4.82 SEK/L) were obtained in those scenarios, where pretreated liquid fraction supplemented with molasses was used as carbon source. In some scenarios, on-site enzyme fermentation was found to be a feasible alternative. PMID:21048869

Process design and economics of on-site cellulase production on various carbon sources in a softwood-based ethanol plant.

PubMed

Barta, Zsolt; Kovacs, Krisztina; Reczey, Kati; Zacchi, Guido

2010-06-28

On-site cellulase enzyme fermentation in a softwood-to-ethanol process, based on SO(2)-catalysed steam pretreatment followed by simultaneous saccharification and fermentation, was investigated from a techno-economic aspect using Aspen Plus© and Aspen Icarus Process Evaluator© softwares. The effect of varying the carbon source of enzyme fermentation, at constant protein and mycelium yields, was monitored through the whole process. Enzyme production step decreased the overall ethanol yield (270 L/dry tonne of raw material in the case of purchased enzymes) by 5-16 L/tonne. Capital cost was found to be the main cost contributor to enzyme fermentation, constituting to 60-78% of the enzyme production cost, which was in the range of 0.42-0.53 SEK/L ethanol. The lowest minimum ethanol selling prices (4.71 and 4.82 SEK/L) were obtained in those scenarios, where pretreated liquid fraction supplemented with molasses was used as carbon source. In some scenarios, on-site enzyme fermentation was found to be a feasible alternative.

Two-step size reduction and post-washing of steam exploded corn stover improving simultaneous saccharification and fermentation for ethanol production.

PubMed

Liu, Zhi-Hua; Chen, Hong-Zhang

2017-01-01

The simultaneous saccharification and fermentation (SSF) of corn stover biomass for ethanol production was performed by integrating steam explosion (SE) pretreatment, hydrolysis and fermentation. Higher SE pretreatment severity and two-step size reduction increased the specific surface area, swollen volume and water holding capacity of steam exploded corn stover (SECS) and hence facilitated the efficiency of hydrolysis and fermentation. The ethanol production and yield in SSF increased with the decrease of particle size and post-washing of SECS prior to fermentation to remove the inhibitors. Under the SE conditions of 1.5MPa and 9min using 2.0cm particle size, glucan recovery and conversion to glucose by enzymes were 86.2% and 87.2%, respectively. The ethanol concentration and yield were 45.0g/L and 85.6%, respectively. With this two-step size reduction and post-washing strategy, the water utilization efficiency, sugar recovery and conversion, and ethanol concentration and yield by the SSF process were improved. Copyright © 2016 Elsevier Ltd. All rights reserved.

Continuous High-solids corn liquefaction and fermentation with stripping of ethanol

USDA-ARS?s Scientific Manuscript database

Removal of ethanol from the fermentor during fermentation can increase productivity and reduce the costs for dewatering the product and coproduct. One approach is to recycle the fermentor contents through a stripping column, where a non-condensable gas removes ethanol to a condenser. Previous resear...

Sequential saccharification of corn fiber and ethanol production by the brown rot fungus Gloeophyllum trabeum.

PubMed

Rasmussen, M L; Shrestha, P; Khanal, S K; Pometto, A L; Hans van Leeuwen, J

2010-05-01

Degradation of lignocellulosic biomass to sugars through a purely biological process is a key to sustainable biofuel production. Hydrolysis of the corn wet-milling co-product-corn fiber-to simple sugars by the brown rot fungus Gloeophyllum trabeum was studied in suspended-culture and solid-state fermentations. Suspended-culture experiments were not effective in producing harvestable sugars from the corn fiber. The fungus consumed sugars released by fungal extracellular enzymes. Solid-state fermentation demonstrated up to 40% fiber degradation within 9days. Enzyme activity assays on solid-state fermentation filtrates confirmed the involvement of starch- and cellulose-degrading enzymes. To reduce fungal consumption of sugars and to accelerate enzyme activity, 2- and 3-d solid-state fermentation biomasses (fiber and fungus) were submerged in buffer and incubated at 37 degrees C without shaking. This anaerobic incubation converted up to almost 11% of the corn fiber into harvestable reducing sugars. Sugars released by G. trabeum were fermented to a maximum yield of 3.3g ethanol/100g fiber. This is the first report, to our knowledge, of G. trabeum fermenting sugar to ethanol. The addition of Saccharomyces cerevisiae as a co-culture led to more rapid fermentation to a maximum yield of 4.0g ethanol/100g fiber. The findings demonstrate the potential for this simple fungal process, requiring no pretreatment of the corn fiber, to produce more ethanol by hydrolyzing and fermenting carbohydrates in this lignocellulosic co-product. Copyright 2010 Elsevier Ltd. All rights reserved.

High Level Ethanol from Sugar Cane Molasses by a New Thermotolerant Saccharomyces cerevisiae Strain in Industrial Scale.

PubMed

Fadel, M; Keera, Abeer A; Mouafi, Foukia E; Kahil, Tarek

2013-01-01

A new local strain of S. cerevisiae F-514, for ethanol production during hot summer season, using Egyptian sugar cane molasses was applied in Egyptian distillery factory. The inouluum was propagated through 300 L, 3 m(3), and 12 m(3) fermenters charged with diluted sugar cane molasses containing 4%-5% sugars. The yeast was applied in fermentation vessels 65 m(3) working volume to study the varying concentrations of urea, DAP, orthophosphoric acid (OPA), and its combinations as well as magnesium sulfate and inoculum size. The fermenter was allowed to stay for a period of 20 hours to give time for maximum conversion of sugars into ethanol. S. cerevisiae F-514 at molasses sugar level of 18% (w/v), inoculum size of 20% (v/v) cell concentration of 3.0 × 10(8)/mL, and combinations of urea, diammonium phosphate (DAP), orthophosphoric acid (OPA), and magnesium sulfate at amounts of 20, 10, 5, and 10 kg/65 m(3) working volume fermenters, respectively, supported maximum ethanol production (9.8%, v/v), fermentation efficiency (FE) 88.1%, and remaining sugars (RS) 1.22%. The fermentation resulted 13.4 g dry yeast/L contained 34.6% crude protein and 8.2% ash. By selecting higher ethanol yielding yeast strain and optimizing, the fermentation parameters both yield and economics of the fermentation process can be improved.

Enhancement of the Initial Rate of Ethanol Fermentation Due to Dysfunction of Yeast Stress Response Components Msn2p and/or Msn4p▿ †

PubMed Central

Watanabe, Daisuke; Wu, Hong; Noguchi, Chiemi; Zhou, Yan; Akao, Takeshi; Shimoi, Hitoshi

2011-01-01

Sake yeasts (strains of Saccharomyces cerevisiae) produce high concentrations of ethanol in sake fermentation. To investigate the molecular mechanisms underlying this brewing property, we compared gene expression of sake and laboratory yeasts in sake mash. DNA microarray and reporter gene analyses revealed defects of sake yeasts in environmental stress responses mediated by transcription factors Msn2p and/or Msn4p (Msn2/4p) and stress response elements (STRE). Furthermore, we found that dysfunction of MSN2 and/or MSN4 contributes to the higher initial rate of ethanol fermentation in both sake and laboratory yeasts. These results provide novel insights into yeast stress responses as major impediments of effective ethanol fermentation. PMID:21131516

Overexpression of pyruvate decarboxylase in the yeast Hansenula polymorpha results in increased ethanol yield in high-temperature fermentation of xylose.

PubMed

Ishchuk, Olena P; Voronovsky, Andriy Y; Stasyk, Oleh V; Gayda, Galina Z; Gonchar, Mykhailo V; Abbas, Charles A; Sibirny, Andriy A

2008-11-01

Improvement of xylose fermentation is of great importance to the fuel ethanol industry. The nonconventional thermotolerant yeast Hansenula polymorpha naturally ferments xylose to ethanol at high temperatures (48-50 degrees C). Introduction of a mutation that impairs ethanol reutilization in H. polymorpha led to an increase in ethanol yield from xylose. The native and heterologous (Kluyveromyces lactis) PDC1 genes coding for pyruvate decarboxylase were expressed at high levels in H. polymorpha under the control of the strong constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH). This resulted in increased pyruvate decarboxylase activity and improved ethanol production from xylose. The introduction of multiple copies of the H. polymorpha PDC1 gene driven by the strong constitutive promoter led to a 20-fold increase in pyruvate decarboxylase activity and up to a threefold elevation of ethanol production.

Fermentation method producing ethanol

DOEpatents

Wang, Daniel I. C.; Dalal, Rajen

1986-01-01

Ethanol is the major end product of an anaerobic, thermophilic fermentation process using a mutant strain of bacterium Clostridium thermosaccharolyticum. This organism is capable of converting hexose and pentose carbohydrates to ethanol, acetic and lactic acids. Mutants of Clostridium thermosaccharolyticum are capable of converting these substrates to ethanol in exceptionally high yield and with increased productivity. Both the mutant organism and the technique for its isolation are provided.

Ethanol yield and volatile compound content in fermentation of agave must by Kluyveromyces marxianus UMPe-1 comparing with Saccharomyces cerevisiae baker's yeast used in tequila production.

PubMed

López-Alvarez, Arnoldo; Díaz-Pérez, Alma Laura; Sosa-Aguirre, Carlos; Macías-Rodríguez, Lourdes; Campos-García, Jesús

2012-05-01

In tequila production, fermentation is an important step. Fermentation determines the ethanol productivity and organoleptic properties of the beverage. In this study, a yeast isolated from native residual agave must was identified as Kluyveromyces marxianus UMPe-1 by 26S rRNA sequencing. This yeast was compared with the baker's yeast Saccharomyces cerevisiae Pan1. Our findings demonstrate that the UMPe-1 yeast was able to support the sugar content of agave must and glucose up to 22% (w/v) and tolerated 10% (v/v) ethanol concentration in the medium with 50% cells survival. Pilot and industrial fermentation of agave must tests showed that the K. marxianus UMPe-1 yeast produced ethanol with yields of 94% and 96% with respect to fermentable sugar content (glucose and fructose, constituting 98%). The S. cerevisiae Pan1 baker's yeast, however, which is commonly used in some tequila factories, showed 76% and 70% yield. At the industrial level, UMPe-1 yeast shows a maximum velocity of fermentable sugar consumption of 2.27g·L(-1)·h(-1) and ethanol production of 1.38g·L(-1)·h(-1), providing 58.78g ethanol·L(-1) at 72h fermentation, which corresponds to 96% yield. In addition, the major and minor volatile compounds in the tequila beverage obtained from UMPe-1 yeast were increased. Importantly, 29 volatile compounds were identified, while the beverage obtained from Pan1-yeast contained fewer compounds and in lower concentrations. The results suggest that the K. marxianus UMPe-1 is a suitable yeast for agave must fermentation, showing high ethanol productivity and increased volatile compound content comparing with a S. cerevisiae baker's yeast used in tequila production. Copyright © 2012 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Flocculating Zymomonas mobilis is a promising host to be engineered for fuel ethanol production from lignocellulosic biomass.

PubMed

Zhao, Ning; Bai, Yun; Liu, Chen-Guang; Zhao, Xin-Qing; Xu, Jian-Feng; Bai, Feng-Wu

2014-03-01

Whereas Saccharomyces cerevisiae uses the Embden-Meyerhof-Parnas pathway to metabolize glucose, Zymomonas mobilis uses the Entner-Doudoroff (ED) pathway. Employing the ED pathway, 50% less ATP is produced, which could lead to less biomass being accumulated during fermentation and an improved yield of ethanol. Moreover, Z. mobilis cells, which have a high specific surface area, consume glucose faster than S. cerevisiae, which could improve ethanol productivity. We performed ethanol fermentations using these two species under comparable conditions to validate these speculations. Increases of 3.5 and 3.3% in ethanol yield, and 58.1 and 77.8% in ethanol productivity, were observed in ethanol fermentations using Z. mobilis ZM4 in media containing ∼100 and 200 g/L glucose, respectively. Furthermore, ethanol fermentation bythe flocculating Z. mobilis ZM401 was explored. Although no significant difference was observed in ethanol yield and productivity, the flocculation of the bacterial species enabled biomass recovery by cost-effective sedimentation, instead of centrifugation with intensive capital investment and energy consumption. In addition, tolerance to inhibitory byproducts released during biomass pretreatment, particularly acetic acid and vanillin, was improved. These experimental results indicate that Z. mobilis, particularly its flocculating strain, is superior to S. cerevisiae as a host to be engineered for fuel ethanol production from lignocellulosic biomass. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Dynamic modeling and analyses of simultaneous saccharification and fermentation process to produce bio-ethanol from rice straw.

PubMed

Ko, Jordon; Su, Wen-Jun; Chien, I-Lung; Chang, Der-Ming; Chou, Sheng-Hsin; Zhan, Rui-Yu

2010-02-01

The rice straw, an agricultural waste from Asians' main provision, was collected as feedstock to convert cellulose into ethanol through the enzymatic hydrolysis and followed by the fermentation process. When the two process steps are performed sequentially, it is referred to as separate hydrolysis and fermentation (SHF). The steps can also be performed simultaneously, i.e., simultaneous saccharification and fermentation (SSF). In this research, the kinetic model parameters of the cellulose saccharification process step using the rice straw as feedstock is obtained from real experimental data of cellulase hydrolysis. Furthermore, this model can be combined with a fermentation model at high glucose and ethanol concentrations to form a SSF model. The fermentation model is based on cybernetic approach from a paper in the literature with an extension of including both the glucose and ethanol inhibition terms to approach more to the actual plants. Dynamic effects of the operating variables in the enzymatic hydrolysis and the fermentation models will be analyzed. The operation of the SSF process will be compared to the SHF process. It is shown that the SSF process is better in reducing the processing time when the product (ethanol) concentration is high. The means to improve the productivity of the overall SSF process, by properly using aeration during the batch operation will also be discussed.

[Effect of phenolic ketones on ethanol fermentation and cellular lipid composition of Pichia stipitis].

PubMed

Yang, Jinlong; Cheng, Yichao; Zhu, Yuanyuan; Zhu, Junjun; Chen, Tingting; Xu, Yong; Yong, Qiang; Yu, Shiyuan

2016-02-01

Lignin degradation products are toxic to microorganisms, which is one of the bottlenecks for fuel ethanol production. We studied the effects of phenolic ketones (4-hydroxyacetophenone, 4-hydroxy-3-methoxy-acetophenone and 4-hydroxy-3,5-dimethoxy-acetophenone) derived from lignin degradation on ethanol fermentation of xylose and cellular lipid composition of Pichia stipitis NLP31. Ethanol and the cellular fatty acid of yeast were analyzed by high performance liquid chromatography (HPLC) and gas chromatography/mass spectrometry (GC/MS). Results indicate that phenolic ketones negatively affected ethanol fermentation of yeast and the lower molecular weight phenolic ketone compound was more toxic. When the concentration of 4-hydroxyacetophenone was 1.5 g/L, at fermentation of 24 h, the xylose utilization ratio, ethanol yield and ethanol concentration decreased by 42.47%, 5.30% and 9.76 g/L, respectively, compared to the control. When phenolic ketones were in the medium, the ratio of unsaturated fatty acids to saturated fatty acids (UFA/SFA) of yeast cells was improved. When 1.5 g/L of three aforementioned phenolic ketones was added to the fermentation medium, the UFA/SFA ratio of yeast cells increased to 3.03, 3.06 and 3.61, respectively, compared to 2.58 of the control, which increased cell membrane fluidity and instability. Therefore, phenolic ketones can reduce the yeast growth, increase the UFA/SFA ratio of yeast and lower ethanol productivity. Effectively reduce or remove the content of lignin degradation products is the key to improve lignocellulose biorefinery.

Simulation and optimization of continuous extractive fermentation with recycle system

NASA Astrophysics Data System (ADS)

Widjaja, Tri; Altway, Ali; Rofiqah, Umi; Airlangga, Bramantyo

2017-05-01

Extractive fermentation is continuous fermentation method which is believed to be able to substitute conventional fermentation method (batch). The recovery system and ethanol refinery will be easier. Continuous process of fermentation will make the productivity increase although the unconverted sugar in continuous fermentation is still in high concentration. In order to make this process more efficient, the recycle process was used. Increasing recycle flow will enhance the probability of sugar to be re-fermented. However, this will make ethanol enter fermentation column. As a result, the accumulated ethanol will inhibit the growth of microorganism. This research aims to find optimum conditions of solvent to broth ratio (S:B) and recycle flow to fresh feed ratio in order to produce the best yield and productivity. This study employed optimization by Hooke Jeeves method using Matlab 7.8 software. The result indicated that optimum condition occured in S: B=2.615 and R: F=1.495 with yield = 50.2439 %.

Production of ethanol from thin stillage by metabolically engineered Escherichia coli.

PubMed

Gonzalez, Ramon; Campbell, Paul; Wong, Matthew

2010-03-01

Thin stillage is a by-product generated in large amounts during the production of ethanol that is rich in carbon sources like glycerol, glucose and maltose. Unfortunately, the fermentation of thin stillage results in a mixture of organic acids and ethanol and minimum utilization of glycerol, the latter a compound that can represent up to 80% of the available substrates in this stream. We report here the efficient production of ethanol from thin stillage by a metabolically engineered strain of Escherichia coli. Simultaneous utilization of glycerol and sugars was achieved by overexpressing either the fermentative or the respiratory glycerol-utilization pathway. However, amplification of the fermentative pathway (encoded by gldA and dhaKLM) led to more efficient consumption of glycerol and promoted the synthesis of reduced products, including ethanol. A previously constructed strain, EH05, containing mutations that prevented the accumulation of competing by-products (i.e. lactate, acetate, and succinate) and overexpressing the fermentative pathway for glycerol utilization [i.e. strain EH05 (pZSKLMgldA)], efficiently converted thin stillage supplemented with only mineral salts to ethanol at yields close to 85% of the theoretical maximum. Ethanol accounted for about 90% (w/w) of the product mixture. These results, along with the comparable performance of strain EH05 (pZSKLMgldA) in 0.5 and 5 l fermenters, indicate a great potential for the adoption of this process by the biofuels industry.

Microbiological and physicochemical characterization of small-scale cocoa fermentations and screening of yeast and bacterial strains to develop a defined starter culture.

PubMed

Pereira, Gilberto Vinícius de Melo; Miguel, Maria Gabriela da Cruz Pedrozo; Ramos, Cíntia Lacerda; Schwan, Rosane Freitas

2012-08-01

Spontaneous cocoa bean fermentations performed under bench- and pilot-scale conditions were studied using an integrated microbiological approach with culture-dependent and culture-independent techniques, as well as analyses of target metabolites from both cocoa pulp and cotyledons. Both fermentation ecosystems reached equilibrium through a two-phase process, starting with the simultaneous growth of the yeasts (with Saccharomyces cerevisiae as the dominant species) and lactic acid bacteria (LAB) (Lactobacillus fermentum and Lactobacillus plantarum were the dominant species), which were gradually replaced by the acetic acid bacteria (AAB) (Acetobacter tropicalis was the dominant species). In both processes, a sequence of substrate consumption (sucrose, glucose, fructose, and citric acid) and metabolite production kinetics (ethanol, lactic acid, and acetic acid) similar to that of previous, larger-scale fermentation experiments was observed. The technological potential of yeast, LAB, and AAB isolates was evaluated using a polyphasic study that included the measurement of stress-tolerant growth and fermentation kinetic parameters in cocoa pulp media. Overall, strains L. fermentum UFLA CHBE8.12 (citric acid fermenting, lactic acid producing, and tolerant to heat, acid, lactic acid, and ethanol), S. cerevisiae UFLA CHYC7.04 (ethanol producing and tolerant to acid, heat, and ethanol), and Acetobacter tropicalis UFLA CHBE16.01 (ethanol and lactic acid oxidizing, acetic acid producing, and tolerant to acid, heat, acetic acid, and ethanol) were selected to form a cocktail starter culture that should lead to better-controlled and more-reliable cocoa bean fermentation processes.

Microbiological and Physicochemical Characterization of Small-Scale Cocoa Fermentations and Screening of Yeast and Bacterial Strains To Develop a Defined Starter Culture

PubMed Central

Pereira, Gilberto Vinícius de Melo; Miguel, Maria Gabriela da Cruz Pedrozo; Ramos, Cíntia Lacerda

2012-01-01

Spontaneous cocoa bean fermentations performed under bench- and pilot-scale conditions were studied using an integrated microbiological approach with culture-dependent and culture-independent techniques, as well as analyses of target metabolites from both cocoa pulp and cotyledons. Both fermentation ecosystems reached equilibrium through a two-phase process, starting with the simultaneous growth of the yeasts (with Saccharomyces cerevisiae as the dominant species) and lactic acid bacteria (LAB) (Lactobacillus fermentum and Lactobacillus plantarum were the dominant species), which were gradually replaced by the acetic acid bacteria (AAB) (Acetobacter tropicalis was the dominant species). In both processes, a sequence of substrate consumption (sucrose, glucose, fructose, and citric acid) and metabolite production kinetics (ethanol, lactic acid, and acetic acid) similar to that of previous, larger-scale fermentation experiments was observed. The technological potential of yeast, LAB, and AAB isolates was evaluated using a polyphasic study that included the measurement of stress-tolerant growth and fermentation kinetic parameters in cocoa pulp media. Overall, strains L. fermentum UFLA CHBE8.12 (citric acid fermenting, lactic acid producing, and tolerant to heat, acid, lactic acid, and ethanol), S. cerevisiae UFLA CHYC7.04 (ethanol producing and tolerant to acid, heat, and ethanol), and Acetobacter tropicalis UFLA CHBE16.01 (ethanol and lactic acid oxidizing, acetic acid producing, and tolerant to acid, heat, acetic acid, and ethanol) were selected to form a cocktail starter culture that should lead to better-controlled and more-reliable cocoa bean fermentation processes. PMID:22636007

Evolved strains of Scheffersomyces stipitis achieving high ethanol productivity on acid- and base-pretreated biomass hydrolyzate at high solids loading

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

Slininger, Patricia J.; Shea-Andersh, Maureen A.; Thompson, Stephanie R.

Lignocellulosic biomass is an abundant, renewable feedstock useful for the production of fuel-grade ethanol via the processing steps of pretreatment, enzyme hydrolysis, and microbial fermentation. Traditional industrial yeasts do not ferment xylose and are not able to grow, survive, or ferment in concentrated hydrolyzates that contain enough sugar to support economical ethanol recovery since they are laden with toxic byproducts generated during pretreatment. Repetitive culturing in two types of concentrated hydrolyzates was applied along with ethanol challenged xylose-fed continuous culture to force targeted evolution of the native pentose fermenting yeast Scheffersomyces (Pichia) stipitis strain NRRL Y-7124 maintained in the ARSmore » Culture Collection, Peoria, IL. Isolates collected from various enriched populations were screened and ranked based on relative xylose uptake rate and ethanol yield. Ranking on hydrolyzates with and without nutritional supplementation was used to identify those isolates with best performance across diverse conditions. Robust S. stipitis strains adapted to perform very well in enzyme hydrolyzates of high solids loading ammonia fiber expansion-pretreated corn stover (18% weight per volume solids) and dilute sulfuric acid-pretreated switchgrass (20% w/v solids) were obtained. Improved features include reduced initial lag phase preceding growth, significantly enhanced fermentation rates, improved ethanol tolerance and yield, reduced diauxic lag during glucose-xylose transition, and ability to accumulate >40 g/L ethanol in <167 h when fermenting hydrolyzate at low initial cell density of 0.5 absorbance units and pH 5 to 6.« less

Evolved strains of Scheffersomyces stipitis achieving high ethanol productivity on acid- and base-pretreated biomass hydrolyzate at high solids loading

DOE PAGES

Slininger, Patricia J.; Shea-Andersh, Maureen A.; Thompson, Stephanie R.; ...

2015-04-09

Lignocellulosic biomass is an abundant, renewable feedstock useful for the production of fuel-grade ethanol via the processing steps of pretreatment, enzyme hydrolysis, and microbial fermentation. Traditional industrial yeasts do not ferment xylose and are not able to grow, survive, or ferment in concentrated hydrolyzates that contain enough sugar to support economical ethanol recovery since they are laden with toxic byproducts generated during pretreatment. Repetitive culturing in two types of concentrated hydrolyzates was applied along with ethanol challenged xylose-fed continuous culture to force targeted evolution of the native pentose fermenting yeast Scheffersomyces (Pichia) stipitis strain NRRL Y-7124 maintained in the ARSmore » Culture Collection, Peoria, IL. Isolates collected from various enriched populations were screened and ranked based on relative xylose uptake rate and ethanol yield. Ranking on hydrolyzates with and without nutritional supplementation was used to identify those isolates with best performance across diverse conditions. Robust S. stipitis strains adapted to perform very well in enzyme hydrolyzates of high solids loading ammonia fiber expansion-pretreated corn stover (18% weight per volume solids) and dilute sulfuric acid-pretreated switchgrass (20% w/v solids) were obtained. Improved features include reduced initial lag phase preceding growth, significantly enhanced fermentation rates, improved ethanol tolerance and yield, reduced diauxic lag during glucose-xylose transition, and ability to accumulate >40 g/L ethanol in <167 h when fermenting hydrolyzate at low initial cell density of 0.5 absorbance units and pH 5 to 6.« less

Comparative behaviour of yeast strains for ethanolic fermentation of culled apple juice.

PubMed

Modi, D R; Garg, S K; Johri, B N

1998-07-01

The culled apple juice contained (% w/v): nitrogen, 0.036; total sugars, 11.6 and was of pH 3.9. Saccharomyces cerevisiae NCIM 3284, Pichia kluyeri and Candida krusei produced more ethanol from culled apple juice at its optimum initial pH 4.5, whereas S. cerevisiae NCIM 3316 did so at pH 5.0. An increase in sugar concentration of apple juice from natural 11.6% to 20% exhibited enhanced ethanol production and improved fermentation efficiency of both the S. cerevisiae strains, whereas P. kluyveri and C. krusei produced high ethanol at 11.6% and 16.0% sugar levels, respectively. Urea was stimulatory for ethanol production as well as fermentation efficiency of the yeast strains under study.

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.

Production of bio ethanol from waste potatoes

NASA Astrophysics Data System (ADS)

Jaber Noufal, Mohamad; Li, Baizhan; Maalla, Zena Ali

2017-03-01

In this research, production of ethanol from waste potatoes fermentation was studied using Saccharomyces cerevisiae. Potato Flour prepared from potato tubers after cooking and drying at 85°C. A homogenous slurry of potato flour prepared in water at solid-liquid ratio 1:10. Liquefaction of potato starch slurry was done with α-amylase at 80°C for 40 min followed by saccharification process which was done with glucoamylase at 65°C for two hr. Fermentation of hydrolysate with Saccharomyces cerevisiae at 35°C for two days resulted in the production of 33 g/l ethanol. The following parameters have been analysed: temperature, time of fermentation and pH. It found that Saccharification process is affected by enzyme Amylase 300 concentration and concentration of 1000μl/100ml gives the efficient effect of the process. The best temperature for fermentation process was found to be about 35°C. Also, it noticed that ethanol production increased as a time of fermentation increased but after 48 hr further growth in fermentation time did not have an appreciable effect. Finally, the optimal value of pH for fermentation process was about 5 to 6.

Lignocellulosic sugar management for xylitol and ethanol fermentation with multiple cell recycling by Kluyveromyces marxianus IIPE453.

PubMed

Dasgupta, Diptarka; Ghosh, Debashish; Bandhu, Sheetal; Adhikari, Dilip K

2017-07-01

Optimum utilization of fermentable sugars from lignocellulosic biomass to deliver multiple products under biorefinery concept has been reported in this work. Alcohol fermentation has been carried out with multiple cell recycling of Kluyveromyces marxianus IIPE453. The yeast utilized xylose-rich fraction from acid and steam treated biomass for cell generation and xylitol production with an average yield of 0.315±0.01g/g while the entire glucose rich saccharified fraction had been fermented to ethanol with high productivity of 0.9±0.08g/L/h. A detailed insight into its genome illustrated the strain's complete set of genes associated with sugar transport and metabolism for high-temperature fermentation. A set flocculation proteins were identified that aided in high cell recovery in successive fermentation cycles to achieve alcohols with high productivity. We have brought biomass derived sugars, yeast cell biomass generation, and ethanol and xylitol fermentation in one platform and validated the overall material balance. 2kg sugarcane bagasse yielded 193.4g yeast cell, and with multiple times cell recycling generated 125.56g xylitol and 289.2g ethanol (366mL). Copyright © 2017 Elsevier GmbH. All rights reserved.

Yeast fermentation affected by homo- and hetero-fermentative Lactobacilli isolated from fuel ethanol distilleries with sugarcane products as substrates

USDA-ARS?s Scientific Manuscript database

The antagonism between by yeast and lactobacilli is largely dependent on the initial population of each organism. While homo-fermentative lactobacillus present higher inhibitory effect upon yeast when in equal cell number, in industrial fuel ethanol conditions where high yeast cell densities prevail...

Resolving Bacterial Contamination of Fuel Ethanol Fermentations with Beneficial Bacteria – an Alternative to Antibiotic Treatment

USDA-ARS?s Scientific Manuscript database

Fuel ethanol fermentations are not performed under aseptic conditions and microbial contamination reduces yields and can lead to costly “stuck fermentations.” Antibiotics are commonly used to combat contaminants, but these may persist in the distillers grains co-product. Among contaminants, it is kn...

Ethanol production from residual wood chips of cellulose industry: acid pretreatment investigation, hemicellulosic hydrolysate fermentation, and remaining solid fraction fermentation by SSF process.

PubMed

Silva, Neumara Luci Conceição; Betancur, Gabriel Jaime Vargas; Vasquez, Mariana Peñuela; Gomes, Edelvio de Barros; Pereira, Nei

2011-04-01

Current research indicates the ethanol fuel production from lignocellulosic materials, such as residual wood chips from the cellulose industry, as new emerging technology. This work aimed at evaluating the ethanol production from hemicellulose of eucalyptus chips by diluted acid pretreatment and the subsequent fermentation of the generated hydrolysate by a flocculating strain of Pichia stipitis. The remaining solid fraction generated after pretreatment was subjected to enzymatic hydrolysis, which was carried out simultaneously with glucose fermentation [saccharification and fermentation (SSF) process] using a strain of Saccharomyces cerevisiae. The acid pretreatment was evaluated using a central composite design for sulfuric acid concentration (1.0-4.0 v/v) and solid to liquid ratio (1:2-1:4, grams to milliliter) as independent variables. A maximum xylose concentration of 50 g/L was obtained in the hemicellulosic hydrolysate. The fermentation of hemicellulosic hydrolysate and the SSF process were performed in bioreactors and the final ethanol concentrations of 15.3 g/L and 28.7 g/L were obtained, respectively.

Simultaneous fermentation of glucose and xylose at elevated temperatures co-produces ethanol and xylitol through overexpression of a xylose-specific transporter in engineered Kluyveromyces marxianus.

PubMed

Zhang, Biao; Zhang, Jia; Wang, Dongmei; Han, Ruixiang; Ding, Rui; Gao, Xiaolian; Sun, Lianhong; Hong, Jiong

2016-09-01

Engineered Kluyveromyces marxianus strains were constructed through over-expression of various transporters for simultaneous co-fermentation of glucose and xylose. The glucose was converted into ethanol, whereas xylose was converted into xylitol which has higher value than ethanol. Over-expressing xylose-specific transporter ScGAL2-N376F mutant enabled yeast to co-ferment glucose and xylose and the co-fermentation ability was obviously improved through increasing ScGAL2-N376F expression. The production of glycerol was blocked and acetate production was reduced by disrupting gene KmGPD1. The obtained K. marxianus YZJ119 utilized 120g/L glucose and 60g/L xylose simultaneously and produced 50.10g/L ethanol and 55.88g/L xylitol at 42°C. The yield of xylitol from consumed xylose was over 98% (0.99g/g). Through simultaneous saccharification and co-fermentation at 42°C, YZJ119 produced a maximal concentration of 44.58g/L ethanol and 32.03g/L xylitol or 29.82g/L ethanol and 31.72g/L xylitol, respectively, from detoxified or non-detoxified diluted acid pretreated corncob. Copyright © 2016 Elsevier Ltd. All rights reserved.

[Continuous ethanol fermentation coupled with recycling of yeast flocs].

PubMed

Wang, Bo; Ge, Xu-Meng; Li, Ning; Bai, Feng-Wu

2006-09-01

A continuous ethanol fermentation system composed of three-stage tanks in series coupled with two sedimentation tanks was established. A self-flocculating yeast strain developed by protoplast fusion from Saccharomyces cerevisiae and Schizosaccharomyces pombe was applied. Two-stage enzymatic hydrolysate of corn powder containing 220g/L of reducing sugar, supplemented with 1.5g/L (NH4)2HPO4 and 2.5g/L KH2PO4, was used as the ethanol fermentation substrate and fed into the first fermentor at the dilution rate of 0.057h(-1). The yeast flocs separated by sedimentation were recycled into the first fermentor as two different models: activation-recycle and direct recycle. The quasi-steady states were obtained for both operation models after the fermentation systems experienced short periods of transitions. Activation process helped enhance the performance of ethanol fermentation at the high dilution rates. The broth containing more than 101g/L ethanol, 3.2g/L residual reducing sugar and 7.7g/L residual total sugar was produced. The ethanol productivity was calculated to be 5.77g/(L x h), which increased by more than 70% compared with that achieved in the same tank in series system without recycling of yeast cells.

Adaptive evolution of Saccharomyces cerevisiae with enhanced ethanol tolerance for Chinese rice wine fermentation.

PubMed

Chen, Shuang; Xu, Yan

2014-08-01

High tolerance towards ethanol is a desirable property for the Saccharomyces cerevisiae strains used in the alcoholic beverage industry. To improve the ethanol tolerance of an industrial Chinese rice wine yeast, a sequential batch fermentation strategy was used to adaptively evolve a chemically mutagenized Chinese rice wine G85 strain. The high level of ethanol produced under Chinese rice wine-like fermentation conditions was used as the selective pressure. After adaptive evolution of approximately 200 generations, mutant G85X-8 was isolated and shown to have markedly increased ethanol tolerance. The evolved strain also showed higher osmotic and temperature tolerances than the parental strain. Laboratory Chinese rice wine fermentation showed that the evolved G85X-8 strain was able to catabolize sugars more completely than the parental G85 strain. A higher level of yeast cell activity was found in the fermentation mash produced by the evolved strain, but the aroma profiles were similar between the evolved and parental strains. The improved ethanol tolerance in the evolved strain might be ascribed to the altered fatty acids composition of the cell membrane and higher intracellular trehalose concentrations. These results suggest that adaptive evolution is an efficient approach for the non-recombinant modification of industrial yeast strains.

Development of a more efficient process for production of fuel ethanol from bamboo.

PubMed

Sun, Zhao-Yong; Wang, Ting; Tan, Li; Tang, Yue-Qin; Kida, Kenji

2015-06-01

A process for production of fuel ethanol from bamboo treated with concentrated sulfuric acid has been previously proposed. To improve efficiency of the process, we tested saccharification with 70 weight% (wt%) sulfuric acid, acid-sugar separation by ion exclusion, addition of nutrients to the ethanol fermentation, and bioconversion of xylose to xylitol. A high efficiency of both sugar recovery (82.5 %) and acid recovery (97.5 %) was achieved in the saccharification process and in the continuous acid-sugar separation using a modified anion exchange resin, respectively. Reduction of the amount of mineral salts added to the saccharified liquid after acid-sugar separation did not negatively affect performance of the continuous ethanol fermentation. The ethanol yield and productivity were 93.7 % and 6 g/l h, respectively, at 35 °C and pH 4.0. And the ethanol yield and productivity were almost the same even at pH 3.5. Moreover, the xylose remaining in the fermented mash was efficiently converted to xylitol in batch fermentation by Candida tropicalis strain 2.1776. These results demonstrate a more efficient process for the production of fuel ethanol from bamboo.

Ensilage and bioconversion of grape pomace into fuel ethanol.

PubMed

Zheng, Yi; Lee, Christopher; Yu, Chaowei; Cheng, Yu-Shen; Simmons, Christopher W; Zhang, Ruihong; Jenkins, Bryan M; VanderGheynst, Jean S

2012-11-07

Two types of grape pomace were ensiled with eight strains of lactic acid bacteria (LAB). Both fresh grape pomace (FrGP) and fermented grape pomace (FeGP) were preserved through alcoholic fermentation but not malolactic conversion. Water leaching prior to storage was used to reduce water-soluble carbohydrates and ethanol from FrGP and FeGP, respectively, to increase malolactic conversion. Leached FeGP had spoilage after 28 days of ensilage, whereas FrGP was preserved. Dilute acid pretreatment was examined for increasing the conversion of pomace to ethanol via Escherichia coli KO11 fermentation. Dilute acid pretreatment doubled the ethanol yield from FeGP, but it did not improve the ethanol yield from FrGP. The ethanol yields from raw pomace were nearly double the yields from the ensiled pomace. For this reason, the recovery of ethanol produced during winemaking from FeGP and ethanol produced during storage of FrGP is critical for the economical conversion of grape pomace to biofuel.

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

PubMed

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

2013-07-01

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

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.

The importance of aeration strategy in fuel alcohol fermentations contaminated with Dekkera/Brettanomyces yeasts.

PubMed

Abbott, D A; Ingledew, W M

2005-11-01

Whole corn mash fermentations infected with industrially-isolated Brettanomyces yeasts were not affected even when viable Brettanomyces yeasts out-numbered Saccharomyces yeasts tenfold at the onset of fermentation. Therefore, aeration, a parameter that is pivotal to the physiology of Dekkera/Brettanomyces yeasts, was investigated in mixed culture fermentations. Results suggest that aeration strategy plays a significant role in Dekkera/Brettanomyces-mediated inhibition of fuel alcohol fermentations. Although growth of Saccharomyces cerevisiae was not impeded, mixed culture fermentations aerated at rates of > or =20 ml air l(-1) mash min(-1) showed decreased ethanol yields and an accumulation of acetic acid. The importance of aeration was examined further in combination with organic acid(s). Growth of Saccharomyces occurred more rapidly than growth of Brettanomyces yeasts in all conditions. The combination of 0.075% (w/v) acetic acid and contamination with Brettanomyces TK 1404W did not negatively impact the final ethanol yield under fermentative conditions. Aeration, however, did prove to be detrimental to final ethanol yields. With the inclusion of aeration in the control condition (no organic acid stress) and in each fermentation containing organic acid(s), the final ethanol yields were decreased. It was therefore concluded that aeration strategy is the key parameter in regards to the negative effects observed in fuel alcohol fermentations infected with Dekkera/Brettanomyces yeasts.

Ethanol from municipal cellulosic wastes

NASA Astrophysics Data System (ADS)

Parker, A. J., Jr.; Timbario, T. J.; Mulloney, J. A., Jr.

This paper addresses the use of municipal cellulosic wastes as a feedstock for producing ethanol fuels, and describes the application of enzymatic hydrolysis technology for their production. The concept incorporates recent process technology developments within the framework of an existing industry familiar with large-scale ethanol fermentation (the brewing industry). Preliminary indications are that the cost of producing ethanol via enzymatic hydrolysis in an existing plant with minimal facility modifications (low capital investment) can be significantly less than that of ethanol from grain fermentation.

75 FR 73017 - Approval and Promulgation of Implementation Plans; Georgia: Prevention of Significant...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-11-29

... produce ethanol through a natural fermentation process from the definition of ``chemical process plants... facilities that produce ethanol through a natural fermentation process from the definition of ``chemical...

75 FR 79300 - Approval and Promulgation of Implementation Plans; Mississippi; Prevention of Significant...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-12-20

... plants'' that produce ethanol through a natural fermentation process. EPA may consider further action for... ethanol by natural fermentation under the North American Industry Classification System (NAICS) codes...

76 FR 55572 - Approval and Promulgation of Implementation Plans; Georgia: Prevention of Significant...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-09-08

...) Exclude facilities that produce ethanol through a natural fermentation process from the definition of... 24060) which exclude facilities that produce ethanol through a natural fermentation process from the...

Evaluation of the fermentation of high gravity thick sugar beet juice worts for efficient bioethanol production

PubMed Central

2013-01-01

Background Sugar beet and intermediates of sugar beet processing are considered to be very attractive feedstock for ethanol production due to their content of fermentable sugars. In particular, the processing of the intermediates into ethanol is considerably facilitated because it does not require pretreatment or enzymatic treatment in contrast to production from starch raw materials. Moreover, the advantage of thick juice is high solid substance and saccharose content which eliminates problems with the storability of this feedstock. Results The objective of this study were to investigate bioethanol production from thick juice worts and the effects of their concentration, the type of mineral supplement, as well as the dose of yeast inoculum on fermentation dynamics and ethanol yield. The obtained results show that to ensure efficient ethanolic fermentation of high gravity thick juice worts, one needs to use a yeast strain with high ethanol tolerance and a large amount of inoculum. The highest ethanol yield (94.9 ± 2.8% of the theoretical yield) and sugars intake of 96.5 ± 2.9% were obtained after the fermentation of wort with an extract content of 250 g/kg supplemented with diammonium hydrogen phosphate (0.3 g/L of wort) and inoculated with 2 g of Ethanol Red dry yeast per L of wort. An increase in extract content in the fermentation medium from 250 g/L to 280 g/kg resulted in decreased efficiency of the process. Also the distillates originating from worts with an extract content of 250 g/kg were characterized by lower acetaldehyde concentration than those obtained from worts with an extract content of 280 g/kg. Conclusions Under the favorable conditions determined in our experiments, 38.9 ± 1.2 L of 100% (v/v) ethyl alcohol can be produced from 100 kg of thick juice. The obtained results show that the selection of process conditions and the yeast for the fermentation of worts with a higher sugar content can improve the economic performance of the alcohol-distilling industry due to more efficient ethanol production, reduced consumption of cooling water, and energy for ethanol distillation, as well as a decreased volume of fermentation stillage. PMID:24206573

Evaluation of the fermentation of high gravity thick sugar beet juice worts for efficient bioethanol production.

PubMed

Dziugan, Piotr; Balcerek, Maria; Pielech-Przybylska, Katarzyna; Patelski, Piotr

2013-11-08

Sugar beet and intermediates of sugar beet processing are considered to be very attractive feedstock for ethanol production due to their content of fermentable sugars. In particular, the processing of the intermediates into ethanol is considerably facilitated because it does not require pretreatment or enzymatic treatment in contrast to production from starch raw materials. Moreover, the advantage of thick juice is high solid substance and saccharose content which eliminates problems with the storability of this feedstock. The objective of this study were to investigate bioethanol production from thick juice worts and the effects of their concentration, the type of mineral supplement, as well as the dose of yeast inoculum on fermentation dynamics and ethanol yield.The obtained results show that to ensure efficient ethanolic fermentation of high gravity thick juice worts, one needs to use a yeast strain with high ethanol tolerance and a large amount of inoculum. The highest ethanol yield (94.9 ± 2.8% of the theoretical yield) and sugars intake of 96.5 ± 2.9% were obtained after the fermentation of wort with an extract content of 250 g/kg supplemented with diammonium hydrogen phosphate (0.3 g/L of wort) and inoculated with 2 g of Ethanol Red dry yeast per L of wort. An increase in extract content in the fermentation medium from 250 g/L to 280 g/kg resulted in decreased efficiency of the process. Also the distillates originating from worts with an extract content of 250 g/kg were characterized by lower acetaldehyde concentration than those obtained from worts with an extract content of 280 g/kg. Under the favorable conditions determined in our experiments, 38.9 ± 1.2 L of 100% (v/v) ethyl alcohol can be produced from 100 kg of thick juice. The obtained results show that the selection of process conditions and the yeast for the fermentation of worts with a higher sugar content can improve the economic performance of the alcohol-distilling industry due to more efficient ethanol production, reduced consumption of cooling water, and energy for ethanol distillation, as well as a decreased volume of fermentation stillage.

Fermentation of lignocellulosic hydrolysate by the alternative industrial ethanol yeast Dekkera bruxellensis.

PubMed

Blomqvist, J; South, E; Tiukova, I; Tiukova, L; Momeni, M H; Hansson, H; Ståhlberg, J; Horn, S J; Schnürer, J; Passoth, V

2011-07-01

Testing the ability of the alternative ethanol production yeast Dekkera bruxellensis to produce ethanol from lignocellulose hydrolysate and comparing it to Saccharomyces cerevisiae. Industrial isolates of D. bruxellensis and S. cerevisiae were cultivated in small-scale batch fermentations of enzymatically hydrolysed steam exploded aspen sawdust. Different dilutions of hydrolysate were tested. None of the yeasts grew in undiluted or 1:2 diluted hydrolysate [final glucose concentration always adjusted to 40 g l⁻¹ (0.22 mol l⁻¹)]. This was most likely due to the presence of inhibitors such as acetate or furfural. In 1:5 hydrolysate, S. cerevisiae grew, but not D. bruxellensis, and in 1:10 hydrolysate, both yeasts grew. An external vitamin source (e.g. yeast extract) was essential for growth of D. bruxellensis in this lignocellulosic hydrolysate and strongly stimulated S. cerevisiae growth and ethanol production. Ethanol yields of 0.42 ± 0.01 g ethanol (g glucose)⁻¹ were observed for both yeasts in 1:10 hydrolysate. In small-scale continuous cultures with cell recirculation, with a gradual increase in the hydrolysate concentration, D. bruxellensis was able to grow in 1:5 hydrolysate. In bioreactor experiments with cell recirculation, hydrolysate contents were increased up to 1:2 hydrolysate, without significant losses in ethanol yields for both yeasts and only slight differences in viable cell counts, indicating an ability of both yeasts to adapt to toxic compounds in the hydrolysate. Dekkera bruxellensis and S. cerevisiae have a similar potential to ferment lignocellulose hydrolysate to ethanol and to adapt to fermentation inhibitors in the hydrolysate. This is the first study investigating the potential of D. bruxellensis to ferment lignocellulosic hydrolysate. Its high competitiveness in industrial fermentations makes D. bruxellensis an interesting alternative for ethanol production from those substrates. © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

Conventional and nonconventional strategies for controlling bacterial contamination in fuel ethanol fermentations.

PubMed

Ceccato-Antonini, Sandra Regina

2018-05-25

Ethanol bio-production in Brazil has some unique characteristics that inevitably lead to bacterial contamination, which results in the production of organic acids and biofilms and flocculation that impair the fermentation yield by affecting yeast viability and diverting sugars to metabolites other than ethanol. The ethanol-producing units commonly give an acid treatment to the cells after each fermentative cycle to decrease the bacterial number, which is not always effective. An alternative strategy must be employed to avoid bacterial multiplication but must be compatible with economic, health and environmental aspects. This review analyzes the issue of bacterial contamination in sugarcane-based fuel ethanol fermentation, and the potential strategies that may be utilized to control bacterial growth besides acid treatment and antibiotics. We have emphasized the efficiency and suitability of chemical products other than acids and those derived from natural sources in industrial conditions. In addition, we have also presented bacteriocins, bacteriophages, and beneficial bacteria as non-conventional antimicrobial agents to mitigate bacterial contamination in the bioethanol industry.

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

Wu, Y.V.; Baghy, M.O.

Sweet potato can yield 1000 gallons of ethanol/acre compared with 250-300 gal/acre for corn. Sweet potatoes of normal, relatively high, and very high dry-matter contents were fermented to ethanol. Pectinase was necessary to decrease viscosity before fermentation for economic processing, especially for varieties of normal and relatively high dry-matter contents. Attained yield of ethanol was 90% of theoretical value. After ethanol was distilled, residual stillage was separated by screening and centrifugation into filter cake, centrifuged solids, and stillage solubles. Filter cake and centrifuged solids had crude protein contents (nitrogen x 6.25, dry basis) of 22-32% and 42-57%, respectively, and accountedmore » for 44-85% and 0-17% of total sweet potato nitrogen. Sweet potatoes and their fermented products had 4.3-7.6 g of lysine/16 g of N and are expected to have good nutritional value. This practical method to ferment sweet potato for ethanol and to recover valuable protein-rich byproducts may have commercial potential. (Refs. 19).« less

High temperature stimulates acetic acid accumulation and enhances the growth inhibition and ethanol production by Saccharomyces cerevisiae under fermenting conditions.

PubMed

Woo, Ji-Min; Yang, Kyung-Mi; Kim, Sae-Um; Blank, Lars M; Park, Jin-Byung

2014-07-01

Cellular responses of Saccharomyces cerevisiae to high temperatures of up to 42 °C during ethanol fermentation at a high glucose concentration (i.e., 100 g/L) were investigated. Increased temperature correlated with stimulated glucose uptake to produce not only the thermal protectant glycerol but also ethanol and acetic acid. Carbon flux into the tricarboxylic acid (TCA) cycle correlated positively with cultivation temperature. These results indicate that the increased demand for energy (in the form of ATP), most likely caused by multiple stressors, including heat, acetic acid, and ethanol, was matched by both the fermentation and respiration pathways. Notably, acetic acid production was substantially stimulated compared to that of other metabolites during growth at increased temperature. The acetic acid produced in addition to ethanol seemed to subsequently result in adverse effects, leading to increased production of reactive oxygen species. This, in turn, appeared to cause the specific growth rate, and glucose uptake rate reduced leading to a decrease of the specific ethanol production rate far before glucose depletion. These results suggest that adverse effects from heat, acetic acid, ethanol, and oxidative stressors are synergistic, resulting in a decrease of the specific growth rate and ethanol production rate and, hence, are major determinants of cell stability and ethanol fermentation performance of S. cerevisiae at high temperatures. The results are discussed in the context of possible applications.

Comparison of single-molecule sequencing and hybrid approaches for finishing the genome of Clostridium autoethanogenum and analysis of CRISPR systems in industrial relevant Clostridia

PubMed Central

2014-01-01

Background Clostridium autoethanogenum strain JA1-1 (DSM 10061) is an acetogen capable of fermenting CO, CO2 and H2 (e.g. from syngas or waste gases) into biofuel ethanol and commodity chemicals such as 2,3-butanediol. A draft genome sequence consisting of 100 contigs has been published. Results A closed, high-quality genome sequence for C. autoethanogenum DSM10061 was generated using only the latest single-molecule DNA sequencing technology and without the need for manual finishing. It is assigned to the most complex genome classification based upon genome features such as repeats, prophage, nine copies of the rRNA gene operons. It has a low G + C content of 31.1%. Illumina, 454, Illumina/454 hybrid assemblies were generated and then compared to the draft and PacBio assemblies using summary statistics, CGAL, QUAST and REAPR bioinformatics tools and comparative genomic approaches. Assemblies based upon shorter read DNA technologies were confounded by the large number repeats and their size, which in the case of the rRNA gene operons were ~5 kb. CRISPR (Clustered Regularly Interspaced Short Paloindromic Repeats) systems among biotechnologically relevant Clostridia were classified and related to plasmid content and prophages. Potential associations between plasmid content and CRISPR systems may have implications for historical industrial scale Acetone-Butanol-Ethanol (ABE) fermentation failures and future large scale bacterial fermentations. While C. autoethanogenum contains an active CRISPR system, no such system is present in the closely related Clostridium ljungdahlii DSM 13528. A common prophage inserted into the Arg-tRNA shared between the strains suggests a common ancestor. However, C. ljungdahlii contains several additional putative prophages and it has more than double the amount of prophage DNA compared to C. autoethanogenum. Other differences include important metabolic genes for central metabolism (as an additional hydrogenase and the absence of a phophoenolpyruvate synthase) and substrate utilization pathway (mannose and aromatics utilization) that might explain phenotypic differences between C. autoethanogenum and C. ljungdahlii. Conclusions Single molecule sequencing will be increasingly used to produce finished microbial genomes. The complete genome will facilitate comparative genomics and functional genomics and support future comparisons between Clostridia and studies that examine the evolution of plasmids, bacteriophage and CRISPR systems. PMID:24655715

Comparative genomic analysis of single-molecule sequencing and hybrid approaches for finishing the Clostridium autoethanogenum JA1-1 strain DSM 10061 genome

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

Brown, Steven D; Nagaraju, Shilpa; Utturkar, Sagar M

Background Clostridium autoethanogenum strain JA1-1 (DSM 10061) is an acetogen capable of fermenting CO, CO2 and H2 (e.g. from syngas or waste gases) into biofuel ethanol and commodity chemicals such as 2,3-butanediol. A draft genome sequence consisting of 100 contigs has been published. Results A closed, high-quality genome sequence for C. autoethanogenum DSM10061 was generated using only the latest single-molecule DNA sequencing technology and without the need for manual finishing. It is assigned to the most complex genome classification based upon genome features such as repeats, prophage, nine copies of the rRNA gene operons. It has a low G +more » C content of 31.1%. Illumina, 454, Illumina/454 hybrid assemblies were generated and then compared to the draft and PacBio assemblies using summary statistics, CGAL, QUAST and REAPR bioinformatics tools and comparative genomic approaches. Assemblies based upon shorter read DNA technologies were confounded by the large number repeats and their size, which in the case of the rRNA gene operons were ~5 kb. CRISPR (Clustered Regularly Interspaced Short Paloindromic Repeats) systems among biotechnologically relevant Clostridia were classified and related to plasmid content and prophages. Potential associations between plasmid content and CRISPR systems may have implications for historical industrial scale Acetone-Butanol-Ethanol (ABE) fermentation failures and future large scale bacterial fermentations. While C. autoethanogenum contains an active CRISPR system, no such system is present in the closely related Clostridium ljungdahlii DSM 13528. A common prophage inserted into the Arg-tRNA shared between the strains suggests a common ancestor. However, C. ljungdahlii contains several additional putative prophages and it has more than double the amount of prophage DNA compared to C. autoethanogenum. Other differences include important metabolic genes for central metabolism (as an additional hydrogenase and the absence of a phophoenolpyruvate synthase) and substrate utilization pathway (mannose and aromatics utilization) that might explain phenotypic differences between C. autoethanogenum and C. ljungdahlii. Conclusions Single molecule sequencing will be increasingly used to produce finished microbial genomes. The complete genome will facilitate comparative genomics and functional genomics and support future comparisons between Clostridia and studies that examine the evolution of plasmids, bacteriophage and CRISPR systems.« less

Effects of sodium meta bisulfite on diffusion fermentation of fodder beets for fuel ethanol production. [Saccharomyces cerevisiae

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

Gibbons, W.R.; Westby, C.A.

1987-01-01

The authors designed and tested a new process for converting fodder beets to ethanol: continuous diffusion-fermentation. This process utilizes the simultaneous diffusion-fermentation concept of the EX-FERM design; however, it overcomes the material handling problems inherent in that system by utilizing a counterflow tubular auger system. This process also eliminates the need for roller mills or presses and dryers which are required for alcohol recovery from solid phase fermentation. The latter is the only other currently feasible procedure for producing distillably worthwhile amounts of ethanol from fodder beets, sweet sorghum, and other similar feedstocks. Results on the use of sodium metamore » bisulfite (SMB) for contamination control with fermenting fodder beet cubes are reported.« less

Direct ethanol production from cellulosic materials by consolidated biological processing using the wood rot fungus Schizophyllum commune.

PubMed

Horisawa, Sakae; Ando, Hiromasa; Ariga, Osamu; Sakuma, Yoh

2015-12-01

In the present study, ethanol production from polysaccharides or wood chips was conducted in a single reactor under anaerobic conditions using the white rot fungus Schizophyllum commune NBRC 4928, which produces enzymes that degrade lignin, cellulose and hemicellulose. The ethanol yields produced from glucose and xylose were 80.5%, and 52.5%, respectively. The absolute yields of ethanol per microcrystalline cellulose (MCC), xylan and arabinogalactan were 0.26g/g-MCC, 0.0419g/g-xylan and 0.0508g/g-arabinogalactan, respectively. By comparing the actual ethanol yields from polysaccharides with monosaccharide fermentation, it was shown that the rate of saccharification was slower than that in fermentation. S. commune NBRC 4928 is concluded to be suitable for CBP because it can produce ethanol from various types of sugar. From the autoclaved cedar chip, only little ethanol was produced by S. commune NBRC 4928 alone but ethanol production was enhanced by combined use of ethanol fermenting and lignin degrading fungi. Copyright © 2015 Elsevier Ltd. All rights reserved.

Yeast selection for fuel ethanol production in Brazil.

PubMed

Basso, Luiz C; de Amorim, Henrique V; de Oliveira, Antonio J; Lopes, Mario L

2008-11-01

Brazil is one of the largest ethanol biofuel producers and exporters in the world and its production has increased steadily during the last three decades. The increasing efficiency of Brazilian ethanol plants has been evident due to the many technological contributions. As far as yeast is concerned, few publications are available regarding the industrial fermentation processes in Brazil. The present paper reports on a yeast selection program performed during the last 12 years aimed at selecting Saccharomyces cerevisiae strains suitable for fermentation of sugar cane substrates (cane juice and molasses) with cell recycle, as it is conducted in Brazilian bioethanol plants. As a result, some evidence is presented showing the positive impact of selected yeast strains in increasing ethanol yield and reducing production costs, due to their higher fermentation performance (high ethanol yield, reduced glycerol and foam formation, maintenance of high viability during recycling and very high implantation capability into industrial fermenters). Results also suggest that the great yeast biodiversity found in distillery environments could be an important source of strains. This is because during yeast cell recycling, selective pressure (an adaptive evolution) is imposed on cells, leading to strains with higher tolerance to the stressful conditions of the industrial fermentation.

Soursop (Annona muricata) vinegar production and its chemical compositions

NASA Astrophysics Data System (ADS)

Ho, Chin Wai; Lazim, Azwan Mat; Fazry, Shazrul; Zaki, Umi Kalsum Hj Hussain; Lim, Seng Joe

2016-11-01

Vinegar is a liquid product that undergoes double fermentations, which are alcoholic and acetous fermentation. Sugar source was converted to ethanol in alcoholic fermentation, meanwhile ethanol was oxidised to acetic acid during acetous fermentation. Soursop (Annona muricata) was the starting material in this study, as it is easily available in Malaysia. Its highly aromatic, juicy and distinctive flavours enables the production of high quality vinegar. The objective of this research is to produce good quality soursop vinegar as an innovative method to preserve and utilise the soursop fruit in Malaysia and to determine its chemical compositions. It was found that the sugar content reduces over time, and it is inversely proportional to the ethanol concentration, due to the production of ethanol from sugar. Acetic acid was also found to increase with increasing fermentation time. pH showed no significant difference (p>0.05) in the reduction of sugar and the production of ethanol. However, significantly higher (p<0.05) production of acetic acid was observed at pH 5.0 and 5.5, compared to that at pH 4.5. There were no significant differences (p > 0.05) in Vitamin C contents in all soursop vinegar samples produced using different treatments.

Transcriptome analysis identifies genes involved in ethanol response of Saccharomyces cerevisiae in Agave tequilana juice.

PubMed

Ramirez-Córdova, Jesús; Drnevich, Jenny; Madrigal-Pulido, Jaime Alberto; Arrizon, Javier; Allen, Kirk; Martínez-Velázquez, Moisés; Alvarez-Maya, Ikuri

2012-08-01

During ethanol fermentation, yeast cells are exposed to stress due to the accumulation of ethanol, cell growth is altered and the output of the target product is reduced. For Agave beverages, like tequila, no reports have been published on the global gene expression under ethanol stress. In this work, we used microarray analysis to identify Saccharomyces cerevisiae genes involved in the ethanol response. Gene expression of a tequila yeast strain of S. cerevisiae (AR5) was explored by comparing global gene expression with that of laboratory strain S288C, both after ethanol exposure. Additionally, we used two different culture conditions, cells grown in Agave tequilana juice as a natural fermentation media or grown in yeast-extract peptone dextrose as artificial media. Of the 6368 S. cerevisiae genes in the microarray, 657 genes were identified that had different expression responses to ethanol stress due to strain and/or media. A cluster of 28 genes was found over-expressed specifically in the AR5 tequila strain that could be involved in the adaptation to tequila yeast fermentation, 14 of which are unknown such as yor343c, ylr162w, ygr182c, ymr265c, yer053c-a or ydr415c. These could be the most suitable genes for transforming tequila yeast to increase ethanol tolerance in the tequila fermentation process. Other genes involved in response to stress (RFC4, TSA1, MLH1, PAU3, RAD53) or transport (CYB2, TIP20, QCR9) were expressed in the same cluster. Unknown genes could be good candidates for the development of recombinant yeasts with ethanol tolerance for use in industrial tequila fermentation.

Process design of SSCF for ethanol production from steam-pretreated, acetic-acid-impregnated wheat straw.

PubMed

Bondesson, Pia-Maria; Galbe, Mats

2016-01-01

Pretreatment is an important step in the production of ethanol from lignocellulosic material. Using acetic acid together with steam pretreatment allows the positive effects of an acid catalyst to be retained, while avoiding the negative environmental effects associated with sulphuric acid. Acetic acid is also formed during the pretreatment and hydrolysis of hemicellulose, and is a known inhibitor that may impair fermentation at high concentrations. The purpose of this study was to improve ethanol production from glucose and xylose in steam-pretreated, acetic-acid-impregnated wheat straw by process design of simultaneous saccharification and co-fermentation (SSCF), using a genetically modified pentose fermenting yeast strain Saccharomyces cerevisiae . Ethanol was produced from glucose and xylose using both the liquid fraction and the whole slurry from pretreated materials. The highest ethanol concentration achieved was 37.5 g/L, corresponding to an overall ethanol yield of 0.32 g/g based on the glucose and xylose available in the pretreated material. To obtain this concentration, a slurry with a water-insoluble solids (WIS) content of 11.7 % was used, using a fed-batch SSCF strategy. A higher overall ethanol yield (0.36 g/g) was obtained at 10 % WIS. Ethanol production from steam-pretreated, acetic-acid-impregnated wheat straw through SSCF with a pentose fermenting S. cerevisiae strain was successfully demonstrated. However, the ethanol concentration was too low and the residence time too long to be suitable for large-scale applications. It is hoped that further process design focusing on the enzymatic conversion of cellulose to glucose will allow the combination of acetic acid pretreatment and co-fermentation of glucose and xylose.

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

PubMed Central

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

2017-01-01

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

DMR (deacetylation and mechanical refining) processing of corn stover achieves high monomeric sugar concentrations (230 g L -1) during enzymatic hydrolysis and high ethanol concentrations (>10% v/v) during fermentation without hydrolysate purification or concentration

DOE PAGES

Chen, Xiaowen; Kuhn, Erik; Jennings, Edward W.; ...

2016-04-01

Distilling and purifying ethanol and other products from second generation lignocellulosic biorefineries adds significant capital and operating costs to biofuel production. The energy usage associated with distillation negatively affects plant gate costs and causes environmental and life-cycle impacts, and the lower titers in fermentation caused by lower sugar concentrations from pretreatment and enzymatic hydrolysis increase energy and water usage and ethanol production costs. In addition, lower ethanol titers increase the volumes required for enzymatic hydrolysis and fermentation vessels increase capital expenditure (CAPEX). Therefore, increasing biofuel titers has been a research focus in renewable biofuel production for several decades. In thismore » work, we achieved approximately 230 g L -1 of monomeric sugars after high solid enzymatic hydrolysis using deacetylation and mechanical refining (DMR) processed corn stover substrates produced at the 100 kg per day scale. The high sugar concentrations and low chemical inhibitor concentrations achieved by the DMR process allowed fermentation to ethanol with titers as high as 86 g L -1, which translates into approximately 10.9% v/v ethanol. To our knowledge, this is the first time that titers greater than 10% v/v ethanol in fermentations derived from corn stover without any sugar concentration or purification steps have been reported. As a result, the potential cost savings from high sugar and ethanol titers achieved by the DMR process are also reported using TEA analysis.« less

DMR (deacetylation and mechanical refining) processing of corn stover achieves high monomeric sugar concentrations (230 g L -1) during enzymatic hydrolysis and high ethanol concentrations (>10% v/v) during fermentation without hydrolysate purification or concentration

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

Chen, Xiaowen; Kuhn, Erik; Jennings, Edward W.

Distilling and purifying ethanol and other products from second generation lignocellulosic biorefineries adds significant capital and operating costs to biofuel production. The energy usage associated with distillation negatively affects plant gate costs and causes environmental and life-cycle impacts, and the lower titers in fermentation caused by lower sugar concentrations from pretreatment and enzymatic hydrolysis increase energy and water usage and ethanol production costs. In addition, lower ethanol titers increase the volumes required for enzymatic hydrolysis and fermentation vessels increase capital expenditure (CAPEX). Therefore, increasing biofuel titers has been a research focus in renewable biofuel production for several decades. In thismore » work, we achieved approximately 230 g L -1 of monomeric sugars after high solid enzymatic hydrolysis using deacetylation and mechanical refining (DMR) processed corn stover substrates produced at the 100 kg per day scale. The high sugar concentrations and low chemical inhibitor concentrations achieved by the DMR process allowed fermentation to ethanol with titers as high as 86 g L -1, which translates into approximately 10.9% v/v ethanol. To our knowledge, this is the first time that titers greater than 10% v/v ethanol in fermentations derived from corn stover without any sugar concentration or purification steps have been reported. As a result, the potential cost savings from high sugar and ethanol titers achieved by the DMR process are also reported using TEA analysis.« less

Potentiality of Yeasts in the Direct Conversion of Starchy Materials to Ethanol and Its Relevance in the New Millennium

NASA Astrophysics Data System (ADS)

Reddy, L. V. A.; Reddy, O. V. S.; Basappa, S. C.

In recent years, the use of renewable and abundantly available starchy and cellulosic materials for industrial production of ethanol is gaining importance, in view of the fact, that ethanol is one of the most prospective future motor fuels, that can be expected to replace fossil fuels, which are fast depleting in the world scenario. Although, the starch and the starchy substrates could be converted successfully to ethanol on industrial scales by the use of commercial amylolytic enzymes and yeast fermentation, the cost of production is rather very high. This is mainly due to the non-enzymatic and enzymatic conversion (gelatinization, liquefaction and saccharification) of starch to sugars, which costs around 20 % of the cost of production of ethanol from starch. In this context, the use of amylolytic yeasts, that can directly convert starch to ethanol by a single step, are potentially suited to reduce the cost of production of ethanol from starch. Research advances made in this direction have shown encouraging results, both in terms of identifying the potentially suited yeasts for the purpose and also their economic ethanol yields. This chapter focuses on the types of starch and starchy substrates and their digestion to fermentable sugars, optimization of fermentation conditions to ethanol from starch, factors that affect starch fermentation, potential amylolytic yeasts which can directly convert starch to ethanol, genetic improvement of these yeasts for better conversion efficiency and their future economic prospects in the new millennium.

Production of ethanol from raw cassava starch by a nonconventional fermentation method

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

Ueda, S.; Zenin, C.T.; Monteiro, D.A.

Raw cassava root starch was transformed into ethanol in a one-step process of fermentation, in which are combined the conventional processes of liquefaction, saccharification, and fermentation to alcohol. Aspergillus awamori NRRL 3112 and Aspergillus niger were cultivated on wheat bran and used as Koji enzymes. Commercial A. niger amyloglucosidase was also used in this experiment. A raw cassava root homogenate-enzymes-yeast mixture fermented optimally at pH 3.5 and 30/degree/C, for five days and produced ethanol. Alcohol yields from raw cassava roots were between 82.3 and 99.6%. Fungal Koji enzymes effectively decreased the viscosity of cassava root fermentation mashes during incubation. Commercialmore » A. niger amyloglucosidase decreased the viscosity slightly. Reduction of viscosity of fermentation mashes was 40, 84, and 93% by commercial amyloglucosidase, A. awamori, and A. niger enzymes, respectively. The reduction of viscosity of fermentation mashes is probably due to the hydrolysis of pentosans by Koji enzymes. 12 refs.« less

Rheology of corn stover slurries during fermentation to ethanol

NASA Astrophysics Data System (ADS)

Ghosh, Sanchari; Epps, Brenden; Lynd, Lee

2017-11-01

In typical processes that convert cellulosic biomass into ethanol fuel, solubilization of the biomass is carried out by saccharolytic enzymes; however, these enzymes require an expensive pretreatment step to make the biomass accessible for solubilization (and subsequent fermentation). We have proposed a potentially-less-expensive approach using the bacterium Clostridium thermocellum, which can initiate fermentation without pretreatment. Moreover, we have proposed a ``cotreatment'' process, in which fermentation and mechanical milling occur alternately so as to achieve the highest ethanol yield for the least milling energy input. In order to inform the energetic requirements of cotreatment, we experimentally characterized the rheological properties of corn stover slurries at various stages of fermentation. Results show that a corn stover slurry is a yield stress fluid, with shear thinning behavior well described by a power law model. Viscosity decreases dramatically upon fermentation, controlling for variables such as solids concentration and particle size distribution. To the authors' knowledge, this is the first study to characterize the changes in the physical properties of biomass during fermentation by a thermophilic bacterium.

Process design considerations for optimal production of ethanol from lignocellulose using available yeasts, including natural pentose-fermenting yeasts, and their derivatives

USDA-ARS?s Scientific Manuscript database

To expand the biomass to fuel ethanol industry, process strategies are needed to foster the production and utilization of microorganisms which can survive and ferment both hexose (C6) and pentose (C5) sugars while exposed to inhibitors (such as ethanol, furfural, and hydroxymethylfurfural, or HMF). ...

Genetically engineered Escherichia coli FBR5: Part II. Ethanol production from xylose and simultaneous product recovery

USDA-ARS?s Scientific Manuscript database

In these studies concentrated xylose solution was fermented to ethanol employing Escherichia coli FBR5 which can ferment both lignocellulosic sugars (hexoses and pentoses). E. coli FBR5 can produce 40-50 gL-1 ethanol from 100 gL-1 xylose in batch reactors. Increasing sugar concentration beyond this...

Production of gluten and germ by ethanol fermentation of raw corn

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

Not Available

1987-01-01

The Illinois ethanol fuel industry has grown to be an important part of our state's economy over the past 10 years. It provides an additional market for Illinois' abundant corn production, provides many industrial jobs, and substitutes a home-grown renewable energy resource for imported oil. More than 30 percent of all gasoline sold in Illinois contains 10 percent ethanol. The economics of producing ethanol from corn is strongly affected by the byproduct value and by the energy required in the production process. This document reports on efforts to research a new microbial process that would improve the ethanol fermentation processmore » in both these areas. The new process allows direct fermentation of corn starch to ethanol without the usual requirement of cooking the corn. This reduces the amount of energy needed for production and recovers the protein-containing gluten and oil-containing germ with all of the original food value intact.« less

Technological properties of indigenous wine yeast strains isolated from wine production regions of Turkey.

PubMed

Bağder Elmacı, Simel; Özçelik, Filiz; Tokatlı, Mehmet; Çakır, İbrahim

2014-05-01

The purpose of this study was to evaluate the important technological and fermentative properties of wine yeast strains previously isolated from different wine producing regions of Turkey. The determination of the following important properties was made: growth at high temperatures; fermentative capability in the presence of high sugar concentration; fermentation rate; hydrogen sulfide production; killer activity; resistance to high ethanol and sulfur dioxide; foam production; and enzymatic profiles. Ten local wine yeast strains belonging to Saccharomyces, and one commercial active dry yeast as a reference strain were evaluated. Fermentation characteristics were evaluated in terms of kinetic parameters, including ethanol yield (YP/S), biomass yield (YX/S), theoretical ethanol yield (%), specific ethanol production rate (qp; g/gh), specific glucose uptake rate (qs; g/gh), and the substrate conversion (%). All tested strains were able to grow at 37 °C and to start fermentation at 30° Brix, and were resistant to high concentrations of sulfur dioxide. 60 % of the strains were weak H2S producers, while the others produced high levels. Foam production was high, and no strains had killer activity. Six of the tested strains had the ability to grow and ferment at concentrations of 14 % ethanol. Except for one strain, all fermented most of the media sugars at a high rate, producing 11.0-12.4 % (v/v) ethanol. Although all but one strain had suitable characteristics for wine production, they possessed poor activities of glycosidase, esterase and proteinase enzymes of oenological interest. Nine of the ten local yeast strains were selected for their good oenological properties and their suitability as a wine starter culture.

Selection of Yeast Strains for Tequila Fermentation Based on Growth Dynamics in Combined Fructose and Ethanol Media.

PubMed

Aldrete-Tapia, J A; Miranda-Castilleja, D E; Arvizu-Medrano, S M; Hernández-Iturriaga, M

2018-02-01

The high concentration of fructose in agave juice has been associated with reduced ethanol tolerance of commercial yeasts used for tequila production and low fermentation yields. The selection of autochthonous strains, which are better adapted to agave juice, could improve the process. In this study, a 2-step selection process of yeasts isolated from spontaneous fermentations for tequila production was carried out based on analysis of the growth dynamics in combined conditions of high fructose and ethanol. First, yeast isolates (605) were screened to identify strains tolerant to high fructose (20%) and to ethanol (10%), yielding 89 isolates able to grow in both conditions. From the 89 isolates, the growth curves under 8 treatments of combined fructose (from 20% to 5%) and ethanol (from 0% to 10%) were obtained, and the kinetic parameters were analyzed with principal component analysis and k-means clustering. The resulting yeast strain groups corresponded to the fast, medium and slow growers. A second clustering of only the fast growers led to the selection of 3 Saccharomyces strains (199, 230, 231) that were able to grow rapidly in 4 out of the 8 conditions evaluated. This methodology differentiated strains phenotypically and could be further used for strain selection in other processes. A method to select yeast strains for fermentation taking into account the natural differences of yeast isolates. This methodology is based on the cell exposition to combinations of sugar and ethanol, which are the most important stress factors in fermentation. This strategy will help to identify the most tolerant strain that could improve ethanol yield and reduce fermentation time. © 2018 Institute of Food Technologists®.

Synergetic effect of yeast cell-surface expression of cellulase and expansin-like protein on direct ethanol production from cellulose

PubMed Central

2013-01-01

Background Numerous studies have examined the direct fermentation of cellulosic materials by cellulase-expressing yeast; however, ethanol productivity in these systems has not yet reached an industrial level. Certain microorganisms, such as the cellulolytic fungus Trichoderma reesei, produce expansin-like proteins, which have a cellulose-loosening effect that may increase the breakdown of cellulose. Here, to improve the direct conversion of cellulose to ethanol, yeast Saccharomyces cerevisiae co-displaying cellulase and expansin-like protein on the cell surface were constructed and examined for direct ethanol fermentation performance. Results The cellulase and expansin-like protein co-expressing strain showed 246 mU/g-wet cell of phosphoric acid swollen cellulose (PASC) degradation activity, which corresponded to 2.9-fold higher activity than that of a cellulase-expressing strain. This result clearly demonstrated that yeast cell-surface expressed cellulase and expansin-like protein act synergistically to breakdown cellulose. In fermentation experiments examining direct ethanol production from PASC, the cellulase and expansin-like protein co-expressing strain produced 3.4 g/L ethanol after 96 h of fermentation, a concentration that was 1.4-fold higher than that achieved by the cellulase-expressing strain (2.5 g/L). Conclusions The PASC degradation and fermentation ability of an engineered yeast strain was markedly improved by co-expressing cellulase and expansin-like protein on the cell surface. To our knowledge, this is the first report to demonstrate the synergetic effect of co-expressing cellulase and expansin-like protein on a yeast cell surface, which may be a promising strategy for constructing direct ethanol fermenting yeast from cellulose. PMID:23835302

Microbial fuel cell treatment of ethanol fermentation process water

DOEpatents

Borole, Abhijeet P [Knoxville, TN

2012-06-05

The present invention relates to a method for removing inhibitor compounds from a cellulosic biomass-to-ethanol process which includes a pretreatment step of raw cellulosic biomass material and the production of fermentation process water after production and removal of ethanol from a fermentation step, the method comprising contacting said fermentation process water with an anode of a microbial fuel cell, said anode containing microbes thereon which oxidatively degrade one or more of said inhibitor compounds while producing electrical energy or hydrogen from said oxidative degradation, and wherein said anode is in electrical communication with a cathode, and a porous material (such as a porous or cation-permeable membrane) separates said anode and cathode.

Efficient xylose fermentation by the brown rot fungus Neolentinus lepideus.

PubMed

Okamoto, Kenji; Kanawaku, Ryuichi; Masumoto, Masaru; Yanase, Hideshi

2012-02-10

The efficient production of bioethanol on an industrial scale requires the use of renewable lignocellulosic biomass as a starting material. A limiting factor in developing efficient processes is identifying microorganisms that are able to effectively ferment xylose, the major pentose sugar found in hemicellulose, and break down carbohydrate polymers without pre-treatment steps. Here, a basidiomycete brown rot fungus was isolated as a new biocatalyst with unprecedented fermentability, as it was capable of converting not only the 6-carbon sugars constituting cellulose, but also the major 5-carbon sugar xylose in hemicelluloses, to ethanol. The fungus was identified as Neolentinus lepideus and was capable of assimilating and fermenting xylose to ethanol in yields of 0.30, 0.33, and 0.34 g of ethanol per g of xylose consumed under aerobic, oxygen-limited, and anaerobic conditions, respectively. A small amount of xylitol was detected as the major by-product of xylose metabolism. N. lepideus produced ethanol from glucose, mannose, galactose, cellobiose, maltose, and lactose with yields ranging from 0.34 to 0.38 g ethanol per g sugar consumed, and also exhibited relatively favorable conversion of non-pretreated starch, xylan, and wheat bran. These results suggest that N. lepideus is a promising candidate for cost-effective and environmentally friendly ethanol production from lignocellulosic biomass. To our knowledge, this is the first report on efficient ethanol fermentation from various carbohydrates, including xylose, by a naturally occurring brown rot fungus. Copyright © 2011 Elsevier Inc. All rights reserved.

Opuntia ficus-indica cladodes as feedstock for ethanol production by Kluyveromyces marxianus and Saccharomyces cerevisiae.

PubMed

Kuloyo, Olukayode O; du Preez, James C; García-Aparicio, Maria del Prado; Kilian, Stephanus G; Steyn, Laurinda; Görgens, Johann

2014-12-01

The feasibility of ethanol production using an enzymatic hydrolysate of pretreated cladodes of Opuntia ficus-indica (prickly pear cactus) as carbohydrate feedstock was investigated, including a comprehensive chemical analysis of the cladode biomass and the effects of limited aeration on the fermentation profiles and sugar utilization. The low xylose and negligible mannose content of the cladode biomass used in this study suggested that the hemicellulose structure of the O. ficus-indica cladode was atypical of hardwood or softwood hemicelluloses. Separate hydrolysis and fermentation and simultaneous saccharification and fermentation procedures using Kluyveromyces marxianus and Saccharomyces cerevisiae at 40 and 35 °C, respectively, gave similar ethanol yields under non-aerated conditions. In oxygen-limited cultures K. marxianus exhibited almost double the ethanol productivity compared to non-aerated cultures, although after sugar depletion utilization of the produced ethanol was evident. Ethanol concentrations of up to 19.5 and 20.6 g l(-1) were obtained with K. marxianus and S. cerevisiae, respectively, representing 66 and 70 % of the theoretical yield on total sugars in the hydrolysate. Because of the low xylan content of the cladode biomass, a yeast capable of xylose fermentation might not be a prerequisite for ethanol production. K. marxianus, therefore, has potential as an alternative to S. cerevisiae for bioethanol production. However, the relatively low concentration of fermentable sugars in the O. ficus-indica cladode hydrolysate presents a technical constraint for commercial exploitation.

Recombinant Zymomonas for pentose fermentation

DOEpatents

Picataggio, S.K.; Min Zhang; Eddy, C.K.; Deanda, K.A.

1998-03-10

The invention relates to microorganisms which normally do not ferment pentose sugar and which are genetically altered to ferment pentose sugar to produce ethanol, and fermentation processes utilizing the same. Examples include Zymomonas mobilis which has been transformed with combinations of E. coli genes for xylose isomerase, xylulokinase, transaldolase, transketolase, L-arabinose isomerase, L-ribulokinase, and L-ribulose-5-phosphate 4-epimerase. Expression of the added genes are under the control of Zymomonas mobilis promoters. These newly created microorganisms are useful for fermenting pentoses and glucose, produced by hydrolysis of hemicellulose and cellulose, to produce ethanol. 7 figs.

Pentose fermentation by recombinant Zymomonas

DOEpatents

Picataggio, S.K.; Zhang, M.; Eddy, C.K.; Deanda, K.A.; Finkelstein, M.; Mohagheghi, A.; Newman, M.M.; McMillan, J.D.

1998-01-27

The invention relates to microorganisms which normally do not ferment pentose sugar and which are genetically altered to ferment pentose sugar to produce ethanol, and fermentation processes utilizing the same. Examples include Zymomonas mobilis which has been transformed with combinations of E. coli genes for xylose isomerase, xylulokinase, transaldolase, transketolase, L-arabinose isomerase, L-ribulokinase, and L-ribulose 5-phosphate 4-epimerase. Expression of the added genes are under the control of Zymomonas mobilis promoters. These newly created microorganisms are useful for fermenting pentoses and glucose, produced by hydrolysis of hemicellulose and cellulose, to produce ethanol. 7 figs.

Pentose fermentation by recombinant zymomonas

DOEpatents

Picataggio, Stephen K.; Zhang, Min; Eddy, Christina K.; Deanda, Kristine A.; Finkelstein, Mark; Mohagheghi, Ali; Newman, Mildred M.; McMillan, James D.

1998-01-01

The invention relates to microorganisms which normally do not ferment pentose sugar and which are genetically altered to ferment pentose sugar to produce ethanol, and fermentation processes utilizing the same. Examples include Zymomonas mobilis which has been transformed with combinations of E. coli genes for xylose isomerase, xylulokinase, transaldolase, transketolase, L-arabinose isomerase, L-ribulokinase, and L-ribulose 5-phosphate 4-epimerase. Expression of the added genes are under the control of Zymomonas mobilis promoters. These newly created microorganisms are useful for fermenting pentoses and glucose, produced by hydrolysis of hemicellulose and cellulose, to produce ethanol.

Recombinant Zymomonas for pentose fermentation

DOEpatents

Picataggio, Stephen K.; Zhang, Min; Eddy, Christina K.; Deanda, Kristine A.

1998-01-01

The invention relates to microorganisms which normally do not ferment pentose sugar and which are genetically altered to ferment pentose sugar to produce ethanol, and fermentation processes utilizing the same. Examples include Zymomonas mobilis which has been transformed with combinations of E. coli genes for xylose isomerase, xylulokinase, transaldolase, transketolase, L-arabinose isomerase, L-ribulokinase, and L-ribulose-5-phosphate 4-epimerase. Expression of the added genes are under the control of Zymomonas mobilis promoters. These newly created microorganisms are useful for fermenting pentoses and glucose, produced by hydrolysis of hemicellulose and cellulose, to produce ethanol.

Increase of ethanol productivity by cell-recycle fermentation of flocculating yeast.

PubMed

Wang, F Z; Xie, T; Hui, M

2011-01-01

Using the recombinant flocculating Angel yeast F6, long-term repeated batch fermentation for ethanol production was performed and a high volumetric productivity resulted from half cells not washed and the optimum opportunity of residual glucose 20 g l(-1) of last medium. The obtained highest productivity was 2.07 g l-(1) h(-1), which was improved by 75.4% compared with that of 1.18 g l(-1) h(-1) in the first batch fermentation. The ethanol concentration reached 8.4% corresponding to the yield of 0.46 g g(-1). These results will contribute greatly to the industrial production of fuel ethanol using the commercial method with the flocculating yeast.

Process simulation of modified dry grind ethanol plant with recycle of pretreated and enzymatically hydrolyzed distillers' grains.

PubMed

Kim, Youngmi; Mosier, Nathan; Ladisch, Michael R

2008-08-01

Distillers' grains (DG), a co-product of a dry grind ethanol process, is an excellent source of supplemental proteins in livestock feed. Studies have shown that, due to its high polymeric sugar contents and ease of hydrolysis, the distillers' grains have potential as an additional source of fermentable sugars for ethanol fermentation. The benefit of processing the distillers' grains to extract fermentable sugars lies in an increased ethanol yield without significant modification in the current dry grind technology. Three different potential configurations of process alternatives in which pretreated and hydrolyzed distillers' grains are recycled for an enhanced overall ethanol yield are proposed and discussed in this paper based on the liquid hot water (LHW) pretreatment of distillers' grains. Possible limitations of each proposed process are also discussed. This paper presents a compositional analysis of distillers' grains, as well as a simulation of the modified dry grind processes with recycle of distillers' grains. Simulated material balances for the modified dry grind processes are established based on the base case assumptions. These balances are compared to the conventional dry grind process in terms of ethanol yield, compositions of its co-products, and accumulation of fermentation inhibitors. Results show that 14% higher ethanol yield is achievable by processing and hydrolyzing the distillers' grains for additional fermentable sugars, as compared to the conventional dry grind process. Accumulation of fermentation by-products and inhibitory components in the proposed process is predicted to be 2-5 times higher than in the conventional dry grind process. The impact of fermentation inhibitors is reviewed and discussed. The final eDDGS (enhanced dried distillers' grains) from the modified processes has 30-40% greater protein content per mass than DDGS, and its potential as a value-added process is also analyzed. While the case studies used to illustrate the process simulation are based on LHW pretreated DG, the process simulation itself provides a framework for evaluation of the impact of other pretreatments.

Recombinant Zymomonas for pentose fermentation

DOEpatents

Picataggio, S.K.; Zhang, M.; Eddy, C.K.; Deanda, K.A.; Finkelstein, M.

1996-05-07

The invention relates to microorganisms which normally do not ferment a pentose sugar and which are genetically altered to ferment this pentose to produce ethanol. A representative example is Zymomonas mobilis which has been transformed with E. coli xylose isomerase, xylulokinase, transaldolase and transketolase genes. Expression of the added genes are under the control of Zymomonas mobilis promoters. This newly created microorganism is useful for fermenting pentoses and glucose, produced by hydrolysis of hemicellulose and cellulose, to produce ethanol. 2 figs.

Recombinant zymomonas for pentose fermentation

DOEpatents

Picataggio, Stephen K.; Zhang, Min; Eddy, Christina K.; Deanda, Kristine A.; Finkelstein, Mark

1996-01-01

The invention relates to microorganisms which normally do not ferment a pentose sugar and which are genetically altered to ferment this pentose to produce ethanol. A representative example is Zymomonas mobilis which has been transformed with E. coli xylose isomerase, xylulokinase, transaldolase and transketolase genes. Expression of the added genes are under the control of Zymomonas mobilis promoters. This newly created microorganism is useful for fermenting pentoses and glucose, produced by hydrolysis of hemicellulose and cellulose, to produce ethanol.

Redox potential driven aeration during very-high-gravity ethanol fermentation by using flocculating yeast.

PubMed

Liu, Chen-Guang; Hao, Xue-Mi; Lin, Yen-Han; Bai, Feng-Wu

2016-05-10

Ethanol fermentation requires oxygen to maintain high biomass and cell viability, especially under very-high-gravity (VHG) condition. In this work, fermentation redox potential (ORP) was applied to drive the aeration process at low dissolved oxygen (DO) levels, which is infeasible to be regulated by a DO sensor. The performance and characteristics of flocculating yeast grown under 300 and 260 g glucose/L conditions were subjected to various aeration strategies including: no aeration; controlled aeration at -150, -100 and -50 mV levels; and constant aeration at 0.05 and 0.2 vvm. The results showed that anaerobic fermentation produced the least ethanol and had the highest residual glucose after 72 h of fermentation. Controlled aerations, depending on the real-time oxygen demand, led to higher cell viability than the no-aeration counterpart. Constant aeration triggered a quick biomass formation, and fast glucose utilization. However, over aeration at 0.2 vvm caused a reduction of final ethanol concentration. The controlled aeration driven by ORP under VHG conditions resulted in the best fermentation performance. Moreover, the controlled aeration could enhance yeast flocculating activity, promote an increase of flocs size, and accelerate yeast separation near the end of fermentation.

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

PubMed

Schlepütz, Tino; Büchs, Jochen

2013-01-01

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

Redox potential driven aeration during very-high-gravity ethanol fermentation by using flocculating yeast

PubMed Central

Liu, Chen-Guang; Hao, Xue-Mi; Lin, Yen-Han; Bai, Feng-Wu

2016-01-01

Ethanol fermentation requires oxygen to maintain high biomass and cell viability, especially under very-high-gravity (VHG) condition. In this work, fermentation redox potential (ORP) was applied to drive the aeration process at low dissolved oxygen (DO) levels, which is infeasible to be regulated by a DO sensor. The performance and characteristics of flocculating yeast grown under 300 and 260 g glucose/L conditions were subjected to various aeration strategies including: no aeration; controlled aeration at −150, −100 and −50 mV levels; and constant aeration at 0.05 and 0.2 vvm. The results showed that anaerobic fermentation produced the least ethanol and had the highest residual glucose after 72 h of fermentation. Controlled aerations, depending on the real-time oxygen demand, led to higher cell viability than the no-aeration counterpart. Constant aeration triggered a quick biomass formation, and fast glucose utilization. However, over aeration at 0.2 vvm caused a reduction of final ethanol concentration. The controlled aeration driven by ORP under VHG conditions resulted in the best fermentation performance. Moreover, the controlled aeration could enhance yeast flocculating activity, promote an increase of flocs size, and accelerate yeast separation near the end of fermentation. PMID:27161047

Enhancement of ethanol fermentation in Saccharomyces cerevisiae sake yeast by disrupting mitophagy function.

PubMed

Shiroma, Shodai; Jayakody, Lahiru Niroshan; Horie, Kenta; Okamoto, Koji; Kitagaki, Hiroshi

2014-02-01

Saccharomyces cerevisiae sake yeast strain Kyokai no. 7 has one of the highest fermentation rates among brewery yeasts used worldwide; therefore, it is assumed that it is not possible to enhance its fermentation rate. However, in this study, we found that fermentation by sake yeast can be enhanced by inhibiting mitophagy. We observed mitophagy in wild-type sake yeast during the brewing of Ginjo sake, but not when the mitophagy gene (ATG32) was disrupted. During sake brewing, the maximum rate of CO2 production and final ethanol concentration generated by the atg32Δ laboratory yeast mutant were 7.50% and 2.12% higher than those of the parent strain, respectively. This mutant exhibited an improved fermentation profile when cultured under limiting nutrient concentrations such as those used during Ginjo sake brewing as well as in minimal synthetic medium. The mutant produced ethanol at a concentration that was 2.76% higher than the parent strain, which has significant implications for industrial bioethanol production. The ethanol yield of the atg32Δ mutant was increased, and its biomass yield was decreased relative to the parent sake yeast strain, indicating that the atg32Δ mutant has acquired a high fermentation capability at the cost of decreasing biomass. Because natural biomass resources often lack sufficient nutrient levels for optimal fermentation, mitophagy may serve as an important target for improving the fermentative capacity of brewery yeasts.

75 FR 21300 - North American Bioproducts Corp.; Filing of Food Additive Petition (Animal Use); Erythromycin...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-04-23

... antimicrobial processing aid in fuel- ethanol fermentations with respect to its consequent presence in by... aid in fuel- ethanol fermentations with respect to its consequent presence in by- product distiller...

75 FR 55798 - North American Bioproducts Corporation; Filing of Food Additive Petition (Animal Use); Penicillin...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-09-14

... antimicrobial processing aid in fuel- ethanol fermentations with respect to its consequent presence in by... antimicrobial processing aid in fuel- ethanol fermentations with respect to its consequent presence in by...

Bioethanol production performance of five recombinant strains of laboratory and industrial xylose-fermenting Saccharomyces cerevisiae.

PubMed

Matsushika, Akinori; Inoue, Hiroyuki; Murakami, Katsuji; Takimura, Osamu; Sawayama, Shigeki

2009-04-01

In this study, five recombinant Saccharomyces cerevisiae strains were compared for their xylose-fermenting ability. The most efficient xylose-to-ethanol fermentation was found by using the industrial strain MA-R4, in which the genes for xylose reductase and xylitol dehydrogenase from Pichia stipitis along with an endogenous xylulokinase gene were expressed by chromosomal integration of the flocculent yeast strain IR-2. The MA-R4 strain rapidly converted xylose to ethanol with a low xylitol yield. Furthermore, the MA-R4 strain had the highest ethanol production when fermenting not only a mixture of glucose and xylose, but also mixed sugars in the detoxified hydrolysate of wood chips. These results collectively suggest that MA-R4 may be a suitable recombinant strain for further study into large-scale ethanol production from mixed sugars present in lignocellulosic hydrolysates.

Metabolic engineering of Saccharomyces cerevisiae ethanol strains PE-2 and CAT-1 for efficient lignocellulosic fermentation.

PubMed

Romaní, Aloia; Pereira, Filipa; Johansson, Björn; Domingues, Lucília

2015-03-01

In this work, Saccharomyces cerevisiae strains PE-2 and CAT-1, commonly used in the Brazilian fuel ethanol industry, were engineered for xylose fermentation, where the first fermented xylose faster than the latter, but also produced considerable amounts of xylitol. An engineered PE-2 strain (MEC1121) efficiently consumed xylose in presence of inhibitors both in synthetic and corn-cob hydrolysates. Interestingly, the S. cerevisiae MEC1121 consumed xylose and glucose simultaneously, while a CEN.PK based strain consumed glucose and xylose sequentially. Deletion of the aldose reductase GRE3 lowered xylitol production to undetectable levels and increased xylose consumption rate which led to higher final ethanol concentrations. Fermentation of corn-cob hydrolysate using this strain, MEC1133, resulted in an ethanol yield of 0.47 g/g of total sugars which is 92% of the theoretical yield. Copyright © 2014 Elsevier Ltd. All rights reserved.

Cost-effective approach to ethanol production and optimization by response surface methodology.

PubMed

Uncu, Oya Nihan; Cekmecelioglu, Deniz

2011-04-01

Food wastes disposed from residential and industrial kitchens have gained attention as a substrate in microbial fermentations to reduce product costs. In this study, the potential of simultaneously hydrolyzing and subsequently fermenting the mixed carbohydrate components of kitchen wastes were assessed and the effects of solid load, inoculum volume of baker's yeast, and fermentation time on ethanol production were evaluated by response surface methodology (RSM). The enzymatic hydrolysis process was complete within 6h. Fermentation experiments were conducted at pH 4.5, a temperature of 30°C, and agitated at 150 rpm without adding the traditional fermentation nutrients. The statistical analysis of the model developed by RSM suggested that linear effects of solid load, inoculum volume, and fermentation time and the quadratic effects of inoculum volume and fermentation time were significant (P<0.05). The verification experiments indicated that the developed model could be successfully used to predict ethanol concentration at >90% accuracy. An optimum ethanol concentration of 32.2g/l giving a yield of 0.40g/g, comparable to yields reported to date, was suggested by the model with 20% solid load, 8.9% inoculum volume, and 58.8h of fermentation. The results indicated that the production costs can be lowered to a large extent by using kitchen wastes having multiple carbohydrate components and eliminating the use of traditional fermentation nutrients from the recipe. Copyright © 2010 Elsevier Ltd. All rights reserved.

Improved ethanol production from xylose in the presence of acetic acid by the overexpression of the HAA1 gene in Saccharomyces cerevisiae.

PubMed

Sakihama, Yuri; Hasunuma, Tomohisa; Kondo, Akihiko

2015-03-01

The hydrolysis of lignocellulosic biomass liberates sugars, primarily glucose and xylose, which are subsequently converted to ethanol by microbial fermentation. The rapid and efficient fermentation of xylose by recombinant Saccharomyces cerevisiae strains is limited by weak acids generated during biomass pretreatment processes. In particular, acetic acid negatively affects cell growth, xylose fermentation rate, and ethanol production. The ability of S. cerevisiae to efficiently utilize xylose in the presence of acetic acid is an essential requirement for the cost-effective production of ethanol from lignocellulosic hydrolysates. Here, an acetic acid-responsive transcriptional activator, HAA1, was overexpressed in a recombinant xylose-fermenting S. cerevisiae strain to yield BY4741X/HAA1. This strain exhibited improved cell growth and ethanol production from xylose under aerobic and oxygen limited conditions, respectively, in the presence of acetic acid. The HAA1p regulon enhanced transcript levels in BY4741X/HAA1. The disruption of PHO13, a p-nitrophenylphosphatase gene, in BY4741X/HAA1 led to further improvement in both yeast growth and the ability to ferment xylose, indicating that HAA1 overexpression and PHO13 deletion act by different mechanisms to enhance ethanol production. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Ethanol production from rice winery waste-rice wine cake by simultaneous saccharification and fermentation without cooking.

PubMed

Vu, Van Hanh; Kim, Keun

2009-10-01

Ethanol production by simultaneous saccharification and fermentation (SSF) of low-value rice wine cake (RWC) without cooking was investigated. RWC is the filtered solid waste of fermented rice wine mash and contains 53% of raw starch. RWC slurry was mixed with raw-starch-digesting enzyme of Rhizopus sp. and yeast for SSF. The yeast strain used was selected from 300 strains for RWC fermentation and identified as Saccharomyces cerevisiae KV25. High efficiency (94%) of ethanol production was achieved at optimal condition of uncooked RWC slurry containing 23.03% of starch. The optimal SSF condition determined was 1.125 unit of raw-starch-digesting enzyme per one gram of RWC, 30 degrees C of fermentation temperature, 4.5 of pH slurry, 36 h-age of seeding culture, initial yeast cell 2 x 10(7) per ml slurry, 17 mM urea as nitrogen additive, 0.25 mM Cu(2+) as metal ion additives, 90 h of fermentation time. In this optimal condition, ethanol production by SSF of uncooked RWC slurry was improved to 16.8% (v/v) from 15.1% (v/v) of pre-optimization.

Acid hydrolysis of Jerusalem artichoke for ethanol fermentation

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

Kim, K.; Hamdy, M.K.

1986-01-01

An excellent substrate for ethanol production is the Jerusalem artichoke (JA) tuber (Helianthus tuberosus). This crop contains a high level of inulin that can be hydrolyzed mainly to D-fructose and has several distinct advantages as an energy source compared to others. The potential ethanol yield of ca. 4678 L/ha on good agricultural land is equivalent to that obtained from sugar beets and twice that of corn. When JA is to be used for ethanol fermentation by conventional yeast, it is first converted to fermentable sugars by enzymes or acids although various strains of yeast were used for the direct fermentationmore » of JA extracts. Fleming and GrootWassink compared various acids (hydrochloric, sulfuric, citric, and phosphoric) and strong cation exchange resin for their effectiveness on inulin hydrolysis and reported that no differences were noted among the acids or resin in their influence on inulin hydrolysis. Undesirable side reactions were noted during acid hydrolysis leading to the formation of HMF and 2-(2-hydroxy acetyl) furan. The HMF at a level of 0.1% is known to inhibit growth and ethanol fermentation by yeast. In this study the authors established optimal conditions for complete acid-hydrolysis of JA with minimum side reactions and maximum sugar-ethanol production. A material balance for the ethanol production was also determined.« less

Current Trends in Bioethanol Production by Saccharomyces cerevisiae: Substrate, Inhibitor Reduction, Growth Variables, Coculture, and Immobilization

PubMed Central

Assefa, Fassil

2014-01-01

Bioethanol is one of the most commonly used biofuels in transportation sector to reduce greenhouse gases. S. cerevisiae is the most employed yeast for ethanol production at industrial level though ethanol is produced by an array of other yeasts, bacteria, and fungi. This paper reviews the current and nonmolecular trends in ethanol production using S. cerevisiae. Ethanol has been produced from wide range of substrates such as molasses, starch based substrate, sweet sorghum cane extract, lignocellulose, and other wastes. The inhibitors in lignocellulosic hydrolysates can be reduced by repeated sequential fermentation, treatment with reducing agents and activated charcoal, overliming, anion exchanger, evaporation, enzymatic treatment with peroxidase and laccase, in situ detoxification by fermenting microbes, and different extraction methods. Coculturing S. cerevisiae with other yeasts or microbes is targeted to optimize ethanol production, shorten fermentation time, and reduce process cost. Immobilization of yeast cells has been considered as potential alternative for enhancing ethanol productivity, because immobilizing yeasts reduce risk of contamination, make the separation of cell mass from the bulk liquid easy, retain stability of cell activities, minimize production costs, enable biocatalyst recycling, reduce fermentation time, and protect the cells from inhibitors. The effects of growth variables of the yeast and supplementation of external nitrogen sources on ethanol optimization are also reviewed. PMID:27379305

Effect of carbon sources on the growth and ethanol production of native yeast Pichia kudriavzevii ITV-S42 isolated from sweet sorghum juice.

PubMed

Díaz-Nava, L E; Montes-Garcia, N; Domínguez, J M; Aguilar-Uscanga, M G

2017-07-01

The importance of non-Saccharomyces yeast species in fermentation processes is widely acknowledged. Within this group, Pichia kudriavzevii ITV-S42 yeast strain shows particularly desirable characteristics for ethanol production. Despite this fact, a thorough study of the metabolic and kinetic characteristics of this strain is currently unavailable. The aim of this work is to study the nutritional requirements of Pichia kudriavzevii ITV-S42 strain and the effect of different carbon sources on the growth and ethanol production. Results showed that glucose and fructose were both assimilated and fermented, achieving biomass and ethanol yields of 0.37 and 0.32 gg -1 , respectively. Glycerol was assimilated but not fermented; achieving a biomass yield of 0.88 gg -1 . Xylose and sucrose were not metabolized by the yeast strain. Finally, the use of a culture medium enriched with salts and yeast extract favored glucose consumption both for growth and ethanol production, improving ethanol tolerance reported for this genre (35 g L -1 ) to 90 g L -1 maximum ethanol concentration (over 100%). Furthermore Pichia kudriavzevii ITV-S42 maintained its fermentative capacity up to 200 g L -1 initial glucose, demonstrating that this yeast is osmotolerant.

Novel DDR Processing of Corn Stover Achieves High Monomeric Sugar Concentrations from Enzymatic Hydrolysis (230 g/L) and High Ethanol Concentration (10% v/v) During Fermentation

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

Chen, Xiaowen; Jennings, Ed; Shekiro, Joe

Distilling and purifying ethanol, butanol, and other products from second and later generation lignocellulosic biorefineries adds significant capital and operating cost for biofuels production. The energy costs associated with distillation affects plant gate and life cycle analysis costs. Lower titers in fermentation due to lower sugar concentrations from pretreatment increase both energy and production costs. In addition, higher titers decrease the volumes required for enzymatic hydrolysis and fermentation vessels. Therefore, increasing biofuels titers has been a research focus in renewable biofuels production for several decades. In this work, we achieved over 200 g/L of monomeric sugars after high solids enzymaticmore » hydrolysis using the novel deacetylation and disc refining (DDR) process on corn stover. The high sugar concentrations and low chemical inhibitor concentrations from the DDR process allowed ethanol titers as high as 82 g/L in 22 hours, which translates into approximately 10 vol% ethanol. To our knowledge, this is the first time that 10 vol% ethanol in fermentation derived from corn stover without any sugar concentration or purification steps has been reported. Techno-economic analysis shows the higher titer ethanol achieved from the DDR process could significantly reduce the minimum ethanol selling price from cellulosic biomass.« less

Fermentation of D-xylose and L-arabinose to ethanol by Erwinia chrysanthemi

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

Tolan, J.S.; Finn, R.K.

1987-09-01

Erwinia spp. are gram-negative facultative anaerobes within the family Enterobacteriacae which possess several desirable traits for the conversion of pentose sugars to ethanol, such as the ability to ferment a broad range of carbohydrates and the ease with which they can be genetically modified. Twenty-eight strains of Erwinia carotovora and E. chrysanthemi were screened for the ability to ferment D-xylose to ethanol. E. chrysanthemi B374 was chosen for further study on the basis of its superior (4%) ethanol tolerance. They have characterized the fermentation of D-xylose and L-arabinose by the wild type and mutants which bear plasmids containing the pyruvatemore » decarboxylase gene from Zymomonas mobilis. Expression of the gene markedly increased the yields of ethanol (from 0.7 up to 1.45 mol/mol of xylose) and decreased the yields of formate, acetate, and lactate. However, the cells with pyruvate decarboxylase grew only one-fourth as fast as the wild type and tolerated only 2% ethanol. Alcohol tolerance was stimulated by the addition of yeast extract to the growth medium. Xylose catabolism was characterized by a high saturation constant K/sub s/ (4.5 mM).« less

Ethanol production from eucalyptus wood hemicellulose hydrolysate by Pichia stipitis.

PubMed

Ferrari, M D; Neirotti, E; Albornoz, C; Saucedo, E

1992-10-05

Ethanol production was evaluated from eucalyptus wood hemicellulose acid hydrolysate using Pichia stipitis NRRL Y-7124. An initial lag phase characterized by flocculation and viability loss of the yeast inoculated was observed. Subsequently, cell regrowth occurred with sequential consumption of sugars and production of ethanol. Polyol formation was detected. Acetic acid present in the hydrolysate was an important inhibitor of the fermentation, reducing the rate and the yield. Its toxic effect was due essentially to its undissociated form. The fermentation was more effective at an oxygen transfer rate between 1.2 and 2.4 mmol/L h and an initial pH of 6.5. The hydrolysate used in the experiences had the following composition (expressed in grams per liter): xylose 30, arabinose 2.8, glucose 1.5, galactose 3.7, mannose 1.0, cellobiose 0.5, acetic acid 10, glucuronic acid 1.5, and galacturonic acid 1.0. The best values obtained were maximum ethanol concentration 12.6 g/L, fermentation time 75 h, fermentable sugar consumption 99% ethanol yield 0.35 g/g sugars consumed, and volumetric ethanol productivity 4 g/L day. ( (c) 1992 John Wiley & Sons, Inc.

Potential of the waste from beer fermentation broth for bio-ethanol production without any additional enzyme, microbial cells and carbohydrates.

PubMed

Ha, Jung Hwan; Shah, Nasrullah; Ul-Islam, Mazhar; Park, Joong Kon

2011-08-10

The potential of the waste from beer fermentation broth (WBFB) for the production of bio-ethanol using a simultaneous saccharification and fermentation process without any extra additions of saccharification enzymes, microbial cells or carbohydrate was tested. The major microbial cells in WBFB were isolated and identified. The variations in compositions of WBFB with stock time were investigated. There was residual activity of starch hydrolyzing enzymes in WBFB. The effects of reaction modes e.g. static and shaking on bio-ethanol production were studied. After 7 days of cultivation using the supernatant of WBFB at 30 °C the ethanol concentration reached 103.8 g/L in shaking culture and 91.5 g/L in static culture. Agitation experiments conducted at a temperature-profile process in which temperature was increased from 25 to 67 °C shortened the simultaneous process time. The original WBFB was more useful than the supernatant of WBFB in getting the higher concentration of ethanol and reducing the fermentation time. From this whole study it was found that WBFB is a cheap and suitable source for bio-ethanol production. Copyright © 2011 Elsevier Inc. All rights reserved.

Effect of acetic acid in recycling water on ethanol production for cassava in an integrated ethanol-methane fermentation process.

PubMed

Yang, Xinchao; Wang, Ke; Zhang, Jianhua; Tang, Lei; Mao, Zhonggui

2016-11-01

Recently, the integrated ethanol-methane fermentation process has been studied to prevent wastewater pollution. However, when the anaerobic digestion reaction runs poorly, acetic acid will accumulate in the recycling water. In this paper, we studied the effect of low concentration of acetic acid (≤25 mM) on ethanol fermentation at different initial pH values (4.2, 5.2 or 6.2). At an initial pH of 4.2, ethanol yields increased by 3.0% and glycerol yields decreased by 33.6% as the acetic acid concentration was increased from 0 to 25 mM. Raising the concentration of acetic acid to 25 mM increased the buffering capacity of the medium without obvious effects on biomass production in the cassava medium. Acetic acid was metabolized by Saccharomyces cerevisiae for the reason that the final concentration of acetic acid was 38.17% lower than initial concentration at pH 5.2 when 25 mM acetic acid was added. These results confirmed that a low concentration of acetic acid in the process stimulated ethanol fermentation. Thus, reducing the acetic acid concentration to a controlled low level is more advantageous than completely removing it.

Conditioning of dilute-acid pretreated corn stover hydrolysate liquors by treatment with lime or ammonium hydroxide to improve conversion of sugars to ethanol.

PubMed

Jennings, Edward W; Schell, Daniel J

2011-01-01

Dilute-acid pretreatment of lignocellulosic biomass enhances the ability of enzymes to hydrolyze cellulose to glucose, but produces many toxic compounds that inhibit fermentation of sugars to ethanol. The objective of this study was to compare the effectiveness of treating hydrolysate liquor with Ca(OH)2 and NH4OH for improving ethanol yields. Corn stover was pretreated in a pilot-scale reactor and then the liquor fraction (hydrolysate) was extracted and treated with various amounts of Ca(OH)2 or NH4OH at several temperatures. Glucose and xylose in the treated liquor were fermented to ethanol using a glucose-xylose fermenting bacteria, Zymomonas mobilis 8b. Sugar losses up to 10% occurred during treatment with Ca(OH)2, but these losses were two to fourfold lower with NH4OH treatment. Ethanol yields for NH4OH-treated hydrolysate were 33% greater than those achieved in Ca(OH)2-treated hydrolysate and pH adjustment to either 6.0 or 8.5 with NH4OH prior to fermentation produced equivalent ethanol yields. Copyright © 2010 Elsevier Ltd. All rights reserved.

Ethanol Cellular Defense Induce Unfolded Protein Response in Yeast

PubMed Central

Pérez-Torrado, Roberto

2016-01-01

Ethanol is a valuable industrial product and a common metabolite used by many cell types. However, this molecule produces high levels of cytotoxicity affecting cellular performance at several levels. In the presence of ethanol, cells must adjust some of their components, such as the membrane lipids to maintain homeostasis. In the case of microorganism as Saccharomyces cerevisiae, ethanol is one of the principal products of their metabolism and is the main stress factor during fermentation. Although, many efforts have been made, mechanisms of ethanol tolerance are not fully understood and very little evidence is available to date for specific signaling by ethanol in the cell. This work studied two S. cerevisiae strains, CECT10094, and Temohaya-MI26, isolated from flor wine and agave fermentation (a traditional fermentation from Mexico) respectively, which differ in ethanol tolerance, in order to understand the molecular mechanisms underlying the ethanol stress response and the reasons for different ethanol tolerance. The transcriptome was analyzed after ethanol stress and, among others, an increased activation of genes related with the unfolded protein response (UPR) and its transcription factor, Hac1p, was observed in the tolerant strain CECT10094. We observed that this strain also resist more UPR agents than Temohaya-MI26 and the UPR-ethanol stress correlation was corroborated observing growth of 15 more strains and discarding UPR correlation with other stresses as thermal or oxidative stress. Furthermore, higher activation of UPR pathway in the tolerant strain CECT10094 was observed using a UPR mCherry reporter. Finally, we observed UPR activation in response to ethanol stress in other S. cerevisiae ethanol tolerant strains as the wine strains T73 and EC1118. This work demonstrates that the UPR pathway is activated under ethanol stress occurring in a standard fermentation and links this response to an enhanced ethanol tolerance. Thus, our data suggest that there is a room for ethanol tolerance improvement by enhancing UPR response. PMID:26925053

Bioconversion of glycerol to ethanol by a mutant Enterobacter aerogenes

PubMed Central

2012-01-01

The main objective of this research is to develop, by adaptive evolution, mutant strains of Enterobacter aerogenes ATCC 13048 that are capable of withstanding high glycerol concentration as well as resisting ethanol-inhibition. The mutant will be used for high ethanol fermentation from glycerol feedstock. Ethanol production from pure (P-) and recovered (R-) glycerol using the stock was evaluated. A six-tube-subculture-generations method was used for developing the mutant. This involved subculturing the organism six consecutive times in tubes containing the same glycerol and ethanol concentrations at the same culture conditions. Then, the glycerol and/or ethanol concentration was increased and the six subculture generations were repeated. A strain capable of growing in 200 g/L glycerol and 30 g/L ethanol was obtained. The ability of this mutant, vis-à-vis the original strain, in utilizing glycerol in a high glycerol containing medium, with the concomitant ethanol yield, was assessed. Tryptic soy broth without dextrose (TSB) was used as the fermentation medium. Fermentation products were analyzed using HPLC. In a 20 g/L glycerol TSB, E. aerogenes ATCC 13048 converted 18.5 g/L P-glycerol and 17.8 g/L R-glycerol into 12 and 12.8 g/L ethanol, respectively. In a 50 g/L P-glycerol TSB, it utilized only 15.6 g/L glycerol; but the new strain used up 39 g/L, yielding 20 g/L ethanol after 120 h, an equivalence of 1.02 mol ethanol/mol-glycerol. This is the highest ethanol yield reported from glycerol bioconversion. The result of this P-glycerol fermentation can be duplicated using the R-glycerol from biodiesel production. PMID:22455837

ADSORPTION AND MEMBRANE SEPARATION MEASUREMENTS WITH MIXTURES OF ETHANOL, ACETIC ACID, AND WATER

EPA Science Inventory

Biomass fermentation produces ethanol and other renewable biofuels. Pervaporation using hydrophobic membranes is potentially a cost-effective means of removing biofuels from fermentation broths for small- to medium-scale applications. Silicalite-filled polydimethylsiloxane (PDMS)...

76 FR 22904 - Ferm Solutions, Inc.; Filing of Food Additive Petition (Animal Use); Erythromycin Thiocyanate

Federal Register 2010, 2011, 2012, 2013, 2014

2011-04-25

...-ethanol fermentations with respect to its consequent presence in byproduct distiller grains used as an... thiocyanate as an antimicrobial processing aid in fuel-ethanol fermentations with respect to its [[Page 22905...

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.

Detoxification and fermentation of pyrolytic sugar for ethanol production.

PubMed

Wang, Hui; Livingston, Darrell; Srinivasan, Radhakrishnan; Li, Qi; Steele, Philip; Yu, Fei

2012-11-01

The sugars present in bio-oil produced by fast pyrolysis can potentially be fermented by microbial organisms to produce cellulosic ethanol. This study shows the potential for microbial digestion of the aqueous fraction of bio-oil in an enrichment medium to consume glucose and produce ethanol. In addition to glucose, inhibitors such as furans and phenols are present in the bio-oil. A pure glucose enrichment medium of 20 g/l was used as a standard to compare with glucose and aqueous fraction mixtures for digestion. Thirty percent by volume of aqueous fraction in media was the maximum additive amount that could be consumed and converted to ethanol. Inhibitors were removed by extraction, activated carbon, air stripping, and microbial methods. After economic analysis, the cost of ethanol using an inexpensive fermentation medium in a large scale plant is approximately $14 per gallon.

Sustaining fermentation in high-gravity ethanol production by feeding yeast to a temperature-profiled multifeed simultaneous saccharification and co-fermentation of wheat straw.

PubMed

Westman, Johan O; Wang, Ruifei; Novy, Vera; Franzén, Carl Johan

2017-01-01

Considerable progress is being made in ethanol production from lignocellulosic feedstocks by fermentation, but negative effects of inhibitors on fermenting microorganisms are still challenging. Feeding preadapted cells has shown positive effects by sustaining fermentation in high-gravity simultaneous saccharification and co-fermentation (SSCF). Loss of cell viability has been reported in several SSCF studies on different substrates and seems to be the main reason for the declining ethanol production toward the end of the process. Here, we investigate how the combination of yeast preadaptation and feeding, cell flocculation, and temperature reduction improves the cell viability in SSCF of steam pretreated wheat straw. More than 50% cell viability was lost during the first 24 h of high-gravity SSCF. No beneficial effects of adding selected nutrients were observed in shake flask SSCF. Ethanol concentrations greater than 50 g L -1 led to significant loss of viability and prevented further fermentation in SSCF. The benefits of feeding preadapted yeast cells were marginal at later stages of SSCF. Yeast flocculation did not improve the viability but simplified cell harvest and improved the feasibility of the cell feeding strategy in demo scale. Cultivation at 30 °C instead of 35 °C increased cell survival significantly on solid media containing ethanol and inhibitors. Similarly, in multifeed SSCF, cells maintained the viability and fermentation capacity when the temperature was reduced from 35 to 30 °C during the process, but hydrolysis yields were compromised. By combining the yeast feeding and temperature change, an ethanol concentration of 65 g L -1 , equivalent to 70% of the theoretical yield, was obtained in multifeed SSCF on pretreated wheat straw. In demo scale, the process with flocculating yeast and temperature profile resulted in 5% (w/w) ethanol, equivalent to 53% of the theoretical yield. Multifeed SSCF was further developed by means of a flocculating yeast and a temperature-reduction profile. Ethanol toxicity is intensified in the presence of lignocellulosic inhibitors at temperatures that are beneficial to hydrolysis in high-gravity SSCF. The counteracting effects of temperature on cell viability and hydrolysis call for more tolerant microorganisms, enzyme systems with lower temperature optimum, or full optimization of the multifeed strategy with temperature profile.

Effects of single and combined cell treatments based on low pH and high concentrations of ethanol on the growth and fermentation of Dekkera bruxellensis and Saccharomyces cerevisiae.

PubMed

Bassi, Ana Paula Guarnieri; da Silva, Jéssica Carolina Gomes; Reis, Vanda Renata; Ceccato-Antonini, Sandra Regina

2013-09-01

The alcoholic fermentation in Brazil displays some peculiarities because the yeast used is recycled in a non-aseptic process. After centrifugation, the cells are treated with acid to control the bacterial growth. However, it is difficult to manage the indigenous yeasts without affecting the main culture of Saccharomyces cerevisiae. This work evaluated how the cell treatment could be modified to combat contaminant yeasts based on the differential sensitivities to low pH and high concentrations of ethanol displayed by an industrial strain of S. cerevisiae and three strains of Dekkera bruxellensis, which are common contaminant yeasts in Brazilian fermentation processes. The tests were initially performed in rich medium with a low pH or a high concentration of ethanol to analyse the yeast growth profile. Then, the single and combined effects of low pH and ethanol concentration on the yeast cell viability were evaluated under non-proliferative conditions. The effects on the fermentation parameters were also verified. S. cerevisiae grew best when not subjected to the stresses, but this yeast and D. bruxellensis had similar growth kinetics when exposed to a low pH or increased ethanol concentrations. However, the combined treatments of low pH (2.0) and ethanol (11 or 13 %) resulted in a decrease of D. bruxellensis cell viability almost three times higher than of S. cerevisiae, which was only slightly affected by all cell treatments. The initial viability of the treated cells was restored within 8 h of growth in sugar cane juice, with the exception of the combined treatment for D. bruxellensis. The ethanol-based cell treatment, in despite of slowing the fermentation, could decrease and maintain D. bruxellensis population under control while S. cerevisiae was taking over the fermentation along six fermentative cycles. These results indicate that it may be possible to control the growth of D. bruxellensis without major effects on S. cerevisiae. The cells could be treated between the fermentation cycles by the parcelled addition of 13 % ethanol to the tanks in which the yeast cream is treated with sulphuric acid at pH 2.0.

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

PubMed Central

Gohel, Vipul; Duan, Gang

2012-01-01

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

Ethanol Fermentation of Various Pretreated and Hydrolyzed Substrates at Low Initial pH

NASA Astrophysics Data System (ADS)

Kádár, Zsófia; Maltha, San Feng; Szengyel, Zsolt; Réczey, Kati; de Laat, Wim

Lignocellulosic materials represent an abundant feedstock for bioethanol production. Because of their complex structure pretreatment is necessary to make it accessible for enzymatic attack. Steam pretreatment with or without acid catalysts seems to be one of the most promising techniques, which has already been applied for large variety of lignocellulosics in order to improve enzymatic digestibility. During this process a range of toxic compounds (lignin and sugar degradation products) are formed which inhibit ethanol fermentation. In this study, the toxicity of hemicellulose hydrolysates obtained in the steam pretreatment of spruce, willow, and corn stover were investigated in ethanol fermentation tests using a yeast strain, which has been previously reported to have a resistance to inhibitory compounds generated during steam pretreatment. To overcome bacterial contamination, fermentations were carried out at low initial pH. The fermentability of hemicellulose hydrolysates of pretreated lignocellulosic substrates at low pH gave promising results with the economically profitable final 5 vol% ethanol concentration corresponding to 85% of theoretical. Adaptation experiments have shown that inhibitor tolerance of yeast strain can be improved by subsequent transfer of the yeast to inhibitory medium.

Enhanced anti-oxidative activity and lignocellulosic ethanol production by biotin addition to medium in Pichia guilliermondii fermentation.

PubMed

Qi, Kai; Xia, Xiao-Xia; Zhong, Jian-Jiang

2015-01-01

Commercialization of lignocellulosic ethanol fermentation requires its high titer, but the reactive oxygen species (ROS) accumulation during the bioprocess damaged the cells and compromised this goal. To improve the cellular anti-oxidative activity during non-detoxified corncob residue hydrolysate fermentation, seed cells were prepared to possess a higher level of intracellular biotin pool (IBP), which facilitated the biosyntheses of catalase and porphyrin. As a result, the catalase activity increased by 1.3-folds compared to control while the ROS level reduced by 50%. Cell viability in high-IBP cells was 1.7-folds of control and the final ethanol titer increased from 31.2 to 41.8 g L(-1) in batch fermentation. The high-IBP cells were further used for repeated-batch fermentation in the non-detoxified lignocellulosic hydrolysate, and the highest titer and average productivity of ethanol reached 63.7 g L(-1) and 1.2 g L(-1)h(-1). The results were favorable to future industrial application of this lignocellulosic bioethanol process. Copyright © 2015 Elsevier Ltd. All rights reserved.

Optoelectronic sensor device for monitoring ethanol concentration in winemaking applications

NASA Astrophysics Data System (ADS)

Jiménez-Márquez, F.; Vázquez, J.; Úbeda, J.; Rodríguez-Rey, J.; Sánchez-Rojas, J. L.

2015-05-01

The supervision of key variables such as sugar, alcohol, released CO2 and microbiological evolution in fermenting grape must is of great importance in the winemaking industry. However, the fermentation kinetics is assessed by monitoring the evolution of the density as it varies during a fermentation, since density is an indicator of the total amount of sugars, ethanol and glycerol. Even so, supervising the fermentation process is an awkward and non-comprehensive task, especially in wine cellars where production rates are massive, and enologists usually measure the density of the extracted samples from each fermentation tank manually twice a day. This work aims at the design of a fast, low-cost, portable and reliable optoelectronic sensor for measuring ethanol concentration in fermenting grape must samples. Different sets of model solutions, which contain ethanol, fructose, glucose, glycerol dissolved in water and emulate the grape must composition at different stages of the fermentation, were prepared both for calibration and validation. The absorption characteristics of these model solutions were analyzed by a commercial spectrophotometer in the NIR region, in order to identify key wavelengths from which valuable information regarding the sample composition can be extracted. Finally, a customized optoelectronic prototype based on absorbance measurements at two wavelengths belonging to the NIR region was designed, fabricated and successfully tested. The system, whose optoelectronics is reduced after a thorough analysis to only two LED lamps and their corresponding paired photodiodes operating at 1.2 and 1.3 μm respectively, calculates the ethanol content by a multiple linear regression.

Continuous Ethanol Fermentation of Pretreated Lignocellulosic Biomasses, Waste Biomasses, Molasses and Syrup Using the Anaerobic, Thermophilic Bacterium Thermoanaerobacter italicus Pentocrobe 411

PubMed Central

Andersen, Rasmus Lund; Jensen, Karen Møller; Mikkelsen, Marie Just

2015-01-01

Lignocellosic ethanol production is now at a stage where commercial or semi-commercial plants are coming online and, provided cost effective production can be achieved, lignocellulosic ethanol will become an important part of the world bio economy. However, challenges are still to be overcome throughout the process and particularly for the fermentation of the complex sugar mixtures resulting from the hydrolysis of hemicellulose. Here we describe the continuous fermentation of glucose, xylose and arabinose from non-detoxified pretreated wheat straw, birch, corn cob, sugar cane bagasse, cardboard, mixed bio waste, oil palm empty fruit bunch and frond, sugar cane syrup and sugar cane molasses using the anaerobic, thermophilic bacterium Thermoanaerobacter Pentocrobe 411. All fermentations resulted in close to maximum theoretical ethanol yields of 0.47–0.49 g/g (based on glucose, xylose, and arabinose), volumetric ethanol productivities of 1.2–2.7 g/L/h and a total sugar conversion of 90–99% including glucose, xylose and arabinose. The results solidify the potential of Thermoanaerobacter strains as candidates for lignocellulose bioconversion. PMID:26295944

Ethanol production by Escherichia coli from Arundo donax biomass under SSF, SHF or CBP process configurations and in situ production of a multifunctional glucanase and xylanase.

PubMed

Loaces, Inés; Schein, Sima; Noya, Francisco

2017-01-01

Diluted acid or liquid hot water (LHW) pretreated Arundo donax biomass was converted into ethanol under separated hydrolysis and fermentation (SHF) or simultaneous saccharification and fermentation (SSF) using Escherichia coli as the fermentative organism. Up to 0.26gL -1 h -1 and 25.0gL -1 of ethanol were obtained with diluted acid pretreated biomass under SSF compared to 0.17gL -1 h -1 and 24gL -1 under SHF. LHW pretreated biomass elicited 25% lower yields on average. Saccharification was carried out with Cellic CTec2 cocktail. Alternatively, under a consolidated bioprocess (CBP) where the ethanologenic bacteria was complemented with a novel multifunctional glucanase and xylanase, ethanol concentration was 7.6gL -1 and 7.2gL -1 after 96h for dilute acid or LHW pretreated biomass, respectively, without any prior saccharification step. According to these results, a bacterial fermentative host combined with in situ enzyme expression can improve ethanol production from A. donax biomass. Copyright © 2016 Elsevier Ltd. All rights reserved.

Sugar and ethanol production from woody biomass via supercritical water hydrolysis in a continuous pilot-scale system using acid catalyst.

PubMed

Jeong, Hanseob; Park, Yong-Cheol; Seong, Yeong-Je; Lee, Soo Min

2017-12-01

The aim of this study were to efficiently produce fermentable sugars by continuous type supercritical water hydrolysis (SCWH) of Quercus mongolica at the pilot scale with varying acid catalyst loading and to use the obtained sugars for ethanol production. The SCWH of biomass was achieved in under one second (380°C, 230bar) using 0.01-0.1% H 2 SO 4 . With 0.05% H 2 SO 4 , 49.8% of sugars, including glucose (16.5% based on biomass) and xylose monomers (10.8%), were liberated from biomass. The hydrolysates were fermented with S. cerevisiae DXSP and D452-2 to estimate ethanol production. To prepare the fermentation medium, the hydrolysates were detoxified using activated charcoal and then concentrated. The ethanol yield of fermentation with S. cerevisiae DXSP was 14.1% (based on biomass). The proposed system has potential for improvement in yield through process optimization. After further development, it is expected to be a competitive alternative to traditional systems for ethanol production from woody biomass. Copyright © 2017 Elsevier Ltd. All rights reserved.

Enhanced photo-fermentative H2 production using Rhodobacter sphaeroides by ethanol addition and analysis of soluble microbial products

PubMed Central

2014-01-01

Background Biological fermentation routes can provide an environmentally friendly way of producing H2 since they use renewable biomass as feedstock and proceed under ambient temperature and pressure. In particular, photo-fermentation has superior properties in terms of achieving high H2 yield through complete degradation of substrates. However, long-term H2 production data with stable performance is limited, and this data is essential for practical applications. In the present work, continuous photo-fermentative H2 production from lactate was attempted using the purple non-sulfur bacterium, Rhodobacter sphaeroides KD131. As a gradual drop in H2 production was observed, we attempted to add ethanol (0.2% v/v) to the medium. Results As continuous operation went on, H2 production was not sustained and showed a negligible H2 yield (< 0.5 mol H2/mol lactateadded) within two weeks. Electron balance analysis showed that the reason for the gradual drop in H2 production was ascribed to the increase in production of soluble microbial products (SMPs). To see the possible effect of ethanol addition, a batch test was first conducted. The presence of ethanol significantly increased the H2 yield from 1.15 to 2.20 mol H2/mol lactateadded, by suppressing the production of SMPs. The analysis of SMPs by size exclusion chromatography showed that, in the later period of fermentation, more than half of the low molecular weight SMPs (< 1 kDa) were consumed and used for H2 production when ethanol had been added, while the concentration of SMPs continuously increased in the absence of ethanol. It was found that the addition of ethanol facilitated the utilization of reducing power, resulting in an increase in the cellular levels of NAD+ and NADP+. In continuous operation, ethanol addition was effective, such that stable H2 production was attained with an H2 yield of 2.5 mol H2/mol lactateadded. Less than 15% of substrate electrons were used for SMP production, whereas 35% were used in the control. Conclusions We have found that SMPs are the key factor in photo-fermentative H2 production, and their production can be suppressed by ethanol addition. However, since external addition of ethanol to the medium represents an extra economic burden, ethanol should be prepared in a cost-effective way. PMID:24883103

Expression of bacteriophage endolysins in Saccharomyces cerevisiae

USDA-ARS?s Scientific Manuscript database

One of the challenges facing the fuel ethanol industry is the management of bacterial contamination during fermentation. Species of Lactobacillus are the predominant contaminants that reduce ethanol yields and cause “stuck” fermentations, decreasing the profitability of biofuel production with expe...

Direct ethanol production from starch, wheat bran and rice straw by the white rot fungus Trametes hirsuta.

PubMed

Okamoto, Kenji; Nitta, Yasuyuki; Maekawa, Nitaro; Yanase, Hideshi

2011-03-07

The white rot fungus Trametes hirsuta produced ethanol from a variety of hexoses: glucose, mannose, cellobiose and maltose, with yields of 0.49, 0.48, 0.47 and 0.47 g/g of ethanol per sugar utilized, respectively. In addition, this fungus showed relatively favorable xylose consumption and ethanol production with a yield of 0.44 g/g. T. hirsuta was capable of directly fermenting starch, wheat bran and rice straw to ethanol without acid or enzymatic hydrolysis. Maximum ethanol concentrations of 9.1, 4.3 and 3.0 g/l, corresponding to 89.2%, 78.8% and 57.4% of the theoretical yield, were obtained when the fungus was grown in a medium containing 20 g/l starch, wheat bran or rice straw, respectively. The fermentation of rice straw pretreated with ball milling led to a small improvement in the ethanol yield: 3.4 g ethanol/20 g ball-milled rice straw. As T. hirsuta is an efficient microorganism capable of hydrolyzing biomass to fermentable sugars and directly converting them to ethanol, it may represent a suitable microorganism in consolidated bioprocessing applications. Copyright © 2010 Elsevier Inc. All rights reserved.

Experimental study of bioethanol production using mixed cassava and durian seed

NASA Astrophysics Data System (ADS)

Seer, Q. H.; Nandong, J.; Shanon, T.

2017-06-01

The production of biofuels using conventional fermentation feedstocks, such as sugar-and starch-based agricultural crops will in the long-term lead to a serious competition with human-animal food consumption. To avoid this competition, it is important to explore various alternative feedstocks especially those from inedible waste materials. Potentially, fruit wastes such as damaged fruits, peels and seeds represent alternative cheap feedstocks for biofuel production. In this work, an experimental study was conducted on ethanol production using mixed cassava and durian seeds through fermentation by Saccharomyces cerevisiae yeast. The effects of pH, temperature and ratio of hydrolyzed cassava to durian seeds on the ethanol yield, substrate consumption and product formation rates were analyzed in the study. In flask-scale fermentation using the mixed cassava-durian seeds, it was found that the highest ethanol yield of 45.9 and a final ethanol concentration of 24.92 g/L were achieved at pH 5.0, temperature 35°C and 50:50 volume ratio of hydrolyzed cassava to durian seeds for a batch period of 48 hours. Additionally, the ethanol, glucose and biomass concentration profiles in a lab-scale bioreactor were examined for the fermentation using the proposed materials under the flask-scale optimum conditions. The ethanol yield of 35.7 and a final ethanol concentration of 14.61 g/L were obtained over a period of 46 hours where the glucose was almost fully consumed. It is worth noting that both pH and temperature have significant impacts on the fermentation process using the mixed cassava-durian seeds.

Reducing bacterial contamination in fuel ethanol fermentations by ozone treatment of uncooked corn mash.

PubMed

Rasmussen, Mary L; Koziel, Jacek A; Jane, Jay-lin; Pometto, Anthony L

2015-06-03

Ozonation of uncooked corn mash from the POET BPX process was investigated as a potential disinfection method for reducing bacterial contamination prior to ethanol fermentation. Corn mash (200 g) was prepared from POET ground corn and POET corn slurry and was ozonated in 250 mL polypropylene bottles. Lactic and acetic acid levels were monitored daily during the fermentation of ozonated, aerated, and nontreated corn mash samples to evaluate bacterial activity. Glycerol and ethanol contents of fermentation samples were checked daily to assess yeast activity. No yeast supplementation, no addition of other antimicrobial agents (such as antibiotics), and spiking with a common lactic acid bacterium found in corn ethanol plants, Lactobacillus plantarum, amplified the treatment effects. The laboratory-scale ozone dosages ranged from 26-188 mg/L, with very low estimated costs of $0.0008-0.006/gal ($0.21-1.6/m(3)) of ethanol. Ozonation was found to decrease the initial pH of ground corn mash samples, which could reduce the sulfuric acid required to adjust the pH prior to ethanol fermentation. Lactic and acetic acid levels tended to be lower for samples subjected to increasing ozone dosages, indicating less bacterial activity. The lower ozone dosages in the range applied achieved higher ethanol yields. Preliminary experiments on ozonating POET corn slurry at low ozone dosages were not as effective as using POET ground corn, possibly because corn slurry samples contained recycled antimicrobials from the backset. The data suggest additional dissolved and suspended organic materials from the backset consumed the ozone or shielded the bacteria.

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.

Stimulation of electro-fermentation in single-chamber microbial electrolysis cells driven by genetically engineered anode biofilms

NASA Astrophysics Data System (ADS)

Awate, Bhushan; Steidl, Rebecca J.; Hamlischer, Thilo; Reguera, Gemma

2017-07-01

Unwanted metabolites produced during fermentations reduce titers and productivity and increase the cost of downstream purification of the targeted product. As a result, the economic feasibility of otherwise attractive fermentations is low. Using ethanol fermentation by the consolidated bioprocessing cellulolytic bacterium Cellulomonas uda, we demonstrate the effectiveness of anodic electro-fermentations at maximizing titers and productivity in a single-chamber microbial electrolysis cell (SCMEC) without the need for metabolic engineering of the fermentative microbe. The performance of the SCMEC platform relied on the genetic improvements of anode biofilms of the exoelectrogen Geobacter sulfurreducens that prevented the oxidation of cathodic hydrogen and improved lactate oxidation. Furthermore, a hybrid bioanode was designed that maximized the removal of organic acids in the fermentation broth. The targeted approach increased cellobiose consumption rates and ethanol titers, yields, and productivity three-fold or more, prevented pH imbalances and reduced batch-to-batch variability. In addition, the sugar substrate was fully consumed and ethanol was enriched in the broth during the electro-fermentation, simplifying its downstream purification. Such improvements and the possibility of scaling up SCMEC configurations highlight the potential of anodic electro-fermentations to stimulate fermentative bacteria beyond their natural capacity and to levels required for industrial implementation.

Dual effect of soluble materials in pretreated lignocellulose on simultaneous saccharification and co-fermentation process for the bioethanol production.

PubMed

Qin, Lei; Li, Xia; Liu, Li; Zhu, Jia-Qing; Guan, Qi-Man; Zhang, Man-Tong; Li, Wen-Chao; Li, Bing-Zhi; Yuan, Ying-Jin

2017-01-01

In this study, wash liquors isolated from ethylenediamine and dry dilute acid pretreated corn stover were used to evaluate the effect of soluble materials in pretreated biomass on simultaneous saccharification and co-fermentation (SSCF) for ethanol production, respectively. Both of the wash liquors had different impacts on enzymatic hydrolysis and fermentation. Enzymatic conversions of glucan and xylan monotonically decreased as wash liquor concentration increased. Whereas, with low wash liquor concentrations, xylose consumption rate, cell viability and ethanol yield were maximally stimulated in fermentation without nutrient supplementary. Soluble lignins were found as the key composition which promoted sugars utilization and cell viability without nutrient supplementary. The dual effects of soluble materials on enzymatic hydrolysis and fermentation resulted in the reduction of ethanol yield as soluble materials increased in SSCF. Copyright © 2016 Elsevier Ltd. All rights reserved.

Fermentation of Acid-pretreated Corn Stover to Ethanol Without Detoxification Using Pichia stipitis

NASA Astrophysics Data System (ADS)

Agbogbo, Frank K.; Haagensen, Frank D.; Milam, David; Wenger, Kevin S.

In this work, the effect of adaptation on P. stipitis fermentation using acidpretreated corn stover hydrolyzates without detoxification was examined. Two different types of adaptation were employed, liquid hydrolyzate and solid state agar adaptation. Fermentation of 12.5% total solids undetoxified acid-pretreated corn stover was performed in shake flasks at different rotation speeds. At low rotation speed (100 rpm), both liquid hydrolyzate and solid agar adaptation highly improved the sugar consumption rate as well as ethanol production rate compared to the wild-type strains. The fermentation rate was higher for solid agar-adapted strains compared to liquid hydrolyzate-adapted strains. At a higher rotation speed (150 rpm), there was a faster sugar consumption and ethanol production for both the liquid-adapted and the wild-type strains. However, improvements in the fermentation rate between the liquid-adapted and wild strains were less pronounced at the high rotation speed.

A grey box model of glucose fermentation and syntrophic oxidation in microbial fuel cells.

PubMed

de Los Ángeles Fernandez, Maria; de Los Ángeles Sanromán, Maria; Marks, Stanislaw; Makinia, Jacek; Gonzalez Del Campo, Araceli; Rodrigo, Manuel; Fernandez, Francisco Jesus

2016-01-01

In this work, the fermentative and oxidative processes taking place in a microbial fuel cell (MFC) fed with glucose were studied and modeled. The model accounting for the bioelectrochemical processes was based on ordinary, Monod-type differential equations. The model parameters were estimated using experimental results obtained from three H-type MFCs operated at open or closed circuits and fed with glucose or ethanol. The experimental results demonstrate that similar fermentation processes were carried out under open and closed circuit operation, with the most important fermentation products being ethanol (with a yield of 1.81molmol(-1) glucose) and lactic acid (with a yield of 1.36molmol(-1) glucose). A peak in the electricity generation was obtained when glucose and fermentation products coexisted in the liquid bulk. However, almost 90% of the electricity produced came from the oxidation of ethanol. Copyright © 2015 Elsevier Ltd. All rights reserved.

Favouring butyrate production for a new generation biofuel by acidogenic glucose fermentation using cells immobilised on γ-alumina.

PubMed

Syngiridis, Kostas; Bekatorou, Argyro; Kandylis, Panagiotis; Larroche, Christian; Kanellaki, Maria; Koutinas, Athanasios A

2014-06-01

The effect of γ-alumina as a fermentation advancing tool and as carrier for culture immobilisation, regarding VFAs and ethanol production during acidogenic fermentation of glucose, was examined at various process conditions (sugar concentration, pH) and operation modes (continuous with and without effluent recirculation and batch). The results showed that at high initial pH (8.9) the continuous acidogenic fermentation of glucose led to high yields of VFAs and favoured the accumulation of butyric acid. The batch process on the other hand at pH 6.5, favoured the ethanol-type fermentation. The results indicate that in the frame of technology development for new generation biofuels, using γ-alumina as a process advancing tool at optimum process conditions (pH, initial glucose concentration and mode of operation), the produced VFAs profile and ethanol concentration may be manipulated. Copyright © 2014. Published by Elsevier Ltd.

Cell recycle batch fermentation of high-solid lignocellulose using a recombinant cellulase-displaying yeast strain for high yield ethanol production in consolidated bioprocessing.

PubMed

Matano, Yuki; Hasunuma, Tomohisa; Kondo, Akihiko

2013-05-01

The aim of this study is to develop a scheme of cell recycle batch fermentation (CRBF) of high-solid lignocellulosic materials. Two-phase separation consisting of rough removal of lignocellulosic residues by low-speed centrifugation and solid-liquid separation enabled effective collection of Saccharomyces cerevisiae cells with decreased lignin and ash. Five consecutive batch fermentation of 200 g/L rice straw hydrothermally pretreated led to an average ethanol titer of 34.5 g/L. Moreover, the display of cellulases on the recombinant yeast cell surface increased ethanol titer to 42.2 g/L. After, five-cycle fermentation, only 3.3 g/L sugar was retained in the fermentation medium, because cellulase displayed on the cell surface hydrolyzed cellulose that was not hydrolyzed by commercial cellulases or free secreted cellulases. Fermentation ability of the recombinant strain was successfully kept during a five-cycle repeated batch fermentation with 86.3% of theoretical yield based on starting biomass. Copyright © 2012 Elsevier Ltd. All rights reserved.

Enhancement of Ethanol Fermentation in Saccharomyces cerevisiae Sake Yeast by Disrupting Mitophagy Function

PubMed Central

Shiroma, Shodai; Jayakody, Lahiru Niroshan; Horie, Kenta; Okamoto, Koji

2014-01-01

Saccharomyces cerevisiae sake yeast strain Kyokai no. 7 has one of the highest fermentation rates among brewery yeasts used worldwide; therefore, it is assumed that it is not possible to enhance its fermentation rate. However, in this study, we found that fermentation by sake yeast can be enhanced by inhibiting mitophagy. We observed mitophagy in wild-type sake yeast during the brewing of Ginjo sake, but not when the mitophagy gene (ATG32) was disrupted. During sake brewing, the maximum rate of CO2 production and final ethanol concentration generated by the atg32Δ laboratory yeast mutant were 7.50% and 2.12% higher than those of the parent strain, respectively. This mutant exhibited an improved fermentation profile when cultured under limiting nutrient concentrations such as those used during Ginjo sake brewing as well as in minimal synthetic medium. The mutant produced ethanol at a concentration that was 2.76% higher than the parent strain, which has significant implications for industrial bioethanol production. The ethanol yield of the atg32Δ mutant was increased, and its biomass yield was decreased relative to the parent sake yeast strain, indicating that the atg32Δ mutant has acquired a high fermentation capability at the cost of decreasing biomass. Because natural biomass resources often lack sufficient nutrient levels for optimal fermentation, mitophagy may serve as an important target for improving the fermentative capacity of brewery yeasts. PMID:24271183

Second-generation ethanol from non-detoxified sugarcane hydrolysate by a rotting wood isolated yeast strain.

PubMed

Bazoti, Suzana F; Golunski, Simone; Pereira Siqueira, Diego; Scapini, Thamarys; Barrilli, Évelyn T; Alex Mayer, Diego; Barros, Katharina O; Rosa, Carlos A; Stambuk, Boris U; Alves, Sérgio L; Valério, Alexsandra; de Oliveira, Débora; Treichel, Helen

2017-11-01

This work aims to evaluate the production of second-generation ethanol from sugarcane bagasse hydrolysate without acetic acid (inhibitor) detoxification. Three isolated yeast strains from lignocellulosic materials were evaluated, and one strain (UFFS-CE-3.1.2), identified using large subunit rDNA sequences as Wickerhamomyces sp., showed satisfactory results in terms of ethanol production without acetic acid removal. A Plackett-Burman design was used to evaluate the influence of hydrolysate composition and nutrients supplementation in the fermentation medium for the second-generation ethanol production. Two fermentation kinetics were performed, with controlled pH at 5.5, or keeping the initial pH at 4.88. The fermentation conducted without pH adjustment and supplementation of nutrients reported the best result in terms of second-generation ethanol production. Wickerhamomyces sp., isolated as UFFS-CE-3.1.2, was considered promising in the production of second-generation ethanol by using crude (non-detoxified) sugarcane hydrolysate. Copyright © 2017 Elsevier Ltd. All rights reserved.

Simultaneous hydrolysis and fermentation of unprocessed food waste into ethanol using thermophilic anaerobic bacteria.

PubMed

Dhiman, Saurabh Sudha; David, Aditi; Shrestha, Namita; Johnson, Glenn R; Benjamin, Kenneth M; Gadhamshetty, Venkataramana; Sani, Rajesh K

2017-11-01

The one-pot CRUDE (Conversion of Raw and Untreated Disposal into Ethanol) process was developed for simultaneous hydrolysis and fermentation of unprocessed food waste into ethanol using thermophilic (growing at 65°C) anaerobic bacteria. Unlike existing waste to energy technologies, the CRUDE process obviates the need for any pre-treatment or enzyme addition. A High-Temperature-High-Pressure (HTHP) distillation technique was also applied that facilitated efficient use of fermentation medium, inoculum recycling, and in-situ ethanol collection. For material balancing of the process, each characterized component was represented in terms of C-mol. Recovery of 94% carbon at the end confirmed the operational efficiency of CRUDE process. The overall energy retaining efficiency calculated from sugars to ethanol was 1262.7kJdryweightkg -1 of volatile solids using HTHP. These results suggest that the CRUDE process can be a starting point for the development of a commercial ethanol production process. Copyright © 2017 Elsevier Ltd. All rights reserved.

Separation, hydrolysis and fermentation of pyrolytic sugars to produce ethanol and lipids.

PubMed

Lian, Jieni; Chen, Shulin; Zhou, Shuai; Wang, Zhouhong; O'Fallon, James; Li, Chun-Zhu; Garcia-Perez, Manuel

2010-12-01

This paper describes a new scheme to convert anhydrosugars found in pyrolysis oils into ethanol and lipids. Pyrolytic sugars were separated from phenols by solvent extraction and were hydrolyzed into glucose using sulfuric acid as a catalyst. Toxicological studies showed that phenols and acids were the main species inhibiting growth of the yeast Saccharomyces cerevisiae. The sulfuric acids, and carboxylic acids from the bio-oils, were neutralized with Ba(OH)(2). The phase rich in sugar was further detoxified with activated carbon. The resulting aqueous phase rich in glucose was fermented with three different yeasts: S. cerevisiae to produce ethanol, and Cryptococcus curvatus and Rhodotorula glutinis to produce lipids. Yields as high as 0.473 g ethanol/g glucose and 0.167 g lipids/g sugar (0.266 g ethanol equivalent/g sugar), were obtained. These results confirm that pyrolytic sugar fermentation to produce ethanol is more efficient than for lipid production. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

Sequential high gravity ethanol fermentation and anaerobic digestion of steam explosion and organosolv pretreated corn stover.

PubMed

Katsimpouras, Constantinos; Zacharopoulou, Maria; Matsakas, Leonidas; Rova, Ulrika; Christakopoulos, Paul; Topakas, Evangelos

2017-11-01

The present work investigates the suitability of pretreated corn stover (CS) to serve as feedstock for high gravity (HG) ethanol production at solids-content of 24wt%. Steam explosion, with and without the addition of H 2 SO 4 , and organosolv pretreated CS samples underwent a liquefaction/saccharification step followed by simultaneous saccharification and fermentation (SSF). Maximum ethanol concentration of ca. 76g/L (78.3% ethanol yield) was obtained from steam exploded CS (SECS) with 0.2% H 2 SO 4 . Organosolv pretreated CS (OCS) also resulted in high ethanol concentration of ca. 65g/L (62.3% ethanol yield). Moreover, methane production through anaerobic digestion (AD) was conducted from fermentation residues and resulted in maximum methane yields of ca. 120 and 69mL/g volatile solids (VS) for SECS and OCS samples, respectively. The results indicated that the implementation of a liquefaction/saccharification step before SSF employing a liquefaction reactor seemed to handle HG conditions adequately. Copyright © 2017 Elsevier Ltd. All rights reserved.

Inhibition of Lactobacillus biofilm growth by Bacillus extracts

USDA-ARS?s Scientific Manuscript database

Industrial ethanol fermentations are not pure cultures, and are expected to contain contaminant bacteria and fungi. These additional organisms deplete the feedstock and lower overall ethanol yield. Severe contamination can lead to “stuck” fermentations, requiring costly shutdowns for cleaning. As La...

Antimicrobials Used in the Fermentation of Fuel Ethanol – Clarification of Jurisdiction

EPA Pesticide Factsheets

EPA has determined that antimicrobials applied to processed food or feed during fermentation of organic material to produce fuel ethanol are outside the scope of EPA’s regulatory authority under FIFRA. The Food and Drug Administration has jurisdiction.

Effective ethanol production from whey powder through immobilized E. coli expressing Vitreoscilla hemoglobin.

PubMed

Sar, Taner; Stark, Benjamin C; Yesilcimen Akbas, Meltem

2017-03-04

Ethanol production from whey powder was investigated by using free as well as alginate immobilized E. coli and E. coli expressing Vitreoscilla hemoglobin (VHb) in both shake flask and fermenter cultures. Media with varying levels of whey (lactose contents of 3%, 5%, 8% or 15%) and yeast extract (0.3% or 0.5%) were evaluated with fermentation times of 48-96 h. Immobilization and VHb expression resulted in higher ethanol production with all media; the increases ranged from 2% to 89% for immobilization and from 2% to 182% for VHb expression. It was determined that growth medium containing 8% lactose with 0.5% yeast extract yielded the highest ethanol production for free or immobilized strains, with or without VHb expression, in both shake flask and fermenter cultures. Immobilization with alginate was found to be a promising process for ethanol production by VHb-expressing ethanologenic E. coli.

Effective ethanol production from whey powder through immobilized E. coli expressing Vitreoscilla hemoglobin

PubMed Central

Sar, Taner; Stark, Benjamin C.; Yesilcimen Akbas, Meltem

2017-01-01

ABSTRACT Ethanol production from whey powder was investigated by using free as well as alginate immobilized E. coli and E. coli expressing Vitreoscilla hemoglobin (VHb) in both shake flask and fermenter cultures. Media with varying levels of whey (lactose contents of 3%, 5%, 8% or 15%) and yeast extract (0.3% or 0.5%) were evaluated with fermentation times of 48–96 h. Immobilization and VHb expression resulted in higher ethanol production with all media; the increases ranged from 2% to 89% for immobilization and from 2% to 182% for VHb expression. It was determined that growth medium containing 8% lactose with 0.5% yeast extract yielded the highest ethanol production for free or immobilized strains, with or without VHb expression, in both shake flask and fermenter cultures. Immobilization with alginate was found to be a promising process for ethanol production by VHb-expressing ethanologenic E. coli. PMID:27579556

Industrial antifoam agents impair ethanol fermentation and induce stress responses in yeast cells.

PubMed

Nielsen, Jens Christian; Senne de Oliveira Lino, Felipe; Rasmussen, Thomas Gundelund; Thykær, Jette; Workman, Christopher T; Basso, Thiago Olitta

2017-11-01

The Brazilian sugarcane industry constitutes one of the biggest and most efficient ethanol production processes in the world. Brazilian ethanol production utilizes a unique process, which includes cell recycling, acid wash, and non-aseptic conditions. Process characteristics, such as extensive CO 2 generation, poor quality of raw materials, and frequent contaminations, all lead to excessive foam formation during fermentations, which is treated with antifoam agents (AFA). In this study, we have investigated the impact of industrial AFA treatments on the physiology and transcriptome of the industrial ethanol strain Saccharomyces cerevisiae CAT-1. The investigated AFA included industrially used AFA acquired from Brazilian ethanol plants and commercially available AFA commonly used in the fermentation literature. In batch fermentations, it was shown that industrial AFA compromised growth rates and glucose uptake rates, while commercial AFA had no effect in concentrations relevant for defoaming purposes. Industrial AFA were further tested in laboratory scale simulations of the Brazilian ethanol production process and proved to decrease cell viability compared to the control, and the effects were intensified with increasing AFA concentrations and exposure time. Transcriptome analysis showed that AFA treatments induced additional stress responses in yeast cells compared to the control, shown by an up-regulation of stress-specific genes and a down-regulation of lipid biosynthesis, especially ergosterol. By documenting the detrimental effects associated with chemical AFA, we highlight the importance of developing innocuous systems for foam control in industrial fermentation processes.

Enzymatic hydrolysis and fermentation of pretreated cashew apple bagasse with alkali and diluted sulfuric Acid for bioethanol production.

PubMed

Rocha, Maria Valderez Ponte; Rodrigues, Tigressa Helena Soares; de Macedo, Gorete Ribeiro; Gonçalves, Luciana R B

2009-05-01

The aim of this work was to optimize the enzymatic hydrolysis of the cellulose fraction of cashew apple bagasse (CAB) after diluted acid (CAB-H) and alkali pretreatment (CAB-OH), and to evaluate its fermentation to ethanol using Saccharomyces cerevisiae. Glucose conversion of 82 +/- 2 mg/g CAB-H and 730 +/- 20 mg/g CAB-OH was obtained when 2% (w/v) of solid and 30 FPU/g bagasse was used during hydrolysis at 45 degrees C, 2-fold higher than when using 15 FPU/g bagasse, 44 +/- 2 mg/g CAB-H, and 450 +/- 50 mg/g CAB-OH, respectively. Ethanol concentration and productivity, achieved after 6 h of fermentation, were 20.0 +/- 0.2 g L(-1) and 3.33 g L(-1) h(-1), respectively, when using CAB-OH hydrolyzate (initial glucose concentration of 52.4 g L(-1)). For CAB-H hydrolyzate (initial glucose concentration of 17.4 g L(-1)), ethanol concentration and productivity were 8.2 +/- 0.1 g L(-1) and 2.7 g L(-1) h(-1) in 3 h, respectively. Hydrolyzates fermentation resulted in an ethanol yield of 0.38 and 0.47 g/g glucose with pretreated CAB-OH and CAB-H, respectively. Ethanol concentration and productivity, obtained using CAB-OH hydrolyzate, were close to the values obtained in the conventional ethanol fermentation of cashew apple juice or sugar cane juice.

Enhanced production of bioethanol from waste of beer fermentation broth at high temperature through consecutive batch strategy by simultaneous saccharification and fermentation.

PubMed

Khattak, Waleed Ahmad; Khan, Taous; Ha, Jung Hwan; Ul-Islam, Mazhar; Kang, Min-Kyung; Park, Joong Kon

2013-10-10

Malt hydrolyzing enzymes and yeast glycolytic and fermentation enzymes in the waste from beer fermentation broth (WBFB) were identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). A new 'one-pot consecutive batch strategy' was developed for efficient bio-ethanol production by simultaneous saccharification and fermentation (SSF) using WBFB without additional enzymes, microbial cells, or carbohydrates. Bio-ethanol production was conducted in batches using WBFB supernatant in the first phase at 25-67°C and 50rpm, followed by the addition of 3% WBFB solid residue to the existing culture broth in the second phase at 67°C. The ethanol production increased from 50 to 102.5g/L when bare supernatant was used in the first phase, and then to 219g ethanol/L in the second phase. The amount of ethanol obtained using this strategy was almost equal to that obtained using the original WBFB containing 25% solid residue at 33°C, and more than double that obtained when bare supernatant was used. Microscopic and gel electrophoresis studies revealed yeast cell wall degradation and secretion of cellular material into the surrounding medium. Scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) supported the existence of enzymes in WBFB involved in bioethanol production at elevated temperatures. The results of this study will provide insight for the development of new strategies for biofuel production. Copyright © 2013 Elsevier Inc. All rights reserved.

Characterization of a recombinant flocculent Saccharomyces cerevisiae strain that co-ferments glucose and xylose: II. influence of pH and acetic acid on ethanol production.

PubMed

Matsushika, Akinori; Sawayama, Shigeki

2012-12-01

The inhibitory effects of pH and acetic acid on the co-fermentation of glucose and xylose in complex medium by recombinant flocculent Saccharomyces cerevisiae MA-R4 were evaluated. In the absence of acetic acid, the fermentation performance of strain MA-R4 was similar between pH 4.0-6.0, but was negatively affected at pH 2.5. The addition of acetic acid to batch cultures resulted in negligible inhibition of several fermentation parameters at pH 6.0, whereas the interactive inhibition of pH and acetic acid on the maximum cell and ethanol concentrations, and rates of sugar consumption and ethanol production were observed at pH levels below 5.4. The inhibitory effect of acetic acid was particularly marked for the consumption rate of xylose, as compared with that of glucose. With increasing initial acetic acid concentration, the ethanol yield slightly increased at pH 5.4 and 6.0, but decreased at pH values lower than 4.7. Notably, ethanol production was nearly completely inhibited under low pH (4.0) and high acetic acid (150-200 mM) conditions. Together, these results indicate that the inhibitory effects of acetic acid and pH on ethanol fermentation by MA-R4 are highly synergistic, although the inhibition can be reduced by increasing the medium pH.

A modified indirect mathematical model for evaluation of ethanol production efficiency in industrial-scale continuous fermentation processes.

PubMed

Canseco Grellet, M A; Castagnaro, A; Dantur, K I; De Boeck, G; Ahmed, P M; Cárdenas, G J; Welin, B; Ruiz, R M

2016-10-01

To calculate fermentation efficiency in a continuous ethanol production process, we aimed to develop a robust mathematical method based on the analysis of metabolic by-product formation. This method is in contrast to the traditional way of calculating ethanol fermentation efficiency, where the ratio between the ethanol produced and the sugar consumed is expressed as a percentage of the theoretical conversion yield. Comparison between the two methods, at industrial scale and in sensitivity studies, showed that the indirect method was more robust and gave slightly higher fermentation efficiency values, although fermentation efficiency of the industrial process was found to be low (~75%). The traditional calculation method is simpler than the indirect method as it only requires a few chemical determinations in samples collected. However, a minor error in any measured parameter will have an important impact on the calculated efficiency. In contrast, the indirect method of calculation requires a greater number of determinations but is much more robust since an error in any parameter will only have a minor effect on the fermentation efficiency value. The application of the indirect calculation methodology in order to evaluate the real situation of the process and to reach an optimum fermentation yield for an industrial-scale ethanol production is recommended. Once a high fermentation yield has been reached the traditional method should be used to maintain the control of the process. Upon detection of lower yields in an optimized process the indirect method should be employed as it permits a more accurate diagnosis of causes of yield losses in order to correct the problem rapidly. The low fermentation efficiency obtained in this study shows an urgent need for industrial process optimization where the indirect calculation methodology will be an important tool to determine process losses. © 2016 The Society for Applied Microbiology.

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

PubMed Central

2011-01-01

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

Screening for new brewing yeasts in the non-Saccharomyces sector with Torulaspora delbrueckii as model.

PubMed

Michel, Maximilian; Kopecká, Jana; Meier-Dörnberg, Tim; Zarnkow, Martin; Jacob, Fritz; Hutzler, Mathias

2016-04-01

This study describes a screening system for future brewing yeasts focusing on non-Saccharomyces yeasts. The aim was to find new yeast strains that can ferment beer wort into a respectable beer. Ten Torulaspora delbrueckii strains were put through the screening system, which included sugar utilization tests, hop resistance tests, ethanol resistance tests, polymerase chain reaction fingerprinting, propagation tests, amino acid catabolism and anabolism, phenolic off-flavour tests and trial fermentations. Trial fermentations were analysed for extract reduction, pH drop, yeast concentration in bulk fluid and fermentation by-products. All investigated strains were able to partly ferment wort sugars and showed high tolerance to hop compounds and ethanol. One of the investigated yeast strains fermented all the wort sugars and produced a respectable fruity flavour and a beer of average ethanol content with a high volatile flavour compound concentration. Two other strains could possibly be used for pre-fermentation as a bio-flavouring agent for beers that have been post-fermented by Saccharomyces strains as a consequence of their low sugar utilization but good flavour-forming properties. Copyright © 2015 John Wiley & Sons, Ltd.

Very high gravity (VHG) ethanolic brewing and fermentation: a research update.

PubMed

Puligundla, Pradeep; Smogrovicova, Daniela; Obulam, Vijaya Sarathi Reddy; Ko, Sanghoon

2011-09-01

There have been numerous developments in ethanol fermentation technology since the beginning of the new millennium as ethanol has become an immediate viable alternative to fast-depleting crude reserves as well as increasing concerns over environmental pollution. Nowadays, although most research efforts are focused on the conversion of cheap cellulosic substrates to ethanol, methods that are cost-competitive with gasoline production are still lacking. At the same time, the ethanol industry has engaged in implementing potential energy-saving, productivity and efficiency-maximizing technologies in existing production methods to become more viable. Very high gravity (VHG) fermentation is an emerging, versatile one among such technologies offering great savings in process water and energy requirements through fermentation of higher concentrations of sugar substrate and, therefore, increased final ethanol concentration in the medium. The technology also allows increased fermentation efficiency, without major alterations to existing facilities, by efficient utilization of fermentor space and elimination of known losses. This comprehensive research update on VHG technology is presented in two main sections, namely VHG brewing, wherein the effects of nutrients supplementation, yeast pitching rate, flavour compound synthesis and foam stability under increased wort gravities are discussed; and VHG bioethanol fermentation studies. In the latter section, aspects related to the role of osmoprotectants and nutrients in yeast stress reduction, substrates utilized/tested so far, including saccharide (glucose, sucrose, molasses, etc.) and starchy materials (wheat, corn, barley, oats, etc.), and mash viscosity issues in VHG bioethanol production are detailed. Thereafter, topics common to both areas such as process optimization studies, mutants and gene level studies, immobilized yeast applications, temperature effect, reserve carbohydrates profile in yeast, and economic aspects are discussed and future prospects are summarized.

Fermentation of lactose to ethanol in cheese whey permeate and concentrated permeate by engineered Escherichia coli.

PubMed

Pasotti, Lorenzo; Zucca, Susanna; Casanova, Michela; Micoli, Giuseppina; Cusella De Angelis, Maria Gabriella; Magni, Paolo

2017-06-02

Whey permeate is a lactose-rich effluent remaining after protein extraction from milk-resulting cheese whey, an abundant dairy waste. The lactose to ethanol fermentation can complete whey valorization chain by decreasing dairy waste polluting potential, due to its nutritional load, and producing a biofuel from renewable source at the same time. Wild type and engineered microorganisms have been proposed as fermentation biocatalysts. However, they present different drawbacks (e.g., nutritional supplements requirement, high transcriptional demand of recombinant genes, precise oxygen level, and substrate inhibition) which limit the industrial attractiveness of such conversion process. In this work, we aim to engineer a new bacterial biocatalyst, specific for dairy waste fermentation. We metabolically engineered eight Escherichia coli strains via a new expression plasmid with the pyruvate-to-ethanol conversion genes, and we carried out the selection of the best strain among the candidates, in terms of growth in permeate, lactose consumption and ethanol formation. We finally showed that the selected engineered microbe (W strain) is able to efficiently ferment permeate and concentrated permeate, without nutritional supplements, in pH-controlled bioreactor. In the conditions tested in this work, the selected biocatalyst could complete the fermentation of permeate and concentrated permeate in about 50 and 85 h on average, producing up to 17 and 40 g/l of ethanol, respectively. To our knowledge, this is the first report showing efficient ethanol production from the lactose contained in whey permeate with engineered E. coli. The selected strain is amenable to further metabolic optimization and represents an advance towards efficient biofuel production from industrial waste stream.

A simple visual ethanol biosensor based on alcohol oxidase immobilized onto polyaniline film for halal verification of fermented beverage samples.

PubMed

Kuswandi, Bambang; Irmawati, Titi; Hidayat, Moch Amrun; Jayus; Ahmad, Musa

2014-01-27

A simple visual ethanol biosensor based on alcohol oxidase (AOX) immobilised onto polyaniline (PANI) film for halal verification of fermented beverage samples is described. This biosensor responds to ethanol via a colour change from green to blue, due to the enzymatic reaction of ethanol that produces acetaldehyde and hydrogen peroxide, when the latter oxidizes the PANI film. The procedure to obtain this biosensor consists of the immobilization of AOX onto PANI film by adsorption. For the immobilisation, an AOX solution is deposited on the PANI film and left at room temperature until dried (30 min). The biosensor was constructed as a dip stick for visual and simple use. The colour changes of the films have been scanned and analysed using image analysis software (i.e., ImageJ) to study the characteristics of the biosensor's response toward ethanol. The biosensor has a linear response in an ethanol concentration range of 0.01%-0.8%, with a correlation coefficient (r) of 0.996. The limit detection of the biosensor was 0.001%, with reproducibility (RSD) of 1.6% and a life time up to seven weeks when stored at 4 °C. The biosensor provides accurate results for ethanol determination in fermented drinks and was in good agreement with the standard method (gas chromatography) results. Thus, the biosensor could be used as a simple visual method for ethanol determination in fermented beverage samples that can be useful for Muslim community for halal verification.

Process design and optimization of novel wheat-based continuous bioethanol production system.

PubMed

Arifeen, Najmul; Wang, Ruohang; Kookos, Ioannis K; Webb, Colin; Koutinas, Apostolis A

2007-01-01

A novel design of a wheat-based biorefinery for bioethanol production, including wheat milling, gluten extraction as byproduct, fungal submerged fermentation for enzyme production, starch hydrolysis, fungal biomass autolysis for nutrient regeneration, yeast fermentation with recycling integrated with a pervaporation membrane for ethanol concentration, and fuel-grade ethanol purification by pressure swing distillation (PSD), was optimized in continuous mode using the equation-based software General Algebraic Modelling System (GAMS). The novel wheat biorefining strategy could result in a production cost within the range of dollars 0.96-0.50 gal(-1) ethanol (dollars 0.25-0.13 L(-1) ethanol) when the production capacity of the plant is within the range of 10-33.5 million gal y(-1) (37.85-126.8 million L y(-1)). The production of value-added byproducts (e.g., bran-rich pearlings, gluten, pure yeast cells) was identified as a crucial factor for improving the economics of fuel ethanol production from wheat. Integration of yeast fermentation with pervaporation membrane could result in the concentration of ethanol in the fermentation outlet stream (up to 40 mol %). The application of a PSD system that consisted of a low-pressure and a high-pressure column and employing heat integration between the high- and low-pressure columns resulted in reduced operating cost (up to 44%) for fuel-grade ethanol production.

Bioethanol Production By Utilizing Cassava Peels Waste Through Enzymatic And Microbiological Hydrolysis

NASA Astrophysics Data System (ADS)

Witantri, R. G.; Purwoko, T.; Sunarto; Mahajoeno, E.

2017-07-01

Cassava peels waste contains, cellulose which is quite high at 43.626%, this is a potential candidate as a raw for bioethanol production. The purpose of this study was to determine the performance of the enzymatic hydrolysis, microbiological (Effective microbe) and fermentation in cassava peel waste is known from the results of quantitative measurement of multiple ethanol parameters (DNS Test, pH, ethanol concentration). This research was carried out in stages, the first stage is hydrolysis with completely randomized design with single factor variation of the catalyst, consisting of three levels ie cellulase enzymes, multienzyme and effective microbial EM4. The second stage is fermentation with factorial randomized block design, consisting of three groups of variations of catalyst, and has two factors: variations of fermipan levels 1, 2, 3% and the duration of fermentation, 2,4,6 days. The parameters in the test is a reducing sugar, pH and concentration of ethanol. The results showed that variation of hydrolysis treatment, fermentation time, and fermipan levels has real effect on the fermentation process. On average the highest ethanol content obtained from the treatment with multienzyme addition, with the addition of 2% fermipan levels and on the 2nd day of fermentation that is equal to 3.76%.

Simultaneous detoxification, saccharification, and ethanol fermentation of weak-acid hydrolyzates

USDA-ARS?s Scientific Manuscript database

Lignocellulosic feedstocks can be prepared for ethanol fermentation by pre-treatment with a dilute mineral acid catalyst that hydrolyzes the hemicellulose and opens up the plant cell wall fibers for subsequent enzymatic saccharification. The acid catalyzed reaction scheme is sequential whereby rele...

Energy efficient recovery and dehydration of ethanol from fermentation broths by Membrane Assisted Vapor Stripping technology

EPA Science Inventory

Distillation combined with molecular sieve dehydration is the current state of the art for fuel grade ethanol production from fermentation broths. To improve the sustainability of bioethanol production, energy efficient separation alternatives are needed, particularly for lower ...

Effect of four pretreatments on enzymatic hydrolysis and ethanol fermentation of wheat straw. Influence of inhibitors and washing.

PubMed

Toquero, Cristina; Bolado, Silvia

2014-04-01

Pretreatment is essential in the production of alcohol from lignocellulosic material. In order to increase enzymatic sugar release and bioethanol production, thermal, dilute acid, dilute basic and alkaline peroxide pretreatments were applied to wheat straw. Compositional changes in pretreated solid fractions and sugars and possible inhibitory compounds released in liquid fractions were analysed. SEM analysis showed structural changes after pretreatments. Enzymatic hydrolysis and fermentation by Pichia stipitis of unwashed and washed samples from each pretreatment were performed so as to compare sugar and ethanol yields. The effect of the main inhibitors found in hydrolysates (formic acid, acetic acid, 5-hydroxymethylfurfural and furfural) was first studied through ethanol fermentations of model media and then compared to real hydrolysates. Hydrolysates of washed alkaline peroxide pretreated biomass provided the highest sugar concentrations, 31.82g/L glucose, and 13.75g/L xylose, their fermentation yielding promising results, with ethanol concentrations reaching 17.37g/L. Copyright © 2014 Elsevier Ltd. All rights reserved.

Ethanol and xylitol production by fermentation of acid hydrolysate from olive pruning with Candida tropicalis NBRC 0618.

PubMed

Mateo, Soledad; Puentes, Juan G; Moya, Alberto J; Sánchez, Sebastián

2015-08-01

Olive tree pruning biomass has been pretreated with pressurized steam, hydrolysed with hydrochloric acid, conditioned and afterwards fermented using the non-traditional yeast Candida tropicalis NBRC 0618. The main aim of this study was to analyse the influence of acid concentration on the hydrolysis process and its effect on the subsequent fermentation to produce ethanol and xylitol. From the results, it could be deduced that both total sugars and d-glucose recovery were enhanced by increasing the acid concentration tested; almost the whole hemicellulose fraction was hydrolysed when 3.77% was used. It has been observed a sequential production first of ethanol, from d-glucose, and then xylitol from d-xylose. The overall ethanol and xylitol yields ranged from 0.27 to 0.38kgkg(-1), and 0.12 to 0.23kgkg(-1) respectively, reaching the highest values in the fermentation of the hydrolysates obtained with hydrochloric acid 2.61% and 1.11%, respectively. Copyright © 2015 Elsevier Ltd. All rights reserved.

Improvement of the fermentability of oxalic acid hydrolysates by detoxification using electrodialysis and adsorption.

PubMed

Jeong, So-Yeon; Trinh, Ly Thi Phi; Lee, Hong-Joo; Lee, Jae-Won

2014-01-01

A two-step detoxification process consisting of electrodialysis and adsorption was performed to improve the fermentability of oxalic acid hydrolysates. The constituents of the hydrolysate differed significantly between mixed hardwood and softwood. Acetic acid and furfural concentrations were high in the mixed hardwood, whereas 5-hydroxymethylfurfural (HMF) concentration was relatively low compared with that of the mixed softwood. The removal efficiency of acetic acid reached 100% by electrodialysis (ED) process in both hydrolysates, while those of furfural and HMF showed very low, due to non-ionizable properties. Most of the remaining inhibitors were removed by XAD-4 resin. In the mixed hardwood hydrolysate without removal of the inhibitors, ethanol fermentation was not completed. Meanwhile, both ED-treated hydrolysates successfully produced ethanol with 0.08 and 0.15 g/Lh ethanol productivity, respectively. The maximum ethanol productivity was attained after fermentation with 0.27 and 0.35 g/Lh of detoxified hydrolysates, which were treated by ED, followed by XAD-4 resin. Copyright © 2013 Elsevier Ltd. All rights reserved.

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).

Bioconversion of paper mill sludge to bioethanol in the presence of accelerants or hydrogen peroxide pretreatment.

PubMed

Gurram, Raghu Nandan; Al-Shannag, Mohammad; Lecher, Nicholas Joshua; Duncan, Shona M; Singsaas, Eric Lawrence; Alkasrawi, Malek

2015-09-01

In this study we investigated the technical feasibility of convert paper mill sludge into fuel ethanol. This involved the removal of mineral fillers by using either chemical pretreatment or mechanical fractionation to determine their effects on cellulose hydrolysis and fermentation to ethanol. In addition, we studied the effect of cationic polyelectrolyte (as accelerant) addition and hydrogen peroxide pretreatment on enzymatic hydrolysis and fermentation. We present results showing that removing the fillers content (ash and calcium carbonate) from the paper mill sludge increases the enzymatic hydrolysis performance dramatically with higher cellulose conversion at faster rates. The addition of accelerant and hydrogen peroxide pretreatment further improved the hydrolysis yields by 16% and 25% (g glucose / g cellulose), respectively with the de-ashed sludge. The fermentation process of produced sugars achieved up to 95% of the maximum theoretical ethanol yield and higher ethanol productivities within 9h of fermentation. Copyright © 2015 Elsevier Ltd. All rights reserved.

Co-fermentation of glucose, xylose and/or cellobiose by yeast

DOEpatents

Jeffries, Thomas W.; Willis, Laura B.; Long, Tanya M.; Su, Yi-Kai

2013-09-10

Provided herein are methods of using yeast cells to produce ethanol by contacting a mixture comprising xylose with a Spathaspora yeast cell under conditions suitable to allow the yeast to ferment at least a portion of the xylose to ethanol. The methods allow for efficient ethanol production from hydrolysates derived from lignocellulosic material and sugar mixtures including at least xylose and glucose or xylose, glucose and cellobiose.

The effect of initial cell concentration on xylose fermentation by Pichia stipitis

Treesearch

Frank K. Agbogbo; Guillermo Coward-Kelly; Mads Torry-Smith; Kevin Wenger; Thomas W. Jeffries

2007-01-01

Xylose was fermented using Pichia stipitis CBS 6054 at different initial cell concentrations. A high initial cell concentration increased the rate of xylose utilization, ethanol formation, and the ethanol yield. The highest ethanol concentration of 41.0 g/L and a yield of 0.38 g/g was obtained using an initial cell concentration of 6.5 g/L. Even though more xylitol was...

Characterization of Candida sp. NY7122, a novel pentose-fermenting soil yeast.

PubMed

Watanabe, Itsuki; Ando, Akira; Nakamura, Toshihide

2012-02-01

Yeasts that ferment both hexose and pentose are important for cost-effective ethanol production. We found that the soil yeast strain NY7122 isolated from a blueberry field in Tsukuba (East Japan) could ferment both hexose and pentose (D-xylose and L-arabinose). NY7122 was closely related to Candida subhashii on the basis of the results of molecular identification using the sequence in the D1/D2 domains of 26S rDNA and 5.8S-internal transcribed spacer region. NY7122 produced at least 7.40 and 3.86 g l⁻¹ ethanol from 20 g l⁻¹ D-xylose and L-arabinose within 24 h. NY7122 could produce ethanol from pentose and hexose sugars at 37°C. The highest ethanol productivity of NY7122 was achieved under a low pH condition (pH 3.5). Fermentation of mixed sugars (50 g l⁻¹ glucose, 20 g l⁻¹ D-xylose, and 10 g l⁻¹ L-arabinose) resulted in a maximum ethanol concentration of 27.3 g l⁻¹ for the NY7122 strain versus 25.1 g l⁻¹ for Scheffersomyces stipitis. This is the first study to report that Candida sp. NY7122 from a soil environment could produce ethanol from both D-xylose and L-arabinose.

Repeated batch fermentation of immobilized E. coli expressing Vitreoscilla hemoglobin for long-term use

PubMed Central

Sar, Taner; Seker, Gamze; Erman, Ayse Gokce; Stark, Benjamin C.; Yesilcimen Akbas, Meltem

2017-01-01

ABSTRACT This study describes an efficient and reusable process for ethanol production from medium containing whey powder, using alginate immobilized ethanologenic E. coli strains either expressing (TS3) or not expressing (FBR5) Vitreoscilla hemoglobin. Reuseabilities of the FBR5 and TS3 strains were investigated regarding their ethanol production capacities over the course of 15 successive 96-h batch fermentations. The ethanol production was fairly stable over the entire duration of the experiment, with strain TS3 maintaining a substantial advantage over strain FBR5. Storage of both strains in 2 different solutions for up to 60 d resulted in only a modest loss of ethanol production, with strain TS3 consistently outperforming strain FBR5 by a substantial amount. Strains stored for 15 or 30 d maintained their abilities to produce ethanol without dimunition over the course of 8 successive batch fermentations; again strain TS3 maintained a substantial advantage over strain FBR5 throughout the entire experiment. Thus, immobilization is a useful strategy to maintain the advantage in ethanol productivity afforded by expression of Vitreoscilla hemoglobin over long periods of time and large numbers of repeated batch fermentations, including, as in this case, using media with food processing wastes as the carbon source. PMID:28394725

Repeated batch fermentation of immobilized E. coli expressing Vitreoscilla hemoglobin for long-term use.

PubMed

Sar, Taner; Seker, Gamze; Erman, Ayse Gokce; Stark, Benjamin C; Yesilcimen Akbas, Meltem

2017-09-03

This study describes an efficient and reusable process for ethanol production from medium containing whey powder, using alginate immobilized ethanologenic E. coli strains either expressing (TS3) or not expressing (FBR5) Vitreoscilla hemoglobin. Reuseabilities of the FBR5 and TS3 strains were investigated regarding their ethanol production capacities over the course of 15 successive 96-h batch fermentations. The ethanol production was fairly stable over the entire duration of the experiment, with strain TS3 maintaining a substantial advantage over strain FBR5. Storage of both strains in 2 different solutions for up to 60 d resulted in only a modest loss of ethanol production, with strain TS3 consistently outperforming strain FBR5 by a substantial amount. Strains stored for 15 or 30 d maintained their abilities to produce ethanol without dimunition over the course of 8 successive batch fermentations; again strain TS3 maintained a substantial advantage over strain FBR5 throughout the entire experiment. Thus, immobilization is a useful strategy to maintain the advantage in ethanol productivity afforded by expression of Vitreoscilla hemoglobin over long periods of time and large numbers of repeated batch fermentations, including, as in this case, using media with food processing wastes as the carbon source.

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

PubMed

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

2014-01-01

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

Effects of pretreatment methods for hazelnut shell hydrolysate fermentation with Pichia Stipitis to ethanol.

PubMed

Arslan, Yeşim; Eken-Saraçoğlu, Nurdan

2010-11-01

In this study, we investigated the use of hazelnut shell as a renewable and low cost lignocellulosic material for bioethanol production for the first time. High lignin content of hazelnut shell is an important obstacle for such a biotransformation. Biomass hydrolysis with acids yields reducing sugar with several inhibitors which limit the fermentability of sugars. The various conditioning methods for biomass and hydrolysate were performed to overcome the toxicity and their effects on the subsequent fermentation of hazelnut shell hydrolysate by Pichia stipitis were evaluated with shaking flasks experiments. Hazelnut shells hydrolysis with 0.7M H(2)SO(4) yielded 49 gl(-1) total reducing sugars and fermentation inhibitors in untreated hydrolysate. First, it was shown that several hydrolysate detoxification methods were solely inefficient in achieving cell growth and ethanol production in the fermentation of hazelnut shell hydrolysates derived from non-delignified biomass. Next, different pretreatments of hazelnut shells were considered for delignification and employed before hydrolysis in conjunction with hydrolysate detoxification to improve alcohol fermentation. Among six delignification methods, the most effective pretreatment regarding to ethanol concentration includes the treatment of shells with 3% (w/v) NaOH at room temperature, which was integrated with sequential hydrolysate detoxification by overliming and then treatment with charcoal twice at 60 degrees C. This treatment brought about a total reduction of 97% in furans and 88.4% in phenolics. Almost all trialed treatments caused significant sugar loss. Under the best assayed conditions, ethanol concentration of 16.79gl(-1) was reached from a hazelnut shell hyrolysate containing initial 50g total reducing sugar l(-1) after partial synthetic xylose supplementation. This value is equal to 91.25% of ethanol concentration that was obtained from synthetic d-xylose under same conditions. The present study demonstrates that Pichia stipitis is able to grow and ferment sugars to ethanol in detoxified hazelnut hydrolysate derived from delignified biomass.

THERMOCHEMICAL CONVERSION OF FERMENTATION-DERIVED OXYGENATES TO FUELS

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

Ramasamy, Karthikeyan K.; Wang, Yong

2013-06-01

At present ethanol generated from renewable resources through fermentation process is the dominant biofuel. But ethanol suffers from undesirable fuel properties such as low energy density and high water solubility. The production capacity of fermentation derived oxygenates are projected to rise in near future beyond the current needs. The conversion of oxygenates to hydrocarbon compounds that are similar to gasoline, diesel and jet fuel is considered as one of the viable option. In this chapter the thermo catalytic conversion of oxygenates generated through fermentation to fuel range hydrocarbons will be discussed.

Effect of phytase application during high gravity (HG) maize mashes preparation on the availability of starch and yield of the ethanol fermentation process.

PubMed

Mikulski, D; Kłosowski, G; Rolbiecka, A

2014-10-01

Phytic acid present in raw materials used in distilling industry can form complexes with starch and divalent cations and thus limit their biological availability. The influence of the enzymatic hydrolysis of phytate complexes on starch availability during the alcoholic fermentation process using high gravity (HG) maize mashes was analyzed. Indicators of the alcoholic fermentation as well as the fermentation activity of Saccharomyces cerevisiae D-2 strain were statistically evaluated. Phytate hydrolysis improved the course of the alcoholic fermentation of HG maize mashes. The final ethanol concentration in the media supplemented with phytase applied either before or after the starch hydrolysis increased by 1.0 and 0.6 % v/v, respectively, as compared to the control experiments. This increase was correlated with an elevated fermentation yield that was higher by 5.5 and 2.0 L EtOH/100 kg of starch, respectively. Phytate hydrolysis resulted also in a statistically significant increase in the initial concentration of fermenting sugars by 14.9 mg/mL of mash, on average, which was a consequence of a better availability of starch for enzymatic hydrolysis. The application of phytase increased the attenuation of HG media fermentation thus improving the economical aspect of the ethanol fermentation process.

Selection of thermotolerant yeasts for simultaneous saccharification and fermentation (SSF) of cellulose to ethanol.

PubMed

Ballesteros, I; Ballesteros, M; Cabañas, A; Carrasco, J; Martín, C; Negro, M J; Saez, F; Saez, R

1991-01-01

A total of 27 yeast strains belonging to the groups Candida, Saccharomyces, and Kluyveromyces were screened for their ability to grow and ferment glucose at temperatures ranging 32-45 degrees C. K. marxianus and K. fragilis were found to be the best ethanol producing organisms at the higher temperature tested and, so, were selected for subsequent simultaneous saccharification and fermentation (SSF) studies. SSF experiments were performed at 42 and 45 degrees C, utilizing Solkafloc (10%) as cellulose substrate and a cellulase loading of 15 FPU/g substrate. Best results were achieved at 42 degrees C with K. marxianus L. G. and K. fragilis L. G., both of which produced close to 38 g/L ethanol and 0.5 ethanol yield, in 78 h.

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

PubMed

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

2017-03-01

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

L-Lactic acid production from glycerol coupled with acetic acid metabolism by Enterococcus faecalis without carbon loss.

PubMed

Murakami, Nao; Oba, Mana; Iwamoto, Mariko; Tashiro, Yukihiro; Noguchi, Takuya; Bonkohara, Kaori; Abdel-Rahman, Mohamed Ali; Zendo, Takeshi; Shimoda, Mitsuya; Sakai, Kenji; Sonomoto, Kenji

2016-01-01

Glycerol is a by-product in the biodiesel production process and considered as one of the prospective carbon sources for microbial fermentation including lactic acid fermentation, which has received considerable interest due to its potential application. Enterococcus faecalis isolated in our laboratory produced optically pure L-lactic acid from glycerol in the presence of acetic acid. Gas chromatography-mass spectrometry analysis using [1, 2-(13)C2] acetic acid proved that the E. faecalis strain QU 11 was capable of converting acetic acid to ethanol during lactic acid fermentation of glycerol. This indicated that strain QU 11 restored the redox balance by oxidizing excess NADH though acetic acid metabolism, during ethanol production, which resulted in lactic acid production from glycerol. The effects of pH control and substrate concentration on lactic acid fermentation were also investigated. Glycerol and acetic acid concentrations of 30 g/L and 10 g/L, respectively, were expected to be appropriate for lactic acid fermentation of glycerol by strain QU 11 at a pH of 6.5. Furthermore, fed-batch fermentation with 30 g/L glycerol and 10 g/L acetic acid wholly exhibited the best performance including lactic acid production (55.3 g/L), lactic acid yield (0.991 mol-lactic acid/mol-glycerol), total yield [1.08 mol-(lactic acid and ethanol)]/mol-(glycerol and acetic acid)], and total carbon yield [1.06 C-mol-(lactic acid and ethanol)/C-mol-(glycerol and acetic acid)] of lactic acid and ethanol. In summary, the strain QU 11 successfully produced lactic acid from glycerol with acetic acid metabolism, and an efficient fermentation system was established without carbon loss. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Single Zymomonas mobilis strain for xylose and arabinose fermentation

DOEpatents

Zhang, M.; Chou, Y.C.; Picataggio, S.K.; Finkelstein, M.

1998-12-01

This invention relates to single microorganisms which normally do not ferment pentose sugars which are genetically altered to ferment the pentose sugars, xylose and arabinose, to produce ethanol, and a fermentation process utilizing the same. Examples include Zymomonas mobilis which has been transformed with a combination of E. coli genes for xylose isomerase, xylulokinase, L-arabinose isomerase, L-ribulokinase, L-ribulose 5-phosphate 4-epimerase, transaldolase and transketolase. Expression of added genes are under the control of Z. mobilis promoters. These newly created microorganisms are useful for fermenting glucose, xylose and arabinose, produced by hydrolysis of hemicellulose and cellulose or starch, to produce ethanol. 6 figs.

Single zymomonas mobilis strain for xylose and arabinose fermentation

DOEpatents

Zhang, Min; Chou, Yat-Chen; Picataggio, Stephen K.; Finkelstein, Mark

1998-01-01

This invention relates to single microorganisms which normally do not ferment pentose sugars which are genetically altered to ferment the pentose sugars, xylose and arabinose, to produce ethanol, and a fermentation process utilizing the same. Examples include Zymomonas mobilis which has been transformed with a combination of E. coli genes for xylose isomerase, xylulokinase, L-arabinose isomerase, L-ribulokinase, L-ribulose 5-phosphate 4-epimerase, transaldolase and transketolase. Expression of added genes are under the control of Z. mobilis promoters. These newly created microorganisms are useful for fermenting glucose, xylose and arabinose, produced by hydrolysis of hemicellulose and cellulose or starch, to produce ethanol.