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Sample records for high-cell-density fed-batch fermentation

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

    Science.gov (United States)

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

    2013-11-01

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

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

    Science.gov (United States)

    Liu, Wan-Cang; Zhu, Ping

    2018-01-01

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

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

    Science.gov (United States)

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

    2015-01-01

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

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

    Science.gov (United States)

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

    2011-05-01

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

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

    Science.gov (United States)

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

    2017-09-01

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

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

    Science.gov (United States)

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

    2002-07-01

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

  7. High solid fed-batch butanol fermentation with simultaneous product recovery: part II - process integration.

    Science.gov (United States)

    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 o...

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

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

    2011-09-01

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

  10. Supervision of Fed-Batch Fermentations

    DEFF Research Database (Denmark)

    Gregersen, Lars; Jørgensen, Sten Bay

    1999-01-01

    Process faults may be detected on-line using existing measurements based upon modelling that is entirely data driven. A multivariate statistical model is developed and used for fault diagnosis of an industrial fed-batch fermentation process. Data from several (25) batches are used to develop...... a model for cultivation behaviour. This model is validated against 13 data sets and demonstrated to explain a significant amount of variation in the data. The multivariate model may directly be used for process monitoring. With this method faults are detected in real time and the responsible measurements...

  11. Systematic optimization of fed-batch simultaneous saccharification and fermentation at high-solid loading based on enzymatic hydrolysis and dynamic metabolic modeling of Saccharomyces cerevisiae.

    Science.gov (United States)

    Unrean, Pornkamol; Khajeeram, Sutamat; Laoteng, Kobkul

    2016-03-01

    An integrative simultaneous saccharification and fermentation (SSF) modeling is a useful guiding tool for rapid process optimization to meet the techno-economic requirement of industrial-scale lignocellulosic ethanol production. In this work, we have developed the SSF model composing of a metabolic network of a Saccharomyces cerevisiae cell associated with fermentation kinetics and enzyme hydrolysis model to quantitatively capture dynamic responses of yeast cell growth and fermentation during SSF. By using model-based design of feeding profiles for substrate and yeast cell in the fed-batch SSF process, an efficient ethanol production with high titer of up to 65 g/L and high yield of 85 % of theoretical yield was accomplished. The ethanol titer and productivity was increased by 47 and 41 %, correspondingly, in optimized fed-batch SSF as compared to batch process. The developed integrative SSF model is, therefore, considered as a promising approach for systematic design of economical and sustainable SSF bioprocessing of lignocellulose.

  12. Optimization of a fed-batch fermentation process for production of ...

    African Journals Online (AJOL)

    user

    2011-02-28

    Feb 28, 2011 ... fermentation. Overall, this study suggested that fed-batch fermentation can be successfully used to ... catalysts that catalyze their own synthesis. Enzymes are .... shows the amount of biomass (g l−1) in the fermentation broth of ...

  13. Expression of recombinant Pseudomonas stutzeri di-heme cytochrome c(4) by high-cell-density fed-batch cultivation of Pseudomonas putida

    DEFF Research Database (Denmark)

    Thuesen, Marianne Hallberg; Nørgaard, Allan; Hansen, Anne Merete

    2003-01-01

    The gene of the di-heme protein cytochrome c(4) from Pseudomonas stutzeri was expressed in Pseudomonas putida. High-yield expression of the protein was achieved by high-cell-density fed-batch cultivation using an exponential glucose feeding strategy. The recombinant cytochrome c(4) protein...

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

    Science.gov (United States)

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

    2013-07-20

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

  15. Improving cellulase productivity of Penicillium oxalicum RE-10 by repeated fed-batch fermentation strategy.

    Science.gov (United States)

    Han, Xiaolong; Song, Wenxia; Liu, Guodong; Li, Zhonghai; Yang, Piao; Qu, Yinbo

    2017-03-01

    Medium optimization and repeated fed-batch fermentation were performed to improve the cellulase productivity by P. oxalicum RE-10 in submerged fermentation. First, Plackett-Burman design (PBD) and central composite design (CCD) were used to optimize the medium for cellulase production. PBD demonstrated wheat bran and NaNO 3 had significant influences on cellulase production. The CCD results showed the maximum filter paper activity (FPA) production of 8.61U/mL could be achieved in Erlenmeyer flasks. The maximal FPA reached 12.69U/mL by submerged batch fermentation in a 7.5-L stirred tank, 1.76-fold higher than that on the original medium. Then, the repeated fed-batch fermentation strategy was performed successfully for increasing the cellulase productivity from 105.75U/L/h in batch fermentation to 158.38U/L/h. The cellulase activity and the glucan conversion of delignined corn cob residue hydrolysis had no significant difference between the enzymes sampled from different cycles of the repeated fed-batch fermentation and that from batch culture. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Optimization of fed-batch fermentation for a staphylokinase-hirudin ...

    African Journals Online (AJOL)

    TUOYO

    2010-08-09

    Aug 9, 2010 ... In this study, the fed-batch fermentation technique was applied to improve the yield of STH, a chimeric protein composed ... Under optimal conditions (GMYT and complex medium), a final STH expression of 1.48 g/l fermentation broth was ... STH production contained the following materials (per L): Sucrose.

  17. The use of fed batch approaches to maximise yields in bacterial fermentation and protein expression

    International Nuclear Information System (INIS)

    McLean, A.

    2001-01-01

    A fermentation facility for the scale up of bacterial and yeast fermentations has been set up at the University of Queensland under the auspices of the ARC Special Research Centre for Functional and Applied Genomics. A major application is the production of recombinant proteins for determination of tertiary structures by X-ray crystallography or nuclear magnetic resonance. For this purpose, large amounts of protein arc needed and the yield from a single fermentation run is crucial to success within constrained laboratory budgets. To achieve maximal yields we are optimising fed batch approaches in bacterial fermentation. Fed batch offers many advantages over batch cultures. Coupled with the ability to monitor online the internal conditions of the fermentation including pH and dissolved oxygen and stirrer cascading functions it is possible to ensure that the nutritional environment of the microorganism is optimised for its growth and or for optimal protein expression. The poster will describe some of our experience in setting up fed batch fermentations and successful applications of fed batches to increasing protein yield. It will also outline services that are available to academic groups outside the University of Queensland For structure determination and functional studies, the production of radiolabelled proteins can also be an advantage. We will describe initial experiments aimed at coupling the principles of fed batch fermentation to the introduction of carbon or nitrogen isotopes into the recombinant protein

  18. Omega-3 production by fermentation of Yarrowia lipolytica: From fed-batch to continuous.

    Science.gov (United States)

    Xie, Dongming; Miller, Edward; Sharpe, Pamela; Jackson, Ethel; Zhu, Quinn

    2017-04-01

    The omega-3 fatty acid, cis-5,8,11,14,17-eicosapentaenoic acid (C20:5; EPA) has wide-ranging benefits in improving heart health, immune function, and mental health. A sustainable source of EPA production through fermentation of metabolically engineered Yarrowia lipolytica has been developed. In this paper, key fed-batch fermentation conditions were identified to achieve 25% EPA in the yeast biomass, which is so far the highest EPA titer reported in the literature. Dynamic models of the EPA fermentation process were established for analyzing, optimizing, and scaling up the fermentation process. In addition, model simulations were used to develop a two-stage continuous process and compare to single-stage continuous and fed- batch processes. The two stage continuous process, which is equipped with a smaller growth fermentor (Stage 1) and a larger production fermentor (Stage 2), was found to be a superior process to achieve high titer, rate, and yield of EPA. A two-stage continuous fermentation experiment with Y. lipolytica strain Z7334 was designed using the model simulation and then tested in a 2 L and 5 L fermentation system for 1,008 h. Compared with the standard 2 L fed-batch process, the two-stage continuous fermentation process improved the overall EPA productivity by 80% and EPA concentration in the fermenter by 40% while achieving comparable EPA titer in biomass and similar conversion yield from glucose. During the long-term experiment it was also found that the Y. lipolytica strain evolved to reduce byproduct and increase lipid production. This is one of the few continuous fermentation examples that demonstrated improved productivity and concentration of a final product with similar conversion yield compared with a fed-batch process. This paper suggests the two-stage continuous fermentation could be an effective process to achieve improved production of omega-3 and other fermentation products where non-growth or partially growth associated kinetics

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

    Science.gov (United States)

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

    2017-11-01

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

  20. Kinetic model-based feed-forward controlled fed-batch fermentation of Lactobacillus rhamnosus for the production of lactic acid from Arabic date juice.

    Science.gov (United States)

    Choi, Minsung; Al-Zahrani, Saeed M; Lee, Sang Yup

    2014-06-01

    Arabic date is overproduced in Arabic countries such as Saudi Arabia and Iraq and is mostly composed of sugars (70-80 wt%). Here we developed a fed-batch fermentation process by using a kinetic model for the efficient production of lactic acid to a high concentration from Arabic date juice. First, a kinetic model of Lactobacillus rhamnosus grown on date juice in batch fermentation was constructed in EXCEL so that the estimation of parameters and simulation of the model can be easily performed. Then, several fed-batch fermentations were conducted by employing different feeding strategies including pulsed feeding, exponential feeding, and modified exponential feeding. Based on the results of fed-batch fermentations, the kinetic model for fed-batch fermentation was also developed. This new model was used to perform feed-forward controlled fed-batch fermentation, which resulted in the production of 171.79 g l(-1) of lactic acid with the productivity and yield of 1.58 and 0.87 g l(-1) h(-1), respectively.

  1. Kinetic study of batch and fed-batch enzymatic saccharification of pretreated substrate and subsequent fermentation to ethanol

    Directory of Open Access Journals (Sweden)

    Gupta Rishi

    2012-03-01

    Full Text Available Abstract Background Enzymatic hydrolysis, the rate limiting step in the process development for biofuel, is always hampered by its low sugar concentration. High solid enzymatic saccharification could solve this problem but has several other drawbacks such as low rate of reaction. In the present study we have attempted to enhance the concentration of sugars in enzymatic hydrolysate of delignified Prosopis juliflora, using a fed-batch enzymatic hydrolysis approach. Results The enzymatic hydrolysis was carried out at elevated solid loading up to 20% (w/v and a comparison kinetics of batch and fed-batch enzymatic hydrolysis was carried out using kinetic regimes. Under batch mode, the actual sugar concentration values at 20% initial substrate consistency were found deviated from the predicted values and the maximum sugar concentration obtained was 80.78 g/L. Fed-batch strategy was implemented to enhance the final sugar concentration to 127 g/L. The batch and fed-batch enzymatic hydrolysates were fermented with Saccharomyces cerevisiae and ethanol production of 34.78 g/L and 52.83 g/L, respectively, were achieved. Furthermore, model simulations showed that higher insoluble solids in the feed resulted in both smaller reactor volume and shorter residence time. Conclusion Fed-batch enzymatic hydrolysis is an efficient procedure for enhancing the sugar concentration in the hydrolysate. Restricting the process to suitable kinetic regimes could result in higher conversion rates.

  2. Kinetic study of batch and fed-batch enzymatic saccharification of pretreated substrate and subsequent fermentation to ethanol

    Science.gov (United States)

    2012-01-01

    Background Enzymatic hydrolysis, the rate limiting step in the process development for biofuel, is always hampered by its low sugar concentration. High solid enzymatic saccharification could solve this problem but has several other drawbacks such as low rate of reaction. In the present study we have attempted to enhance the concentration of sugars in enzymatic hydrolysate of delignified Prosopis juliflora, using a fed-batch enzymatic hydrolysis approach. Results The enzymatic hydrolysis was carried out at elevated solid loading up to 20% (w/v) and a comparison kinetics of batch and fed-batch enzymatic hydrolysis was carried out using kinetic regimes. Under batch mode, the actual sugar concentration values at 20% initial substrate consistency were found deviated from the predicted values and the maximum sugar concentration obtained was 80.78 g/L. Fed-batch strategy was implemented to enhance the final sugar concentration to 127 g/L. The batch and fed-batch enzymatic hydrolysates were fermented with Saccharomyces cerevisiae and ethanol production of 34.78 g/L and 52.83 g/L, respectively, were achieved. Furthermore, model simulations showed that higher insoluble solids in the feed resulted in both smaller reactor volume and shorter residence time. Conclusion Fed-batch enzymatic hydrolysis is an efficient procedure for enhancing the sugar concentration in the hydrolysate. Restricting the process to suitable kinetic regimes could result in higher conversion rates. PMID:22433563

  3. Fed-batch fermentation dealing with nitrogen limitation in microbial transglutaminase production by Streptoverticillium mobaraense

    NARCIS (Netherlands)

    Rinzema, A; Tramper, J; de Bruin, E; Bol, J

    In the later stages of a batch fermentation for microbial transglutaminase production by Streptoverticillium mobaraense the availability of a nitrogen source accessible to the microorganism becomes critical. Fed-batch fermentation is investigated with the aim of avoiding this substrate limitation.

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

    Science.gov (United States)

    Callewaert, Raf; De Vuyst, Luc

    2000-01-01

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

  5. Butanol production from wheat straw by simultaneous saccharification and fermentation using Clostridium beijerinckii: Part II-Fed-batch fermentation

    International Nuclear Information System (INIS)

    Qureshi, Nasib; Saha, Badal C.; Cotta, Michael A.

    2008-01-01

    In these studies, Clostridium beijerinckii P260 was used to produce butanol (acetone-butanol-ethanol, or ABE) from wheat straw (WS) hydrolysate in a fed-batch reactor. It has been demonstrated that simultaneous hydrolysis of WS to achieve 100% hydrolysis to simple sugars (to the extent achievable under present conditions) and fermentation to butanol is possible. In addition to WS, the reactor was fed with a sugar solution containing glucose, xylose, arabinose, galactose, and mannose. The culture utilized all of the above sugars. It was noticed that near the end of fermentation (286-533 h), the culture had difficulties utilizing xylose. As a result of supplemental sugar feed to the reactor, ABE productivity was improved by 16% as compared with previous studies. In our previous experiment on simultaneous saccharification of WS and fermentation to butanol, a productivity of 0.31 g L -1 h -1 was observed, while in the present studies a productivity of 0.36 g L -1 h -1 was observed. It should be noted that a productivity of 0.77 g L -1 h -1 was observed when the culture was highly active. The fed-batch fermentation was operated for 533 h. It should be noted that C. beijerinckii P260 can be used to produce butanol from WS in integrated fermentations

  6. CONVERSION OF PINEAPPLE JUICE WASTE INTO LACTIC ACID IN BATCH AND FEDBATCH FERMENTATION SYSTEMS

    Directory of Open Access Journals (Sweden)

    Abdullah Mochamad Busairi

    2012-01-01

    Full Text Available Pineapple juice waste contains valuable components, which are mainly sucrose, glucose, and fructose. Recently, lactic acid has been considered to be an important raw material for the production of biodegradable lactide polymer. The fermentation experiments were carried out in a 3 litres fermentor (Biostat B Model under anaerobic condition with stirring speed of 50 rpm, temperature at 40oC, and pH of 6.00. Effect of feed concentration on lactic acid production, bacterial growth, substrate utilisation and productivity was studied. The results obtained from fed- batch culture fermentation showed that the maximum lactic acid productivity was 0.44 g/L.h for feed concentration of 90 g/L at 48 hours. Whereas the lactic acid productivity obtained from fed-batch culture was twice and half fold higher than that of batch culture productivity.  Buangan jus nanas mengandung komponen yang berharga terutama sukrosa, glukosa, dan fruktosa. Asam laktat adalah bahan baku yang terbaru dan penting untuk dibuat sebagai polimer laktat yang dapat terdegradasi oleh lingkungan. Percobaan dilakukan pada fermentor 3 liter (Model Biostat B di bawah kondisi anaerob dengan kecepatan pengadukan 50 rpm, temperatur 40oC, dan pH 6,00. Pengaruh konsentrasi umpan terhadap produksi asam laktat, pertumbuhan mikroba, pengggunaan substrat dan produktivitas telah dipelajari. Hasil yang didapatkan pada fermentasi dengan menggunakan sistem fed-batch menunjukkan bahwa produktivitas asam laktat maksimum adalah 0.44 g/L,jam dengan konsentrasi umpan, 90 g/L pada waktu 48 jam. Bahkan produktivitas asam laktat yang didapat pada kultur fed-batch lebih tinggi 2,5 kali dari pada proses menggunakan sistem batch

  7. Nonlinear impulsive system of fed-batch culture in fermentative production and its properties

    International Nuclear Information System (INIS)

    Gao Caixia; Li Kezan; Feng Enmin; Xiu Zhilong

    2006-01-01

    In this study, the nonlinear dynamical system of fed-batch fermentation is investigated in the process of bio-dissimilation of glycerol to 1,3-propanediol by Klebsiella pneumoniae. Considering the abrupt increase of glycerol in fed-batch culture, this paper proposes a nonlinear impulsive system of the culture process, which is fit for formulating the factual fermentation better than the continuous models in being. We study the questions of existence and properties of mild solutions for the system and the continuous dependence of solutions on initial values and the controllable variable. Finally, the numerical simulations show that the errors between experimental and computational values using the impulsive system are less than those using the previous continuous system

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

    Science.gov (United States)

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

    2011-09-01

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

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

    Science.gov (United States)

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

    2014-02-01

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

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

    Science.gov (United States)

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

    2010-10-01

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

  11. A high-yielding, generic fed-batch process for recombinant antibody production of GS-engineered cell lines

    DEFF Research Database (Denmark)

    Fan, Li; Zhao, Liang; Sun, Yating

    2009-01-01

    An animal component-free and chemically defined fed-batch process for GS-engineered cell lines producing recombinant antibodies has been developed. The fed-batch process relied on supplying sufficient nutrients to match their consumption, simultaneously minimizing the accumulation of byproducts....... This generic and high-yielding fed-batch process would shorten development time, and ensure process stability, thereby facilitating the manufacture of therapeutic antibodies by GS-engineered cell lines....

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

    Science.gov (United States)

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

    2015-01-10

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

  13. Fructose production by Zymomonas mobilis in fed-batch culture with minimal sorbitol formation

    Energy Technology Data Exchange (ETDEWEB)

    Edye, L A; Johns, M R; Ewings, K N

    1989-08-01

    Fed-batch cultures of Zymomonas mobilis (UQM 2864), a mutant unable to metabolise fructose, grown on diluted sugar cane syrup (200 g/l sucrose) achieved yields of 90.5 g/l fructose and 48.3 g/l ethanol with minimal sorbitol formation and complete utilization of the substrate. The effect of inoculum size on sorbitol formation in the batch stage of fed-batch fermentation are reported. Fermentation of sucrose (350 g/l) supplemented with nutrients yielded 142 g/l fructose and 76.5 g/l ethanol. Some fructose product loss at high fructose concentrations was observed. The fed-batch fermentation process offers a method for obtaining high concentrations of fructose and ethanol from sucrose materials. (orig.).

  14. Ethanol production from Sorghum bicolor using both separate and simultaneous saccharification and fermentation in batch and fed batch systems

    DEFF Research Database (Denmark)

    Mehmood, Sajid; Gulfraz, M.; Rana, N. F.

    2009-01-01

    The objective of this work was to find the best combination of different experimental conditions during pre-treatment, enzymatic saccharification, detoxification of inhibitors and fermentation of Sorghum bicolor straw for ethanol production. The optimization of pre-treatment using different...... were used in order to increase the monomeric sugar during enzymatic hydrolysis and it has been observed that the addition of these surfactants contributed significantly in cellulosic conversion but no effect was shown on hemicellulosic hydrolysis. Fermentability of hydrolyzate was tested using...... Saccharomyces cerevisiae Ethanol Red (TM) and it was observed that simultaneous saccharification and fermentation ( SSF) with both batch and fed batch resulted in better ethanol yield as compared to separate hydrolysis and fermentation ( SHF). Detoxification of furan during SHF facilitated reduction...

  15. Batch and multi-step fed-batch enzymatic saccharification of Formiline-pretreated sugarcane bagasse at high solid loadings for high sugar and ethanol titers.

    Science.gov (United States)

    Zhao, Xuebing; Dong, Lei; Chen, Liang; Liu, Dehua

    2013-05-01

    Formiline pretreatment pertains to a biomass fractionation process. In the present work, Formiline-pretreated sugarcane bagasse was hydrolyzed with cellulases by batch and multi-step fed-batch processes at 20% solid loading. For wet pulp, after 144 h incubation with cellulase loading of 10 FPU/g dry solid, fed-batch process obtained ~150 g/L glucose and ~80% glucan conversion, while batch process obtained ~130 g/L glucose with corresponding ~70% glucan conversion. Solid loading could be further increased to 30% for the acetone-dried pulp. By fed-batch hydrolysis of the dried pulp in pH 4.8 buffer solution, glucose concentration could be 247.3±1.6 g/L with corresponding 86.1±0.6% glucan conversion. The enzymatic hydrolyzates could be well converted to ethanol by a subsequent fermentation using Saccharomices cerevisiae with ethanol titer of 60-70 g/L. Batch and fed-batch SSF indicated that Formiline-pretreated substrate showed excellent fermentability. The final ethanol concentration was 80 g/L with corresponding 82.7% of theoretical yield. Copyright © 2012 Elsevier Ltd. All rights reserved.

  16. A Genetic Algorithm for Feeding Trajectory Optimisation of Fed-batch Fermentation Processes

    Directory of Open Access Journals (Sweden)

    Stoyan Tzonkov

    2009-03-01

    Full Text Available In this work a genetic algorithm is proposed with the purpose of the feeding trajectory optimization during a fed-batch fermentation of E. coli. The feed rate profiles are evaluated based on a number of objective functions. Optimization results obtained for different feeding trajectories demonstrate that the genetic algorithm works well and shows good computational performance. Developed optimal feed profiles meet the defined criteria. The ration of the substrate concentration and the difference between actual cell concentration and theoretical maximum cell concentration is defined as the most appropriate objective function. In this case the final cell concentration of 43 g·l-1 and final product concentration of 125 g·l-1 are achieved and there is not significant excess of substrate.

  17. Optimization of cyclosporin A production by Beauveria nivea in continuous fed-batch fermentation

    Directory of Open Access Journals (Sweden)

    Dong Huijun

    2011-01-01

    Full Text Available To develop the effective control method for fed-batch culture of cyclosporin A production, we chose fructose, L-valine and (NH42HPO4 as feeding nutrients and compared their productivities in relation to different concentrations. The feeding rate of three kinds of feeding materials was controlled to maintain the suitable residual concentration. The fed-batch fermentation results indicated that the optimal concentrations of fructose, L-valine and (NH42HPO4 were about 20 g/L, 0.5 g/L and 0.6 g/L for cyclosporin A production, respectively. The cultivation of Beauveria nivea could produce cyclosporin A up to 6.2 g/L for 240 hrs through a continuous feeding-rate-controlled-batch process under the optimal feeding conditions.

  18. Production of ethanol in batch and fed-batch fermentation of soluble sugar

    International Nuclear Information System (INIS)

    Chaudhary, M.Y.; Shah, M.A.; Shah, F.H.

    1991-01-01

    Keeping in view of the demand and need for alternate energy source, especially liquid fuels and the availability of raw materials in Pakistan, we have carried out biochemical and technological studies for ethanol through fermentation of renewable substrates. Molasses and sugar cane have been used as substrate for yeast fermentation. Selected yeast were used in both batch and semi continuous fermentation of molasses. Clarified dilute molasses were fermented with different strains of Saccharomyces cerevisiae. Ethanol concentration after 64 hours batch fermentation reached 9.4% with 90% yield based on sugar content. During feed batch system similar results were obtained after a fermentation cycle of 48 hours resulting in higher productivity. Similarly carbohydrates in fruit juices and hydro lysates of biomass can be economically fermented to ethanol to be used as feed stock for other chemicals. (author)

  19. Fed batch fermentation scale up in the production of recombinant streptokinase

    Directory of Open Access Journals (Sweden)

    Salvador Losada-Nerey

    2017-01-01

    Full Text Available Due to the high international demand of the recombinant streptokinase (Skr produced at the National Center for Bioproducts (BioCen, it was necessary to increase the production capacity of the drug, since the current production volume does not cover the demand. A scale up of the process of fermentation of the recombinant streptokinase was made using a fed batch culture, from the bank scale towards a 300L fermenter. The scaling criteria used were: the intensive variables of the process, the relationships of volumes of the fermentation medium and inoculum, the volumetric coefficient of oxygen transfer and air volume to liquid flow relationship which were kept constant. With this scale up procedure it was possible to reproduce the results obtained at the bank scale of and to double the biomass production volume with the same equipment, fulfilling all the quality requirements of the product and to cover the current demand of the market. Techno-economic indicators demonstrated the feasibility of this option.

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

    Science.gov (United States)

    Wang, Tao; Sun, Jibin; Yuan, Jingqi

    2015-04-01

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

  1. High-titer and productivity of l-(+)-lactic acid using exponential fed-batch fermentation with Bacillus coagulans arr4, a new thermotolerant bacterial strain.

    Science.gov (United States)

    Coelho, Luciana Fontes; Beitel, Susan Michelz; Sass, Daiane Cristina; Neto, Paulo Marcelo Avila; Contiero, Jonas

    2018-04-01

    Bacillus coagulans arr4 is a thermotolerant microorganism with great biotechnological potential for l-(+)-lactic acid production from granulated sugar and yeast extract. The highest l-(+)-lactic acid production was obtained with Ca(OH) 2 . The maximum production of l-(+)-lactic acid (206.81 g/L) was observed in exponential feeding using granulated sugar solution (900 g/L) and yeast extract (1%) at 50 °C, pH 6.5, and initial granulated sugar concentration of 100 g/L at 39 h. 5.3 g/L h productivity and 97% yield were observed, and no sugar remained. Comparing the simple batch with exponential fed-batch fermentation, the l(+) lactic acid production was improved in 133.22% and dry cell weight was improved in 83.29%, using granulated sugar and yeast extract. This study presents the highest productivity of lactic acid ever observed in the literature, on the fermentation of thermotolerant Bacillus sp. as well as an innovative and high-efficiency purification technology, using low-cost substances as Celite and charcoal. The recovery of lactic acid was 86%, with 100% protein removal, and the fermentation medium (brown color) became a colorless solution.

  2. Comparing a Dynamic Fed-Batch and a Continuous Steady-State Simulation of Ethanol Fermentation in a Distillery to a Stoichiometric Conversion Simulation

    OpenAIRE

    Fonseca, G.C.; Costa, C.B.B.; Cruz, A.J.G.

    2017-01-01

    Abstract An autonomous sugarcane bioethanol plant was simulated in EMSO software, an equation oriented process simulator. Three types of fermentation units were simulated: a six parallel fed-batch reactor system, a set of four CSTR in steady state and one consisting of a single stoichiometric reactor. Stoichiometric models are less accurate than kinetic-based fermentation models used for fed-batch and continuous fermenter simulations, since they do not account for inhibition effects and depen...

  3. Comparing a Dynamic Fed-Batch and a Continuous Steady-State Simulation of Ethanol Fermentation in a Distillery to a Stoichiometric Conversion Simulation

    Directory of Open Access Journals (Sweden)

    G.C. Fonseca

    Full Text Available Abstract An autonomous sugarcane bioethanol plant was simulated in EMSO software, an equation oriented process simulator. Three types of fermentation units were simulated: a six parallel fed-batch reactor system, a set of four CSTR in steady state and one consisting of a single stoichiometric reactor. Stoichiometric models are less accurate than kinetic-based fermentation models used for fed-batch and continuous fermenter simulations, since they do not account for inhibition effects and depend on a known conversion rate of reactant to be specified instead. On the other hand, stoichiometric models are faster and simpler to converge. In this study it was found that the conversion rates of sugar for the fermentation systems analyzedwere predictable from information on the composition of the juice stream. Those rates were used in the stoichiometric model, which accurately reproduced the results from both the fed-batch and the continuous fermenter system.

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

    Science.gov (United States)

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

    2017-05-01

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

  5. Cloning, multicopy expression and fed-batch production of Rhodotorula araucariae epoxide hydrolase in yarrowia lipolytica

    CSIR Research Space (South Africa)

    Ramduth, D

    2008-05-01

    Full Text Available demonstrated a 4 fold enhanced EH activity over the transformant. The transformant was then evaluated in batch and fed batch fermentations, where the batch fermentations resulted in - 50% improved EH activity from flask evaluations. In fed batch fermentations...

  6. Continuous fed-batch vacuum fermentation system for glycerol from molasses by the sulfite process

    Energy Technology Data Exchange (ETDEWEB)

    Kalle, G.P.; Naik, S.C.

    1985-01-01

    A continuous fed-batch vacuum fermentation system has been described for the production of glycerol from cane molasses (and juice) by a conventional sulfite process. A glycerol concentration of 80 g/l was achieved with a productivity of 30 g/l/day at a dilution rate of 0.4/day which is twice that from a vacuum batch process (15 g/l/day) or four times that obtained without vacuum (8 g/l/day). 8 references.

  7. Modelling of Fed-batch Fermentation Process with Droppings for L-lysine Production

    Directory of Open Access Journals (Sweden)

    Velitchka Ivanova

    2006-04-01

    Full Text Available The aim of the article is the development of dynamic unstructured model of L-lysine fed-batch fermentation process with droppings. This approach includes the following procedures: description of the process by generalized stoichiometric equations; preliminary data processing; identification of the specific rates (growth rate (mu , substrate utilization rate (nu, production rate (rho; establishment and optimization of the dynamic model of the process; simulation researches.

  8. Exponential fed-batch strategy for enhancing biosurfactant production by Bacillus subtilis.

    Science.gov (United States)

    Amin, G A

    2014-01-01

    Surfactin produced by Bacillus subtilis BDCC-TUSA-3 from Maldex-15 was used as a growth-associated product in a conventional batch process. Maldex-15 is a cheap industrial by-product recovered during manufacturing of high fructose syrup from corn starch. Surfactin production was greatly improved in exponential fed-batch fermentation. Maldex-15 and other nutrients were exponentially fed into the culture based on the specific growth rate of the bacterium. In order to maximize surfactin yield and productivity, conversion of different quantities of Maldex-15 into surfactin was investigated in five different fermentation runs. In all runs, most of the Maldex-15 was consumed and converted into surfactin and cell biomass with appreciable efficiencies. The best results were obtained with the fermentation run supplied with 204 g Maldex-15. Up to 36.1 g l(-1) of surfactin and cell biomass of 31.8 g l(-1) were achieved in 12 h. Also, a marked substrate yield of 0.272 g g(-1) and volumetric reactor productivity of 2.58 g 1(-1) h(-1) were obtained, confirming the establishment of a cost-effective commercial surfactin production.

  9. Cheese whey-induced high-cell-density production of recombinant proteins in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Neubauer Peter

    2003-04-01

    Full Text Available Abstract Background Use of lactose-rich concentrates from dairy processes for the induction of recombinant gene's expression has not received much attention although they are interesting low cost substrates for production of recombinant enzymes. Applicability of dairy waste for induction of recombinant genes in Escherichia coli was studied. Clones expressing Lactobacillus phage muramidase and Lactobacillus alcohol dehydrogenase were used for the experiments. Results Shake flask cultivations in mineral salt medium showed that cheese whey or deproteinised whey induced gene expression as efficiently as IPTG (isopropyl-β-D-thiogalactopyranoside or pure lactose. Addition of yeast extract or proteolytically degraded whey proteins did not improve the recombinant protein yield. In contrast, addition of yeast extract to the well-balanced mineral salt medium decreased the product yield. Feeding with glycerol provided sufficient amount of easily assimilable carbon source during the induction period without preventing lactose intake and induction by lactose. High-cell-density fed-batch cultivations showed that product yields comparable to IPTG-induction can be achieved by feeding bacteria with a mixture of glycerol and concentrated whey permeate during the induction. Conclusion Whey and concentrated whey permeate can be applied as an alternative inducer in recombinant high-cell-density fed-batch fermentations. The yield of the recombinant product was comparable to fermentations induced by IPTG. In low-cell-density shake flask experiments the yield was higher with whey or whey permeate than with IPTG.

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

    Science.gov (United States)

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

    2018-05-21

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

  11. Cell recycle batch fermentation of high-solid lignocellulose using a recombinant cellulase-displaying yeast strain for high yield ethanol production in consolidated bioprocessing.

    Science.gov (United States)

    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.

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

    Science.gov (United States)

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

    2018-03-24

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

  13. Improving lactic acid productivity from wheat straw hydrolysates by membrane integrated repeated batch fermentation under non-sterilized conditions

    DEFF Research Database (Denmark)

    Zhang, Yuming; Chen, Xiangrong; Qi, Benkun

    2014-01-01

    to eliminate the sequential utilization of mixed sugar and feedback inhibition during batch fermentation, membrane integrated repeated batch fermentation (MIRB) was used to improve LA productivity. With MIRB, a high cell density was obtained and the simultaneous fermentation of glucose, xylose and arabinose...

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

    Science.gov (United States)

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

    2018-01-01

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

  15. [Effects of different methanol feeding strategy on hirudin production in high-density fermentation by recombinant Pichia pastoris].

    Science.gov (United States)

    Zhou, Xiang-Shan; Fan, Wei-Min; Zhang, Yuan-Xing

    2002-05-01

    Four different methanol feeding modes were evaluated in the hirudin production in high-density fermentation by Pichia pastoris. It was difficult to avoid methanol excessive in the broth with the feeding strategy only based on DO level. On the other hand, the fluctuation in methanol concentration was observed with methanol feeding strategy by off-line gas chromatography. However, the stable methanol concentration was perfectly achieved by the on-line monitoring with methanol sensor. The supply of energy was improved by feeding glycerol at a limited rate as well as methanol in the induction phase. Therefore, the high cell dry weight (162 g/L) and high hirudin activity (2.4 x 10(4) ATU/mL or 1.7 g/L) was obtained in the fed-batch fermentation of recombinant Pichia pastoris by methanol-glycerol mixed feeding.

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

    Science.gov (United States)

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

    2015-07-30

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

  17. One Approach for Dynamic L-lysine Modelling of Repeated Fed-batch Fermentation

    Directory of Open Access Journals (Sweden)

    Kalin Todorov

    2007-03-01

    Full Text Available This article deals with establishment of dynamic unstructured model of variable volume fed-batch fermentation process with intensive droppings for L-lysine production. The presented approach of the investigation includes the following main procedures: description of the process by generalized stoichiometric equations; preliminary data processing and calculation of specific rates for main kinetic variables; identification of the specific rates as a second-order non-linear dynamic models; establishment and optimisation of dynamic model of the process; simulation researches. MATLAB is used as a research environment.

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

    Science.gov (United States)

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

    2012-07-01

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

  19. Fed-batch CHO cell culture for lab-scale antibody production

    DEFF Research Database (Denmark)

    Fan, Yuzhou; Ley, Daniel; Andersen, Mikael Rørdam

    2017-01-01

    Fed-batch culture is the most commonly used upstream process in industry today for recombinant monoclonal antibody production using Chinese hamster ovary cells. Developing and optimizing this process in the lab is crucial for establishing process knowledge, which enable rapid and predictable tech......-transfer to manufacturing scale. In this chapter, we will describe stepwise how to carry out fed-batch CHO cell culture for lab-scale antibody production....

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

    Directory of Open Access Journals (Sweden)

    Ranjan Singh

    2012-09-01

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

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

    Science.gov (United States)

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

    2010-09-30

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

  2. Simulation of Feedforward-Feedback Control of Dissolved Oxygen of Microbial Repeated Fed-batch Culture

    Directory of Open Access Journals (Sweden)

    Ling Gao

    2016-09-01

    Full Text Available Fed-batch culture is often used in industry, and dissolved oxygen (DO concentration control is important in fermentation process control. DO control is often applied by using feedback (FB control strategy. But, feedforward-feedback (FF-FB control has the advantage in dealing with the time-varying characteristics resulted from the cell growth during the fermentation process. Mathematical modeling and computer simulation is a useful tool in analysis of the control system.  In this research, the FF-FB DO control and FB substrate control of repeated fed-batch culture process is modeled and simulated. The results showed the feasibility of the control strategy. These results are useful for control system development and process analyses and optimization.

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

    Science.gov (United States)

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

    2016-04-01

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

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

    OpenAIRE

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

    2012-01-01

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

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

    Science.gov (United States)

    Kuwae, Shinobu; Miyakawa, Ichiko; Doi, Tomohiro

    2018-01-11

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

  6. A fed-batch strategy to produce high poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-4-hydroxybutyrate) terpolymer yield with enhanced mechanical properties in bioreactor.

    Science.gov (United States)

    Aziz, Nursolehah Abd; Huong, Kai-Hee; Sipaut, Coswald Stephen; Amirul, A A

    2017-11-01

    This study reports an efficient fed-batch strategy to improve poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-4-hydroxybutyrate) [P(3HB-co-3HV-co-4HB)] terpolymer production by Cupriavidus sp. USMAA2-4 with enhanced mechanical properties in bioreactor. The cultivations have been performed by combining oleic acid with γ-butyrolactone at different concentration ratios with 1-pentanol at a fixed concentration. The batch and fed-batch fermentations have resulted in P(3HB-co-3HV-co-4HB) with compositions of 9-35 mol% 3HV and 4-24 mol% 4HB monomers. The DO-stat fed-batch fermentation strategies have significantly improved the production with a maximum 4.4-fold increment of cell dry weight (CDW). Besides, appropriate feeding of the substrates has resulted in an increment of terpolymer productivity from 0.086-0.347 g/L/h, with a significantly shortened cultivation time. The bacterial growth and terpolymer formation have been found to be affected by the concentration of carbon sources supplied. Characterization of P(3HB-co-3HV-co-4HB) has demonstrated that incorporation of 3HV and 4HB monomer has significantly improved the physical and thermodynamic properties of the polymers, by reducing the polymer's crystallinity. The tensile strength, Young's modulus of the terpolymer has been discovered to increase with the increase of M w . The fed-batch fermentation strategies employed in this study have resulted in terpolymers with a range of flexible materials having improved tensile strength and Young's modulus as compared to the terpolymer produced from batch fermentation. Possession of lower melting temperature indicates an enhanced thermal stability which broadens the polymer processing window.

  7. Mathematical modeling of fed-batch fermentation of Schizochytrium sp. FJU-512 growth and DHA production using a shift control strategy.

    Science.gov (United States)

    Zhang, Mingliang; Wu, Weibin; Guo, Xiaolei; Weichen, You; Qi, Feng; Jiang, Xianzhang; Huang, Jianzhong

    2018-03-01

    To obtain high-cell-density cultures of Schizochytrium sp. FJU-512 for DHA production, two stages of fermentation strategy were used and carbon/nitrogen ratio, DO and temperature were controlled at different levels. The final dry cell weight, total lipid production and DHA yield in 15 l bioreactor reached 103.9, 37.2 and 16.0 g/l, respectively. For the further study of microbial growth and DHA production dynamics, we established a set of kinetic models for the fed-batch production of DHA by Schizochytrium sp. FJU-512 in 15 and 100 l fermenters and a compensatory parameter n was integrated into the model in order to find the optimal mathematical equations. A modified Logistic model was proposed to fit the cell growth data and the following kinetic parameters were obtained: µ m  = 0.0525/h, X m  = 100 g/l and n  = 4.1717 for the 15 l bioreactor, as well as µ m  = 0.0382/h, X m  = 107.4371 g/l and n  = 10 for the 100 l bioreactor. The Luedeking-Piret equations were utilized to model DHA production, yielding values of α  = 0.0648 g/g and β  = 0.0014 g/g/h for the 15 l bioreactor, while the values of α and β obtained for the 100 l fermentation were 0.0209 g/g and 0.0030 g/g/h. The predicted results compared with experimental data showed that the established models had a good fitting precision and were able to exactly depict the dynamic features of the DHA production process.

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

    Science.gov (United States)

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

    2014-09-01

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

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

    Science.gov (United States)

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

    2015-01-01

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

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

    Science.gov (United States)

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

    2015-01-01

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

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

    Science.gov (United States)

    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).

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

    DEFF Research Database (Denmark)

    Mears, Lisa; Stocks, Stuart M.; Albaek, Mads O.

    2017-01-01

    A novel model-based control strategy has been developed for filamentous fungal fed-batch fermentation processes. The system of interest is a pilot scale (550 L) filamentous fungus process operating at Novozymes A/S. In such processes, it is desirable to maximize the total product achieved...... is recursively updated using on-line measurements. The model was applied in order to predict the current system states, including the biomass concentration, and to simulate the expected future trajectory of the system until a specified end time. In this way, the desired feed rate is updated along the progress...

  13. Selection of chemically defined media for CHO cell fed-batch culture processes

    NARCIS (Netherlands)

    Pan, X.; Streefland, M.; Dalm, C.; Wijffels, R.H.; Martens, D.E.

    2017-01-01

    Two CHO cell clones derived from the same parental CHOBC cell line and producing the same monoclonal antibody (BC-G, a low producing clone; BC-P, a high producing clone) were tested in four basal media in all possible combinations with three feeds (=12 conditions) in fed-batch cultures.
    Higher

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

    Science.gov (United States)

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

    2017-01-01

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

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

    Science.gov (United States)

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

    2014-11-01

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

  16. A Genetic Algorithms Based Approach for Identification of Escherichia coli Fed-batch Fermentation

    Directory of Open Access Journals (Sweden)

    Olympia Roeva

    2004-10-01

    Full Text Available This paper presents the use of genetic algorithms for identification of Escherichia coli fed-batch fermentation process. Genetic algorithms are a directed random search technique, based on the mechanics of natural selection and natural genetics, which can find the global optimal solution in complex multidimensional search space. The dynamic behavior of considered process has known nonlinear structure, described with a system of deterministic nonlinear differential equations according to the mass balance. The parameters of the model are estimated using genetic algorithms. Simulation examples for demonstration of the effectiveness and robustness of the proposed identification scheme are included. As a result, the model accurately predicts the process of cultivation of E. coli.

  17. Fed batch enzymatic saccharification of food waste improves the sugar concentration in the hydrolysates and eventually the ethanol fermentation by Saccharomyces cerevisiae H058

    Directory of Open Access Journals (Sweden)

    Shoubao Yan

    2012-04-01

    Full Text Available The enzymatic hydrolysis of food waste by commercially available enzymes and the subsequent ethanol fermentation of the hydrolysates by Saccharomyces cerecisiae H058 were studied in this work. The optimum batch enzymatic conditions were found to be saccharification pH of 4.5, temperature of 55!, glucoamylase concentration of 120 u/g, α-amylase concentration of 10 u/g, solid-liquid ratio of 1: 0.75 (w/w. Fed batch hydrolysis process was started with a solid-liquid ratio of 1: 1 (w/w, with solid food waste added at time lapse of 2 h to get a final solid-liquid ratio of 1: 0.5 (w/w. After 4 h of reaction, the reducing sugar concentration reached 194.43 g/L with a enzymatic digestibility of 93.12%. Further fermentation of the batch and fed batch enzymatic hydrolysates, which contained reducing sugar concentration of 131.41 and 194.43 g/L respectively, was performed using Saccharomyces cerevisiae H058, 62.93 and 90.72 g/L ethanol was obtained within 48 h.

  18. Modelling of L-valine Repeated Fed-batch Fermentation Process Taking into Account the Dissolved Oxygen Tension

    Directory of Open Access Journals (Sweden)

    Tzanko Georgiev

    2009-03-01

    Full Text Available This article deals with synthesis of dynamic unstructured model of variable volume fed-batch fermentation process with intensive droppings for L-valine production. The presented approach of the investigation includes the following main procedures: description of the process by generalized stoichiometric equations; preliminary data processing and calculation of specific rates for main kinetic variables; identification of the specific rates takes into account the dissolved oxygen tension; establishment and optimisation of dynamic model of the process; simulation researches. MATLAB is used as a research environment.

  19. User-friendly optimization approach of fed-batch fermentation conditions for the production of iturin A using artificial neural networks and support vector machine

    Directory of Open Access Journals (Sweden)

    Fudi Chen

    2015-07-01

    Conclusion: According to the modeling results, the GRNN is considered as the most suitable ANN model for the design of the fed-batch fermentation conditions for the production of iturin A because of its high robustness and precision, and the SVM is also considered as a very suitable alternative model. Under the tolerance of 30%, the prediction accuracies of the GRNN and SVM are both 100% respectively in repeated experiments.

  20. High cell density cultures produced by internal retention: application in continuous ethanol fermentation

    Directory of Open Access Journals (Sweden)

    Berta Carola Pérez

    2004-07-01

    Full Text Available Ethanol has provoked great interest due to its potential as an alternative fuel. Nevertheless, fermentation processes must be developed by increasing the low volumetric productivity achieved in conventional cultures (batch or continuous to make this product become economically competitive. This can be achieved by using techniques leading to high cell concentration and reducing inhibition by the end-product. One of the frequently employed methods involves cell recycling. This work thus developed a membrane reactor incorporating a filtration module with 5 u,m stainless steel tubular units inside a 3L stirred jar fermenter for investigating its application in continuous ethanol production. The effects of cell concentration and transmembrane pressure difference on permeate flux were evaluated for testing the filtration module's performance. The internal cell retention system was operated in Saccharomyces cerevisiae continuous culture derived from sucrose, once fermentation conditions had been selected (30 °C, 1.25 -1.75 vvm, pH 4.5. Filter unit permeability was maintained by applying pulses of air. More than 97% of the grown cells were retained in the fermenter, reaching 51 g/L cell concentration and 8.51 g/L.h average ethanol productivity in culture with internal cell retention; this was twice that obtained in a conventional continuous culture. Key words: Membrane reactor, Saccharomyces cerevisiae, alcoholic fermentation, cell recycling.

  1. Production of Medium-Chain-Length Poly(3-Hydroxyalkanoates from Saponified Palm Kernel Oil by Pseudomonas putida: Kinetics of Batch and Fed-Batch Fermentations

    Directory of Open Access Journals (Sweden)

    Annuar, M. S. M.

    2006-01-01

    Full Text Available The kinetics of medium-chain-length poly(3-hydroxyalkanoates, PHAMCL production by Pseudomonas putida PGA1 in batch and fed-batch fermentations were studied. With saponified palm kernel oil (SPKO supplying the free fatty acids mixture as the sole carbon and energy source, PHAMCL accumulation is encouraged under ammonium-limited condition, which is a nitrogen stress environment. The amount of PHAMCL accumulated and its specific production rate, qPHA were influenced by the residual ammonium concentration level in the culture medium. It was observed that in both fermentation modes, when the residual ammonium was exhausted (< 0.05 gL-1, the PHAMCL accumulation (11.9% and qPHA (0.0062 h-1 were significantly reduced. However, this effect can be reversed by feeding low amount of ammonium to the culture, resulting in significantly improved PHAMCL yield (71.4% and specific productivity (0.6 h-1. It is concluded that the feeding of low ammonium concentration to the culture medium during the PHAMCL accumulation has a positive effect on sustaining the PHAMCL biosynthetic capability of the organism. It was also found that increasing SPKO concentration in the medium significantly reduced (up to 50% the volumetric oxygen transfer coefficient (KLa of the fermentation system.

  2. Fed-batch and perfusion culture processes: economic, environmental, and operational feasibility under uncertainty.

    Science.gov (United States)

    Pollock, James; Ho, Sa V; Farid, Suzanne S

    2013-01-01

    This article evaluates the current and future potential of batch and continuous cell culture technologies via a case study based on the commercial manufacture of monoclonal antibodies. The case study compares fed-batch culture to two perfusion technologies: spin-filter perfusion and an emerging perfusion technology utilizing alternating tangential flow (ATF) perfusion. The operational, economic, and environmental feasibility of whole bioprocesses based on these systems was evaluated using a prototype dynamic decision-support tool built at UCL encompassing process economics, discrete-event simulation and uncertainty analysis, and combined with a multi-attribute decision-making technique so as to enable a holistic assessment. The strategies were compared across a range of scales and titres so as to visualize how their ranking changes in different industry scenarios. The deterministic analysis indicated that the ATF perfusion strategy has the potential to offer cost of goods savings of 20% when compared to conventional fed-batch manufacturing processes when a fivefold increase in maximum viable cell densities was assumed. Savings were also seen when the ATF cell density dropped to a threefold increase over the fed-batch strategy for most combinations of titres and production scales. In contrast, the fed-batch strategy performed better in terms of environmental sustainability with a lower water and consumable usage profile. The impact of uncertainty and failure rates on the feasibility of the strategies was explored using Monte Carlo simulation. The risk analysis results demonstrated the enhanced robustness of the fed-batch process but also highlighted that the ATF process was still the most cost-effective option even under uncertainty. The multi-attribute decision-making analysis provided insight into the limited use of spin-filter perfusion strategies in industry. The resulting sensitivity spider plots enabled identification of the critical ratio of weightings of

  3. Biosynthesis of Citric Acid from Glycerol by Acetate Mutants of Yarrowia lipolytica in Fed-Batch Fermentation

    Directory of Open Access Journals (Sweden)

    Anita Rywińska

    2009-01-01

    Full Text Available Pure and crude glycerol from biodiesel production have been used as substrates for citric acid production by acetate-negative mutants of Yarrowia lipolytica in fed-batch fermentation. Both the final concentration and the yield of the product were the highest when Y. lipolytica Wratislavia AWG7 strain was used in the culture with pure or crude glycerol. With a medium containing 200 g/L of glycerol, production reached a maximum of citric acid of 139 g/L after 120 h. This high yield of the product (up to 0.69 g of citric acid per gram of glycerol consumed was achieved with both pure and crude glycerol. Lower yield of citric acid in the culture with Y. lipolytica Wratislavia K1 strain (about 0.45 g/g resulted from increased erythritol concentrations (up to 40 g/L, accumulated simultaneously with the citric acid. The concentration of isocitric acid, a by-product in this fermentation, was very low, in the range from 2.6 to 4.6 g/L.

  4. Lipid production in batch and fed-batch cultures of Rhodosporidium toruloides from 5 and 6 carbon carbohydrates

    Directory of Open Access Journals (Sweden)

    Wiebe Marilyn G

    2012-05-01

    Full Text Available Abstract Background Microbial lipids are a potential source of bio- or renewable diesel and the red yeast Rhodosporidium toruloides is interesting not only because it can accumulate over 50% of its dry biomass as lipid, but also because it utilises both five and six carbon carbohydrates, which are present in plant biomass hydrolysates. Methods R. toruloides was grown in batch and fed-batch cultures in 0.5 L bioreactors at pH 4 in chemically defined, nitrogen restricted (C/N 40 to 100 media containing glucose, xylose, arabinose, or all three carbohydrates as carbon source. Lipid was extracted from the biomass using chloroform-methanol, measured gravimetrically and analysed by GC. Results Lipid production was most efficient with glucose (up to 25 g lipid L−1, 48 to 75% lipid in the biomass, at up to 0.21 g lipid L−1 h−1 as the sole carbon source, but high lipid concentrations were also produced from xylose (36 to 45% lipid in biomass. Lipid production was low (15–19% lipid in biomass with arabinose as sole carbon source and was lower than expected (30% lipid in biomass when glucose, xylose and arabinose were provided simultaneously. The presence of arabinose and/or xylose in the medium increased the proportion of palmitic and linoleic acid and reduced the proportion of oleic acid in the fatty acids, compared to glucose-grown cells. High cell densities were obtained in both batch (37 g L−1, with 49% lipid in the biomass and fed-batch (35 to 47 g L−1, with 50 to 75% lipid in the biomass cultures. The highest proportion of lipid in the biomass was observed in cultures given nitrogen during the batch phase but none with the feed. However, carbohydrate consumption was incomplete when the feed did not contain nitrogen and the highest total lipid and best substrate consumption were observed in cultures which received a constant low nitrogen supply. Conclusions Lipid production in R. toruloides was lower from arabinose and mixed

  5. Untargeted GC-MS Metabolomics Reveals Changes in the Metabolite Dynamics of Industrial Scale Batch Fermentations of Streptoccoccus thermophilus Broth

    DEFF Research Database (Denmark)

    Khakimov, Bekzod; Christiansen, Lene D.; Heins, Anna-Lena

    2017-01-01

    An industrial scale biomass production using batch or fed-batch fermentations usually optimized by selection of bacterial strains, tuning fermentation media, feeding strategy, and temperature. However, in-depth investigation of the biomass metabolome during the production may reveal new knowledge...... shows that in-depth metabolic analysis of fermentation broth provides a new tool for advanced optimization of high-volume-low-cost biomass production by lowering the cost, increase the yield, and augment the product quality....... for better optimization. In this study, for the first time, the authors investigated seven fermentation batches performed on five Streptoccoccus thermophilus strains during the biomass production at Chr. Hansen (Denmark) in a real life large scale fermentation process. The study is designed to investigate...

  6. Bioethanol production from starchy biomass by direct fermentation using saccharomyces diastaticus in batch free and immobilized cell systems

    Energy Technology Data Exchange (ETDEWEB)

    Kilonzo, P.M.; Margaritis, A. [University of Western Ontario, London, ON (Canada). Dept. of Chemical and Biochemical Engineering; Yu, J.; Ye, Q. [East China Univ. of Science and Technology, Shanghai (China). Biochemical Engineering Research Inst. and State Key Lab

    2006-07-01

    The feasibility of using amylolytic yeasts for the direct fermentation of starchy biomass to ethanol was discussed. Although amylolytic yeasts such as Saccharomycopsis, Lipomyces, and Schwaniomyces secrete both {alpha}-amylase and glucoamylase enzymes that synergistically enhance starch degradation, they are not suitable for industrial bio-ethanol production because of low tolerance for ethanol and slow fermentation rate. For that reason, this study examined the direct ethanol fermentation of soluble starch or dextrin with the amylolytic yeast Saccharomyces diastaticus in batch free and immobilized cells systems. Saccharomyces diastaticus secretes glucoamylase and can therefore assimilate and ferment starch and starch-like biomass. The main focus of the study was on parameters leading to higher ethanol yields from high concentration of dextrin and soluble starch using batch cultures. A natural attachment method was proposed in which polyurethane foam sheets were used as the carrier for amylolytic yeasts immobilization in ethanol fermentations. The support was chosen because it was inexpensive, autoclavable, pliable and could be tailored to suit process requirements regarding net surface charge, shape and size. It was found that Saccharomyces diastaticus was very efficient in terms of fermentation of high initial concentrations of dextrin or soluble starch. Higher concentrations of ethanol were produced. In batch fermentations, the cells fermented high dextrin concentrations more efficiently. In particular, in batch fermentation, more than 92 g-L of ethanol was produced from 240 g-L of dextrin, at conversion efficiency of 90 per cent. The conversion efficiency decreased to 60 per cent but a higher final ethanol concentration of 147 g/L was attained with a medium containing 500 g/L of dextrin. In an immobilized cell bioreactor, Saccharomyces diastaticus produced 83 g/L of ethanol from 240 g/L of dextrin, corresponding to ethanol volumetric productivity of 9.1 g

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

    Science.gov (United States)

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

    2017-02-01

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

  8. Fed-batch methanol feeding strategy for recombinant protein production by Pichia pastoris in the presence of co-substrate sorbitol.

    Science.gov (United States)

    Celik, Eda; Calik, Pinar; Oliver, Stephen G

    2009-09-01

    Batch-wise sorbitol addition as a co-substrate at the induction phase of methanol fed-batch fermentation by Pichia pastoris (Mut(+)) was proposed as a beneficial recombinant protein production strategy and the metabolic responses to methanol feeding rate in the presence of sorbitol was systematically investigated. Adding sorbitol batch-wise to the medium provided the following advantages over growth on methanol alone: (a) eliminating the long lag-phase for the cells and reaching 'high cell density production' at t = 24 h of the process (C(X) = 70 g CDW/l); (b) achieving 1.8-fold higher recombinant human erythropoietin (rHuEPO) (at t = 18 h); (c) reducing specific protease production 1.2-fold; (d) eliminating the lactic acid build-up period; (e) lowering the oxygen uptake rate two-fold; and (f) obtaining 1.4-fold higher overall yield coefficients. The maximum specific alcohol oxidase activity was not affected in the presence of sorbitol, and it was observed that sorbitol and methanol were utilized simultaneously. Thus, in the presence of sorbitol, 130 mg/l rHuEPO was produced at t = 24 h, compared to 80 mg/l rHuEPO (t = 24 h) on methanol alone. This work demonstrates not only the ease and efficiency of incorporating sorbitol to fermentations by Mut(+) strains of P. pastoris for the production of any bio-product, but also provides new insights into the metabolism of the methylotrophic yeast P. pastoris.

  9. ON-LINE MONITORING OF BIOMASS CONCENTRATION BASED ON A CAPACITANCE SENSOR: ASSESSING THE METHODOLOGY FOR DIFFERENT BACTERIA AND YEAST HIGH CELL DENSITY FED-BATCH CULTURES

    Directory of Open Access Journals (Sweden)

    A. C. L. Horta

    2015-12-01

    Full Text Available Abstract The performance of an in-situ capacitance sensor for on-line monitoring of biomass concentration was evaluated for some of the most important microorganisms in the biotechnology industry: Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris and Bacillus megaterium. A total of 33 batch and fed-batch cultures were carried out in a bench-scale bioreactor and biomass formation trends were followed by dielectric measurements during the growth phase as well as the induction phase, for 5 recombinant E. coli strains. Permittivity measurements and viable cellular concentrations presented a linear correlation for all the studied conditions. In addition, the permittivity signal was further used for inference of the cellular growth rate. The estimated specific growth rates mirrored the main trends of the metabolic states of the different cells and they can be further used for setting-up control strategies in fed-batch cultures.

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

    Science.gov (United States)

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

    2015-04-01

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

  11. Production of tea vinegar by batch and semicontinuous fermentation

    OpenAIRE

    Kaur, Pardeep; Kocher, G. S.; Phutela, R. P.

    2010-01-01

    The fermented tea vinegar combines the beneficial properties of tea and vinegar. The complete fermentation takes 4 to 5 weeks in a batch culture and thus can be shortened by semi continuous/ continuous fermentation using immobilized bacterial cells. In the present study, alcoholic fermentation of 1.0 and 1.5% tea infusions using Saccharomyces cerevisae G was carried out that resulted in 84.3 and 84.8% fermentation efficiency (FE) respectively. The batch vinegar fermentation of these wines wit...

  12. Monitoring and robust adaptive control of fed-batch cultures of microorganisms exhibiting overflow metabolism [abstract

    Directory of Open Access Journals (Sweden)

    Vande Wouwer, A.

    2010-01-01

    Full Text Available Overflow metabolism characterizes cells strains that are likely to produce inhibiting by-products resulting from an excess of substrate feeding and a saturated respiratory capacity. The critical substrate level separating the two different metabolic pathways is generally not well defined. Monitoring of this kind of cultures, going from model identification to state estimation, is first discussed. Then, a review of control techniques which all aim at maximizing the cell productivity of fed-batch fermentations is presented. Two main adaptive control strategies, one using an estimation of the critical substrate level as set-point and another regulating the by-product concentration, are proposed. Finally, experimental investigations of an adaptive RST control scheme using the observer polynomial for the regulation of the ethanol concentration in Saccharomyces cerevisiae fed-batch cultures ranging from laboratory to industrial scales, are also presented.

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

    Science.gov (United States)

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

    2017-06-01

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

  14. Ethanol production by repeated batch and continuous fermentations of blackstrap molasses using immobilized yeast cells on thin-shell silk cocoons

    International Nuclear Information System (INIS)

    Rattanapan, Anuchit; Limtong, Savitree; Phisalaphong, Muenduen

    2011-01-01

    Highlights: → Thin-shell silk cocoons for immobilization of Saccharomycescerevisiae. → Advantages: high mechanical strength, light weight, biocompatibility and high surface area. → Enhanced cell stability and ethanol productivity by the immobilization system. -- Abstract: A thin-shell silk cocoon (TSC), a residual from the silk industry, is used as a support material for the immobilization of Saccharomyces cerevisiae M30 in ethanol fermentation because of its properties such as high mechanical strength, light weight, biocompatibility and high surface area. In batch fermentation with blackstrap molasses as the main fermentation substrate, an optimal ethanol concentration of 98.6 g/L was obtained using a TSC-immobilized cell system at an initial reducing sugar concentration of 240 g/L. The ethanol concentration produced by the immobilized cells was 11.5% higher than that produced by the free cells. Ethanol production in five-cycle repeated batch fermentation demonstrated the enhanced stability of the immobilized yeast cells. Under continuous fermentation in a packed-bed reactor, a maximum ethanol productivity of 19.0 g/(L h) with an ethanol concentration of 52.8 g/L was observed at a 0.36 h -1 dilution rate.

  15. Perfusion cell culture decreases process and product heterogeneity in a head-to-head comparison with fed-batch.

    Science.gov (United States)

    Walther, Jason; Lu, Jiuyi; Hollenbach, Myles; Yu, Marcella; Hwang, Chris; McLarty, Jean; Brower, Kevin

    2018-05-30

    In this study, we compared the impacts of fed-batch and perfusion platforms on process and product attributes for IgG1- and IgG4-producing cell lines. A "plug-and-play" approach was applied to both platforms at bench scale, using commercially available basal and feed media, a standard feed strategy for fed-batch, and ATF filtration for perfusion. Product concentration in fed-batch was 2.5 times greater than perfusion, while average productivity in perfusion was 7.5 times greater than fed-batch. PCA revealed more variability in the cell environment and metabolism during the fed-batch run. LDH measurements showed that exposure of product to cell lysate was 7-10 times greater in fed-batch. Product analysis shows larger abundances of neutral species in perfusion, likely due to decreased bioreactor residence times and extracellular exposure. The IgG1 perfusion product also had higher purity and lower half-antibody. Glycosylation was similar across both culture modes. The first perfusion harvest slice for both product types showed different glycosylation than subsequent harvests, suggesting that product quality lags behind metabolism. In conclusion, process and product data indicate that intra-lot heterogeneity is decreased in perfusion cultures. Additional data and discussion is required to understand the developmental, clinical and commercial implications, and in what situations increased uniformity would be beneficial. This article is protected by copyright. All rights reserved.

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

    Science.gov (United States)

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

    2017-07-01

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

  17. High-level expression of a fungal pyranose oxidase in high cell-density fed-batch cultivations of Escherichia coli using lactose as inducer

    Czech Academy of Sciences Publication Activity Database

    Kotík, Michael; Kočanová, Marcela; Marešová, Helena; Kyslík, Pavel

    2004-01-01

    Roč. 36, - (2004), s. 61-69 ISSN 1046-5928 Institutional research plan: CEZ:AV0Z5020903 Keywords : fed-batch * lactose * inclusion bodies Subject RIV: EE - Microbiology, Virology Impact factor: 1.336, year: 2004

  18. Concentrated fed-batch cell culture increases manufacturing capacity without additional volumetric capacity.

    Science.gov (United States)

    Yang, William C; Minkler, Daniel F; Kshirsagar, Rashmi; Ryll, Thomas; Huang, Yao-Ming

    2016-01-10

    Biomanufacturing factories of the future are transitioning from large, single-product facilities toward smaller, multi-product, flexible facilities. Flexible capacity allows companies to adapt to ever-changing pipeline and market demands. Concentrated fed-batch (CFB) cell culture enables flexible manufacturing capacity with limited volumetric capacity; it intensifies cell culture titers such that the output of a smaller facility can rival that of a larger facility. We tested this hypothesis at bench scale by developing a feeding strategy for CFB and applying it to two cell lines. CFB improved cell line A output by 105% and cell line B output by 70% compared to traditional fed-batch (TFB) processes. CFB did not greatly change cell line A product quality, but it improved cell line B charge heterogeneity, suggesting that CFB has both process and product quality benefits. We projected CFB output gains in the context of a 2000-L small-scale facility, but the output was lower than that of a 15,000-L large-scale TFB facility. CFB's high cell mass also complicated operations, eroded volumetric productivity, and showed our current processes require significant improvements in specific productivity in order to realize their full potential and savings in manufacturing. Thus, improving specific productivity can resolve CFB's cost, scale-up, and operability challenges. Copyright © 2015 Elsevier B.V. All rights reserved.

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

    DEFF Research Database (Denmark)

    Fan, Yuzhou; Jimenez Del Val, Ioscani; Müller, Christian

    2015-01-01

    optimization, especially media optimization. Gaining knowledge on their interrelations could provide insight for obtaining higher immunoglobulin G (IgG) titer and better controlling glycosylationrelated product quality. In this work, different fed-batch processes with two chemically defined proprietary media......Fed-batch Chinese hamster ovary (CHO) cell culture is the most commonly used process for IgG production in the biopharmaceutical industry. Amino acid and glucose consumption, cell growth, metabolism, antibody titer, and N-glycosylation patterns are always the major concerns during upstream process...... and glutamine concentrations and uptake rates were positively correlated with intracellular UDP-Gal availability. All these findings are important for optimization of fed-batch culture for improving IgG production and directing glycosylation quality....

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

    Science.gov (United States)

    Wannawilai, Siwaporn; Sirisansaneeyakul, Sarote; Chisti, Yusuf

    2015-01-20

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

  1. Characterization of starter kimchi fermented with Leuconostoc kimchii GJ2 and its cholesterol-lowering effects in rats fed a high-fat and high-cholesterol diet.

    Science.gov (United States)

    Jo, Se Yeon; Choi, Eun A; Lee, Jae Joon; Chang, Hae Choon

    2015-10-01

    The hypocholesterolemic effects of lactic acid bacteria and kimchi have been demonstrated previously. However, the kimchi fermentation process still relies on naturally present microorganisms. To obtain functional kimchi with consistent quality, we validated the capacity of Leuconostoc kimchii GJ2 as a starter culture to control kimchi fermentation. Moreover, cholesterol-lowering effects of starter kimchi as a health-promoting product were explored. Bacteriocin production by Lc. kimchii GJ2 was highly enhanced in the presence of 5% Lactobacillus sakei NJ1 cell fractions. When kimchi was fermented with bacteriocin-enhanced Lc. kimchii GJ2, Lc. kimchii GJ2 became overwhelmingly predominant (98.3%) at the end of fermentation and maintained its dominance (up to 82%) for 84 days. Growing as well as dead cells of Lc. kimchii GJ2 showed high cholesterol assimilation (in vitro). Rats were fed a high-fat and high-cholesterol diet supplemented with starter kimchi. The results showed that feeding of starter kimchi significantly reduced serum total cholesterol, triglyceride and low-density lipoprotein cholesterol levels. Additionally, atherogenic index, cardiac risk factor and triglyceride and total cholesterol levels in liver and epididymal adipose tissue decreased significantly in rats fed starter kimchi. Kimchi fermented with Lc. kimchii GJ2 as a starter culture has efficient cholesterol-lowering effects. © 2014 Society of Chemical Industry.

  2. COMPARISON OF VACUUM AND HIGH PRESSURE EVAPORATED WOOD HYDROLYZATE FOR ETHANOL PRODUCTION BY REPEATED FED-BATCH USING FLOCCULATING SACCHAROMYCES CEREVISIAE

    Directory of Open Access Journals (Sweden)

    Anahita Dehkhoda

    2009-02-01

    Full Text Available With the aim of increasing the sugars concentration in dilute-acid ligno-cellulosic hydrolyzate to more than 100 g/l for industrial applications, the hydrolyzate from spruce was concentrated about threefold by high-pressure or vacuum evaporations. It was then fermented by repeated fed-batch cultivation using flocculating Saccharomyces cerevisiae with no prior detoxification. The sugars and inhibitors concentrations in the hydrolyzates were compared after the evaporations and also fermenta-tion. The evaporations were carried out either under vacuum (VEH at 0.5 bar and 80°C or with 1.3 bar pressure (HPEH at 107.5°C, which resulted in 153.3 and 164.6 g/l total sugars, respectively. No sugar decomposition occurred during either of the evaporations, while more than 96% of furfural and to a lesser extent formic and acetic acids disappeared from the hydrolyzates. However, HMF and levulinic acid remained in the hydrolyzates and were concentrated proportionally. The concentrated hydrolyzates were then fermented in a 4 l bioreactor with 12-22 g/l yeast and 0.14-0.22 h-1 initial dilute rates (ID. More than 84% of the fermentable sugars present in the VEH were fermented by fed-batch cultivation using 12 g/l yeast and initial dilution rate (ID of 0.22 h-1, and resulted in 0.40±0.01 g/g ethanol from the fermentable sugars in one cycle of fermentation. Fermentation of HPEH was as successful as VEH and resulted in more than 86% of the sugar consumption under the corresponding conditions. By lowering the initial dilution rate to 0.14 h-1, more than 97% of the total fermentable sugars were consumed, and ethanol yield was 0.44±0.01 g/g in one cycle of fermentation. The yeast was able to convert or assimilate HMF, levulinic, acetic, and formic acids by 96, 30, 43, and 74%, respectively.

  3. Fed-Batch Biomolecule Production by Bacillus subtilis: A State of the Art Review.

    Science.gov (United States)

    Ÿztürk, Sibel; Ÿalık, Pınar; Ÿzdamar, Tunçer H

    2016-04-01

    Bacillus subtilis is a highly promising production system for various biomolecules. This review begins with the algorithm of fed-batch operations (FBOs) and then illustrates the approaches to design the initial production medium and/or feed stream. Additionally, the feeding strategies developed with or without feedback control for fed-batch B. subtilis fermentations were compiled with a special emphasis on recombinant protein (r-protein) production. For biomolecule production by wild-type B. subtilis, due to the different intracellular production patterns, no consensus exists on the FBO strategy that gives the maximum productivity, whereas for r-protein production appropriate feeding strategies vary depending on the promoter used. Thus, we conclude that the B. subtilis community is still seeking an approved strong promoter and generalized FBO strategies. Copyright © 2015 Elsevier Ltd. All rights reserved.

  4. Batch-batch stable microbial community in the traditional fermentation process of huyumei broad bean pastes.

    Science.gov (United States)

    Zhu, Linjiang; Fan, Zihao; Kuai, Hui; Li, Qi

    2017-09-01

    During natural fermentation processes, a characteristic microbial community structure (MCS) is naturally formed, and it is interesting to know about its batch-batch stability. This issue was explored in a traditional semi-solid-state fermentation process of huyumei, a Chinese broad bean paste product. The results showed that this MCS mainly contained four aerobic Bacillus species (8 log CFU per g), including B. subtilis, B. amyloliquefaciens, B. methylotrophicus, and B. tequilensis, and the facultative anaerobe B. cereus with a low concentration (4 log CFU per g), besides a very small amount of the yeast Zygosaccharomyces rouxii (2 log CFU per g). The dynamic change of the MCS in the brine fermentation process showed that the abundance of dominant species varied within a small range, and in the beginning of process the growth of lactic acid bacteria was inhibited and Staphylococcus spp. lost its viability. Also, the MCS and its dynamic change were proved to be highly reproducible among seven batches of fermentation. Therefore, the MCS naturally and stably forms between different batches of the traditional semi-solid-state fermentation of huyumei. Revealing microbial community structure and its batch-batch stability is helpful for understanding the mechanisms of community formation and flavour production in a traditional fermentation. This issue in a traditional semi-solid-state fermentation of huyumei broad bean paste was firstly explored. This fermentation process was revealed to be dominated by a high concentration of four aerobic species of Bacillus, a low concentration of B. cereus and a small amount of Zygosaccharomyces rouxii. Lactic acid bacteria and Staphylococcus spp. lost its viability at the beginning of fermentation. Such the community structure was proved to be highly reproducible among seven batches. © 2017 The Society for Applied Microbiology.

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

    Science.gov (United States)

    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.

  6. Efficient soluble expression of disulfide bonded proteins in the cytoplasm of Escherichia coli in fed-batch fermentations on chemically defined minimal media.

    Science.gov (United States)

    Gąciarz, Anna; Khatri, Narendar Kumar; Velez-Suberbie, M Lourdes; Saaranen, Mirva J; Uchida, Yuko; Keshavarz-Moore, Eli; Ruddock, Lloyd W

    2017-06-15

    The production of recombinant proteins containing disulfide bonds in Escherichia coli is challenging. In most cases the protein of interest needs to be either targeted to the oxidizing periplasm or expressed in the cytoplasm in the form of inclusion bodies, then solubilized and re-folded in vitro. Both of these approaches have limitations. Previously we showed that soluble expression of disulfide bonded proteins in the cytoplasm of E. coli is possible at shake flask scale with a system, known as CyDisCo, which is based on co-expression of a protein of interest along with a sulfhydryl oxidase and a disulfide bond isomerase. With CyDisCo it is possible to produce disulfide bonded proteins in the presence of intact reducing pathways in the cytoplasm. Here we scaled up production of four disulfide bonded proteins to stirred tank bioreactors and achieved high cell densities and protein yields in glucose fed-batch fermentations, using an E. coli strain (BW25113) with the cytoplasmic reducing pathways intact. Even without process optimization production of purified human single chain IgA 1 antibody fragment reached 139 mg/L and hen avidin 71 mg/L, while purified yields of human growth hormone 1 and interleukin 6 were around 1 g/L. Preliminary results show that human growth hormone 1 was also efficiently produced in fermentations of W3110 strain and when glucose was replaced with glycerol as the carbon source. Our results show for the first time that efficient production of high yields of soluble disulfide bonded proteins in the cytoplasm of E. coli with the reducing pathways intact is feasible to scale-up to bioreactor cultivations on chemically defined minimal media.

  7. Perfusion seed cultures improve biopharmaceutical fed-batch production capacity and product quality.

    Science.gov (United States)

    Yang, William C; Lu, Jiuyi; Kwiatkowski, Chris; Yuan, Hang; Kshirsagar, Rashmi; Ryll, Thomas; Huang, Yao-Ming

    2014-01-01

    Volumetric productivity and product quality are two key performance indicators for any biopharmaceutical cell culture process. In this work, we showed proof-of-concept for improving both through the use of alternating tangential flow perfusion seed cultures coupled with high-seed fed-batch production cultures. First, we optimized the perfusion N-1 stage, the seed train bioreactor stage immediately prior to the production bioreactor stage, to minimize the consumption of perfusion media for one CHO cell line and then successfully applied the optimized perfusion process to a different CHO cell line. Exponential growth was observed throughout the N-1 duration, reaching >40 × 10(6) vc/mL at the end of the perfusion N-1 stage. The cultures were subsequently split into high-seed (10 × 10(6) vc/mL) fed-batch production cultures. This strategy significantly shortened the culture duration. The high-seed fed-batch production processes for cell lines A and B reached 5 g/L titer in 12 days, while their respective low-seed processes reached the same titer in 17 days. The shortened production culture duration potentially generates a 30% increase in manufacturing capacity while yielding comparable product quality. When perfusion N-1 and high-seed fed-batch production were applied to cell line C, higher levels of the active protein were obtained, compared to the low-seed process. This, combined with correspondingly lower levels of the inactive species, can enhance the overall process yield for the active species. Using three different CHO cell lines, we showed that perfusion seed cultures can optimize capacity utilization and improve process efficiency by increasing volumetric productivity while maintaining or improving product quality. © 2014 American Institute of Chemical Engineers.

  8. Development of bioprocess for high density cultivation yield of the probiotic Bacillus coagulans and its spores

    Directory of Open Access Journals (Sweden)

    Kavita R. Pandey

    2016-09-01

    Full Text Available Bacillus coagulans is a spore forming lactic acid bacterium. Spore forming bacteria, have been extensively studied and commercialized as probiotics. Probiotics are produced by fermentation technology. There is a limitation to biomass produced by conventional modes of fermentation. With the great demand generated by range of probiotic products, biomass is becoming very valuable for several pharmaceutical, dairy and probiotic companies. Thus, there is a need to develop high cell density cultivation processes for enhanced biomass accumulation. The bioprocess development was carried out in 6.6 L bench top lab scale fermentor. Four different cultivation strategies were employed to develop a bioprocess for higher growth and sporulation efficiencies of probiotic B. coagulans. Batch fermentation of B. coagulans yielded 18 g L-1 biomass (as against 8.0 g L-1 productivity in shake flask with 60% spore efficiency. Fed-batch cultivation was carried out for glucose, which yielded 25 g L-1 of biomass. C/N ratio was very crucial in achieving higher spore titres. Maximum biomass yield recorded was 30 g L-1, corresponding to 3.8 × 1011 cells mL-1 with 81% of cells in sporulated stage. The yield represents increment of 85 times the productivity and 158 times the spore titres relative to the highest reported values for high density cultivation of B. coagulans.

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

    Science.gov (United States)

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

    2015-06-12

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

  10. Fed-batch production of concentrated fructose syrup and ethanol using Saccharomyces cerevisiae ATCC 36859

    Energy Technology Data Exchange (ETDEWEB)

    Koren, D W [CANMET, Ottawa, ON (Canada); Duvnjak, Z [Univ. of Ottawa, ON (Canada). Dept. of Chemical Engineering

    1992-01-01

    A fed-batch process is used for the production of concentrated pure fructose syrup and ethanol from various glucose/fructose mixtures by S.cerevisiae ATCC 36859. Applying this technique, glucose-free fructose syrups with over 250 g/l of this sugar were obtained using High Fructose Corn Syrup and hydrolyzed Jerusalem artichoke juice. Bey encouraging ethanol evaporation from the reactor and condensing it, a separate ethanol product with a concentration of up to 350 g/l was also produced. The rates of glucose consumption and ethanol production were higher than in classical batch ethanol fermentation processes. (orig.).

  11. Ethanol production potential from fermented rice noodle wastewater treatment using entrapped yeast cell sequencing batch reactor

    Science.gov (United States)

    Siripattanakul-Ratpukdi, Sumana

    2012-03-01

    Fermented rice noodle production generates a large volume of starch-based wastewater. This study investigated the treatment of the fermented rice noodle wastewater using entrapped cell sequencing batch reactor (ECSBR) compared to traditional sequencing batch reactor (SBR). The yeast cells were applied because of their potential to convert reducing sugar in the wastewater to ethanol. In present study, preliminary treatment by acid hydrolysis was performed. A yeast culture, Saccharomyces cerevisiae, with calcium alginate cell entrapment was used. Optimum yeast cell loading in batch experiment and fermented rice noodle treatment performances using ECSBR and SBR systems were examined. In the first part, it was found that the cell loadings (0.6-2.7 × 108 cells/mL) did not play an important role in this study. Treatment reactions followed the second-order kinetics with the treatment efficiencies of 92-95%. In the second part, the result showed that ECSBR performed better than SBR in both treatment efficiency and system stability perspectives. ECSBR maintained glucose removal of 82.5 ± 10% for 5-cycle treatment while glucose removal by SBR declined from 96 to 40% within the 5-cycle treatment. Scanning electron microscopic images supported the treatment results. A number of yeast cells entrapped and attached onto the matrix grew in the entrapment matrix.

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

    Science.gov (United States)

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

    2014-01-01

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

  13. Fed-batch bioreactor performance and cell line stability evaluation of the artificial chromosome expression technology expressing an IgG1 in Chinese hamster ovary cells.

    Science.gov (United States)

    Combs, Rodney G; Yu, Erwin; Roe, Susanna; Piatchek, Michele Bailey; Jones, Heather L; Mott, John; Kennard, Malcolm L; Goosney, Danika L; Monteith, Diane

    2011-01-01

    The artificial chromosome expression (ACE) technology system uses an engineered artificial chromosome containing multiple site-specific recombination acceptor sites for the rapid and efficient construction of stable cell lines. The construction of Chinese hamster ovary(CHO) cell lines expressing an IgG1 monoclonal antibody (MAb) using the ACE system has been previously described (Kennard et al., Biotechnol Bioeng. 2009;104:540-553). To further demonstrate the manufacturing feasibility of the ACE system, four CHO cell lines expressing the human IgG1 MAb 4A1 were evaluated in batch and fed-batch shake flasks and in a 2-L fed-batch bioreactor. The batch shake flasks achieved titers between 0.7 and 1.1 g/L, whereas the fed-batch shake flask process improved titers to 2.5–3.0 g/L. The lead 4A1 ACE cell line achieved titers of 4.0 g/L with an average specific productivity of 40 pg/(cell day) when cultured in a non optimized 2-L fed-batch bioreactor using a completely chemically defined process. Generational stability characterization of the lead 4A1-expressing cell line demonstrated that the cell line was stable for up to 75 days in culture. Product quality attributes of the 4A1 MAb produced by the ACE system during the stability evaluation period were unchanged and also comparable to existing expression technologies such as the CHO-dhfr system. The results of this evaluation demonstrate that a clonal, stable MAb-expressing CHO cell line can be produced using ACE technology that performs competitively using a chemically defined fed-batch bioreactor process with comparable product quality attributes to cell lines generated by existing technologies.

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

    Science.gov (United States)

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

    2015-10-10

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

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

    Science.gov (United States)

    Wang, Juan; Sun, Qingying; Feng, Enmin

    2012-11-01

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

  16. Targeting population heterogeneity in Saccharomyces cerevisiae batch fermentation for optimal cell factories

    DEFF Research Database (Denmark)

    Heins, Anna-Lena; Lencastre Fernandes, Rita; Lundin, L.

    )). Significant gradients of e.g. dissolved oxygen, substrates, and pH are typically observed in many industrial scale fermentation processes. Consequently, the microbial cells experience rapid changes in environmental conditions as they circulate throughout the reactor, which might pose stress on the cells...... and affect their metabolism and consequently affect the heterogeneity level of the population. To further investigate these phenomena and gain a deeper understanding of population heterogeneity, Saccharomyces cerevisiae growth reporter strains based on the expression of green fluorescent protein (GFP) were...... environmental factors on heterogeneity level and amount of living cells. A highly dynamic behavior with regard to subpopulation distribution during the different growth stages was seen for the batch cultivations. Moreover, it could be demonstrated that the glucose concentration had a clear influence...

  17. Secretory expression of functional barley limit dextrinase by Pichia pastoris using high cell-density fermentation

    DEFF Research Database (Denmark)

    Vester-Christensen, Malene Bech; Abou Hachem, Maher; Næsted, Henrik

    2010-01-01

    biosynthesis by trimming of intermediate branched alpha-glucan structures. Highly active barley LD is obtained by secretory expression during high cell-density fermentation of Pichia pastoris. The LD encoding gene fragment without signal peptide was subcloned in-frame with the Saccharomyces cerevisiae alpha...

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    Gahlawat, Geeta; Srivastava, Ashok K

    2013-06-01

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

  20. Vapor-fed bio-hybrid fuel cell.

    Science.gov (United States)

    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

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

    Science.gov (United States)

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

    2012-10-01

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

  2. Degradation of toluene and trichloroethylene by Burkholderia cepacia G4 in growth-limited fed-batch culture

    NARCIS (Netherlands)

    Mars, Astrid E.; Houwing, Joukje; Dolfing, Jan; Janssen, Dick B.

    Burkholderia (Pseudomonas) cepacia G4 was cultivated in a fed-batch bioreactor on either toluene or toluene plus trichloroethylene (TCE), The culture was allowed to reach a constant cell density under conditions in which the amount of toluene supplied equals the maintenance energy demand of the

  3. Plasmid fermentation process for DNA immunization applications.

    Science.gov (United States)

    Carnes, Aaron E; Williams, James A

    2014-01-01

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

  4. Effects of pre-fermentation and pulsed-electric-field treatment of primary sludge in microbial electrochemical cells.

    Science.gov (United States)

    Ki, Dongwon; Parameswaran, Prathap; Popat, Sudeep C; Rittmann, Bruce E; Torres, César I

    2015-11-01

    The aim of this study was to investigate the combination of two technologies - pulsed electric field (PEF) pre-treatment and semi-continuous pre-fermentation of primary sludge (PS) - to produce volatile fatty acids (VFAs) as the electron donor for microbial electrolysis cells (MECs). Pre-fermentation with a 3-day solids retention time (SRT) led to the maximum generation of VFAs, with or without pretreatment of the PS through pulsed-electric-fields (PEF). PEF treatment before fermentation enhanced the accumulation of the preferred VFA, acetate, by 2.6-fold. Correspondingly, MEC anodes fed with centrate from 3-day pre-fermentation of PEF-treated PS had a maximum current density ∼3.1 A/m(2), which was 2.4-fold greater than the control pre-fermented centrate. Over the full duration of batch MEC experiments, using pre-fermented centrate led to successful performance in terms of Coulombic efficiency (95%), Coulombic recovery (80%), and COD-removal efficiency (85%). Copyright © 2015 Elsevier Ltd. All rights reserved.

  5. Optimal Control of a Fed-Batch Fermentation Involving Multiple Feeds

    Directory of Open Access Journals (Sweden)

    Chongyang Liu

    2012-01-01

    Full Text Available A nonlinear dynamical system, in which the feed rates of glycerol and alkali are taken as the control functions, is first proposed to formulate the fed-batch culture of 1,3-propanediol (1,3-PD production. To maximize the 1,3-PD concentration at the terminal time, a constrained optimal control model is then presented. A solution approach is developed to seek the optimal feed rates based on control vector parametrization method and improved differential evolution algorithm. The proposed methodology yielded an increase by 32.17% of 1,3-PD concentration at the terminal time.

  6. High-concentration sugars production from corn stover based on combined pretreatments and fed-batch process.

    Science.gov (United States)

    Yang, Maohua; Li, Wangliang; Liu, Binbin; Li, Qiang; Xing, Jianmin

    2010-07-01

    In this paper, high-concentration sugars were produced from pretreated corn stover. The raw corn stover was pretreated in a process combining steam explosion and alkaline hydrogen-peroxide. The hemicellulose and lignin were removed greatly. The cellulose content increased to 73.2%. Fed-batch enzymatic hydrolysis was initiated with 12% (w/v) solids loading and 20 FPU/g solids. Then, 6% solids were fed consecutively at 12, 36 and 60 h. After 144 h, the final concentrations of reducing sugar, glucose, cellobiose and xylose reached 220, 175, 22 and 20 g/L, respectively. The final total biomass conversion was 60% in fed-batch process. Copyright 2009 Elsevier Ltd. All rights reserved.

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

    Science.gov (United States)

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

    2015-03-01

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

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

    Science.gov (United States)

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

    2015-07-01

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

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

    Science.gov (United States)

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

    2014-01-01

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

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

    Science.gov (United States)

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

    2017-01-01

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

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

    Science.gov (United States)

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

    2015-03-01

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

  12. Mechanistic Models for Process Development and Optimization of Fed-batch Fermentation Systems

    DEFF Research Database (Denmark)

    Mears, Lisa; Stocks, Stuart M.; Albæk, Mads O.

    2016-01-01

    This work discusses the application of mechanistic models to pilot scale filamentous fungal fermentation systems operated at Novozymes A/S. For on-line applications, a state estimator model is developed based on a stoichiometric balance in order to predict the biomass and product concentration....... This is based on on-line gas measurements and ammonia addition flow rate measurements. Additionally, a mechanistic model is applied offline as a tool for batch planning, based on definition of the process back pressure, aeration rate and stirrer speed. This allows the batch starting fill to be planned, taking...... into account the oxygen transfer conditions, as well as the evaporation rates of the system. Mechanistic models are valuable tools which are applicable for both process development and optimization. The state estimator described will be a valuable tool for future work as part of control strategy development...

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

    Science.gov (United States)

    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.

  14. Enhancing Ethanol Production by Fermentation Using Saccharomyces cereviseae under Vacuum Condition in Batch Operation

    Directory of Open Access Journals (Sweden)

    A Abdullah

    2012-02-01

    Full Text Available Ethanol is one of renewable energy, which considered being an excellent alternative clean-burning fuel to replaced gasoline. In fact, the application of ethanol as fuel still blended with gasoline. The advantages of using ethanol as fuel are that the raw material mostly from renewable resources and the product has low emission which means environmental friendly. Ethanol can be produced by fermentation of sugars (glucose/fructose. The constraint in the ethanol fermentation batch or continuous process is the ethanol product inhibition. Inhibition in ethanol productivity and cell growth can be overcome by taking the product continuously from the fermentor. The process can be done by using a vacuum fermentation. The objective of this research is to investigate the effect of pressure and glucose concentration in ethanol fermentation. The research was conducted in laboratory scale and batch process. Equipment consists of fermentor with vacuum system. The observed responses were dried cells of yeast, concentration of glucose, and concentration of ethanol. Observations were made every 4 hours during a day of experiment. The results show that the formation of ethanol has a growth-associated product characteristic under vacuum operation. Vacuum condition can increase the cell formation productivity and the ethanol formation, as it is compared with fermentation under atmospheric condition. The maximum cells productivity and ethanol formation in batch operation under vacuum condition was reached at 166.6 mmHg of pressure. The maximum numbers of cells and ethanol formation was reached at 141.2 mm Hg of pressure. High initial glucose concentration significantly can affect the productivity and the yield of ethanol.

  15. High cell density cultivation of Escherichia coli K4 in a microfiltration bioreactor: a step towards improvement of chondroitin precursor production

    Directory of Open Access Journals (Sweden)

    De Rosa Mario

    2011-02-01

    Full Text Available Abstract Background The bacteria Escherichia coli K4 produces a capsular polysaccharide (K4 CPS whose backbone is similar to the non sulphated chondroitin chain. The chondroitin sulphate is one of the major components of the extra-cellular matrix of the vertebrate connective tissues and a high value molecule, widely employed as active principle in the treatment of osteoarthritis. It is usually obtained by extraction from animal tissues, but the risk of virus contaminations, as well as the scarceness of raw material, makes this productive process unsafe and unable to satisfy the growing market demand. In previous studies a new biotechnological process to produce chondroitin from Escherichia coli K4 capsular polysaccharide was investigated and a 1.4 g·L-1 K4 CPS concentration was reached using fed-batch fermentation techniques. In this work, on the trail of these results, we exploited new fermentation strategies to further improve the capsular polysaccharide production. Results The inhibitory effect of acetate on the bacterial cells growth and K4 CPS production was studied in shake flask conditions, while a new approach, that combined the optimization of the feeding profiles, the improvement of aeration conditions and the use of a microfiltration bioreactor, was investigated in three different types of fermentation processes. High polysaccharide concentrations (4.73 ± 0.2 g·L-1, with corresponding average yields (0.13 ± 0.006 gK4 CPS·gcdw-1, were obtained; the increase of K4 CPS titre, compared to batch and fed-batch results, was of 16-fold and 3.3-fold respectively, while average yield was almost 3.5 and 1.4 fold higher. Conclusion The increase of capsular polysaccharide titre confirmed the validity of the proposed fermentation strategy and opened the way to the use of the microfiltration bioreactor for the biotechnological production of chondroitin.

  16. Biodegradable poly-ε-caprolactone microcarriers for efficient production of human mesenchymal stromal cells and secreted cytokines in batch and fed-batch bioreactors.

    Science.gov (United States)

    Lam, Alan Tin-Lun; Li, Jian; Toh, Jessica Pei-Wen; Sim, Eileen Jia-Hui; Chen, Allen Kuan-Liang; Chan, Jerry Kok-Yen; Choolani, Mahesh; Reuveny, Shaul; Birch, William R; Oh, Steve Kah-Weng

    2017-03-01

    Large numbers of human mesenchymal stromal cells (MSCs) used for a variety of applications in tissue engineering and cell therapy can be generated by scalable expansion in a bioreactor using microcarriers (MCs) systems. However, the enzymatic digestion process needed to detach cells from the growth surface can affect cell viability and potentially the potency and differentiation efficiency. Thus, the main aim of our study was to develop biocompatible and biodegradable MCs that can support high MSC yields while maintaining their differentiation capability and potency. After cell expansion, the cells that covered MCs can be directly implanted in vivo without the need for cell harvesting or use of scaffold. Poly-ε-caprolactone (PCL) is known as a biocompatible and biodegradable material. However, it cannot be used for generation of MCs because its high density (1.14 g/cm 3 ) would exclude its applicability for suspension MCs in stirred reactors. In this article, we describe expansion and potency of MSCs propagated on low-density (1.06 g/cm 3 ) porous PCL MCs coated with extracellular matrices (LPCLs) in suspended stirred reactors. Using these LPCLs, cell yields of about 4 × 10 4 cells/cm 2 and 7- to 10-fold increases were obtained using four different MSC lines (bone marrow, cord blood, fetal and Wharton's jelly). These yields were comparable with those obtained using non-degradable MCs (Cytodex 3) and higher than two-dimensional monolayer (MNL) cultures. A fed-batch process, which demonstrated faster cell expansion (4.5 × 10 4 cells/cm 2 in 5 days as compared with 7 days in batch culture) and about 70% reduction in growth media usage, was developed and scaled up from 100-mL spinner flask to 1-L controlled bioreactor. Surface marker expression, trilineage differentiation and clonogenic potential of the MSCs expanded on LPCL were not affected. Cytokine secretion kinetics, which occurred mostly during late logarithmic phase, was usually comparable with

  17. Microbial nar-GFP cell sensors reveal oxygen limitations in highly agitated and aerated laboratory-scale fermentors

    Directory of Open Access Journals (Sweden)

    Rao Govind

    2009-01-01

    Full Text Available Abstract Background Small-scale microbial fermentations are often assumed to be homogeneous, and oxygen limitation due to inadequate micromixing is often overlooked as a potential problem. To assess the relative degree of micromixing, and hence propensity for oxygen limitation, a new cellular oxygen sensor has been developed. The oxygen responsive E. coli nitrate reductase (nar promoter was used to construct an oxygen reporter plasmid (pNar-GFPuv which allows cell-based reporting of oxygen limitation. Because there are greater than 109 cells in a fermentor, one can outfit a vessel with more than 109 sensors. Our concept was tested in high density, lab-scale (5 L, fed-batch, E. coli fermentations operated with varied mixing efficiency – one verses four impellers. Results In both cases, bioreactors were maintained identically at greater than 80% dissolved oxygen (DO during batch phase and at approximately 20% DO during fed-batch phase. Trends for glucose consumption, biomass and DO showed nearly identical behavior. However, fermentations with only one impeller showed significantly higher GFPuv expression than those with four, indicating a higher degree of fluid segregation sufficient for cellular oxygen deprivation. As the characteristic time for GFPuv expression (approx 90 min. is much larger than that for mixing (approx 10 s, increased specific fluorescence represents an averaged effect of oxygen limitation over time and by natural extension, over space. Conclusion Thus, the pNar-GFPuv plasmid enabled bioreactor-wide oxygen sensing in that bacterial cells served as individual recirculating sensors integrating their responses over space and time. We envision cell-based oxygen sensors may find utility in a wide variety of bioprocessing applications.

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

    Science.gov (United States)

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

    2016-03-01

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

  19. Design of methanol Feed control in Pichia pastoris fermentations based upon a growth model.

    Science.gov (United States)

    Zhang, Wenhui; Smith, Leonard A; Plantz, Bradley A; Schlegel, Vicki L; Meagher, Michael M

    2002-01-01

    The methylotrophic yeast Pichia pastoris is an effective system for recombinant protein productions that utilizes methanol as an inducer, and also as carbon and energy source for a Mut(+) (methanol utilization plus) strain. Pichia fermentation is conducted in a fed-batch mode to obtain a high cell density for a high productivity. An accurate methanol control is required in the methanol fed-batch phase (induction phase) in the fermentation. A simple "on-off" control strategy is inadequate for precise control of methanol concentrations in the fermentor. In this paper we employed a PID (proportional, integral and derivative) control system for the methanol concentration control and designed the PID controller settings on the basis of a Pichia growth model. The closed-loop system was built with four components: PID controller, methanol feed pump, fermentation process, and methanol sensor. First, modeling and transfer functions for all components were derived, followed by frequency response analysis, a powerful method for calculating the optimal PID parameters K(c) (controller gain), tau(I) (controller integral time constant), and tau(D) (controller derivative time constant). Bode stability criteria were used to develop the stability diagram for evaluating the designed settings during the entire methanol fed-batch phase. Fermentations were conducted using four Pichia strains, each expressing a different protein, to verify the control performance with optimal PID settings. The results showed that the methanol concentration matched the set point very well with only small overshoot when the set point was switched, which indicated that a very good control performance was achieved. The method developed in this paper is robust and can serve as a framework for the design of other PID feedback control systems in biological processes.

  20. Fed-batch culture for the direct conversion of cellulosic substrates to acetic acid/ethanol by Fusarium oxysporum

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, P.K.R.; Singh, A.; Schuegerl, K. (Hannover Univ. (Germany). Inst. fuer Technische Chemie)

    1991-01-01

    The production of acetic acid/ethanol and hydrolytic enzymes from potato waste (cellulosic waste from potato starch industries) by Fusarium oxysporum 841 was improved considerably by using fed-batch culture. In this, two types of feed policies were adopted consisting of different substrate concentrations and feeding times. In fed-batch culture, the enzymes tested, namely avicelase, CMCase, cellobiase and xylanase, showed significant improvements over batch fermentations with regard to enzyme titres and productivities. The maximum concentration, yield and productivity of acetic acid were 22.5 g litre{sup -1}, 0.38 g (g {sub strate}){sup -1} and 0.09 g litre{sup -1} h{sup -1}, respectively, and these values for ethanol were 5.7 g litre{sup -1}, 0.1 g (g substrate){sup -1} and 0.03 g litre{sup -1}h{sup -1}, respectively. (author).

  1. Batch-to-batch uniformity of bacterial community succession and flavor formation in the fermentation of Zhenjiang aromatic vinegar.

    Science.gov (United States)

    Wang, Zong-Min; Lu, Zhen-Ming; Yu, Yong-Jian; Li, Guo-Quan; Shi, Jin-Song; Xu, Zheng-Hong

    2015-09-01

    Solid-state fermentation of traditional Chinese vinegar is a mixed-culture refreshment process that proceeds for many centuries without spoilage. Here, we investigated bacterial community succession and flavor formation in three batches of Zhenjiang aromatic vinegar using pyrosequencing and metabolomics approaches. Temporal patterns of bacterial succession in the Pei (solid-state vinegar culture) showed no significant difference (P > 0.05) among three batches of fermentation. In all the batches investigated, the average number of community operational taxonomic units (OTUs) decreased dramatically from 119 ± 11 on day 1 to 48 ± 16 on day 3, and then maintained in the range of 61 ± 9 from day 5 to the end of fermentation. We confirmed that, within a batch of fermentation process, the patterns of bacterial diversity between the starter (took from the last batch of vinegar culture on day 7) and the Pei on day 7 were similar (90%). The relative abundance dynamics of two dominant members, Lactobacillus and Acetobacter, showed high correlation (coefficient as 0.90 and 0.98 respectively) among different batches. Furthermore, statistical analysis revealed dynamics of 16 main flavor metabolites were stable among different batches. The findings validate the batch-to-batch uniformity of bacterial community succession and flavor formation accounts for the quality of Zhenjiang aromatic vinegar. Based on our understanding, this is the first study helps to explain the rationality of age-old artistry from a scientific perspective. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Improved fed-batch production of high-purity PHB (poly-3 hydroxy butyrate) by Cupriavidus necator (MTCC 1472) from sucrose-based cheap substrates under response surface-optimized conditions.

    Science.gov (United States)

    Dey, Pinaki; Rangarajan, Vivek

    2017-10-01

    Experimental investigations were carried out for Cupriavidus necator (MTCC 1472)-based improved production of poly-3 hydroxy butyrate (PHB) through induced nitrogen limiting fed-batch cultivation strategies. Initially Plackett-Burman design and response surface methodology were implemented to optimize most influencing process parameters. With optimized process parameter values, continuous feeding strategies ware applied in a 5-l fermenter with table sugar concentration of 100 g/l, nitrogen concentration of 0.12 g/l for fed-batch fermentation with varying dilution rates of 0.02 and 0.046 1/h. To get enriched production of PHB, concentration of the sugar was further increased to 150 and 200 g/l in feeding. Maximum concentrations of PHB achieved were 22.35 and 23.07 g/l at those dilution rates when sugar concentration maintains at 200 g/l in feeding. At maximum concentration of PHB (23.07 g/l), productivity of 0.58 g/l h was achieved with maximum PHB accumulation efficiency up to 64% of the dry weight of biomass. High purity of PHB, close to medical grade was achieved after surfactant hypochlorite extraction method, and it was further confirmed by SEM, EDX, and XRD studies.

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

    Energy Technology Data Exchange (ETDEWEB)

    Huang, Y.; Yang, S.T. [Ohio State Univ., Columbus, OH (United States). Dept. of Chemical Engineering

    1998-11-20

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

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

    Science.gov (United States)

    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.

  5. A Single-Batch Fermentation System to Simulate Human Colonic Microbiota for High-Throughput Evaluation of Prebiotics

    Science.gov (United States)

    Sasaki, Daisuke; Fukuda, Itsuko; Tanaka, Kosei; Yoshida, Ken-ichi; Kondo, Akihiko; Osawa, Ro

    2016-01-01

    We devised a single-batch fermentation system to simulate human colonic microbiota from fecal samples, enabling the complex mixture of microorganisms to achieve densities of up to 1011 cells/mL in 24 h. 16S rRNA gene sequence analysis of bacteria grown in the system revealed that representatives of the major phyla, including Bacteroidetes, Firmicutes, and Actinobacteria, as well as overall species diversity, were consistent with those of the original feces. On the earlier stages of fermentation (up to 9 h), trace mixtures of acetate, lactate, and succinate were detectable; on the later stages (after 24 h), larger amounts of acetate accumulated along with some of propionate and butyrate. These patterns were similar to those observed in the original feces. Thus, this system could serve as a simple model to simulate the diversity as well as the metabolism of human colonic microbiota. Supplementation of the system with several prebiotic oligosaccharides (including fructo-, galacto-, isomalto-, and xylo-oligosaccharides; lactulose; and lactosucrose) resulted in an increased population in genus Bifidobacterium, concomitant with significant increases in acetate production. The results suggested that this fermentation system may be useful for in vitro, pre-clinical evaluation of the effects of prebiotics prior to testing in humans. PMID:27483470

  6. Optimization of simultaneous saccharification and fermentation conditions with amphipathic lignin derivatives for concentrated bioethanol production.

    Science.gov (United States)

    Cheng, Ningning; Koda, Keiichi; Tamai, Yutaka; Yamamoto, Yoko; Takasuka, Taichi E; Uraki, Yasumitsu

    2017-05-01

    Amphipathic lignin derivatives (A-LDs) prepared from the black liquor of soda pulping of Japanese cedar are strong accelerators for bioethanol production under a fed-batch simultaneous enzymatic saccharification and fermentation (SSF) process. To improve the bioethanol production concentration, conditions such as reaction temperature, stirring program, and A-LDs loadings were optimized in both small scale and large scale fed-batch SSF. The fed-batch SSF in the presence of 3.0g/L A-LDs at 38°C gave the maximum ethanol production and a high enzyme recovery rate. Furthermore, a jar-fermenter equipped with a powerful mechanical stirrer was designed for 1.5L-scale fed-batch SSF to achieve rigorous mixing during high substrate loading. Finally, the 1.5L fed-batch SSF with a substrate loading of 30% (w/v) produced a high ethanol concentration of 87.9g/L in the presence of A-LDs under optimized conditions. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  7. Facilitating control of fed-batch fermentation processes by monitoring the growth rates of saccharomyces cerevisiae

    NARCIS (Netherlands)

    Keulers, M.L.B.; Ariaans, L.J.J.M.; Soeterboek, R.; Giuseppin, M.

    1994-01-01

    In this paper we present a growth rate controller for a fed-batch bioprocess. An observer estimates the growth rate. The observer is based on knowledge about the stoichiometric relations of the process. Furthermore, the observer needs online measurements of the oxygen uptake rate and the

  8. Increasing the production of desulfurizing biocatalysts by means of fed - batch culture

    International Nuclear Information System (INIS)

    Berdugo, C I; Mena, J A; Acero, J R; Mogollon, L

    2001-01-01

    Over the past years, environmental regulations have driven a lot of effort for the development of new technologies for the upgrading of fossil fuels. Biotechnology offers an alternative way to process fossil fuels by means of a biodesulfurization technology where the production of the biocatalyst is one of the key topics. Traditionally, the production is carried out in batch culture where the maximum cellular concentration is restricted by inherent limitations of the culture type and the microorganism growth rate. This work addresses the production of two desulfurizing microorganisms: Rhodococcus erythropolis IGTS8 and gordona rubropertinctus ICP172 using fed-batch culture. Fed-batch cultures were conducted in a 12 L fermentor using ICP 4 medium containing glucose and DMSO as carbon and sulfur sources. As a result, cell concentration was increased 1.5 and 3 times with fed-batch cultures using constant and exponential flow respectively, achieving a maximum cell concentration of 7.3 g DCW/L of biocatalyst igts8 and 12.85 gGDCW/L of the new biocatalyst ICP172. Both biocatalysts presented biodesulfurization activity in a spiked matrix DBT/HXD and in diesel matrix with the detection of 2-HBP which is the end-product of DBT degradation pathway

  9. From Fed-batch to Continuous Enzymatic Biodiesel Production

    DEFF Research Database (Denmark)

    Price, Jason Anthony; Nordblad, Mathias; Woodley, John M.

    2015-01-01

    In this this paper, we use mechanistic modelling to guide the development of acontinuous enzymatic process that is performed as a fed-batch operation. In this workwe use the enzymatic biodiesel process as a case study. A mechanistic model developedin our previous work was used to determine...... measured components (triglycerides, diglycerides, monoglycerides, free fatty acid and fatty acid methyl esters(biodiesel)) much better than using fed-batch data alone given the smaller residuals. We also observe a reduction in the correlation between the parameters.The model was then used to predict that 5...... reactors are required (with a combined residence time of 30 hours) to reach a final biodiesel concentration within 2 % of the95.6 mass % achieved in a fed-batch operation, for 24 hours....

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

    Science.gov (United States)

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

    2010-01-01

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

  11. High Cell Density Process for Constitutive Production of a Recombinant Phytase in Thermotolerant Methylotrophic Yeast Ogataea thermomethanolica Using Table Sugar as Carbon Source.

    Science.gov (United States)

    Charoenrat, Theppanya; Antimanon, Sompot; Kocharin, Kanokarn; Tanapongpipat, Sutipa; Roongsawang, Niran

    2016-12-01

    The yeast Ogataea thermomethanolica has recently emerged as a potential host for heterologous protein expression at elevated temperature. To evaluate the feasibility of O. thermomethanolica as heterologous host in large-scale fermentation, constitutive production of fungal phytase was investigated in fed-batch fermentation. The effect of different temperatures, substrate feeding strategies, and carbon sources on phytase production was investigated. It was found that O. thermomethanolica can grow in the temperature up to 40 °C and optimal at 34 °C. However, the maximum phytase production was observed at 30 °C and slightly decreased at 34 °C. The DOT stat control was the most efficient feeding strategy to obtain high cell density and avoid by-product formation. The table sugar can be used as an alternative substrate for phytase production in O. thermomethanolica. The highest phytase activity (134 U/mL) was obtained from table sugar at 34 °C which was 20-fold higher than batch culture (5.7 U/mL). At a higher cultivation temperature of 38 °C, table sugar can be used as a low-cost substrate for the production of phytase which was expressed with an acceptable yield (85 U/mL). Lastly, the results from this study reveal the industrial favorable benefits of employing O. thermomethanolica as a host for heterologous protein production.

  12. Mechanistic simulation of batch acetone-butanol-ethanol (ABE) fermentation with in situ gas stripping using Aspen Plus™.

    Science.gov (United States)

    Darkwah, Kwabena; Nokes, Sue E; Seay, Jeffrey R; Knutson, Barbara L

    2018-05-22

    Process simulations of batch fermentations with in situ product separation traditionally decouple these interdependent steps by simulating a separate "steady state" continuous fermentation and separation units. In this study, an integrated batch fermentation and separation process was simulated for a model system of acetone-butanol-ethanol (ABE) fermentation with in situ gas stripping, such that the fermentation kinetics are linked in real-time to the gas stripping process. A time-dependent cell growth, substrate utilization, and product production is translated to an Aspen Plus batch reactor. This approach capitalizes on the phase equilibria calculations of Aspen Plus to predict the effect of stripping on the ABE fermentation kinetics. The product profiles of the integrated fermentation and separation are shown to be sensitive to gas flow rate, unlike separate steady state fermentation and separation simulations. This study demonstrates the importance of coupled fermentation and separation simulation approaches for the systematic analyses of unsteady state processes.

  13. Strategies for improving production performance of probiotic Pediococcus acidilactici viable cell by overcoming lactic acid inhibition.

    Science.gov (United States)

    Othman, Majdiah; Ariff, Arbakariya B; Wasoh, Helmi; Kapri, Mohd Rizal; Halim, Murni

    2017-11-27

    Lactic acid bacteria are industrially important microorganisms recognized for fermentative ability mostly in their probiotic benefits as well as lactic acid production for various applications. Fermentation conditions such as concentration of initial glucose in the culture, concentration of lactic acid accumulated in the culture, types of pH control strategy, types of aeration mode and different agitation speed had influenced the cultivation performance of batch fermentation of Pediococcus acidilactici. The maximum viable cell concentration obtained in constant fed-batch fermentation at a feeding rate of 0.015 L/h was 6.1 times higher with 1.6 times reduction in lactic acid accumulation compared to batch fermentation. Anion exchange resin, IRA 67 was found to have the highest selectivity towards lactic acid compared to other components studied. Fed-batch fermentation of P. acidilactici coupled with lactic acid removal system using IRA 67 resin showed 55.5 and 9.1 times of improvement in maximum viable cell concentration compared to fermentation without resin for batch and fed-batch mode respectively. The improvement of the P. acidilactici growth in the constant fed-batch fermentation indicated the use of minimal and simple process control equipment is an effective approach for reducing by-product inhibition. Further improvement in the cultivation performance of P. acidilactici in fed-bath fermentation with in situ addition of anion-exchange resin significantly helped to enhance the growth of P. acidilactici by reducing the inhibitory effect of lactic acid and thus increasing probiotic production.

  14. Mathematical modeling of lipase and protease production by Penicillium restrictum in a batch fermenter.

    Science.gov (United States)

    Freire, D M; Sant'Anna, G L; Alves, T L

    1999-01-01

    This work presents a mathematical model that describes time course variations of extracellular lipase and protease activities for the batch fermentation of the fungus Penicillium restrictum, a new and promising strain isolated from soil and wastes of a Brazilian babassu coconut oil industry. The fermentation process was modeled by an unstructured model, which considered the following dependent variables: cells, fat acid, dissolved oxygen concentrations, lipase and protease activities, and cell lysate concentration. The last variable represents the amount of cells that has been lysed by the shear stress and natural cell death. Proteases released to the medium, as consequence of this process, enhance lipase inactivation. The model is able to predict the effects of some operation variables such as air flow rate and agitation speed. The mathematical model was validated against batch-fermentation data obtained under several operating conditions. Because substrate concentration has antagonistic effects on lipase activity, a typical optimization scheme should be developed in order to minimize these deleterious effects while maximizing lipase activity.

  15. Combined age and segregated kinetic model for industrial-scale penicillin fed-batch cultivation

    NARCIS (Netherlands)

    Wang, Z.F.; Lauwerijssen, M.J.C.; Yuan, J.Q.

    2005-01-01

    This paper proposes a cell age model forPenicillium chrysogenum fed-batch cultivation to supply a qualitative insight into morphology-associated dynamics. The average ages of the segregated cell populations, such as growing cells, non-growing cells and intact productive cells, were estimated by this

  16. On-line monitoring of important organoleptic methyl-branched aldehydes during batch fermentation of starter culture Staphylococcus xylosus reveal new insight into their production in a model fermentation

    DEFF Research Database (Denmark)

    de Vos Petersen, Christian; Beck, Hans Christian; Lauritsen, Frants R

    2004-01-01

    A small fermentor (55 mL) was directly interfaced to a membrane inlet mass spectrometer for continuous on-line monitoring of oxygen and volatile metabolites during batch fermentations of the starter culture Staphylococcus xylosus. Using this technique, we were able to correlate production...... of a longer aerobic growth period. Growing S. xylosus under conditions resembling those in a fermented sausage revealed that NaCl (5%) increased aldehyde production considerably, whereas KNO(3) (0.03%) or NaNO(2) (0.03%) had little effect. A lowering of pH from 7.2 to 6.0 reduced cell density, but had a minor...

  17. Effect of auxotrophies on yeast performance in aerated fed-batch reactor

    Energy Technology Data Exchange (ETDEWEB)

    Landi, Carmine; Paciello, Lucia [Dept. Ingegneria Industriale, Universita di Salerno, Via Ponte Don Melillo, 84084 Fisciano, Salerno (Italy); Alteriis, Elisabetta de [Dept. Biologia Strutturale e Funzionale, Universita degli Studi di Napoli ' Federico II' , Via Cinthia, 80100 Napoli (Italy); Brambilla, Luca [Dept. Biotecnologie e Bioscienze, Universita Milano-Bicocca, Piazza della Scienza, 20126 Milano (Italy); Parascandola, Palma, E-mail: pparascandola@unisa.it [Dept. Ingegneria Industriale, Universita di Salerno, Via Ponte Don Melillo, 84084 Fisciano, Salerno (Italy)

    2011-10-28

    Highlights: Black-Right-Pointing-Pointer The paper contributes to fill the gap existing between the basic and applied research. Black-Right-Pointing-Pointer Mathematical model sheds light on the physiology of auxotrophic yeast strains. Black-Right-Pointing-Pointer Yeast behavior in fed-batch is influenced by biological and environmental determinants. Black-Right-Pointing-Pointer Process optimization would make possible the production of heterologous proteins which are not yet on the market. -- Abstract: A systematic investigation on the effects of auxotrophies on the performance of yeast in aerated fed-batch reactor was carried out. Six isogenic strains from the CEN.PK family of Saccharomyces cerevisiae, one prototroph and five auxotrophs, were grown in aerated fed-batch reactor using the same operative conditions and a proper nutritional supplementation. The performance of the strains, in terms of final biomass decreased with increasing the number of auxotrophies. Auxotrophy for leucine exerted a profound negative effect on the performance of the strains. Accumulation of reactive oxygen species (ROS) in the cells of the strain carrying four auxotrophies and its significant viability loss, were indicative of an oxidative stress response induced by exposure of cells to the environmental conditions. The mathematical model was fundamental to highlight how the carbon flux, depending on the number and type of auxotrophies, was diverted towards the production of increasingly large quantities of energy for maintenance.

  18. Effect of auxotrophies on yeast performance in aerated fed-batch reactor

    International Nuclear Information System (INIS)

    Landi, Carmine; Paciello, Lucia; Alteriis, Elisabetta de; Brambilla, Luca; Parascandola, Palma

    2011-01-01

    Highlights: ► The paper contributes to fill the gap existing between the basic and applied research. ► Mathematical model sheds light on the physiology of auxotrophic yeast strains. ► Yeast behavior in fed-batch is influenced by biological and environmental determinants. ► Process optimization would make possible the production of heterologous proteins which are not yet on the market. -- Abstract: A systematic investigation on the effects of auxotrophies on the performance of yeast in aerated fed-batch reactor was carried out. Six isogenic strains from the CEN.PK family of Saccharomyces cerevisiae, one prototroph and five auxotrophs, were grown in aerated fed-batch reactor using the same operative conditions and a proper nutritional supplementation. The performance of the strains, in terms of final biomass decreased with increasing the number of auxotrophies. Auxotrophy for leucine exerted a profound negative effect on the performance of the strains. Accumulation of reactive oxygen species (ROS) in the cells of the strain carrying four auxotrophies and its significant viability loss, were indicative of an oxidative stress response induced by exposure of cells to the environmental conditions. The mathematical model was fundamental to highlight how the carbon flux, depending on the number and type of auxotrophies, was diverted towards the production of increasingly large quantities of energy for maintenance.

  19. Performance of the auxotrophic Saccharomyces cerevisiae BY4741 as host for the production of IL-1β in aerated fed-batch reactor: role of ACA supplementation, strain viability, and maintenance energy

    Directory of Open Access Journals (Sweden)

    Zueco Jesus

    2009-12-01

    Full Text Available Abstract Background Saccharomyces cerevisiae BY4741 is an auxotrophic commonly used strain. In this work it has been used as host for the expression and secretion of human interleukin-1β (IL1β, using the cell wall protein Pir4 as fusion partner. To achieve high cell density and, consequently, high product yield, BY4741 [PIR4-IL1β] was cultured in an aerated fed-batch reactor, using a defined mineral medium supplemented with casamino acids as ACA (auxotrophy-complementing amino acid source. Also the S. cerevisiae mutant BY4741 Δyca1 [PIR4-IL1β], carrying the deletion of the YCA1 gene coding for a caspase-like protein involved in the apoptotic response, was cultured in aerated fed-batch reactor and compared to the parental strain, to test the effect of this mutation on strain robustness. Viability of the producer strains was examined during the runs and a mathematical model, which took into consideration the viable biomass present in the reactor and the glucose consumption for both growth and maintenance, was developed to describe and explain the time-course evolution of the process for both, the BY4741 parental and the BY4741 Δyca1 mutant strain. Results Our results show that the concentrations of ACA in the feeding solution, corresponding to those routinely used in the literature, are limiting for the growth of S. cerevisiae BY4741 [PIR4-IL1β] in fed-batch reactor. Even in the presence of a proper ACA supplementation, S. cerevisiae BY4741 [PIR4-IL1β] did not achieve a high cell density. The Δyca1 deletion did not have a beneficial effect on the overall performance of the strain, but it had a clear effect on its viability, which was not impaired during fed-batch operations, as shown by the kd value (0.0045 h-1, negligible if compared to that of the parental strain (0.028 h-1. However, independently of their robustness, both the parental and the Δyca1 mutant ceased to grow early during fed-batch runs, both strains using most of the

  20. Enhancing Ethanol Production by Fermentation Using Saccharomyces cereviseae under Vacuum Condition in Batch Operation

    Directory of Open Access Journals (Sweden)

    A Abdullah

    2012-04-01

    Full Text Available Ethanol is one of renewable energy, which considered being an excellent alternativeclean-burning fuel to replaced gasoline. In fact, the application of ethanol as fuel still blended withgasoline. The advantages of using ethanol as fuel are that the raw material mostly from renewableresources and the product has low emission which means environmental friendly. Ethanol can beproduced by fermentation of sugars (glucose/fructose. The constraint in the ethanol fermentationbatch or continuous process is the ethanol product inhibition. Inhibition in ethanol productivityand cell growth can be overcome by taking the product continuously from the fermentor. Theprocess can be done by using a vacuum fermentation. The objective of this research is toinvestigate the effect of pressure and glucose concentration in ethanol fermentation. The researchwas conducted in laboratory scale and batch process. Equipment consists of fermentor withvacuum system. The observed responses were dried cells of yeast, concentration of glucose, andconcentration of ethanol. Observations were made every 4 hours during a day of experiment. Theresults show that the formation of ethanol has a growth-associated product characteristic undervacuum operation. Vacuum condition can increase the cell formation productivity and the ethanolformation, as it is compared with fermentation under atmospheric condition. The maximum cellsproductivity and ethanol formation in batch operation under vacuum condition was reached at166.6 mmHg of pressure. The maximum numbers of cells and ethanol formation was reached at141.2 mm Hg of pressure. High initial glucose concentration significantly can affect the productivityand the yield of ethanol.

  1. Development of a two-stage feeding strategy based on the kind and level of feeding nutrients for improving fed-batch production of L-threonine by Escherichia coli.

    Science.gov (United States)

    Liu, Shuwen; Liang, Yong; Liu, Qian; Tao, Tongtong; Lai, Shujuan; Chen, Ning; Wen, Tingyi

    2013-01-01

    Fed-batch fermentation is the predominant method for industrial production of amino acids. In this study, we comprehensively investigated the effects of four kinds of feeding nutrients and developed an accurate optimization strategy for fed-batch production of L-threonine. The production of L-threonine was severely inhibited when cell growth ceased in the bath culture. Similarly, L-threonine production was also associated with cell growth in the carbon-, phosphate-, and sulfate-limited fed-batch cultures, but the accumulation of L-threonine was markedly increased because of the extended production time in the growth stage. Interestingly, auxotrophic amino acid (L-isoleucine)-limited feeding promoted L-threonine production over the non-growth phase. Metabolite analysis indicates that substantial production of acetate and glutamate and the resulting accumulation of ammonium may lead to the inhibition of L-threonine production. During the growth phase, the levels of L-isoleucine were accurately optimized by balancing cell growth and production with Pontryagin's maximum principle, basing on the relationship between the specific growth rate μ and specific production rate ρ. Furthermore, the depletion of L-isoleucine and phosphate at the end of the growth phase favored the synthesis of L-threonine in the subsequent non-growth phase. Combining the two-stage feeding profiles, the final L-threonine concentration and conversion rate were increased by 5.9- and 2.1-fold, respectively, compared to batch processes without feeding control. The identification of efficient feeding nutrient and the development of accurate feeding strategies provide potential guidelines for microbial production of amino acids.

  2. Coproduction of acetic acid and electricity by application of microbial fuel cell technology to vinegar fermentation.

    Science.gov (United States)

    Tanino, Takanori; Nara, Youhei; Tsujiguchi, Takuya; Ohshima, Takayuki

    2013-08-01

    The coproduction of a useful material and electricity via a novel application of microbial fuel cell (MFC) technology to oxidative fermentation was investigated. We focused on vinegar production, i.e., acetic acid fermentation, as an initial and model useful material that can be produced by oxidative fermentation in combination with MFC technology. The coproduction of acetic acid and electricity by applying MFC technology was successfully demonstrated by the simultaneous progress of acetic acid fermentation and electricity generation through a series of repeated batch fermentations. Although the production rate of acetic acid was very small, it increased with the number of repeated batch fermentations that were conducted. We obtained nearly identical (73.1%) or larger (89.9%) acetic acid yields than that typically achieved by aerated fermentation (75.8%). The open-cycle voltages measured before and after fermentation increased with the total fermentation time and reached a maximum value of 0.521 V prior to the third batch fermentation. The maximum current and power densities measured in this study (19.1 μA/cm² and 2.47 μW/cm², respectively) were obtained after the second batch fermentation. Copyright © 2013 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  3. Production of Medium Chain Length Polyhydroxyalkanoates From Oleic Acid Using Pseudomonas putida PGA1 by Fed Batch Culture

    Directory of Open Access Journals (Sweden)

    Sidik Marsudi

    2010-10-01

    Full Text Available Bacterial polyhydroxyalkanoates (PHAs are a class of p0lymers currently receiving much attention because of their potential as renewable and biodegradable plastics. A wide variety of bacteria has been reported to produce PHAs including Pseudomonas strains. These strains are known as versatile medium chain length PHAs (PHAs-mcl producers using fatty acids as carbon source. Oleic acid was used to produce PHAs-mcl using Pseudomonas putida PGA 1 by continuous feeding of both nitrogen and carbon source, in a fed batch culture. During cell growth, PHAs also accumulated, indicating that PHA production in this organism is growth associated. Residual cell increased until the nitrogen source was depleted. At the end of fermentation, final cell concentration, PHA content, and roductivity were 30.2 g/L, 44.8 % of cell dry weight, and 0.188 g/l/h, respectively.

  4. Glycoprofiling effects of media additives on IgG produced by CHO cells in fed-batch bioreactors

    DEFF Research Database (Denmark)

    Kildegaard, Helene Faustrup; Fan, Yuzhou; Wagtberg Sen, Jette

    2016-01-01

    Therapeutic monoclonal antibodies (mAbs) are mainly produced by heterogonous expression in Chinese hamster ovary (CHO) cells. The glycosylation profile of the mAbs has major impact on the efficacy and safety of the drug and is therefore an important parameter to control during production. In this......Therapeutic monoclonal antibodies (mAbs) are mainly produced by heterogonous expression in Chinese hamster ovary (CHO) cells. The glycosylation profile of the mAbs has major impact on the efficacy and safety of the drug and is therefore an important parameter to control during production....... In this study, the effect on IgG N-glycosylation from feeding CHO cells with eight glycosylation precursors during cultivation was investigated. The study was conducted in fed-batch mode in bioreactors with biological replicates to obtain highly controlled and comparable conditions. We assessed charge...

  5. Simulation of kefiran production of Lactobacillus kefiranofaciens JCM6985 in fed-batch reactor

    Directory of Open Access Journals (Sweden)

    Benjamas Cheirsilp

    2006-09-01

    Full Text Available Kinetics of kefiran production by Lactobacillus kefiranofaciens JCM6985 has been investigated. A mathematical model taking into account the mechanism of exopolysaccharides production has been developed. Experiments were carried out in batch mode in order to obtain kinetic model parameters that were further applied to simulate fed-batch processes. A simplification of parameter fitting was also introduced for complicated model. The fed-batch mode allows more flexibility in the control of the substrate concentration as well as product concentration in the culture medium. Based on the batch mathematical model, a fed-batch model was developed and simulations were done. Simulation study in fed-batch reactor resulted that substrate concentration should be controlled at 20 g L-1 to soften the product inhibition and also to stimulate utilization of substrate and its hydrolysate. From simulation results of different feeding techniques, it was found that constant feeding at 0.01 L h-1 was most practically effective feeding profile for exopolysaccharides production in fed-batch mode.

  6. Stochastic growth logistic model with aftereffect for batch fermentation process

    Energy Technology Data Exchange (ETDEWEB)

    Rosli, Norhayati; Ayoubi, Tawfiqullah [Faculty of Industrial Sciences and Technology, Universiti Malaysia Pahang, Lebuhraya Tun Razak, 26300 Gambang, Pahang (Malaysia); Bahar, Arifah; Rahman, Haliza Abdul [Department of Mathematical Sciences, Faculty of Science, Universiti Teknologi Malaysia, 81310 Johor Bahru, Johor (Malaysia); Salleh, Madihah Md [Department of Biotechnology Industry, Faculty of Biosciences and Bioengineering, Universiti Teknologi Malaysia, 81310 Johor Bahru, Johor (Malaysia)

    2014-06-19

    In this paper, the stochastic growth logistic model with aftereffect for the cell growth of C. acetobutylicum P262 and Luedeking-Piret equations for solvent production in batch fermentation system is introduced. The parameters values of the mathematical models are estimated via Levenberg-Marquardt optimization method of non-linear least squares. We apply Milstein scheme for solving the stochastic models numerically. The effciency of mathematical models is measured by comparing the simulated result and the experimental data of the microbial growth and solvent production in batch system. Low values of Root Mean-Square Error (RMSE) of stochastic models with aftereffect indicate good fits.

  7. Stochastic growth logistic model with aftereffect for batch fermentation process

    Science.gov (United States)

    Rosli, Norhayati; Ayoubi, Tawfiqullah; Bahar, Arifah; Rahman, Haliza Abdul; Salleh, Madihah Md

    2014-06-01

    In this paper, the stochastic growth logistic model with aftereffect for the cell growth of C. acetobutylicum P262 and Luedeking-Piret equations for solvent production in batch fermentation system is introduced. The parameters values of the mathematical models are estimated via Levenberg-Marquardt optimization method of non-linear least squares. We apply Milstein scheme for solving the stochastic models numerically. The effciency of mathematical models is measured by comparing the simulated result and the experimental data of the microbial growth and solvent production in batch system. Low values of Root Mean-Square Error (RMSE) of stochastic models with aftereffect indicate good fits.

  8. Stochastic growth logistic model with aftereffect for batch fermentation process

    International Nuclear Information System (INIS)

    Rosli, Norhayati; Ayoubi, Tawfiqullah; Bahar, Arifah; Rahman, Haliza Abdul; Salleh, Madihah Md

    2014-01-01

    In this paper, the stochastic growth logistic model with aftereffect for the cell growth of C. acetobutylicum P262 and Luedeking-Piret equations for solvent production in batch fermentation system is introduced. The parameters values of the mathematical models are estimated via Levenberg-Marquardt optimization method of non-linear least squares. We apply Milstein scheme for solving the stochastic models numerically. The effciency of mathematical models is measured by comparing the simulated result and the experimental data of the microbial growth and solvent production in batch system. Low values of Root Mean-Square Error (RMSE) of stochastic models with aftereffect indicate good fits

  9. Estimation of the Maximum Theoretical Productivity of Fed-Batch Bioreactors

    Energy Technology Data Exchange (ETDEWEB)

    Bomble, Yannick J [National Renewable Energy Laboratory (NREL), Golden, CO (United States); St. John, Peter C [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Crowley, Michael F [National Renewable Energy Laboratory (NREL), Golden, CO (United States)

    2017-10-18

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

  10. High-level ethanol production from starch by a flocculent Saccharomyces cerevisiae strain displaying cell-surface glucoamylase

    Energy Technology Data Exchange (ETDEWEB)

    Kondo, A.; Shigechi, H.; Abe, M.; Uyama, K. [Dept. of Chemical Science and Engineering, Kobe Univ., Nadaku, Kobe (Japan); Matsumoto, T.; Fukuda, H. [Div. of Molecular Science, Kobe Univ., Nadaku, Kobe (Japan); Takahashi, S.; Ueda, M.; Tanaka, A. [Dept. of Synthetic Chemistry and Biological Chemistry, Kyoto Univ., Sakyoku, Kyoto (Japan); Kishimoto, M. [Dept. of Biotechnology, Osaka Univ., Osaka (Japan)

    2002-07-01

    A Strain of host yeast YF207, which is a tryptophan auxotroph and shows strong flocculation ability, was obtained from Saccharomyces diastaticus ATCC60712 and S. cerevisiae W303-1B by tetrad analysis. The plasmid pGA11, which is a multicopy plasmid for cell-surface expression of the Rhyzopus oryzae glucoamylase/{alpha}-agglutinin fusion protein, was then introduced into this flocculent yeast strain (YF207/pGA11). Yeast YF207/pGA11 grew rapidly under aerobic condition (dissolved oxygen 2.0 ppm), using soluble starch. The harvested cells were used for batch fermentation of soluble starch to ethanol under anaerobic condition and showed high ethanol production rates (0.71 g h{sup -1} I{sup -1}) without a time lag, because glucoamylase was immobilized on the yeast cell surface. During repeated utilization of cells for fermentation, YF207/pGA11 maintained high ethanol production rates over 300 h. Moreover, in fed-batch fermentation with YF207/pGA11 for approximately 120 h, the ethanol concentration reached up to 50 g I{sup -1}. In conclusion, flocculent yeast cells displaying cell-surface glucoamylase are considered to be very effective for the direct fermentation of soluble starch to ethanol. (orig.)

  11. Simultaneous Saccharification and Fermentation of Lactic Acid from Empty Fruit Bunch at High Solids Loading

    Directory of Open Access Journals (Sweden)

    Nursia Hassan

    2016-03-01

    Full Text Available The production of value-added chemicals from the bioconversion of lignocellulose biomass has been considered a promising venture. In this study, microwave, alkali-pretreated empty fruit bunch (EFB was used as the substrate, utilizing pelletized filamentous Rhizopus oryzae NRRL 395 and cellulolytic enzymes for lactic acid production in a fed-batch simultaneous saccharification and fermentation (SSF process. Insoluble solids generally do not affect the SSF process until a certain concentration is exceeded. To achieve a high lactic acid concentration in the broth, a high solids loading was required to allow a higher rate of glucose conversion. However, the results revealed a decrease in the final lactic acid yield when running SSF at a massive insoluble solids level. High osmotic pressure in the medium led to poor cellular performance and caused the Rhizopus oryzae pellets to break down, affecting the lactic acid production. To improve the process performance, a fed-batch operation mode was used. The fed-batch operation was shown to facilitate higher lactic acid yield, compared with the SSF batch mode. Enzyme feeding, as well as substrate feeding, was also investigated as a means of enabling a higher dry matter content, with a high glucose conversion in SSF of cellulose-rich EFB.

  12. Direct alcohol fuel cells: toward the power densities of hydrogen-fed proton exchange membrane fuel cells.

    Science.gov (United States)

    Chen, Yanxin; Bellini, Marco; Bevilacqua, Manuela; Fornasiero, Paolo; Lavacchi, Alessandro; Miller, Hamish A; Wang, Lianqin; Vizza, Francesco

    2015-02-01

    A 2 μm thick layer of TiO2 nanotube arrays was prepared on the surface of the Ti fibers of a nonwoven web electrode. After it was doped with Pd nanoparticles (1.5 mgPd  cm(-2) ), this anode was employed in a direct alcohol fuel cell. Peak power densities of 210, 170, and 160 mW cm(-2) at 80 °C were produced if the cell was fed with 10 wt % aqueous solutions of ethanol, ethylene glycol, and glycerol, respectively, in 2 M aqueous KOH. The Pd loading of the anode was increased to 6 mg cm(-2) by combining four single electrodes to produce a maximum peak power density with ethanol at 80 °C of 335 mW cm(-2) . Such high power densities result from a combination of the open 3 D structure of the anode electrode and the high electrochemically active surface area of the Pd catalyst, which promote very fast kinetics for alcohol electro-oxidation. The peak power and current densities obtained with ethanol at 80 °C approach the output of H2 -fed proton exchange membrane fuel cells. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. High cell density strategy for poly(3-hydroxybutyrate production by Cupriavidus necator

    Directory of Open Access Journals (Sweden)

    J. L. Ienczak

    2011-12-01

    Full Text Available Poly(3-hydroxybutyrate (P(3HB is a carbon and intracellular storage source for different microorganisms and its production can achieve high productivities by means of high cell density cultures. The aim of this study was to propose a high cell density strategy for P(3HB production by Cupriavidus necator. The exponential growth phase demands an accurate control of the oxygen transfer system in the bioreactor, due to maximum specific growth rate (µXr, and, consequently, a maximum specific oxygen uptake rate (QO2, in addition to significant residual biomass (Xr growth in high cell density cultures. In this context, this work investigated the strategy for obtaining high cell density, with the inclusion of a linear growth phase for P(3HB production by C. necator in a fed-batch culture. The linear growth phase was included between the exponential growth phase and the P(3HB production phase as a strategy to reduce the specific growth rate (µXr and specific oxygen uptake rate (QO2, with constant residual biomass growth rate (d(V.Xr/dt = k = constant and linear increase of biomass. Three strategies of culture were performed. The results showed that a high residual biomass concentration (30 gXr.L-1 can be reached by the inclusion of the linear growth strategy and specific growth rates (µXr between 0.08 and 0.05 h-1, at the beginning of the production phase, are necessary to attain a high P(3HB productivity.

  14. Evolving Microbial Communities in Cellulose-Fed Microbial Fuel Cell

    Directory of Open Access Journals (Sweden)

    Renata Toczyłowska-Mamińska

    2018-01-01

    Full Text Available The abundance of cellulosic wastes make them attractive source of energy for producing electricity in microbial fuel cells (MFCs. However, electricity production from cellulose requires obligate anaerobes that can degrade cellulose and transfer electrons to the electrode (exoelectrogens, and thus most previous MFC studies have been conducted using two-chamber systems to avoid oxygen contamination of the anode. Single-chamber, air-cathode MFCs typically produce higher power densities than aqueous catholyte MFCs and avoid energy input for the cathodic reaction. To better understand the bacterial communities that evolve in single-chamber air-cathode MFCs fed cellulose, we examined the changes in the bacterial consortium in an MFC fed cellulose over time. The most predominant bacteria shown to be capable electron generation was Firmicutes, with the fermenters decomposing cellulose Bacteroidetes. The main genera developed after extended operation of the cellulose-fed MFC were cellulolytic strains, fermenters and electrogens that included: Parabacteroides, Proteiniphilum, Catonella and Clostridium. These results demonstrate that different communities evolve in air-cathode MFCs fed cellulose than the previous two-chamber reactors.

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

    Science.gov (United States)

    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.

  16. A simple Pichia pastoris fermentation and downstream processing strategy for making recombinant pandemic Swine Origin Influenza a virus Hemagglutinin protein.

    Science.gov (United States)

    Athmaram, T N; Singh, Anil Kumar; Saraswat, Shweta; Srivastava, Saurabh; Misra, Princi; Kameswara Rao, M; Gopalan, N; Rao, P V L

    2013-02-01

    The present Influenza vaccine manufacturing process has posed a clear impediment to initiation of rapid mass vaccination against spreading pandemic influenza. New vaccine strategies are therefore needed that can accelerate the vaccine production. Pichia offers several advantages for rapid and economical bulk production of recombinant proteins and, hence, can be attractive alternative for producing an effective influenza HA based subunit vaccine. The recombinant Pichia harboring the transgene was subjected to fed-batch fermentation at 10 L scale. A simple fermentation and downstream processing strategy is developed for high-yield secretory expression of the recombinant Hemagglutinin protein of pandemic Swine Origin Influenza A virus using Pichia pastoris via fed-batch fermentation. Expression and purification were optimized and the expressed recombinant Hemagglutinin protein was verified by sodium dodecyl sulfate polyacrylamide gel electrophoresis, Western blot and MALDI-TOF analysis. In this paper, we describe a fed-batch fermentation protocol for the secreted production of Swine Influenza A Hemagglutinin protein in the P. pastoris GS115 strain. We have shown that there is a clear relationship between product yield and specific growth rate. The fed-batch fermentation and downstream processing methods optimized in the present study have immense practical application for high-level production of the recombinant H1N1 HA protein in a cost effective way using P. pastoris.

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

    Science.gov (United States)

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

    2018-03-01

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

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

    KAUST Repository

    Ren, Lijiao

    2014-07-01

    Power production of four hydraulically connected microbial fuel cells (MFCs) was compared with the reactors operated using individual electrical circuits (individual), and when four anodes were wired together and connected to four cathodes all wired together (combined), in fed-batch or continuous flow conditions. Power production under these different conditions could not be made based on a single resistance, but instead required polarization tests to assess individual performance relative to the combined MFCs. Based on the power curves, power produced by the combined MFCs (2.12 ± 0.03 mW, 200 ω) was the same as the summed power (2.13 mW, 50 ω) produced by the four individual reactors in fed-batch mode. With continuous flow through the four MFCs, the maximum power (0.59 ± 0.01 mW) produced by the combined MFCs was slightly lower than the summed maximum power of the four individual reactors (0.68 ± 0.02 mW). There was a small parasitic current flow from adjacent anodes and cathodes, but overall performance was relatively unaffected. These findings demonstrate that optimal power production by reactors hydraulically and electrically connected can be predicted from performance by individual reactors. © 2013 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

    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.

  20. Open and continuous fermentation: products, conditions and bioprocess economy.

    Science.gov (United States)

    Li, Teng; Chen, Xiang-bin; Chen, Jin-chun; Wu, Qiong; Chen, Guo-Qiang

    2014-12-01

    Microbial fermentation is the key to industrial biotechnology. Most fermentation processes are sensitive to microbial contamination and require an energy intensive sterilization process. The majority of microbial fermentations can only be conducted over a short period of time in a batch or fed-batch culture, further increasing energy consumption and process complexity, and these factors contribute to the high costs of bio-products. In an effort to make bio-products more economically competitive, increased attention has been paid to developing open (unsterile) and continuous processes. If well conducted, continuous fermentation processes will lead to the reduced cost of industrial bio-products. To achieve cost-efficient open and continuous fermentations, the feeding of raw materials and the removal of products must be conducted in a continuous manner without the risk of contamination, even under 'open' conditions. Factors such as the stability of the biological system as a whole during long cultivations, as well as the yield and productivity of the process, are also important. Microorganisms that grow under extreme conditions such as high or low pH, high osmotic pressure, and high or low temperature, as well as under conditions of mixed culturing, cell immobilization, and solid state cultivation, are of interest for developing open and continuous fermentation processes. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Fed-batch production of hydrophobin RodB from Aspergillus fumigatus in host Pichia pastoris

    DEFF Research Database (Denmark)

    Pedersen, Mona Højgaard; Borodina, Irina; Frisvad, Jens Christian

    was dependent on the methanol-induced AOX1 promoter. Later production was scaled up to a 2 L fed-batch fermentor. Protein production was analyzed by SDS-PAGE, coomassie and silver-stained, as well as western blotting using an anti-his detection antibody. RodB was purified using His-select Nickel Affinity gel....... The emulsifying property of rRodB was investigated using olive oil stained with Sudan black suspended in tris-buffer. The stability of oil micelles were studied by light microscopy. Results: Protein bands of expected size were detected by SDS-PAGE and western blotting in both the fermentation broth and excess...

  2. A biochemically structured model for ethanol fermentation by Kluyveromyces marxianus: A batch fermentation and kinetic study

    DEFF Research Database (Denmark)

    Sansonetti, Sascha; Hobley, Timothy John; Calabrò, V.

    2011-01-01

    Anaerobic batch fermentations of ricotta cheese whey (i.e. containing lactose) were performed under different operating conditions. Ethanol concentrations of ca. 22gL−1 were found from whey containing ca. 44gL−1 lactose, which corresponded to up to 95% of the theoretical ethanol yield within 15h......, lactose, biomass and glycerol during batch fermentation could be described within a ca. 6% deviation, as could the yield coefficients for biomass and ethanol produced on lactose. The model structure confirmed that the thermodynamics considerations on the stoichiometry of the system constrain the metabolic...... coefficients within a physically meaningful range thereby providing valuable and reliable insight into fermentation processes....

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

    Science.gov (United States)

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

    2006-11-01

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

  4. Application of simple fed-batch technique to high-level secretory production of insulin precursor using Pichia pastoris with subsequent purification and conversion to human insulin

    Directory of Open Access Journals (Sweden)

    Chugh Dipti

    2010-05-01

    Full Text Available Abstract Background The prevalence of diabetes is predicted to rise significantly in the coming decades. A recent analysis projects that by the year 2030 there will be ~366 million diabetics around the world, leading to an increased demand for inexpensive insulin to make this life-saving drug also affordable for resource poor countries. Results A synthetic insulin precursor (IP-encoding gene, codon-optimized for expression in P. pastoris, was cloned in frame with the Saccharomyces cerevisiae α-factor secretory signal and integrated into the genome of P. pastoris strain X-33. The strain was grown to high-cell density in a batch procedure using a defined medium with low salt and high glycerol concentrations. Following batch growth, production of IP was carried out at methanol concentrations of 2 g L-1, which were kept constant throughout the remaining production phase. This robust feeding strategy led to the secretion of ~3 gram IP per liter of culture broth (corresponding to almost 4 gram IP per liter of cell-free culture supernatant. Using immobilized metal ion affinity chromatography (IMAC as a novel approach for IP purification, 95% of the secreted product was recovered with a purity of 96% from the clarified culture supernatant. Finally, the purified IP was trypsin digested, transpeptidated, deprotected and further purified leading to ~1.5 g of 99% pure recombinant human insulin per liter of culture broth. Conclusions A simple two-phase cultivation process composed of a glycerol batch and a constant methanol fed-batch phase recently developed for the intracellular production of the Hepatitis B surface antigen was adapted to secretory IP production. Compared to the highest previously reported value, this approach resulted in an ~2 fold enhancement of IP production using Pichia based expression systems, thus significantly increasing the efficiency of insulin manufacture.

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

    Science.gov (United States)

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

    2009-02-01

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

  6. High level expression of Glomerella cingulata cutinase in dense cultures of Pichia pastoris grown under fed-batch conditions.

    Science.gov (United States)

    Seman, W M K Wan; Bakar, S A; Bukhari, N A; Gaspar, S M; Othman, R; Nathan, S; Mahadi, N M; Jahim, J; Murad, A M A; Bakar, F D Abu

    2014-08-20

    A Pichia pastoris transformant carrying the cutinase cDNA of Glomerella cingulata was over-expressed in a 5L bioreactor (2.0L working volume) under fed-batch conditions. Bioreactor experiments rely on varying selected parameters in repeated rounds of optimisation: here these included duration of induction, pH and temperature. Highest cell densities (320gL(-1) wet cell weight) with a cutinase production of 3800mgL(-1) and an activity of 434UmL(-1) were achieved 24h after induction with methanol in basal salt medium (at pH 5 and 28°C). Characterisation of the cutinase showed that it was stable between pH 6 and pH 11, had an optimum pH of 8.0 and retained activity for 30min at 50°C (optimum temperature 25°C).The preferred substrates of G. cingulata cutinase were the medium- to long-chain ρ-nitrophenyl esters of ρ-nitrophenylcaprylate (C8), ρ-nitrophenyllaurate (C12) and ρ-nitrophenylmyristate (C14), with the highest catalytic efficiency, kcat/Km of 7.7±0.7mM(-1)s(-1) for ρ-nitrophenylcaprylate. Microscopic analyses showed that the G. cingulata cutinase was also capable of depolymerising the high molecular weight synthetic polyester, polyethylene terephthalate. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Fed-batch production of vanillin by Bacillus aryabhattai BA03.

    Science.gov (United States)

    Paz, Alicia; Outeiriño, David; Pinheiro de Souza Oliveira, Ricardo; Domínguez, José Manuel

    2018-01-25

    Bacillus aryabhattai BA03, a strain isolated in our laboratory, has interesting properties related to the production of natural aromas and flavors. Specifically, we have found that it was able to produce vanillin from ferulic acid (FA). Furthermore, this strain produces high amounts of 4-vinylguaiacol in only 14h, this being the only intermediate metabolite observed in the process. FA is an inexpensive feedstock for the production of natural value-added compounds when extracted from lignocellulosic wastes. In this study, we optimized the operational conditions (temperature, pH and agitation), medium composition and bioconversion technology (batch or fed-batch) to produce vanillin. In a fed-batch process conducted with just one additional supplementation after 24h, the maximal concentration of vanillin (147.1±0.9mg/L) was observed after 216h (Q V =0.681mg/Lh; Y V/fFA =0.082mg/mg) after degrading 90.3% FA. In view of our data, we postulate that Bacillus aryabhattai BA03 carries out a decarboxylation of ferulic acid as a metabolic pathway. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Glucoamylase production in batch, chemostat and fed-batch cultivations by an industrial strain of Aspergillus niger

    DEFF Research Database (Denmark)

    Pedersen, Henrik; Beyer, Michael; Nielsen, Jens

    2000-01-01

    The Aspergillus niger strain BO-1 was grown in batch, continuous (chemostat) and fed-batch cultivations in order to study the production of the extracellular enzyme glucoamylase under different growth conditions. In the pH range 2.5-6.0, the specific glucoamylase productivity and the specific...

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

    Science.gov (United States)

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

    2007-06-01

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

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

    KAUST Repository

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

    2014-01-01

    together (combined), in fed-batch or continuous flow conditions. Power production under these different conditions could not be made based on a single resistance, but instead required polarization tests to assess individual performance relative

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

    Science.gov (United States)

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

    2017-11-01

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

  12. Enhanced fed-batch production of pyrroloquinoline quinine in Methylobacillus sp. CCTCC M2016079 with a two-stage pH control strategy.

    Science.gov (United States)

    Si, Zhenjun; Machaku, David; Wei, Peilian; Huang, Lei; Cai, Jin; Xu, Zhinan

    2017-06-01

    The effects of pH control strategy and fermentative operation modes on the biosynthesis of pyrroloquinoline quinine (PQQ) were investigated systematically with Methylobacillus sp. CCTCC M2016079 in the present work. Firstly, the shake-flask cultivations and benchtop fermentations at various pH values ranging from 5.3 to 7.8 were studied. Following a kinetic analysis of specific cell growth rate (μ x ) and specific PQQ formation rate (μ p ), the discrepancy in optimal pH values between cell growth and PQQ biosynthesis was observed, which stimulated us to develop a novel two-stage pH control strategy. During this pH-shifted process, the pH in the broth was controlled at 6.8 to promote the cell growth for the first 48 h and then shifted to 5.8 to enhance the PQQ synthesis until the end of fermentation. By applying this pH-shifted control strategy, the maximum PQQ production was improved to 158.61 mg/L in the benchtop fermenter, about 44.9% higher than that under the most suitable constant pH fermentation. Further fed-batch study showed that PQQ production could be improved from 183.38 to 272.21 mg/L by feeding of methanol at the rate of 11.5 mL/h in this two-stage pH process. Meanwhile, the productivity was also increased from 2.02 to 2.84 mg/L/h. In order to support cell growth during the shifted pH stage, the combined feeding of methanol and yeast extract was carried out, which brought about the highest concentration (353.28 mg/L) and productivity (3.27 mg/L/h) of PQQ. This work has revealed the potential of our developed simple and economical strategy for the large-scale production of PQQ.

  13. Production of high-density Chlorella culture grown in fermenters

    Czech Academy of Sciences Publication Activity Database

    Doucha, Jiří; Lívanský, Karel

    2012-01-01

    Roč. 24, č. 1 (2012), s. 35-43 ISSN 0921-8971 R&D Projects: GA MŠk OE09025 Institutional support: RVO:61388971 Keywords : Chlorella vulgaris * Heterotrophic culture * Fed-batch Subject RIV: EE - Microbiology, Virology Impact factor: 2.326, year: 2012

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

    Science.gov (United States)

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

    2015-01-01

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

  15. Software sensor for primary metabolite production case of alcoholic fermentation

    Energy Technology Data Exchange (ETDEWEB)

    Roux, G.; Dahhou, B.; Queinnec, I. [Centre National de la Recherche Scientifique (CNRS), 31 - Toulouse (France)]|[Institut National des Sciences Appliquees (INSA), 31 - Toulouse (France); Goma, G. [Institut National des Sciences Appliquees (INSA), 31 - Toulouse (France)

    1995-12-31

    This paper investigate the application of an observer for state and parameter estimation to batch, continuous and fed batch fermentations for alcohol production taken as model for a primary metabolite production. This observer is provided to palliate the lack of suitable sensors for on-line biomass and ethanol concentrations measurements and to estimate the time varying specific growth rate. Estimates are obtained from an interlaced structure filter based on a `modified extended Kalman filter` by using on-line measurements of carbon dioxide outflow rate and substrate concentration. The filter algorithm was tested during batch, continuous and fed batch fermentation processes. The filter behaviour observed in the experiments gives good results with an agreement theory/practice. (authors) 18 refs.

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

    Science.gov (United States)

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

    2018-01-01

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

  17. Comparison of high-titer lactic acid fermentation from NaOH- and NH3-H2O2-pretreated corncob by Bacillus coagulans using simultaneous saccharification and fermentation

    Science.gov (United States)

    Zhang, Zhenting; Xie, Yuejiao; He, Xiaolan; Li, Xinli; Hu, Jinlong; Ruan, Zhiyong; Zhao, Shumiao; Peng, Nan; Liang, Yunxiang

    2016-01-01

    Lignocellulose is one of the most abundant renewable feedstocks that has attracted considerable attention as a substrate for biofuel and biochemical production. One such biochemical product, lactic acid, is an important fermentation product because of its great potential for the production of biodegradable and biocompatible polylactic acid. High-titer lactic acid production from lignocellulosic materials has been achieved recently; however, it requires biodetoxification or results in large amounts of waste washing water. In this study, we employed two alkaline pretreatment methods and compared their effects on lactic acid fermentation of pretreated corncob by Bacillus coagulans LA204 using fed-batch simultaneous saccharification and fermentation under non-sterile conditions. The lactic acid titer, yield, and productivity from 16% (w/w) NaOH-pretreated and washed corncob were 122.99 g/L, 0.77 g/g corncob, and 1.37 g/L/h, respectively, and from 16% NH3-H2O2-pretreated and washed corncob were 118.60 g/L, 0.74 g/g corncob, and 1.32 g/L/h, respectively. Importantly, the lactic acid titer, yield, and productivity from 18.4% NH3-H2O2-pretreated and unwashed corncob by using fed-batch simultaneous saccharification and fermentation reached 79.47 g/L, 0.43 g/g corncob, and 1.10 g/L/h, respectively, demonstrating that this method is possible for industrial applications and saves washing water. PMID:27853308

  18. Comparison of high-titer lactic acid fermentation from NaOH- and NH3-H2O2-pretreated corncob by Bacillus coagulans using simultaneous saccharification and fermentation.

    Science.gov (United States)

    Zhang, Zhenting; Xie, Yuejiao; He, Xiaolan; Li, Xinli; Hu, Jinlong; Ruan, Zhiyong; Zhao, Shumiao; Peng, Nan; Liang, Yunxiang

    2016-11-17

    Lignocellulose is one of the most abundant renewable feedstocks that has attracted considerable attention as a substrate for biofuel and biochemical production. One such biochemical product, lactic acid, is an important fermentation product because of its great potential for the production of biodegradable and biocompatible polylactic acid. High-titer lactic acid production from lignocellulosic materials has been achieved recently; however, it requires biodetoxification or results in large amounts of waste washing water. In this study, we employed two alkaline pretreatment methods and compared their effects on lactic acid fermentation of pretreated corncob by Bacillus coagulans LA204 using fed-batch simultaneous saccharification and fermentation under non-sterile conditions. The lactic acid titer, yield, and productivity from 16% (w/w) NaOH-pretreated and washed corncob were 122.99 g/L, 0.77 g/g corncob, and 1.37 g/L/h, respectively, and from 16% NH 3 -H 2 O 2 -pretreated and washed corncob were 118.60 g/L, 0.74 g/g corncob, and 1.32 g/L/h, respectively. Importantly, the lactic acid titer, yield, and productivity from 18.4% NH 3 -H 2 O 2 -pretreated and unwashed corncob by using fed-batch simultaneous saccharification and fermentation reached 79.47 g/L, 0.43 g/g corncob, and 1.10 g/L/h, respectively, demonstrating that this method is possible for industrial applications and saves washing water.

  19. Computer evaluation of the results of batch fermentations

    Energy Technology Data Exchange (ETDEWEB)

    Nyeste, L; Sevella, B

    1980-01-01

    A useful aid to the mathematical modeling of fermentation systems, for the kinetic evaluation of batch fermentations, is described. The generalized logistic equation may be used to describe the growth curves, substrate consumption, and product formation. A computer process was developed to fit the equation to experimental points, automatically determining the equation constants on the basis of the iteration algorithm of the method of non-linear least squares. By fitting the process to different master programs of various fermentations, the complex kinetic evaluation of fermentations becomes possible. Based on the analysis easily treatable generalized logistic equation, it is possible to calculate by computer different kinetic characteristics, e.g. rates, special rates, yields, etc. The possibility of committing subjective errors was reduced to a minimum. Employment of the method is demonstrated on some fermentation processes and problems arising in the course of application are discussed.

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

    Directory of Open Access Journals (Sweden)

    Viktor Konakovsky

    2016-01-01

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

  1. Inexpensive on-line alcohol sensor for fermentation monitoring and control

    Energy Technology Data Exchange (ETDEWEB)

    Birch, S W; Turner, A P.F.; Ashby, R E

    1987-01-01

    An inorganic electrochemical fuel cell sensor was interfaced to a microcomputer and used to measure on-line the alcohol concentration in the off-gas of a fermentor. A calibration curve was obtained for methanol (linear range 0-9 g/l) and ethanol (linear range 0-6 g/l) to relate the alcohol concentration in the fermentor liquid with that in the off-gas. The consumption of methanol in a batch fermentation of the methylotroph Ps.BB1 was monitored (sampling frequency of 5 minutes) and compared with samples taken for off-line analysis by GLC. On-line control of the methanol concentration in a fed-batch fermentation was achieved by proportional and integral control. 24 references.

  2. Integrated production of lactic acid and biomass on distillery stillage.

    Science.gov (United States)

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

    2013-09-01

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

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

    Science.gov (United States)

    Shen, Jiacheng; Agblevor, Foster A

    2010-03-01

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

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

    Science.gov (United States)

    Sriyudthsak, Kansuporn; Shiraishi, Fumihide

    2010-11-01

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

  5. Comparison of different options for harvest of a therapeutic protein product from high cell density yeast fermentation broth.

    Science.gov (United States)

    Wang, Alice; Lewus, Rachael; Rathore, Anurag S

    2006-05-05

    Recovery of therapeutic protein from high cell density yeast fermentations at commercial scale is a challenging task. In this study, we investigate and compare three different harvest approaches, namely centrifugation followed by depth filtration, centrifugation followed by filter-aid enhanced depth filtration, and microfiltration. This is achieved by presenting a case study involving recovery of a therapeutic protein from Pichia pastoris fermentation broth. The focus of this study is on performance of the depth filtration and the microfiltration steps. The experimental data has been fitted to the conventional models for cake filtration to evaluate specific cake resistance and cake compressibility. In the case of microfiltration, the experimental data agrees well with flux predicted by shear induced diffusion model. It is shown that, under optimal conditions, all three options can deliver the desired product recovery ( >80%), harvest time ( making a final decision on a harvesting approach.

  6. Improvement of Coenzyme Q10 Production: Mutagenesis Induced by High Hydrostatic Pressure Treatment and Optimization of Fermentation Conditions

    Directory of Open Access Journals (Sweden)

    Yahong Yuan

    2012-01-01

    Full Text Available Coenzyme Q10 (CoQ10, ubiquinone, a potent antioxidative dietary supplement, was produced by submerged fermentation using Agrobacterium tumefaciens instead of chemical synthesis or solvent extraction. Agrobacterium tumefaciens 1.2554 was subjected to mutagenesis using a series of treatments including high hydrostatic pressure (HHP treatment, UV irradiation, and diethyl sulfate (DES treatment to obtain mutant strains showing higher CoQ10 production than wild-type strains. A mutant strain PK38 with four genetic markers was isolated: the specific CoQ10 content of the mutant strain increased by 52.83% compared with the original strain. Effects of carbon and nitrogen sources on CoQ10 production with PK38 were studied. Sucrose at concentration of 30 g/l was tested as the best carbon source, and yeast extract at concentration of 30 g/l supplemented with 10 g/l of ammonium sulfate was identified to be the most favorable for CoQ10 production using PK38. Fed-batch culture strategy was then used for increasing production of CoQ10 in 5-l fermentor. Using the exponential feeding fed-batch culture of sucrose, cell growth and CoQ10 formation were significantly improved. With this strategy, the final cell biomass, CoQ10 production, and specific CoQ10 production increased by 126.11, 173.12, and 22.76%, respectively, compared to those of batch culture.

  7. Modeling of Clostridium tyrobutyricum for Butyric Acid Selectivity in Continuous Fermentation

    Directory of Open Access Journals (Sweden)

    Jianjun Du

    2014-04-01

    Full Text Available A mathematical model was developed to describe batch and continuous fermentation of glucose to organic acids with Clostridium tyrobutyricum. A modified Monod equation was used to describe cell growth, and a Luedeking-Piret equation was used to describe the production of butyric and acetic acids. Using the batch fermentation equations, models predicting butyric acid selectivity for continuous fermentation were also developed. The model showed that butyric acid production was a strong function of cell mass, while acetic acid production was a function of cell growth rate. Further, it was found that at high acetic acid concentrations, acetic acid was metabolized to butyric acid and that this conversion could be modeled. In batch fermentation, high butyric acid selectivity occurred at high initial cell or glucose concentrations. In continuous fermentation, decreased dilution rate improved selectivity; at a dilution rate of 0.028 h−1, the selectivity reached 95.8%. The model and experimental data showed that at total cell recycle, the butyric acid selectivity could reach 97.3%. This model could be used to optimize butyric acid production using C. tyrobutyricum in a continuous fermentation scheme. This is the first study that mathematically describes batch, steady state, and dynamic behavior of C. tyrobutyricum for butyric acid production.

  8. Study on Molasses Concentration from Sugarcanne Bagasse for Biohydrogen Production using Enriched Granular Activated Carbon (GAC) Immobilised Cells by Repeated Batch Cultivation

    Science.gov (United States)

    Idris, Norfatiha; Aminah Lutpi, Nabilah; Ruhaizul Che Ridzuan, Che Mohd; Shian, Wong Yee; Nuraiti Tengku Izhar, Tengku

    2018-03-01

    Repeated batch cultivation is known as most attractive method in improving hydrogen productivity, due to the facts that this approach could minimize the reuse of the cell and the inoculum preparation. In addition, with the combination of attach growth system during the fermentation processes to produce biohydrogen, the density of cells will be increased and the cell washout could be avoided. Therefore, this study aimed to examine the effectiveness of repeated batch cultivation for enrichment of anaerobic mixed culture onto granular activated carbon (GAC) and investigate the effect of molasses concentration during immobilization of mixed culture onto the GAC. The molasses concentration using 50 %, 40 %, 30 %, 20 % and 10 % of diluted molasses were used as feedstock in the fermentation process. The maximum hydrogen production of 60 ml was obtained at 30 % of molasses concentration with 831 ppm of hydrogen concentration. Thus, the kinetic parameter obtained from the batch profiling based on modified Gompertz equation are, Hm= 58 ml for the maximum hydrogen production and Rm= 2.02 ml/h representing the hydrogen production rate.

  9. Increase of ethanol productivity by cell-recycle fermentation of flocculating yeast.

    Science.gov (United States)

    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.

  10. Isoflavone and Protein Constituents of Lactic Acid-Fermented Soy Milk Combine to Prevent Dyslipidemia in Rats Fed a High Cholesterol Diet

    OpenAIRE

    Kobayashi, Maki; Egusa, Shintaro; Fukuda, Mitsuru

    2014-01-01

    A high cholesterol diet induces dyslipidemia. This study investigated whether isoflavone aglycones in lactic acid-fermented soy milk (LFS) improve lipid metabolism in rats fed a high cholesterol diet. Male Sprague-Dawley rats aged seven weeks were fed an AIN-93G diet, a 1% cholesterol diet (a high cholesterol diet), a high-cholesterol diet containing 4% isoflavone extract of LFS (LFS extract diet), a high-cholesterol diet containing 19.4% ethanol-washed LFS (ethanol-washed LFS diet, isoflavon...

  11. Bioprocess Intensification of Beer Fermentation Using Immobilised Cells

    Science.gov (United States)

    Verbelen, Pieter J.; Nedović, Viktor A.; Manojlović, Verica; Delvaux, Freddy R.; Laskošek-Čukalović, Ida; Bugarski, Branko; Willaert, Ronnie

    Beer production with immobilised yeast has been the subject of research for approximately 30 years but has so far found limited application in the brewing industry, due to engineering problems, unrealised cost advantages, microbial contaminations and an unbalanced beer flavor (Linko et al. 1998; Brányik et al. 2005; Willaert and Nedović 2006). The ultimate aim of this research is the production of beer of desired quality within 1-3 days. Traditional beer fermentation systems use freely suspended yeast cells to ferment wort in an unstirred batch reactor. The primary fermentation takes approximately 7 days with a subsequent secondary fermentation (maturation) of several weeks. A batch culture system employing immobilization could benefit from an increased rate of fermentation. However, it appears that in terms of increasing productivity, a continuous fermentation system with immobilization would be the best method (Verbelen et al. 2006). An important issue of the research area is whether beer can be produced by immobilised yeast in continuous culture with the same characteristic as the traditional method.

  12. Time delay and noise explaining the behaviour of the cell growth in fermentation process

    Energy Technology Data Exchange (ETDEWEB)

    Ayuobi, Tawfiqullah; Rosli, Norhayati [Faculty of Industrial Sciences and Technology, Universiti Malaysia Pahang, Lebuhraya Tun Razak, 26300 Gambang, Pahang (Malaysia); Bahar, Arifah [Department of Mathematical Sciences, Faculty of Science, Universiti Teknologi Malaysia, 81310 Johor Bahru, Johor (Malaysia); Salleh, Madihah Md [Department of Biotechnology Industry, Faculty of Biosciences and Bioengineering, Universiti Teknologi Malaysia, 81310 Johor Bahru, Johor (Malaysia)

    2015-02-03

    This paper proposes to investigate the interplay between time delay and external noise in explaining the behaviour of the microbial growth in batch fermentation process. Time delay and noise are modelled jointly via stochastic delay differential equations (SDDEs). The typical behaviour of cell concentration in batch fermentation process under this model is investigated. Milstein scheme is applied for solving this model numerically. Simulation results illustrate the effects of time delay and external noise in explaining the lag and stationary phases, respectively for the cell growth of fermentation process.

  13. Time delay and noise explaining the behaviour of the cell growth in fermentation process

    Science.gov (United States)

    Ayuobi, Tawfiqullah; Rosli, Norhayati; Bahar, Arifah; Salleh, Madihah Md

    2015-02-01

    This paper proposes to investigate the interplay between time delay and external noise in explaining the behaviour of the microbial growth in batch fermentation process. Time delay and noise are modelled jointly via stochastic delay differential equations (SDDEs). The typical behaviour of cell concentration in batch fermentation process under this model is investigated. Milstein scheme is applied for solving this model numerically. Simulation results illustrate the effects of time delay and external noise in explaining the lag and stationary phases, respectively for the cell growth of fermentation process.

  14. Time delay and noise explaining the behaviour of the cell growth in fermentation process

    International Nuclear Information System (INIS)

    Ayuobi, Tawfiqullah; Rosli, Norhayati; Bahar, Arifah; Salleh, Madihah Md

    2015-01-01

    This paper proposes to investigate the interplay between time delay and external noise in explaining the behaviour of the microbial growth in batch fermentation process. Time delay and noise are modelled jointly via stochastic delay differential equations (SDDEs). The typical behaviour of cell concentration in batch fermentation process under this model is investigated. Milstein scheme is applied for solving this model numerically. Simulation results illustrate the effects of time delay and external noise in explaining the lag and stationary phases, respectively for the cell growth of fermentation process

  15. Long-term cathode performance and the microbial communities that develop in microbial fuel cells fed different fermentation endproducts.

    Science.gov (United States)

    Kiely, Patrick D; Rader, Geoffrey; Regan, John M; Logan, Bruce E

    2011-01-01

    To better understand how cathode performance and substrates affected communities that evolved in these reactors over long periods of time, microbial fuel cells were operated for more than 1 year with individual endproducts of lignocellulose fermentation (acetic acid, formic acid, lactic acid, succinic acid, or ethanol). Large variations in reactor performance were primarily due to the specific substrates, with power densities ranging from 835 ± 21 to 62 ± 1mW/m(3). Cathodes performance degraded over time, as shown by an increase in power of up to 26% when the cathode biofilm was removed, and 118% using new cathodes. Communities that developed on the anodes included exoelectrogenic families, such as Rhodobacteraceae, Geobacteraceae, and Peptococcaceae, with the Deltaproteobacteria dominating most reactors. Pelobacter propionicus was the predominant member in reactors fed acetic acid, and it was abundant in several other MFCs. These results provide valuable insights into the effects of long-term MFC operation on reactor performance. Copyright © 2010 Elsevier Ltd. All rights reserved.

  16. Long-term cathode performance and the microbial communities that develop in microbial fuel cells fed different fermentation endproducts

    KAUST Repository

    Kiely, Patrick D.

    2011-01-01

    To better understand how cathode performance and substrates affected communities that evolved in these reactors over long periods of time, microbial fuel cells were operated for more than 1year with individual endproducts of lignocellulose fermentation (acetic acid, formic acid, lactic acid, succinic acid, or ethanol). Large variations in reactor performance were primarily due to the specific substrates, with power densities ranging from 835±21 to 62±1mW/m3. Cathodes performance degraded over time, as shown by an increase in power of up to 26% when the cathode biofilm was removed, and 118% using new cathodes. Communities that developed on the anodes included exoelectrogenic families, such as Rhodobacteraceae, Geobacteraceae, and Peptococcaceae, with the Deltaproteobacteria dominating most reactors. Pelobacter propionicus was the predominant member in reactors fed acetic acid, and it was abundant in several other MFCs. These results provide valuable insights into the effects of long-term MFC operation on reactor performance. © 2010 Elsevier Ltd.

  17. An integral term adaptive neural control of fed-batch fermentation biotechnological process; Control neuronal adaptable con termino integral para un proceso biotecnologico de fermentacion por lote alimentado

    Energy Technology Data Exchange (ETDEWEB)

    Baruch, Ieroham; Hernandez, Luis Alberto; Barrera Cortes, Josefina [Centro de Investigacion y de Estudios Avanzados, Instituto Politecnico Nacional, Mexico D.F. (Mexico)

    2005-07-15

    A nonlinear mathematical model of aerobic biotechnological process of a fed-batch fermentation system is derived using ordinary differential equations. A neurocontrol is applied using Recurrent Trainable Neural Network (RTNN) plus integral term; the first network performs an approximation of the plant's output; the second network generates the control signal so that the biomass concentration could be regulated by the nutrient influent flow rate into the bioreactor. [Spanish] Un modelo matematico no lineal de un proceso biotecnologico aerobio de un sistema de fermentacion por lote alimentado es presentado mediante ecuaciones diferenciales ordinarias. Es propuesto un control utilizando dos redes neuronales recurrentes entrenables (RNRE) con la adicion de un termino integral; la primera red representa un aproximador de la salida de la planta y la segunda genera la senal de control tal que la concentracion de la biomasa pueda ser regulada mediante la alimentacion de un flujo con nutrientes al biorreactor.

  18. The use of date waste for lactic acid production by a fed-batch culture using Lactobacillus casei subsp. rhamnosus.

    Science.gov (United States)

    Nancib, Aicha; Nancib, Nabil; Boubendir, Abdelhafid; Boudrant, Joseph

    2015-01-01

    The production of lactic acid from date juice by Lactobacillus caseisubsp. rhamnosus in batch and fed-batch cultures has been investigated. The fed-batch culture system gave better results for lactic acid production and volumetric productivity. The aim of this work is to determine the effects of the feeding rate and the concentration of the feeding medium containing date juice glucose on the cell growth, the consumption of glucose and the lactic acid production by Lactobacillus casei subsp. rhamnosus in fed-batch cultures. For this study, two concentrations of the feeding medium (62 and 100 g/L of date juice glucose) were tested at different feeding rates (18, 22, 33, 75 and 150 mL/h). The highest volumetric productivity (1.3 g/L.h) and lactic acid yield (1.7 g/g) were obtained at a feeding rate of 33 mL/h and a date juice glucose concentration of 62 g/L in the feeding medium. As a result, most of the date juice glucose was completely utilised (residual glucose 1 g/L), and a maximum lactic acid production level (89.2 g/L) was obtained.

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

    Science.gov (United States)

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

    2006-01-01

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

  20. EFFECT OF FERMENTED Sauropus androgynus LEAVES ON BLOOD LIPID FRACTION AND HAEMATOLOGICAL PROFILE IN BROILER CHICKENS

    Directory of Open Access Journals (Sweden)

    U. Santoso

    2015-12-01

    Full Text Available This study was conducted to evaluate effect of fermented Sauropus androgynus leaves on blood lipid fractions and haematological profiles in broilers. One hundred and twelve broilers were distributed to 7 treatment groups. One group was fed diets without Sauropus androgynus leaves as the control, and other six groups were fed Sauropus androgynus leaves fermented by Neurospora crassa, Lactobacillus sp. or Saccharomyces cerevisiae at level of 25 g or 50 g/kg diet. Experimental results showed that the treatments had no effect on cholesterol, high-density lipoprotein cholesterol (HDL-c, low-density lipoprotein cholesterol (LDL-c and atherogenic index, very low-density lipoprotein cholesterol (VLDL-c and triglyceride concentration (P>0.05. It was shown that fermented Sauropus androgynus leaves significantly affected red blood count (RBC, white blood count (WBC, packed cell volume (PCV, trombosit dan erythrocyte sedimentation rate (ESR (P

  1. Uneven batch data alignment with application to the control of batch end-product quality.

    Science.gov (United States)

    Wan, Jian; Marjanovic, Ognjen; Lennox, Barry

    2014-03-01

    Batch processes are commonly characterized by uneven trajectories due to the existence of batch-to-batch variations. The batch end-product quality is usually measured at the end of these uneven trajectories. It is necessary to align the time differences for both the measured trajectories and the batch end-product quality in order to implement statistical process monitoring and control schemes. Apart from synchronizing trajectories with variable lengths using an indicator variable or dynamic time warping, this paper proposes a novel approach to align uneven batch data by identifying short-window PCA&PLS models at first and then applying these identified models to extend shorter trajectories and predict future batch end-product quality. Furthermore, uneven batch data can also be aligned to be a specified batch length using moving window estimation. The proposed approach and its application to the control of batch end-product quality are demonstrated with a simulated example of fed-batch fermentation for penicillin production. Copyright © 2013 ISA. Published by Elsevier Ltd. All rights reserved.

  2. Acquisition of data from on-line laser turbidimeter and calculation of some kinetic variables in computer-coupled automated fed-batch culture

    International Nuclear Information System (INIS)

    Kadotani, Y.; Miyamoto, K.; Mishima, N.; Kominami, M.; Yamane, T.

    1995-01-01

    Output signals of a commercially available on-line laser turbidimeter exhibit fluctuations due to air and/or CO 2 bubbles. A simple data processing algorithm and a personal computer software have been developed to smooth the noisy turbidity data acquired, and to utilize them for the on-line calculations of some kinetic variables involved in batch and fed-batch cultures of uniformly dispersed microorganisms. With this software, about 10 3 instantaneous turbidity data acquired over 55 s are averaged and convert it to dry cell concentration, X, every minute. Also, volume of the culture broth, V, is estimated from the averaged output data of weight loss of feed solution reservoir, W, using an electronic balance on which the reservoir is placed. Then, the computer software is used to perform linear regression analyses over the past 30 min of the total biomass, VX, the natural logarithm of the total biomass, ln(VX), and the weight loss, W, in order to calculate volumetric growth rate, d(VX)/dt, specific growth rate, μ [ = dln(VX)/dt] and the rate of W, dW/dt, every minute in a fed-batch culture. The software used to perform the first-order regression analyses of VX, ln(VX) and W was applied to batch or fed-batch cultures of Escherichia coli on minimum synthetic or natural complex media. Sample determination coefficients of the three different variables (VX, ln(VX) and W) were close to unity, indicating that the calculations are accurate. Furthermore, growth yield, Y x/s , and specific substrate consumption rate, q sc , were approximately estimated from the data, dW/dt and in a ‘balanced’ fed-batch culture of E. coli on the minimum synthetic medium where the computer-aided substrate-feeding system automatically matches well with the cell growth. (author)

  3. Application of the fuzzy theory to simulation of batch fermentation

    Energy Technology Data Exchange (ETDEWEB)

    Filev, D P; Kishimoto, M; Sengupta, S; Yoshida, T; Taguchi, H

    1985-12-01

    A new approach for system identification with a linguistic model of batch fermentation processes is proposed. The fuzzy theory was applied in order to reduce the uncertainty of quantitative description of the processes by use of qualitative characteristics. An example of fuzzy modeling was illustrated in the simulation of batch ethanol production from molasses after interpretation of the new method, and extension of the fuzzy model was also discussed for several cases of different measurable variables.

  4. Parallel steady state studies on a milliliter scale accelerate fed-batch bioprocess design for recombinant protein production with Escherichia coli.

    Science.gov (United States)

    Schmideder, Andreas; Cremer, Johannes H; Weuster-Botz, Dirk

    2016-11-01

    In general, fed-batch processes are applied for recombinant protein production with Escherichia coli (E. coli). However, state of the art methods for identifying suitable reaction conditions suffer from severe drawbacks, i.e. direct transfer of process information from parallel batch studies is often defective and sequential fed-batch studies are time-consuming and cost-intensive. In this study, continuously operated stirred-tank reactors on a milliliter scale were applied to identify suitable reaction conditions for fed-batch processes. Isopropyl β-d-1-thiogalactopyranoside (IPTG) induction strategies were varied in parallel-operated stirred-tank bioreactors to study the effects on the continuous production of the recombinant protein photoactivatable mCherry (PAmCherry) with E. coli. Best-performing induction strategies were transferred from the continuous processes on a milliliter scale to liter scale fed-batch processes. Inducing recombinant protein expression by dynamically increasing the IPTG concentration to 100 µM led to an increase in the product concentration of 21% (8.4 g L -1 ) compared to an implemented high-performance production process with the most frequently applied induction strategy by a single addition of 1000 µM IPGT. Thus, identifying feasible reaction conditions for fed-batch processes in parallel continuous studies on a milliliter scale was shown to be a powerful, novel method to accelerate bioprocess design in a cost-reducing manner. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1426-1435, 2016. © 2016 American Institute of Chemical Engineers.

  5. Propionic acid production in a plant fibrous-bed bioreactor with immobilized Propionibacterium freudenreichii CCTCC M207015.

    Science.gov (United States)

    Chen, Fei; Feng, Xiaohai; Xu, Hong; Zhang, Dan; Ouyang, Pingkai

    2012-12-15

    A plant fibrous-bed bioreactor (PFB) was constructed for propionic acid production. Sugar cane bagasse was applied to the PFB as immobilizing material. Starting at a concentration of 80g/L of glucose, Propionibacterium freudenreichii CCTCC M207015 produced 41.20±2.03g/L of propionic acid at 108h in the PFB. The value was 21.07% higher than that produced by free cell fermentation. Intermittent and constant fed-batch fermentations were performed in the PFB to optimize the fermentation results. The highest propionic acid concentration obtained from constant fed-batch fermentation was 136.23±6.77g/L, which is 1.40 times higher than the highest concentration (97.00g/L) previously reported. Scanning electron microscopy analysis showed that cells exhibited striking changes in morphology after PFB domestication. Compared with free cell fermentation, the fluxes of propionic acid synthesis and the pentose phosphate pathway in PFB fermentation increased by 84.65% and 227.62%, respectively. On the other hand, a decrease in succinic and acetic acid fluxes was also observed. The metabolic flux distributions of the two PFB fed-batch fermentation strategies also demonstrated that constant fed-batch fermentation is a more beneficial method for the immobilized production of propionic acid. The relevant key enzyme activities and metabolic flux variations of the batch cultures showed good consistency. These results suggest that the PFB was effective in high-concentration propionic acid production. Crown Copyright © 2012. Published by Elsevier B.V. All rights reserved.

  6. De novo biosynthesis of biodiesel by Escherichia coli in optimized fed-batch cultivation.

    Directory of Open Access Journals (Sweden)

    Yangkai Duan

    Full Text Available Biodiesel is a renewable alternative to petroleum diesel fuel that can contribute to carbon dioxide emission reduction and energy supply. Biodiesel is composed of fatty acid alkyl esters, including fatty acid methyl esters (FAMEs and fatty acid ethyl esters (FAEEs, and is currently produced through the transesterification reaction of methanol (or ethanol and triacylglycerols (TAGs. TAGs are mainly obtained from oilseed plants and microalgae. A sustainable supply of TAGs is a major bottleneck for current biodiesel production. Here we report the de novo biosynthesis of FAEEs from glucose, which can be derived from lignocellulosic biomass, in genetically engineered Escherichia coli by introduction of the ethanol-producing pathway from Zymomonas mobilis, genetic manipulation to increase the pool of fatty acyl-CoA, and heterologous expression of acyl-coenzyme A: diacylglycerol acyltransferase from Acinetobacter baylyi. An optimized fed-batch microbial fermentation of the modified E. coli strain yielded a titer of 922 mg L(-1 FAEEs that consisted primarily of ethyl palmitate, -oleate, -myristate and -palmitoleate.

  7. Models for high cell density bioreactors must consider biomass volume fraction: Cell recycle example.

    Science.gov (United States)

    Monbouquette, H G

    1987-06-01

    Intrinsic models, which take into account biomass volume fraction, must be formulated for adequate simulation of high-biomass-density fermentations with cell recycle. Through comparison of corresponding intrinsic and non-intrinsic models in dimensionless form, constraints for non-intrinsic model usage in terms of biokinetic and fermenter operating parameters can be identified a priori. Analysis of a simple product-inhibition model indicates that the non-intrinsic approach is suitable only when the attainable biomass volume fraction in the fermentation broth is less than about 0.10. Inappropriate application of a non-intrinsic model can lead to gross errors in calculated substrate and product concentrations, substrate conversion, and volumetric productivity.

  8. Models for high cell density bioreactors must consider biomass volume fraction: cell recycle example

    Energy Technology Data Exchange (ETDEWEB)

    Monbouquette, H.G.

    1987-06-01

    Intrinsic models, which take into account biomass volume fraction, must be formulated for adequate simulation of high-biomass-density fermentations with cell recycle. Through comparison of corresponding intrinsic and non-intrinsic models in dimensionless form, constraints for non-intrinsic model usage in terms of biokinetic and fermenter operating parameters can be identified a priori. Analysis of a simple product-inhibition model indicates that the non-intrinsic approach is suitable only when the attainable biomass volume fraction in the fermentation broth is less than about 0.10. Inappropriate application of a non-intrinsic model can lead to gross errors in calculated substrate and product concentrations, substrate conversion, and volumetric productivity. (Refs. 14).

  9. Bioelectricity generation from coconut husk retting wastewater in fed batch operating microbial fuel cell by phenol degrading microorganism

    International Nuclear Information System (INIS)

    Jayashree, C.; Arulazhagan, P.; Adish Kumar, S.; Kaliappan, S.; Yeom, Ick Tae; Rajesh Banu, J.

    2014-01-01

    Dual chamber microbial fuel cell (MFC) operated at fed batch mode for the treatment of retting wastewater has potently achieved both current generation and phenol removal. Hydraulic retention time (HRT) of the reactor was varied from 40 days to 10 days. COD (chemical oxygen demand) removal was 91% at 40 days HRT, with an initial COD concentration of 530 ± 50 g m −3 . Retting wastewater with an initial phenol concentration of 320 ± 60 g m −3 procured a highest phenol removal of 93% at 40 days HRT of the microbial fuel cell. Maximum power density of 362 mW m −2 was achieved using retting wastewater at HRT of 20 days with an internal resistance of 150 Ω in a dual chambered MFC. The bacterial strains in anode region, reported to be responsible for potential phenol removal, were identified as Ochrobactrum sp. RA1 (KJ408266), Ochrobactrum sp. RA2 (KJ408267) and Pesudomonas aeruginosa RA3 (KJ408268) using phylogenetic analysis. The study reveals that, dual chambered MFC effectively removed the phenol from retting wastewater along with power generation. - Highlights: • Maximum power density of 362 mW m −2 (150 Ω) was achieved at HRT of 20 days. • 91% COD removal and 93% phenol removal was observed at HRT of 40 days. • 25% coulombic efficiency was achieved in treatment of retting wastewater with MFC. • Phylogenetic analysis detect phenol degrading Ochrobactrum sp.RA1 in anode biofilm. • In addition, Ochrobactrum sp.RA2 and Pseudomonas aeruginosa RA3 were also isolated

  10. Fermentation and purification strategies for the production of betulinic acid and its lupane-type precursors in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Czarnotta, Eik; Dianat, Mariam; Korf, Marcel

    2017-01-01

    from the bark of plane tree or birch. Here, we reengineered the reported betulinic acid pathway into S. cerevisiae and used this novel strain to develop efficient fermentation and product purification methods. Fed-batch cultivations with ethanol excess, using either an ethanol-pulse feed or controlling...... a constant ethanol concentration in the fermentation medium, significantly enhanced production of betulinic acid and its triterpenoid precursors. The beneficial effect of excess ethanol was further exploited in nitrogen-limited resting cell fermentations, yielding betulinic acid concentrations of 182 mg...

  11. Measurement processing for state estimation and fault identification in batch fermentations

    Directory of Open Access Journals (Sweden)

    R. Dondo

    2004-09-01

    Full Text Available This work describes an application of maximum likelihood identification and statistical detection techniques for determining the presence and nature of abnormal behaviors in batch fermentations. By appropriately organizing these established techniques, a novel algorithm that is able to detect and isolate faults in nonlinear and uncertain processes was developed. The technique processes residuals from a nonlinear filter based on the assumed model of fermentation. This information is combined with mass balances to conduct statistical tests that are used as the core of the detection procedure. The approach uses a sliding window to capture the present statistical properties of filtering and mass-balance residuals. In order to avoid divergence of the nonlinear monitor filter, the maximum likelihood states and parameters are periodically estimated. The maximum likelihood parameters are used to update the kinetic parameter values of the monitor filter. If the occurrence of a fault is detected, alternative faulty model structures are evaluated statistically through the use of log-likelihood function values and chi2 detection tests. Simulation obtained for xanthan gum batch fermentations are encouraging.

  12. Two-stage conversion of crude glycerol to energy using dark fermentation linked with microbial fuel cell or microbial electrolysis cell.

    Science.gov (United States)

    Chookaew, Teera; Prasertsan, Poonsuk; Ren, Zhiyong Jason

    2014-03-25

    Crude glycerol is a main byproduct of the biodiesel industry, and the beneficial use of waste glycerol has been a major challenge. This study characterises the conversion of crude glycerol into bioenergy such as H2 and electricity using a two-stage process linking dark fermentation with a microbial fuel cell (MFC) or microbial electrolysis cell (MEC). The results showed that fermentation achieved a maximum H2 rate of 332 mL/L and a yield of 0.55 mol H2/mol glycerol, accompanied by 20% of organic removal. Fed with the raw fermentation products with an initial COD of 7610 mg/L, a two-chamber MFC produced 92 mW/m(2) in power density and removed 50% of COD. The Columbic efficiency was 14%. When fed with 50% diluted fermentation product, a similar power output (90m W/m(2)) and COD removal (49%) were obtained, but the CE doubled to 27%. Similar substrates were used to produce H2 in two-chamber MECs, and the diluted influent had a higher performance, with the highest yield at 106 mL H2/g COD and a CE of 24%. These results demonstrate that dark fermentation linked with MFC/MEC can be a feasible option for conversion of waste glycerol into bioenergy. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. Construction and fed-batch cultivation of Candida famata with enhanced riboflavin production.

    Science.gov (United States)

    Dmytruk, Kostyantyn; Lyzak, Oleksy; Yatsyshyn, Valentyna; Kluz, Maciej; Sibirny, Vladimir; Puchalski, Czeslaw; Sibirny, Andriy

    2014-02-20

    Riboflavin (vitamin B2) is an essential nutrition component serving as a precursor of coenzymes FMN and FAD that are involved mostly in reactions of oxidative metabolism. Riboflavin is produced in commercial scale and is used in feed and food industries, and in medicine. The yeast Candida famata (Candida flareri) belongs to the group of so called "flavinogenic yeasts" which overproduce riboflavin under iron limitation. Three genes SEF1, RIB1 and RIB7 coding for a putative transcription factor, GTP cyclohydrolase II and riboflavin synthase, respectively were simultaneously overexpressed in the background of a non-reverting riboflavin producing mutant AF-4, obtained earlier in our laboratory using methods of classical selection (Dmytruk et al. (2011), Metabolic Engineering 13, 82-88). Cultivation conditions of the constructed strain were optimized for shake-flasks and bioreactor cultivations. The constructed strain accumulated up to 16.4g/L of riboflavin in optimized medium in a 7L laboratory bioreactor during fed-batch fermentation. Copyright © 2013 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

    2017-01-01

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

  15. Quantitative evaluation of yeast's requirement for glycerol formation in very high ethanol performance fed-batch process

    Directory of Open Access Journals (Sweden)

    Nevoigt Elke

    2010-05-01

    Full Text Available Abstract Background Glycerol is the major by-product accounting for up to 5% of the carbon in Saccharomyces cerevisiae ethanolic fermentation. Decreasing glycerol formation may redirect part of the carbon toward ethanol production. However, abolishment of glycerol formation strongly affects yeast's robustness towards different types of stress occurring in an industrial process. In order to assess whether glycerol production can be reduced to a certain extent without jeopardising growth and stress tolerance, the yeast's capacity to synthesize glycerol was adjusted by fine-tuning the activity of the rate-controlling enzyme glycerol 3-phosphate dehydrogenase (GPDH. Two engineered strains whose specific GPDH activity was significantly reduced by two different degrees were comprehensively characterized in a previously developed Very High Ethanol Performance (VHEP fed-batch process. Results The prototrophic strain CEN.PK113-7D was chosen for decreasing glycerol formation capacity. The fine-tuned reduction of specific GPDH activity was achieved by replacing the native GPD1 promoter in the yeast genome by previously generated well-characterized TEF promoter mutant versions in a gpd2Δ background. Two TEF promoter mutant versions were selected for this study, resulting in a residual GPDH activity of 55 and 6%, respectively. The corresponding strains were referred to here as TEFmut7 and TEFmut2. The genetic modifications were accompanied to a strong reduction in glycerol yield on glucose; the level of reduction compared to the wild-type was 61% in TEFmut7 and 88% in TEFmut2. The overall ethanol production yield on glucose was improved from 0.43 g g-1 in the wild type to 0.44 g g-1 measured in TEFmut7 and 0.45 g g-1 in TEFmut2. Although maximal growth rate in the engineered strains was reduced by 20 and 30%, for TEFmut7 and TEFmut2 respectively, strains' ethanol stress robustness was hardly affected; i.e. values for final ethanol concentration (117 ± 4 g

  16. Analytical solution of Luedeking-Piret equation for a batch fermentation obeying Monod growth kinetics.

    Science.gov (United States)

    Garnier, Alain; Gaillet, Bruno

    2015-12-01

    Not so many fermentation mathematical models allow analytical solutions of batch process dynamics. The most widely used is the combination of the logistic microbial growth kinetics with Luedeking-Piret bioproduct synthesis relation. However, the logistic equation is principally based on formalistic similarities and only fits a limited range of fermentation types. In this article, we have developed an analytical solution for the combination of Monod growth kinetics with Luedeking-Piret relation, which can be identified by linear regression and used to simulate batch fermentation evolution. Two classical examples are used to show the quality of fit and the simplicity of the method proposed. A solution for the combination of Haldane substrate-limited growth model combined with Luedeking-Piret relation is also provided. These models could prove useful for the analysis of fermentation data in industry as well as academia. © 2015 Wiley Periodicals, Inc.

  17. Application of Multivariate Analysis Tools to Industrial Scale Fermentation Data

    DEFF Research Database (Denmark)

    Mears, Lisa; Nørregård, Rasmus; Stocks, Stuart M.

    . 2014). This may be the case for fed-batch fermentation processes, where mechanistic modelling is challenging due to non-linear dynamics, and non-steady state operation. There is also a lack of sensors for key parameters which are considered to define the quality of the batch, such as product...

  18. Comparison of the effectivities of two-phase and single-phase anaerobic sequencing batch reactors during dairy wastewater treatment

    Energy Technology Data Exchange (ETDEWEB)

    Goebloes, Sz.; Portoero, P.; Bordas, D.; Kalman, M.; Kiss, I. [Institute for Biotechnology, Bay Zoltan Foundation for Applied Research, H-6726 Szeged (Hungary)

    2008-05-15

    The performances of anaerobic sequencing batch reactors fed with two different substrates were studied. The substrates were raw acid whey and acid whey fermented with Kluyveromyces lactis in order to investigate the suitability of ethanol for biogas production. The organic loading rates (OLRs) during the experiment ranged from 1.6 to 12.8 g COD dm{sup -3} d{sup -1} and the corresponding decreasing hydraulic retention times from 40 to 5 days for both reactor systems. The efficiency of each system depended on the OLR: the highest COD removal rate was observed at the lowest OLR applied (about 100% in both systems), and at maximum OLR the COD removal efficiency was 68% for the reactors fed with the raw whey and 80% for those fed with the pre-fermented whey. Under the same high OLR conditions the methane yield was 0.122 dm{sup -3} CH{sub 4} g{sup -1} COD{sub degraded} for the anaerobic digesters fed with the untreated whey, and 0.197 dm{sup -3} CH{sub 4} g{sup -1} COD{sub degraded} for those fed with the pre-fermented whey. The digesters functioned without pH control. At the maximum OLR the pH in the reactors fed with the raw acid whey was 5.1, while in those fed with the pre-fermented whey it was 7.15. The results demonstrate that the use of the pre-fermented acid whey as substrate for anaerobic digestion without pH control is feasible, especially at high OLR levels. This substrate is preferable to the raw acid whey, because of the ethanol formed as a non-acidic fermentation product of the yeast. (author)

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

    Science.gov (United States)

    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.

  20. A multi-pronged investigation into the effect of glucose starvation and culture duration on fed-batch CHO cell culture

    DEFF Research Database (Denmark)

    Fan, Yuzhou; Jimenez Del Val, Ioscani; Müller, Christian

    2015-01-01

    to the interplay between the dilution effect associated with change in specific productivity of mAbs and the changed nucleotide sugar metabolism. Herein, we also show and discuss that increased cell culture duration negatively affect the maturation of glycans. In addition, comparative proteomics analysis of cells......In this study, omics-based analysis tools were used to explore the effect of glucose starvation and culture duration on monoclonal antibody (mAb) production in fed-batch CHO cell culture to gain better insight into how these parameters can be controlled to ensure optimal mAb productivity...... and quality. Titer and N-glycosylation of mAbs, as well as proteomic signature and metabolic status of the production cells in the culture were assessed. We found that the impact of glucose starvation on the titer and N-glycosylation of mAbs was dependent on the degree of starvation during early stationary...

  1. Fermentation of Saccharomyces cerevisiae - Combining kinetic modeling and optimization techniques points out avenues to effective process design.

    Science.gov (United States)

    Scheiblauer, Johannes; Scheiner, Stefan; Joksch, Martin; Kavsek, Barbara

    2018-09-14

    A combined experimental/theoretical approach is presented, for improving the predictability of Saccharomyces cerevisiae fermentations. In particular, a mathematical model was developed explicitly taking into account the main mechanisms of the fermentation process, allowing for continuous computation of key process variables, including the biomass concentration and the respiratory quotient (RQ). For model calibration and experimental validation, batch and fed-batch fermentations were carried out. Comparison of the model-predicted biomass concentrations and RQ developments with the corresponding experimentally recorded values shows a remarkably good agreement for both batch and fed-batch processes, confirming the adequacy of the model. Furthermore, sensitivity studies were performed, in order to identify model parameters whose variations have significant effects on the model predictions: our model responds with significant sensitivity to the variations of only six parameters. These studies provide a valuable basis for model reduction, as also demonstrated in this paper. Finally, optimization-based parametric studies demonstrate how our model can be utilized for improving the efficiency of Saccharomyces cerevisiae fermentations. Copyright © 2018 Elsevier Ltd. All rights reserved.

  2. Hypoxia-elicited impairment of cell wall integrity, glycosylation precursor synthesis, and growth in scaled-up high-cell density fed-batch cultures of Saccharomyces cerevisiae.

    Science.gov (United States)

    Aon, Juan C; Sun, Jianxin; Leighton, Julie M; Appelbaum, Edward R

    2016-08-15

    In this study we examine the integrity of the cell wall during scale up of a yeast fermentation process from laboratory scale (10 L) to industrial scale (10,000 L). In a previous study we observed a clear difference in the volume fraction occupied by yeast cells as revealed by wet cell weight (WCW) measurements between these scales. That study also included metabolite analysis which suggested hypoxia during scale up. Here we hypothesize that hypoxia weakens the yeast cell wall during the scale up, leading to changes in cell permeability, and/or cell mechanical resistance, which in turn may lead to the observed difference in WCW. We tested the cell wall integrity by probing the cell wall sensitivity to Zymolyase. Also exometabolomics data showed changes in supply of precursors for the glycosylation pathway. The results show a more sensitive cell wall later in the production process at industrial scale, while the sensitivity at early time points was similar at both scales. We also report exometabolomics data, in particular a link with the protein glycosylation pathway. Significantly lower levels of Man6P and progressively higher GDP-mannose indicated partially impaired incorporation of this sugar nucleotide during co- or post-translational protein glycosylation pathways at the 10,000 L compared to the 10 L scale. This impairment in glycosylation would be expected to affect cell wall integrity. Although cell viability from samples obtained at both scales were similar, cells harvested from 10 L bioreactors were able to re-initiate growth faster in fresh shake flask media than those harvested from the industrial scale. The results obtained help explain the WCW differences observed at both scales by hypoxia-triggered weakening of the yeast cell wall during the scale up.

  3. Ethanol Production from Whey by Kluyveromyces marxianus in Batch Fermentation System: Kinetics Parameters Estimation

    Directory of Open Access Journals (Sweden)

    Dessy Ariyanti

    2013-03-01

    Full Text Available Whey is the liquid remaining after milk has been curdled and strained. It is a by-product of the manufacture of cheese or casein and has several commercial uses. In environmental point of view, whey is kind of waste which has high pollution level due to it’s contain high organic compound with BOD and COD value 50 and 80 g/L respectively. On the other side, whey also contain an amount of lactose (4.5%-5%; lactose can be used as carbon source and raw material for producing ethanol via fermentation using yeast strain Kluyveromyces marxianus. The objective of this research is to investigate the ethanol production kinetics from crude whey through fermentation using Kluyveromyces marxianus and to predict the model kinetics parameter. The yeast was able to metabolize most of the lactose within 16 h to give 8.64 g/L ethanol, 4.43 g/L biomass, and remain the 3.122 g/L residual lactose. From the results presented it also can be concluded that common kinetic model for microbial growth, substrate consumption, and product formation is a good alternative to describe an experimental batch fermentation of Kluyveromyces marxianus grown on a medium composed of whey. The model was found to be capable of reflecting all batch culture phases to a certain degree of accuracy, giving the parameter value: μmax, Ks, YX/S, α, β : 0.32, 10.52, 0.095, 1.52, and 0.11 respectively. © 2013 BCREC UNDIP. All rights reserved(Selected Paper from International Conference on Chemical and Material Engineering (ICCME 2012Received: 27th September 2012; Revised: 29th November 2012; Accepted: 7th December 2012[How to Cite: D. Ariyanti, H. Hadiyanto, (2013. Ethanol Production from Whey by Kluyveromyces marxianus in Batch Fermentation System: Kinetics Parameters Estimation. Bulletin of Chemical Reaction Engineering & Catalysis, 7 (3: 179-184. (doi:10.9767/bcrec.7.3.4044.179-184][Permalink/DOI: http://dx.doi.org/10.9767/bcrec.7.3.4044.179-184 ] View in  |

  4. The morphology of Ganoderma lucidum mycelium in a repeated-batch fermentation for exopolysaccharide production

    Directory of Open Access Journals (Sweden)

    Wan Abd Al Qadr Imad Wan-Mohtar

    2016-09-01

    Full Text Available The morphology of Ganoderma lucidum BCCM 31549 mycelium in a repeated-batch fermentation (RBF was studied for exopolysaccharide (EPS production. RBF was optimised for time to replace and volume to replace. G. lucidum mycelium showed the ability to self-immobilise and exhibited high stability for repeated use in RBF with engulfed pellets. Furthermore, the ovoid and starburst-like pellet morphology was disposed to EPS production in the shake flask and bioreactor, respectively. Seven RBF could be carried out in 500 mL flasks, and five repeated batches were performed in a 2 L bioreactor. Under RBF conditions, autolysis of pellet core in the shake flask and shaving off of the outer hairy region in the bioreactor were observed at the later stages of RBF (R4 for the shake flask and R6 for the bioreactor. The proposed strategy showed that the morphology of G. lucidum mycelium can withstand extended fermentation cycles.

  5. Estudio comparativo de cuatro sistemas de propagación de levadura cervecera por lote alimentado Comparative study of four fed-batch propagation systems of beer yeast

    Directory of Open Access Journals (Sweden)

    Hoyos H.

    1998-06-01

    Full Text Available

    Se realizó la propagación por lotes de levadura cervecera y con base en estos resultados se ajustó un modelo matemático. Se llevaron a cabo cuatro fermentaciones con diferentes técnicas de alimentación y se confrontaron los resultados mediante simulación. Las técnicas estudiadas fueron alimentación puntual, alimentación continua-puntual y de alta densidad. La mayor concentración celular y factor de propagación se presentó con el cultivo de alta densidad. La simulación demostró que la alimentación continua o puntual afecta el comportamiento celular, ya sea sobre la velocidad específica y/o el factor estequiométrico Yx/s . Se encontró que la técnica de alta densidad aumenta el factor estequiométrico Yx/s.

    Beer yeast was propagated using batch culture, and a mathematical model was fitted to the resulting data. Intermittent, continuousintermittent, and high-density fed-batch techniques were used. The highest cell yield was found using the high density technique. Simulation also unveiled an effect of the feeding technique on cellular growth rate and yield. The high density technique increased the stoichiometric factor Yx/s.

  6. Biogas production from protein-rich biomass: fed-batch anaerobic fermentation of casein and of pig blood and associated changes in microbial community composition.

    Directory of Open Access Journals (Sweden)

    Etelka Kovács

    Full Text Available It is generally accepted as a fact in the biogas technology that protein-rich biomass substrates should be avoided due to inevitable process inhibition. Substrate compositions with a low C/N ratio are considered difficult to handle and may lead to process failure, though protein-rich industrial waste products have outstanding biogas generation potential. This common belief has been challenged by using protein-rich substrates, i.e. casein and precipitated pig blood protein in laboratory scale continuously stirred mesophilic fed-batch biogas fermenters. Both substrates proved suitable for sustained biogas production (0.447 L CH4/g protein oDM, i.e. organic total solids in high yield without any additives, following a period of adaptation of the microbial community. The apparent key limiting factors in the anaerobic degradation of these proteinaceous materials were the accumulation of ammonia and hydrogen sulfide. Changes in time in the composition of the microbiological community were determined by next-generation sequencing-based metagenomic analyses. Characteristic rearrangements of the biogas-producing community upon protein feeding and specific differences due to the individual protein substrates were recognized. The results clearly demonstrate that sustained biogas production is readily achievable, provided the system is well-characterized, understood and controlled. Biogas yields (0.45 L CH4/g oDM significantly exceeding those of the commonly used agricultural substrates (0.25-0.28 L CH4/g oDM were routinely obtained. The results amply reveal that these high-energy-content waste products can be converted to biogas, a renewable energy carrier with flexible uses that can replace fossil natural gas in its applications. Process control, with appropriate acclimation of the microbial community to the unusual substrate, is necessary. Metagenomic analysis of the microbial community by next-generation sequencing allows a precise determination of the

  7. Biogas Production from Protein-Rich Biomass: Fed-Batch Anaerobic Fermentation of Casein and of Pig Blood and Associated Changes in Microbial Community Composition

    Science.gov (United States)

    Kovács, Etelka; Wirth, Roland; Maróti, Gergely; Bagi, Zoltán; Rákhely, Gábor; Kovács, Kornél L.

    2013-01-01

    It is generally accepted as a fact in the biogas technology that protein-rich biomass substrates should be avoided due to inevitable process inhibition. Substrate compositions with a low C/N ratio are considered difficult to handle and may lead to process failure, though protein-rich industrial waste products have outstanding biogas generation potential. This common belief has been challenged by using protein-rich substrates, i.e. casein and precipitated pig blood protein in laboratory scale continuously stirred mesophilic fed-batch biogas fermenters. Both substrates proved suitable for sustained biogas production (0.447 L CH4/g protein oDM, i.e. organic total solids) in high yield without any additives, following a period of adaptation of the microbial community. The apparent key limiting factors in the anaerobic degradation of these proteinaceous materials were the accumulation of ammonia and hydrogen sulfide. Changes in time in the composition of the microbiological community were determined by next-generation sequencing-based metagenomic analyses. Characteristic rearrangements of the biogas-producing community upon protein feeding and specific differences due to the individual protein substrates were recognized. The results clearly demonstrate that sustained biogas production is readily achievable, provided the system is well-characterized, understood and controlled. Biogas yields (0.45 L CH4/g oDM) significantly exceeding those of the commonly used agricultural substrates (0.25-0.28 L CH4/g oDM) were routinely obtained. The results amply reveal that these high-energy-content waste products can be converted to biogas, a renewable energy carrier with flexible uses that can replace fossil natural gas in its applications. Process control, with appropriate acclimation of the microbial community to the unusual substrate, is necessary. Metagenomic analysis of the microbial community by next-generation sequencing allows a precise determination of the alterations in

  8. Enhanced acarbose production by Streptomyces M37 using a two-stage fermentation strategy.

    Directory of Open Access Journals (Sweden)

    Fei Ren

    Full Text Available In this work, we investigated the effect of pH on Streptomyces M37 growth and its acarbose biosynthesis ability. We observed that low pH was beneficial for cell growth, whereas high pH favored acarbose synthesis. Moreover, addition of glucose and maltose to the fermentation medium after 72 h of cultivation promoted acarbose production. Based on these results, a two-stage fermentation strategy was developed to improve acarbose production. Accordingly, pH was kept at 7.0 during the first 72 h and switched to 8.0 after that. At the same time, glucose and maltose were fed to increase acarbose accumulation. With this strategy, we achieved an acarbose titer of 6210 mg/L, representing an 85.7% increase over traditional batch fermentation without pH control. Finally, we determined that the increased acarbose production was related to the high activity of glutamate dehydrogenase and glucose 6-phosphate dehydrogenase.

  9. Batch Fermentative Biohydrogen Production Process Using Immobilized Anaerobic Sludge from Organic Solid Waste

    Directory of Open Access Journals (Sweden)

    Patrick T. Sekoai

    2016-12-01

    Full Text Available This study examined the potential of organic solid waste for biohydrogen production using immobilized anaerobic sludge. Biohydrogen was produced under batch mode at process conditions of 7.9, 30.3 °C and 90 h for pH, temperature and fermentation time, respectively. A maximum biohydrogen fraction of 48.67%, which corresponded to a biohydrogen yield of 215.39 mL H2/g Total Volatile Solids (TVS, was achieved. Therefore, the utilization of immobilized cells could pave the way for a large-scale biohydrogen production process.

  10. A parametric study ot protease production in batch and fed-batch cultures of Bacillus firmus.

    Science.gov (United States)

    Moon, S H; Parulekar, S J

    1991-03-05

    Proteolytic enzymes produced by Bacillus species find a wide variety of applications in brewing, detergent, food, and leather industries. Owing to significant differences normally observed in culture conditions promoting cell growth and those promoting production of metabolites such as enzymes, for increased efficacy of bioreactor operations it is essential to identify these sets of conditions (including medium formulation). This study is focused on formulation of a semidefined medium that substantially enhances synthesis and secretion of an alkaline protease in batch cultures of Bacillus firmus NRS 783, a known superior producer of this enzyme. The series of experiments conducted to identify culture conditions that lead to improved protease production also enables investigation of the regulatory effects of important culture parameters including pH, dissolved oxygen, and concentrations of nitrogen and phosphorous sources and yeast extract in the medium on cell growth, synthesis and secretion of protease, and production of two major nonbiomass products, viz., acetic acid and ethanol. Cell growth and formation of the three nonbiomass products are hampered significantly under nitrogen, phosphorous, or oxygen limitation, with the cells being unable to grow in an oxygen-free environment. Improvement in protease production is achieved with respect to each culture parameter, leading in the process to 80% enhancement in protease activity over that attained using media reported in the literature. Results of a few fed-batch experiments with constant feed rate, conducted to examine possible enhancement in protease production and to further investigate repression of protease synthesis by excess of the principal carbon and nitrogen sources, are also discussed. The detailed investigation of stimulatory and repressory effects of simple and complex nutrients on protease production and metabolism of Bacillus firmus conducted in this study will provide useful guidelines for design

  11. Production of enzymatically active recombinant full-length barley high pI alpha-glucosidase of glycoside family 31 by high cell-density fermentation of Pichia pastoris and affinity purification

    DEFF Research Database (Denmark)

    Næsted, Henrik; Kramhøft, Birte; Lok, F.

    2006-01-01

    Recombinant barley high pI alpha-glucosidase was produced by high cell-density fermentation of Pichia pastoris expressing the cloned full-length gene. The gene was amplified from a genomic clone and exons (coding regions) were assembled by overlap PCR. The resulting cDNA was expressed under contr...... nM x s(-1), and 85 s(-1) using maltose as substrate. This work presents the first production of fully active recombinant alpha-glucosidase of glycoside hydrolase family 31 from higher plants. (c) 2005 Elsevier Inc. All rights reserved....

  12. Effects of Yogurt Containing Fermented Pepper Juice on the Body Fat and Cholesterol Level in High Fat and High Cholesterol Diet Fed Rat.

    Science.gov (United States)

    Yeon, Su-Jung; Hong, Go-Eun; Kim, Chang-Kyu; Park, Woo Joon; Kim, Soo-Ki; Lee, Chi-Ho

    2015-01-01

    This experiment investigated whether yogurt containing fermented pepper juice (FPJY) affects cholesterol level in high fat and high cholesterol diet (HFCD) fed rat. Twenty five Sprague-Dawley male rats of 7 wk were divided into 5 groups, and fed following diets for 9 wk; CON (control diet), HFCD (HFCD), PY (HFCD supplemented with 2% of plain yogurt), LFY (HFCD supplemented with 2% of FPJY), and HFY (HFCD supplemented with 5% of FPJY). In the LFY group, hepatic total lipid level decreased significantly compared to the HFCD group (p0.05). In HFY group, body weight and hepatic total lipid level significantly decreased over the HFCD group (p0.05). Liver weight decreased as FPJY content was increased. Results suggested FPJY would inhibit organ hypertrophy and accumulation of body fat, hepatic lipid, and cholesterol in HFCD fed rat.

  13. Semi-scale production of PHAs from waste frying oil by Pseudomonas fluorescens S48

    Directory of Open Access Journals (Sweden)

    Rawia F. Gamal

    2013-01-01

    Full Text Available The present study aimed at developing a strategy to improve the volumetric production of PHAs by Pseudomonas fluorescens S48 using waste frying oil (WFO as the sole carbon source. For this purpose, several cultivations were set up to steadily improve nutrients supply to attain high cell density and high biopolymer productivity. The production of PHAs was examined in a 14 L bioreactor as one-stage batch, two-stage batch, and high-cell-density fed-batch cultures. The highest value of polymer content in one-stage bioreactor was obtained after 60 h (33.7%. Whereas, the two-stage batch culture increased the polymer content to 50.1% after 54 h. High-cell-density (0.64 g/L at continuous feeding rate 0.55 mL/l/h of WFO recorded the highest polymer content after 54 h (55.34%. Semi-scale application (10 L working volume increased the polymer content in one-stage batch, two-stage batch and high cell density fed-batch cultures by about 12.3%, 5.8% and 11.3%, respectively, as compared with that obtained in 2 L fermentation culture. Six different methods for biopolymer extraction were done to investigate their efficiency for optimum polymer recovery. The maximum efficiency of solvent recovery of PHA was attained by chloroform-hypochlorite dispersion extraction. Gas chromatography (GC analysis of biopolymer produced by Pseudomonas fluorescens S48 indicated that it solely composed of 3-hydrobutyric acid (98.7%. A bioplastic film was prepared from the obtained PHB. The isolate studied shares the same identical sequence, which is nearly the complete 16S rRNA gene. The identity of this sequence to the closest pseudomonads strains is about 98-99%. It was probably closely related to support another meaningful parsiomony analysis and construction of a phylogenetic tree. The isolate is so close to Egyptian strain named EG 639838.

  14. Stable acetate production in extreme-thermophilic (70°C) mixed culture fermentation by selective enrichment of hydrogenotrophic methanogens

    Science.gov (United States)

    Zhang, Fang; Zhang, Yan; Ding, Jing; Dai, Kun; van Loosdrecht, Mark C. M.; Zeng, Raymond J.

    2014-06-01

    The control of metabolite production is difficult in mixed culture fermentation. This is particularly related to hydrogen inhibition. In this work, hydrogenotrophic methanogens were selectively enriched to reduce the hydrogen partial pressure and to realize efficient acetate production in extreme-thermophilic (70°C) mixed culture fermentation. The continuous stirred tank reactor (CSTR) was stable operated during 100 days, in which acetate accounted for more than 90% of metabolites in liquid solutions. The yields of acetate, methane and biomass in CSTR were 1.5 +/- 0.06, 1.0 +/- 0.13 and 0.4 +/- 0.05 mol/mol glucose, respectively, close to the theoretical expected values. The CSTR effluent was stable and no further conversion occurred when incubated for 14 days in a batch reactor. In fed-batch experiments, acetate could be produced up to 34.4 g/L, significantly higher than observed in common hydrogen producing fermentations. Acetate also accounted for more than 90% of soluble products formed in these fed-batch fermentations. The microbial community analysis revealed hydrogenotrophic methanogens (mainly Methanothermobacter thermautotrophicus and Methanobacterium thermoaggregans) as 98% of Archaea, confirming that high temperature will select hydrogenotrophic methanogens over aceticlastic methanogens effectively. This work demonstrated a potential application to effectively produce acetate as a value chemical and methane as an energy gas together via mixed culture fermentation.

  15. Solid phase fermentation of Helianthus tuberosus for ethanol

    Energy Technology Data Exchange (ETDEWEB)

    Baerwald, G.; Hamad, S.H.

    1989-01-01

    The direct fermentation of pure inulin and hammer mill crushed Helianthus tuberosus tubers (topinambur, Jerusalem artichoke) was studied using two heat-tolerant yeasts, namely Kluyveromyces marxianus and Candida kefyr. A Saccharomyces cerevisiae was included in the study so as to compare the yields of these two yeasts with that of a commercial distiller's yeast. The inulin fermentation was carried out in an 18-L bioreactor using the fed-batch and the batch-fermentation methods. The final ethanol concentration was 6.1% (L/L) which represents 82% of the theoretical yield. Commercial scale experiments with hammer mill crushed tubers gave yields lower than those found in the laboratory: 69% of the theoretical yield for direct fermentation without enzyme addition, and about 91% when cellolytic enzymes were added.

  16. Construction of Halomonas bluephagenesis capable of high cell density growth for efficient PHA production.

    Science.gov (United States)

    Ren, Yilin; Ling, Chen; Hajnal, Ivan; Wu, Qiong; Chen, Guo-Qiang

    2018-05-01

    High-cell-density cultivation is an effective way to improve the productivity of microbial fermentations and in turn reduce the cost of the final products, especially in the case of intracellular products. Halomonas bluephagenesis TD01 is a halophilic platform bacterium for the next generation of industrial biotechnology with a native PHA synthetic pathway, able to grow under non-sterile continuous fermentation conditions. A selection strategy for mutant strains that can grow to a high cell density was developed. Based on an error-prone DNA polymerase III ε subunit, a genome-wide random mutagenesis system was established and used in conjunction with an artificial high cell density culture environment during the selection process. A high-cell-density H. bluephagenesis TDHCD-R3 obtained after 3 rounds of selection showed an obvious enhancement of resistance to toxic metabolites including acetate, formate, lactate and ethanol compared to wild-type. H. bluephagenesis TDHCD-R3-8-3 constructed from H. bluephagenesis TDHCD-R3 by overexpressing an optimized phaCAB operon was able to grow to 15 g/L cell dry weight (CDW) containing 94% PHA in shake flask studies. H. bluephagenesis TDHCD-R3-8-3 was grown to more than 90 g/L CDW containing 79% PHA compared with only 81 g/L with 70% PHA by the wild type when incubated in a 7-L fermentor under the same conditions.

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

    Science.gov (United States)

    Namfon, Panjanapongchai; Ratchanok, Sahaworarak; Chalida, Daengbussade

    2017-03-01

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

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

    Science.gov (United States)

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

    2017-02-08

    The halophilic bacterium Chromohalobacter salexigens has been proposed as promising cell factory for the production of the compatible solutes ectoine and hydroxyectoine. This bacterium has evolved metabolic adaptations to efficiently grow under high salt concentrations by accumulating ectoines as compatible solutes. However, metabolic overflow, which is a major drawback for the efficient conversion of biological feedstocks, occurs as a result of metabolic unbalances during growth and ectoines production. Optimal production of ectoines is conditioned by the interplay of carbon and nitrogen metabolisms. In this work, we set out to determine how nitrogen supply affects the production of ectoines. Chromohalobacter salexigens was challenged to grow in media with unbalanced carbon/nitrogen ratio. In C. salexigens, overflow metabolism and ectoines production are a function of medium composition. At low ammonium conditions, the growth rate decreased importantly, up to 80%. Shifts in overflow metabolism were observed when changing the C/N ratio in the culture medium. 13 C-NMR analysis of ectoines labelling revealed a high metabolic rigidity, with almost constant flux ratios in all conditions assayed. Unbalanced C/N ratio led to pyruvate accumulation, especially upon N-limitation. Analysis of an ect - mutant demonstrated the link between metabolic overflow and ectoine biosynthesis. Under non ectoine synthesizing conditions, glucose uptake and metabolic overflow decreased importantly. Finally, in fed-batch cultures, biomass yield was affected by the feeding scheme chosen. High growth (up to 42.4 g L -1 ) and volumetric ectoine yields (up to 4.21 g L -1 ) were obtained by minimizing metabolite overflow and nutrient accumulation in high density cultures in a low nitrogen fed-batch culture. Moreover, the yield coefficient calculated for the transformation of glucose into biomass was 30% higher in fed-batch than in the batch culture, demonstrating that the metabolic

  19. Co-culturing of Lactobacillus paracasei subsp. paracasei with a Lactobacillus delbrueckii subsp. delbrueckii mutant to make high cell density for increased lactate productivity from cassava bagasse hydrolysate.

    Science.gov (United States)

    John, Rojan Pappy; Nampoothiri, K Madhavan

    2011-03-01

    To increase the productivity of lactic acid, a co-culture of lactobacilli was made by mixing 1:1 ratio of Lactobacillus paracasei subsp. paracasei and a fast growing L. delbrueckii subsp. delbrueckii mutant. The culture was embedded on to polyurethane foam (PUF) cubes as a biofilm and used for fermentation. In order to prevent the cell leakage, the PUF cubes were further entrapped in calcium cross-linked alginate. The maximum lactic acid production using a high cell density free culture was >38 g l(-1) from ~40 g l(-1) of reducing sugar within 12 h of fermentation. Using PUF biofilms, the same yield of lactic acid attained after 24 h. When the cubes were further coated with alginate it took 36 h for the maximum yield. Even though, the productivity is slightly lesser with the alginate coating, cell leakage was decreased and cubes were reused without much decrease in production in repeated batches. Using a conventional control inoculum (3%, w/v), it took 120 h to yield same amount of lactic acid.

  20. Enzyme controlled glucose auto-delivery for high cell density cultivations in microplates and shake flasks

    Directory of Open Access Journals (Sweden)

    Casteleijn Marco G

    2008-11-01

    Full Text Available Abstract Background Here we describe a novel cultivation method, called EnBase™, or enzyme-based-substrate-delivery, for the growth of microorganisms in millilitre and sub-millilitre scale which yields 5 to 20 times higher cell densities compared to standard methods. The novel method can be directly applied in microwell plates and shake flasks without any requirements for additional sensors or liquid supply systems. EnBase is therefore readily applicable for many high throughput applications, such as DNA production for genome sequencing, optimisation of protein expression, production of proteins for structural genomics, bioprocess development, and screening of enzyme and metagenomic libraries. Results High cell densities with EnBase are obtained by applying the concept of glucose-limited fed-batch cultivation which is commonly used in industrial processes. The major difference of the novel method is that no external glucose feed is required, but glucose is released into the growth medium by enzymatic degradation of starch. To cope with the high levels of starch necessary for high cell density cultivation, starch is supplied to the growing culture suspension by continuous diffusion from a storage gel. Our results show that the controlled enzyme-based supply of glucose allows a glucose-limited growth to high cell densities of OD600 = 20 to 30 (corresponding to 6 to 9 g l-1 cell dry weight without the external feed of additional compounds in shake flasks and 96-well plates. The final cell density can be further increased by addition of extra nitrogen during the cultivation. Production of a heterologous triosphosphate isomerase in E. coli BL21(DE3 resulted in 10 times higher volumetric product yield and a higher ratio of soluble to insoluble product when compared to the conventional production method. Conclusion The novel EnBase method is robust and simple-to-apply for high cell density cultivation in shake flasks and microwell plates. The

  1. Novel strategies for control of fermentation processes

    DEFF Research Database (Denmark)

    Mears, Lisa; Stocks, Stuart; Sin, Gürkan

    Bioprocesses are inherently sensitive to fluctuations in processing conditions and must be tightly regulated to maintain cellular productivity. Industrial fermentations are often difficult to replicate across production sites or between facilities as the small operating differences in the equipment...... of a fermentation. Industrial fermentation processes are typically operated in fed batch mode, which also poses specific challenges for process monitoring and control. This is due to many reasons including non-linear behaviour, and a relatively poor understanding of the system dynamics. It is therefore challenging...

  2. Isoflavone and Protein Constituents of Lactic Acid-Fermented Soy Milk Combine to Prevent Dyslipidemia in Rats Fed a High Cholesterol Diet

    Science.gov (United States)

    Kobayashi, Maki; Egusa, Shintaro; Fukuda, Mitsuru

    2014-01-01

    A high cholesterol diet induces dyslipidemia. This study investigated whether isoflavone aglycones in lactic acid-fermented soy milk (LFS) improve lipid metabolism in rats fed a high cholesterol diet. Male Sprague-Dawley rats aged seven weeks were fed an AIN-93G diet, a 1% cholesterol diet (a high cholesterol diet), a high-cholesterol diet containing 4% isoflavone extract of LFS (LFS extract diet), a high-cholesterol diet containing 19.4% ethanol-washed LFS (ethanol-washed LFS diet, isoflavone-poor diet), or a high cholesterol diet containing 23.2% intact LFS (intact LFS diet) for five weeks. The plasma total cholesterol (TC) level was increased in the rats fed the LFS extract diet compared with those fed the high cholesterol diet. The TC level was decreased by the intact LFS and ethanol-washed LFS diets. The cholesterol-lowering effect was stronger in the rats fed the intact LFS diet than those fed the ethanol-washed LFS diet. The plasma triglyceride (TG) level was unchanged in the rats fed the LFS extract diet, but it decreased in rats fed the intact LFS and ethanol-washed LFS diets. Although, compared with the high cholesterol diet, the LFS extract and ethanol-washed LFS diets did not reduce hepatic cholesterol and TG, both levels were remarkably lowered by the intact LFS diet. These results suggest that the improvement in lipid metabolism of rats fed a high-cholesterol diet containing LFS isoflavone aglycones is not due to an independent effect but due to a cooperative effect with soy protein. PMID:25514389

  3. Isoflavone and Protein Constituents of Lactic Acid-Fermented Soy Milk Combine to Prevent Dyslipidemia in Rats Fed a High Cholesterol Diet

    Directory of Open Access Journals (Sweden)

    Maki Kobayashi

    2014-12-01

    Full Text Available A high cholesterol diet induces dyslipidemia. This study investigated whether isoflavone aglycones in lactic acid-fermented soy milk (LFS improve lipid metabolism in rats fed a high cholesterol diet. Male Sprague-Dawley rats aged seven weeks were fed an AIN-93G diet, a 1% cholesterol diet (a high cholesterol diet, a high-cholesterol diet containing 4% isoflavone extract of LFS (LFS extract diet, a high-cholesterol diet containing 19.4% ethanol-washed LFS (ethanol-washed LFS diet, isoflavone-poor diet, or a high cholesterol diet containing 23.2% intact LFS (intact LFS diet for five weeks. The plasma total cholesterol (TC level was increased in the rats fed the LFS extract diet compared with those fed the high cholesterol diet. The TC level was decreased by the intact LFS and ethanol-washed LFS diets. The cholesterol-lowering effect was stronger in the rats fed the intact LFS diet than those fed the ethanol-washed LFS diet. The plasma triglyceride (TG level was unchanged in the rats fed the LFS extract diet, but it decreased in rats fed the intact LFS and ethanol-washed LFS diets. Although, compared with the high cholesterol diet, the LFS extract and ethanol-washed LFS diets did not reduce hepatic cholesterol and TG, both levels were remarkably lowered by the intact LFS diet. These results suggest that the improvement in lipid metabolism of rats fed a high-cholesterol diet containing LFS isoflavone aglycones is not due to an independent effect but due to a cooperative effect with soy protein.

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

    KAUST Repository

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

    2014-01-01

    Larger scale microbial fuel cells (MFCs) require compact architectures to efficiently treat wastewater. We examined how anode-brush diameter, number of anodes, and electrode spacing affected the performance of the MFCs operated in fed-batch and continuous flow mode. All anodes were initially tested with the brush core set at the same distance from the cathode. In fed-batch mode, the configuration with three larger brushes (25 mm diameter) produced 80% more power (1240 mW m-2) than reactors with eight smaller brushes (8 mm) (690 mW m-2). The higher power production by the larger brushes was due to more negative and stable anode potentials than the smaller brushes. The same general result was obtained in continuous flow operation, although power densities were reduced. However, by moving the center of the smaller brushes closer to the cathode (from 16.5 to 8 mm), power substantially increased from 690 to 1030 mW m-2 in fed batch mode. In continuous flow mode, power increased from 280 to 1020 mW m-2, resulting in more power production from the smaller brushes than the larger brushes (540 mW m-2). These results show that multi-electrode MFCs can be optimized by selecting smaller anodes, placed as close as possible to the cathode. © 2013 Elsevier B.V. All rights reserved.

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

    KAUST Repository

    Lanas, Vanessa

    2014-02-01

    Larger scale microbial fuel cells (MFCs) require compact architectures to efficiently treat wastewater. We examined how anode-brush diameter, number of anodes, and electrode spacing affected the performance of the MFCs operated in fed-batch and continuous flow mode. All anodes were initially tested with the brush core set at the same distance from the cathode. In fed-batch mode, the configuration with three larger brushes (25 mm diameter) produced 80% more power (1240 mW m-2) than reactors with eight smaller brushes (8 mm) (690 mW m-2). The higher power production by the larger brushes was due to more negative and stable anode potentials than the smaller brushes. The same general result was obtained in continuous flow operation, although power densities were reduced. However, by moving the center of the smaller brushes closer to the cathode (from 16.5 to 8 mm), power substantially increased from 690 to 1030 mW m-2 in fed batch mode. In continuous flow mode, power increased from 280 to 1020 mW m-2, resulting in more power production from the smaller brushes than the larger brushes (540 mW m-2). These results show that multi-electrode MFCs can be optimized by selecting smaller anodes, placed as close as possible to the cathode. © 2013 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

    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.

  7. A Novel Application for Low Frequency Electrochemical Impedance Spectroscopy as an Online Process Monitoring Tool for Viable Cell Concentrations

    Directory of Open Access Journals (Sweden)

    Christoph Slouka

    2016-11-01

    Full Text Available New approaches in process monitoring during industrial fermentations are not only limited to classical pH, dO2 and offgas analysis, but use different in situ and online sensors based on different physical principles to determine biomass, product quality, lysis and far more. One of the very important approaches is the in situ accessibility of viable cell concentration (VCC. This knowledge provides increased efficiency in monitoring and controlling strategies during cultivations. Electrochemical impedance spectroscopy—EIS—is used to monitor biomass in a fermentation of E. coli BL21(DE3, producing a recombinant protein using a fed batch-based approach. Increases in the double layer capacitance (Cdl, determined at frequencies below 1 kHz, are proportional to the increase of biomass in the batch and fed batch phase, monitored in offline and online modes for different cultivations. A good correlation of Cdl with cell density is found and in order to get an appropriate verification of this method, different state-of-the-art biomass measurements are performed and compared. Since measurements in this frequency range are largely determined by the double layer region between the electrode and media, rather minor interferences with process parameters (aeration, stirring are to be expected. It is shown that impedance spectroscopy at low frequencies is a powerful tool for cultivation monitoring.

  8. Fed-Batch Production of Bacterial Ghosts Using Dielectric Spectroscopy for Dynamic Process Control

    Directory of Open Access Journals (Sweden)

    Andrea Meitz

    2016-03-01

    Full Text Available The Bacterial Ghost (BG platform technology evolved from a microbiological expression system incorporating the ϕX174 lysis gene E. E-lysis generates empty but structurally intact cell envelopes (BGs from Gram-negative bacteria which have been suggested as candidate vaccines, immunotherapeutic agents or drug delivery vehicles. E-lysis is a highly dynamic and complex biological process that puts exceptional demands towards process understanding and control. The development of a both economic and robust fed-batch production process for BGs required a toolset capable of dealing with rapidly changing concentrations of viable biomass during the E-lysis phase. This challenge was addressed using a transfer function combining dielectric spectroscopy and soft-sensor based biomass estimation for monitoring the rapid decline of viable biomass during the E-lysis phase. The transfer function was implemented to a feed-controller, which followed the permittivity signal closely and was capable of maintaining a constant specific substrate uptake rate during lysis phase. With the described toolset, we were able to increase the yield of BG production processes by a factor of 8–10 when compared to currently used batch procedures reaching lysis efficiencies >98%. This provides elevated potentials for commercial application of the Bacterial Ghost platform technology.

  9. Batch pervaporative fermentation with coupled membrane and its influence on energy consumption in permeate recovery and distillation stage

    International Nuclear Information System (INIS)

    Leon, Juan A.; Palacios-Bereche, Reynaldo; Nebra, Silvia A.

    2016-01-01

    In the ethanol production process from sugarcane molasses, the distillation process is a high-energy demand stage. The distillation energy efficiency is strongly associated with the alcoholic fermentation performance in the process. The final ethanol concentration in the alcoholic wines has a direct impact on consumption of thermal energy in ethanol separation. In this paper, ethanol production with a H-SBMF (Hybrid-Simple Batch Membrane Fermenter) using PDMS (polydimethylsiloxane) pervaporation membrane was modelled and simulated, in order to determine its influence on energy consumption in distillation. Steam in distillation and electrical energy needs in permeate recovery were mainly influenced by membrane adaptation. The H-SBMF achieved a higher ethanol production in the range of 10–13% compared to the conventional batch fermenter, and an increase in productivity of 150%. The distillation system consisted of two sets of columns: the ethanol recovery column and the rectification column. The permeate recovery system (i.e. vacuum and compression) was regarded in order to evaluate the electrical energy requirement, and the thermal energy demand was evaluated. A decrease in steam consumption was evidenced by the adaptation of the membrane to the fermenter. Higher energy efficiencies were achieved in distillation with larger membrane areas, achieving almost 17% steam reduction. - Highlights: • Higher and faster ethanol productions were achieved by fermenter hybridization. • Multi-stage permeate compression and inter-stage heat recovery were assumed. • Energy demand was studied based on an integrated fermentation and distillation scheme. • High-energy efficiency was attained in the distillation to produce hydrated alcohol.

  10. In Vivo Hypocholesterolemic Effect of MARDI Fermented Red Yeast Rice Water Extract in High Cholesterol Diet Fed Mice

    Directory of Open Access Journals (Sweden)

    Swee Keong Yeap

    2014-01-01

    Full Text Available Fermented red yeast rice has been traditionally consumed as medication in Asian cuisine. This study aimed to determine the in vivo hypocholesterolemic and antioxidant effects of fermented red yeast rice water extract produced using Malaysian Agricultural Research and Development Institute (MARDI Monascus purpureus strains in mice fed with high cholesterol diet. Absence of monacolin-k, lower level of γ-aminobutyric acid (GABA, higher content of total amino acids, and antioxidant activities were detected in MARDI fermented red yeast rice water extract (MFRYR. In vivo MFRYR treatment on hypercholesterolemic mice recorded similar lipid lowering effect as commercial red yeast rice extract (CRYR as it helps to reduce the elevated serum liver enzyme and increased the antioxidant levels in liver. This effect was also associated with the upregulation of apolipoproteins-E and inhibition of Von Willebrand factor expression. In summary, MFRYR enriched in antioxidant and amino acid without monacolin-k showed similar hypocholesterolemic effect as CRYR that was rich in monacolin-k and GABA.

  11. In Vivo Hypocholesterolemic Effect of MARDI Fermented Red Yeast Rice Water Extract in High Cholesterol Diet Fed Mice

    Science.gov (United States)

    Beh, Boon Kee; Kong, Joan; Ho, Wan Yong; Mohd Yusof, Hamidah; Hussin, Aminuddin bin; Jaganath, Indu Bala; Alitheen, Noorjahan Banu; Jamaluddin, Anisah

    2014-01-01

    Fermented red yeast rice has been traditionally consumed as medication in Asian cuisine. This study aimed to determine the in vivo hypocholesterolemic and antioxidant effects of fermented red yeast rice water extract produced using Malaysian Agricultural Research and Development Institute (MARDI) Monascus purpureus strains in mice fed with high cholesterol diet. Absence of monacolin-k, lower level of γ-aminobutyric acid (GABA), higher content of total amino acids, and antioxidant activities were detected in MARDI fermented red yeast rice water extract (MFRYR). In vivo MFRYR treatment on hypercholesterolemic mice recorded similar lipid lowering effect as commercial red yeast rice extract (CRYR) as it helps to reduce the elevated serum liver enzyme and increased the antioxidant levels in liver. This effect was also associated with the upregulation of apolipoproteins-E and inhibition of Von Willebrand factor expression. In summary, MFRYR enriched in antioxidant and amino acid without monacolin-k showed similar hypocholesterolemic effect as CRYR that was rich in monacolin-k and GABA. PMID:25031606

  12. Repeated batch fermentation of immobilized E. coli expressing Vitreoscilla hemoglobin for long-term use.

    Science.gov (United States)

    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.

  13. Amphipathic lignin derivatives to accelerate simultaneous saccharification and fermentation of unbleached softwood pulp for bioethanol production.

    Science.gov (United States)

    Cheng, Ningning; Yamamoto, Yoko; Koda, Keiichi; Tamai, Yutaka; Uraki, Yasumitsu

    2014-12-01

    Amphipathic lignin derivatives (A-LDs) were already demonstrated to improve enzymatic saccharification of lignocellulose. Based on this knowledge, two kinds of A-LDs prepared from black liquor of soda pulping of Japanese cedar were applied to a fed-batch simultaneous saccharification and fermentation (SSF) process for unbleached soda pulp of Japanese cedar to produce bioethanol. Both lignin derivatives slightly accelerated yeast fermentation of glucose but not inhibited it. In addition, ethanol yields based on the theoretical maximum ethanol production in the fed-batch SSF process was increased from 49% without A-LDs to 64% in the presence of A-LDs. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Change in hyphal morphology of Aspergillus Oryzae during fed-batch cultivation

    DEFF Research Database (Denmark)

    Haack, Martin Brian; Olsson, Lisbeth; Hansen, K

    2006-01-01

    the batch phase from 2.8-2.9 up to 4.0-4.4 mu m. The diameter of the hyphal elements remained constant, around 4 mu m, after the feed was started. However, the diameter of the immediate hyphal tip, where the enzyme secretion is thought to take place, increased dramatically with up to a factor 2.5 during......Industrial enzymes are often produced by filamentous fungi in fed-batch cultivations. During cultivation, the different morphological forms displayed by the fungi have an impact on the overall production. The morphology of a recombinant lipase producing Aspergillus oryzae strain was investigated...

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

    Science.gov (United States)

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

    2017-08-01

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

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

    Science.gov (United States)

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

    2017-07-01

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

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

    Science.gov (United States)

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

    2018-01-01

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

  18. Effect of skim milk and dahi (yogurt) on blood glucose, insulin, and lipid profile in rats fed with high fructose diet.

    Science.gov (United States)

    Yadav, Hariom; Jain, Shalini; Sinha, P R

    2006-01-01

    In the present study, the effect of skim milk and the fermented milk product named dahi (yogurt) on plasma glucose, insulin, and lipid levels as well as on liver glycogen and lipid contents in rats fed with high fructose diet has been investigated. Rats were fed with high fructose diet (21%) supplemented with skim milk, dahi (10 g/day each), or no milk product (control group) for 6 weeks. After 6 weeks of high fructose diet administration, the plasma glucose became significantly higher in control animals (246 mg/dL), whereas it was lower in skim milk (178 mg/dL)- and dahi (143 mg/dL)-fed rats. The glucose tolerance became impaired at the third week of feeding of high fructose diet in control animals, whereas in skim milk- and dahi-fed animals achievement of glucose intolerance was delayed until the fourth and fifth week, respectively. Blood glycosylated hemoglobin and plasma insulin were significantly lower in skim milk (10% and 34%, respectively)- and dahi (17%, and 48%, respectively)-fed animals than those of the control group. Plasma total cholesterol, triglycerides, low-density lipoprotein-cholesterol, and very-low-density lipoprotein-cholesterol and blood free fatty acids were significantly lower in skim milk (13%, 14%, 14%, 19%, and 14%, respectively)- and dahi (22%, 33%, 30%, 33%, and 29%, respectively)-fed animals as compared with control animals. Moreover, the total cholesterol, triglyceride, and glycogen contents in liver tissues were also lower in skim milk (55%, 50%, and 36%, respectively)- and dahi (64%, 27%, and 4%, respectively)-fed animals as compared with control animals. In contrast, high-density lipoprotein-cholesterol in plasma was higher in skim milk (14%)- and dahi (29%)-fed animals as compared with control animals. These results indicate that skim milk and its fermented milk product, dahi, delay the progression of fructose-induced diabetes and dyslipidemia in rats and that these may be useful as antidiabetic food supplements that can be

  19. Degradation of chlorophenol mixtures in a fed-batch system by two ...

    African Journals Online (AJOL)

    This work was undertaken to investigate the effect of variations of the feed rate on a fed-batch set-up used to degrade xenobiotics. The mixture of substrates was composed of PCP, 2,4,6 TCP and 2,3,5,6 TeCP (pentachlorophenol, 2,4,6 trichlorophenol and 2,3,5,6 tetrachlorophenol respectively). Two acclimated bacteria ...

  20. Preferences based Control Design of Complex Fed-batch Cultivation Process

    Directory of Open Access Journals (Sweden)

    Yuri Pavlov

    2009-08-01

    Full Text Available In the paper is presented preferences based control design and stabilization of the growth rate of fed-batch cultivation processes. The control is based on an enlarged Wang-Monod-Yerusalimsky kinetic model. Expected utility theory is one of the approaches for utilization of conceptual information (expert preferences. In the article is discussed utilization of stochastic machine learning procedures for evaluation of expert utilities as criteria for optimization.

  1. THE EFFECT OF THE ADDITION OF INVERT SUGAR ON THE PRODUCTION OF CEPHALOSPORIN C IN A FED-BATCH BIOREACTOR

    Directory of Open Access Journals (Sweden)

    A.S. Silva

    1998-12-01

    Full Text Available Cephalosporin C, a b -lactam antibiotic, is the starting molecule for industrial production of semi-synthetic cephalosporins. The bioprocess for its production is carried out in batch stirred and aerated tank reactors utilizing strains of the filamentous fungus Cephalosporium acremonium. In this work a comparison was made between the processes of production of cephalosporin C in a conventional batch bioreactor, with synthetic medium containing glucose and sucrose, and in a fed-batch reactor at several flowrates of supplementary medium containing invert sucrose. In general, the fed-batch process was shown to be more efficient than the conventional batch one, and the process in which the lowest supplementation flowrate was used presented an antibiotic production significantly higher than those obtained under the other conditions.

  2. Optimization of the Production of Polygalacturonase from Aspergillus kawachii Cloned in Saccharomyces cerevisiae in Batch and Fed-Batch Cultures

    Directory of Open Access Journals (Sweden)

    Diego Jorge Baruque

    2011-01-01

    Full Text Available Polygalacturonases (PG; EC 3.2.1.15 catalyze the hydrolysis of pectin and/or pectic acid and are useful for industrial applications such as juice clarification and pectin extraction. Growth and heterologous expression of recombinant Saccharomyces cerevisiae which expresses an acidic PG from Aspergillus kawachii has been studied in batch and fed-batch cultures. Kinetics and stoichiometric parameters of the recombinant yeast were determined in batch cultures in a synthetic medium. In these cultures, the total biomass concentration, protein concentration, and enzyme activity achieved were 2.2 g/L, 10 mg/L, and 3 U/mL, respectively, to give a productivity of 0.06 U/(mL·h. In fed-batch cultures, various strategies for galactose feeding were used: (i after a glucose growth phase, the addition of a single pulse of galactose which gave a productivity of 0.19 U/(mL·h; (ii after a glucose growth phase, a double pulse of galactose at the same final concentration was added, resulting in a productivity of 0.21 U/(mL·h; (iii a simultaneous feeding of glucose and galactose, yielding a productivity of 1.32 U/(mL·h. Based on these results, the simultaneous feeding of glucose and galactose was by far the most suitable strategy for the production of this enzyme. Moreover, some biochemical characteristics of the recombinant enzyme such as a molecular mass of ~60 kDa, an isoelectric point of 3.7 and its ability to hydrolyze polygalacturonic acid at pH=2.5 were determined.

  3. Study on Batch Culture Growth Model for Lactococcus lactis IO-1

    OpenAIRE

    Ishizaki, Ayaaki; Ohta, Tomomi; Kobayashi, Genta; 石崎, 文彬; 太田, 智美; 小林, 元太

    1991-01-01

    L-lactate fermentation employing Lactncoccus lactis IO-1 demonstrated a typical end product inhibition. By numerical analysis of fermentation results of the batch culture of this microorganism, the specific rates for cell growth, substrate consumption and product formation were clearly expressed by the end product inhibition formulae. All constants for those formulae were determined by the fermentation results. A mathematical model for batch culture growth of this microorganism in which the n...

  4. Fermentation and microflora of plaa-som, a Thai fermented fish product prepared with different salt concentrations

    DEFF Research Database (Denmark)

    Paludan-Müller, Christine; Madsen, M.; Sophanodora, P.

    2002-01-01

    % salt (w/w) as well as two high-salt batches, containing 9% and 11% salt. pH decreased rapidly from 6 to 4.5 in low-salt batches, whereas in high-salt batches, a slow or no decrease in pH was found. Lactic acid bacteria (LAB) and yeasts were isolated as the dominant microorganisms during fermentation....... LAB counts increased to 108-109 cfu g-1 and yeast counts to 107-5 x 107 cfu g-1 in all batches, except in the 11% salt batch, where counts were 1-2 log lower. Phenotypic tests, ITS-PCR, carbohydrate fermentations and 16S rRNA gene sequencing identified LAB isolates as Pediococcus pentosaceus......Plaa-som is a Thai fermented fish product prepared from snakehead fish, salt, palm syrup and sometimes roasted rice. We studied the effects of different salt concentrations on decrease in pH and on microflora composition during fermentation. Two low-salt batches were prepared, containing 6% and 7...

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

    Science.gov (United States)

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

    2013-01-10

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

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

    Science.gov (United States)

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

    2014-01-01

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

  7. Production and storage of biohydrogen during sequential batch fermentation of Spirogyra hydrolyzate by Clostridium butyricum

    International Nuclear Information System (INIS)

    Ortigueira, Joana; Pinto, Tiago; Gouveia, Luísa; Moura, Patrícia

    2015-01-01

    The biological hydrogen production from Spirogyra sp. biomass was studied in a SBR (sequential batch reactor) equipped with a biogas collecting and storage system. Two acid hydrolysis pre-treatments (1N and 2N H 2 SO 4 ) were applied to the Spirogyra biomass and the subsequent fermentation by Clostridium butyricum DSM 10702 was compared. The 1N and 2N hydrolyzates contained 37.2 and 40.8 g/L of total sugars, respectively, and small amounts of furfural and HMF (hydroxymethylfurfural). These compounds did not inhibit the hydrogen production from crude Spirogyra hydrolyzates. The fermentation was scaled up to a batch operated bioreactor coupled with a collecting system that enabled the subsequent characterization and storage of the biogas produced. The cumulative hydrogen production was similar for both 1N and 2N hydrolyzate, but the hydrogen production rates were 438 and 288 mL/L.h, respectively, suggesting that the 1N hydrolyzate was more suitable for sequential batch fermentation. The SBR with 1N hydrolyzate was operated continuously for 13.5 h in three consecutive batches and the overall hydrogen production rate and yield reached 324 mL/L.h and 2.59 mol/mol, respectively. This corresponds to a potential daily production of 10.4 L H 2 /L Spirogyra hydrolyzate, demonstrating the excellent capability of C. butyricum to produce hydrogen from microalgal biomass. - Highlights: • Production of biohydrogen from crude Spirogyra hydrolyzates. • Set-up of a collecting and storage system for continuous biogas sampling. • The hydrogen production rate is 324 mL/L.h in the SBR (sequential batch reactor). • The SBR produces daily an equivalent to 10.4 L H 2 /L of crude Spirogyra hydrolyzate

  8. Genetic Algorithmic Optimization of PHB Production by a Mixed Culture in an Optimally Dispersed Fed-batch Bioreactor

    Directory of Open Access Journals (Sweden)

    Pratap R. Patnaik

    2009-10-01

    Full Text Available Poly-β-hydroxybutyrate (PHB is an energy-storage polymer whose properties are similar to those of chemical polymers such as polyethylene and polypropylene. Moreover, PHB is biodegradable, absorbed by human tissues and less energy-consuming than synthetic polymers. Although Ralstonia eutropha is widely used to synthesize PHB, it is inefficient in utilizing glucose and similar sugars. Therefore a co-culture of R. eutropha and Lactobacillus delbrueckii is preferred since the latter can convert glucose to lactate, which R. eutropha can metabolize easily. Tohyama et al. [24] maximized PHB production in a well-mixed fed-batch bioreactor with glucose and (NH42SO4 as the primary substrates. Since production-scale bioreactors often deviate from ideal laboratory-scale reactors, a large bioreactor was simulated by means of a dispersion model with the kinetics determined by Tohyama et al. [24] and dispersion set at an optimum Peclet number of 20 [32]. The time-dependent feed rates of the two substrates were determined through a genetic algorithm (GA to maximize PHB production. This bioreactor produced 22.2% more PHB per liter and 12.8% more cell mass than achieved by Tohyama et al. [24]. These results, and similar observations with other fermentations, indicate the feasibility of enhancing the efficiency of large nonideal bioreactors through GA optimizations.

  9. Enzymatic hydrolysis of rice straw and glucose fermentation using a Vertical Ball Mill Bioreactor (VBMB): Impact of operational conditions

    DEFF Research Database (Denmark)

    Castro, Rafael C.A.; Mussatto, Solange I.; Roberto, Inês C.

    ). This bioreactor was equipped with adjustable flat round plate impellers, allowing its operation with glass spheres as shear agent. For enzymatic hydrolysis, the spheres were the only variable with significant impact on the results, being achieved 87% cellulose conversion after 24 h when using the highest level...... saccharification and fermentation, in batch or fed-batch configurations, and with possibilities of operating at high solids content. Acknowledgments: FAPESP (2013/13953-6 and 2015/24813-6) and CNPq....

  10. Switching the mode of sucrose utilization by Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Miletti Luiz C

    2008-02-01

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

  11. Switching the mode of sucrose utilization by Saccharomyces cerevisiae.

    Science.gov (United States)

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

    2008-02-27

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

  12. Boosted structured additive regression for Escherichia coli fed-batch fermentation modeling.

    Science.gov (United States)

    Melcher, Michael; Scharl, Theresa; Luchner, Markus; Striedner, Gerald; Leisch, Friedrich

    2017-02-01

    The quality of biopharmaceuticals and patients' safety are of highest priority and there are tremendous efforts to replace empirical production process designs by knowledge-based approaches. Main challenge in this context is that real-time access to process variables related to product quality and quantity is severely limited. To date comprehensive on- and offline monitoring platforms are used to generate process data sets that allow for development of mechanistic and/or data driven models for real-time prediction of these important quantities. Ultimate goal is to implement model based feed-back control loops that facilitate online control of product quality. In this contribution, we explore structured additive regression (STAR) models in combination with boosting as a variable selection tool for modeling the cell dry mass, product concentration, and optical density on the basis of online available process variables and two-dimensional fluorescence spectroscopic data. STAR models are powerful extensions of linear models allowing for inclusion of smooth effects or interactions between predictors. Boosting constructs the final model in a stepwise manner and provides a variable importance measure via predictor selection frequencies. Our results show that the cell dry mass can be modeled with a relative error of about ±3%, the optical density with ±6%, the soluble protein with ±16%, and the insoluble product with an accuracy of ±12%. Biotechnol. Bioeng. 2017;114: 321-334. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  13. Transcriptional response of P. pastoris in fed-batch cultivations to Rhizopus oryzae lipase production reveals UPR induction

    Directory of Open Access Journals (Sweden)

    Valero Francisco

    2007-07-01

    Full Text Available Abstract Background The analysis of transcriptional levels of the genes involved in protein synthesis and secretion is a key factor to understand the host organism's responses to recombinant protein production, as well as their interaction with the cultivation conditions. Novel techniques such as the sandwich hybridization allow monitoring quantitatively the dynamic changes of specific RNAs. In this study, the transcriptional levels of some genes related to the unfolded protein response (UPR and central metabolism of Pichia pastoris were analysed during batch and fed-batch cultivations using an X-33-derived strain expressing a Rhizopus oryzae lipase under control of the formaldehyde dehydrogenase promoter (FLD1, namely the alcohol oxidase gene AOX1, the formaldehyde dehydrogenase FLD1, the protein disulfide isomerase PDI, the KAR2 gene coding for the BiP chaperone, the 26S rRNA and the R. oryzae lipase gene ROL. Results The transcriptional levels of the selected set of genes were first analysed in P. pastoris cells growing in shake flask cultures containing different carbon and nitrogen sources combinations, glycerol + ammonium, methanol + methylamine and sorbitol + methylamine. The transcriptional levels of the AOX1 and FLD1 genes were coherent with the known regulatory mechanism of C1 substrates in P. pastoris, whereas ROL induction lead to the up-regulation of KAR2 and PDI transcriptional levels, thus suggesting that ROL overexpression triggers the UPR. This was further confirmed in fed-batch cultivations performed at different growth rates. Transcriptional levels of the analysed set of genes were generally higher at higher growth rates. Nevertheless, when ROL was overexpressed in a strain having the UPR constitutively activated, significantly lower relative induction levels of these marker genes were detected. Conclusion The bead-based sandwich hybridization assay has shown its potential as a reliable instrument for quantification of

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

    Science.gov (United States)

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

    2005-03-01

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

  15. Biological treatment of potato processing wastewater for red pigment production by immobilized cells of UV-irradiated monascus sp. in repeated batch

    International Nuclear Information System (INIS)

    Khalaf, S.A.

    2004-01-01

    Potato processing wastewater (PPW) was collected and analyzed for biological oxygen demand (BOD), chemical oxygen demand (COD), total suspended solids (TSS), total nitrogen and starch content. A fungal strain isolated from PPW identified as Monascus sp. PPW was evaluated for its ability to grow and produce red pigment, biomass and reduce the starch content of the ,PPW. Active UV-irradiated isolate of the above strain was obtained by exposing the parent strain to UV-radiation and coded Monascus. sp. PPW-UV7 and used as immobilized cell system for PPW treatment process in repeated batch fermentation. The immobilized cells (in sponge cubes) were able to reduce COD by about 85.7 %, with biomass production of 9.22 gl+ l and over productivity of red pigment of 2.6 gl+ 1 after 8 days fermentation (2 batches). The immobilized cells showed stability and viability for 8 batches (32 days) during the process treatment

  16. A fast approach to determine a fed batch feeding profile for recombinant Pichia pastoris strains

    Directory of Open Access Journals (Sweden)

    Herwig Christoph

    2011-10-01

    Full Text Available Abstract Background The microorganism Pichia pastoris is a commonly used microbial host for the expression of recombinant proteins in biotechnology and biopharmaceutical industry. To speed up process development, a fast methodology to determine strain characteristic parameters, which are needed to subsequently set up fed batch feeding profiles, is required. Results Here, we show the general applicability of a novel approach to quantify a certain minimal set of bioprocess-relevant parameters, i.e. the adaptation time of the culture to methanol, the specific substrate uptake rate during the adaptation phase and the maximum specific substrate uptake rate, based on fast and easy-to-do batch cultivations with repeated methanol pulses in a batch culture. A detailed analysis of the adaptation of different P. pastoris strains to methanol was conducted and revealed that each strain showed very different characteristics during adaptation, illustrating the need of individual screenings for an optimal parameter definition during this phase. Based on the results obtained in batch cultivations, dynamic feeding profiles based on the specific substrate uptake rate were employed for different P. pastoris strains. In these experiments the maximum specific substrate uptake rate, which had been defined in batch experiments, also represented the upper limit of methanol uptake, underlining the validity of the determined process-relevant parameters and the overall experimental strategy. Conclusion In this study, we show that a fast approach to determine a minimal set of strain characteristic parameters based on easy-to-do batch cultivations with methanol pulses is generally applicable for different P. pastoris strains and that dynamic fed batch strategies can be designed on the specific substrate uptake rate without running the risk of methanol accumulation.

  17. Kinetic Modeling of Ethanol Batch Fermentation by Escherichia Coli FBWHR Using Hot-Water Sugar Maple Wood Extract Hydrolyzate as Substrate

    Directory of Open Access Journals (Sweden)

    Yang Wang

    2014-12-01

    Full Text Available A recombinant strain of Escherichia coli FBWHR was used for ethanol fermentation from hot-water sugar maple wood extract hydrolyzate in batch experiments. Kinetic studies of cell growth, sugar utilization and ethanol production were investigated at different initial total sugar concentrations of wood extract hydrolyzate. The highest ethanol concentration of 24.05 g/L was obtained using an initial total sugar concentration of 70.30 g/L. Unstructured models were developed to describe cell growth, sugar utilization and ethanol production and validated by comparing the predictions of model and experimental data. The results from this study could be expected to provide insights into the process performance, optimize the process and aid in the design of processes for large-scale production of ethanol fermentation from woody biomass.

  18. Achieving precision in high density batch mode micro-electro-discharge machining

    International Nuclear Information System (INIS)

    Richardson, Mark T; Gianchandani, Yogesh B

    2008-01-01

    This paper reports a parametric study of batch mode micro-electro-discharge machining (µEDM) of high density features in stainless steel. Lithographically fabricated copper tools with single cross, parallel line and 8 × 8 circle/square array features of 5–100 µm width and 5–75 µm spacing were used to quantify trends in machining tolerance and the impact of debris accumulation. As the tool feature density is increased, debris accumulation effects begin to dominate, eventually degrading both tool and workpiece. Two independent techniques for mitigating this debris buildup are separately investigated. The first is a passivation coating which suppresses spurious discharges triggered from the sidewalls of the machining tool. By this method, the mean tool wear rate decreases from a typical of about 34% to 1.7% and machining non-uniformity reduces from 4.9 µm to 1.1 µm across the workpiece. The second technique involves a two-step machining process that enhances the hydrodynamic removal of machining debris compared to standard methods. This improves surface and edge finish, machining time and tool wear

  19. Xylitol production by Candida parapsilosis under fed-batch culture

    Directory of Open Access Journals (Sweden)

    Sandra A. Furlan

    2001-06-01

    Full Text Available Xylitol production by Candida parapsilosis was investigated under fed-batch cultivation, using single (xylose or mixed (xylose and glucose sugars as substrates. The presence of glucose in the medium induced the production of ethanol as secondary metabolite and improved specific rates of growth, xylitol formation and substrate consumption. Fractionated supply of the feed medium at constant sugar concentration did not promote any increase on the productivity compared to the single batch cultivation.A produção de xylitol por Candida parapsilosis foi investigada em regime de batelada alimentada, usando substratos açucarados de composição simples (xilose ou composta (xilose e glicose. A presença de glicose no meio induziu a formação de etanol como metabólito secundário. A suplementação fracionada do meio de alimentação numa concentração fixa de açúcar não resultou em aumento da produtividade em relação àquela alcançada em batelada simples.

  20. In silico aided metabolic engineering of Klebsiella oxytoca and fermentation optimization for enhanced 2,3-butanediol production.

    Science.gov (United States)

    Park, Jong Myoung; Song, Hyohak; Lee, Hee Jong; Seung, Doyoung

    2013-09-01

    Klebsiella oxytoca naturally produces a large amount of 2,3-butanediol (2,3-BD), a promising bulk chemical with wide industrial applications, along with various byproducts. In this study, the in silico gene knockout simulation of K. oxytoca was carried out for 2,3-BD overproduction by inhibiting the formation of byproducts. The knockouts of ldhA and pflB genes were targeted with the criteria of maximization of 2,3-BD production and minimization of byproducts formation. The constructed K. oxytoca ΔldhA ΔpflB strain showed higher 2,3-BD yields and higher final concentrations than those obtained from the wild-type and ΔldhA strains. However, the simultaneous deletion of both genes caused about a 50 % reduction in 2,3-BD productivity compared with K. oxytoca ΔldhA strain. Based on previous studies and in silico investigation that the agitation speed during 2,3-BD fermentation strongly affected cell growth and 2,3-BD synthesis, the effect of agitation speed on 2,3-BD production was investigated from 150 to 450 rpm in 5-L bioreactors containing 3-L culture media. The highest 2,3-BD productivity (2.7 g/L/h) was obtained at 450 rpm in batch fermentation. Considering the inhibition of acetoin for 2,3-BD production, fed-batch fermentations were performed using K. oxytoca ΔldhA ΔpflB strain to enhance 2,3-BD production. Altering the agitation speed from 450 to 350 rpm at nearly 10 g/L of acetoin during the fed-batch fermentation allowed for the production of 113 g/L 2,3-BD, with a yield of 0.45 g/g, and for the production of 2.1 g/L/h of 2,3-BD.

  1. Fermentation and recovery of L-glutamic acid from cassava starch hydrolysate by ion-exchange resin column

    Directory of Open Access Journals (Sweden)

    Nampoothiri K. Madhavan

    1999-01-01

    Full Text Available Investigations were carried out with the aim of producing L-glutamic acid from Brevibacterium sp. by utilizing a locally available starchy substrate, cassava (Manihot esculenta Crantz. Initial studies were carried out in shake flasks, which showed that even though the yield was high with 85-90 DE (Dextrose Equivalent value, the maximum conversion yield (~34% was obtained by using only partially digested starch hydrolysate, i.e. 45-50 DE. Fermentations were carried out in batch mode in a 5 L fermenter, using suitably diluted cassava starch hydrolysate, using a 85-90 DE value hydrolysate. Media supplemented with nutrients resulted in an accumulation of 21 g/L glutamic acid with a fairly high (66.3% conversation yield of glucose to glutamic acid (based on glucose consumed and on 81.74% theoretical conversion rate. The bioreactor conditions most conducive for maximum production were pH 7.5, temperature 30°C and an agitation of 180 rpm. When fermentation was conducted in fed-batch mode by keeping the residual reducing sugar concentration at 5% w/v, 25.0 g/L of glutamate was obtained after 40 h fermentation (16% more the batch mode. Chromatographic separation by ion-exchange resin was used for the recovery and purification of glutamic acid. It was further crystallized and separated by making use of its low solubility at the isoelectric point (pH 3.2.

  2. Batch fermentation of whey ultra filtrate by Lactobacillus helveticus for lactic acid production

    Energy Technology Data Exchange (ETDEWEB)

    Roy, D; Goulet, J; Le Duy, Q

    1986-06-01

    Cheese whey ultrafiltrate (WU) was used as the carbon source for the production of lactic acid by batch fermentation with Lactobacillus helveticus strain milano. The fermentation was conducted in a 400 ml fermentor at an agitation rate of 200 revolutions per minute and under conditions of controlled temperature (42 degrees C) and pH. In the whey ultrafiltrate-corn steep liquor (WU-CSL) medium, the optimal pH for fermentation was 5.9. Inoculum propagated in skim milk (SM) medium or in lactose synthetic (LS) medium resulted in the best performance in fermentation (in terms of growth, lactic acid production, lactic acid yield and maximum productivity of lactic acid), as compared to that propagated in glucose synthetic (GS) medium. The yeast extract ultrafiltrate (YEU) used as the nitrogen/growth factor source in the WU medium at 1.5% (w/v) gave the highest maximum productivity of lactic acid of 2.70 g/l-h, as compared to the CSL and the tryptone ultrafiltrate (TU). 27 references.

  3. Dynamics of chemical elements in the fermentation process of ethanol production

    International Nuclear Information System (INIS)

    Nepomuceno, N.; Fernandes, E.A.N.; Bacchi, M.A.

    1997-01-01

    Brazil has become the largest producer of biomass ethanol derived from sugar cane. The industrial production is based on the fermentation of sugar cane juice by yeast, inside of large volume vats, in a fed-batch process that recycles yeast cells. To study the dynamics of chemical elements in each operating cycle, five stages of the fermentation process were considered: must, yeast suspension, wine, non-yeast wine and yeast cream. For this, a mass balance of the terrigenous elements, Ce, Co, Cs, Eu, Fe, Hf, La, Na, Sc, Sm, and Th, and the sugar cane plant elements, Br, K, Rb, and Zn, were established in fermentation vats of an industrial scale unit, with sampling undertaken during different climatic conditions (dry and rainy periods). A similar distribution of the sugar cane characteristics elements was found for the stages analysed, while for the terrigenous elements a trend of accumulation in the yeast cream was observed. Preferential absorption of Br, K, Rb, and Zn by yeast cells was indicated by the smaller concentrations observed in yeast suspension than in yeast cream. (author)

  4. Enhanced anti-oxidative activity and lignocellulosic ethanol production by biotin addition to medium in Pichia guilliermondii fermentation.

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

  6. Improving wood hydrolyzate fermentation by using schizosaccharomycetes

    Energy Technology Data Exchange (ETDEWEB)

    Kalyuzhnyi, M Ya; Ustinova, V I; Petrushko, G I

    1967-01-01

    The development of Schizosaccharomycetes (I) in wood hydrolyzates is not observed when fermentation is carried out by the convetional batch process, evidently because of the highly inhibitory action of the medium. More recently, with the introduction of continuous fermentation of wood and other hydrolyzates, the occurrence of I has been frequently reported, and in some hydrolysis plants, I became predominant, eliminating the budding yeast strains. The phenomenon can be attributed to higher temperatures employed in continuous fermentation, and to a more favorable medium, as the hydrolyzate is diluted with spent fermentation liquor (the flow of fresh medium constitutes about 20% of the fermentation-vat volume). The I cells, when grown under favorable conditions, have a high fermenting power, adapt easily to the fermentation of galactose, and give higher yields of ethanol than the budding yeast. As observed at plants using I, however, the cells are sensitive to variations in the fermentation process, and are inactivated upon storage. This is usually attributed to their inability to store polysaccharides, and especially glycogen. An experimental study undertaken to determine conditions under which reserve polysaccharides accumulate in I cells showed that the important factor is the quality of the medium in which the cells are grown and the conditions of storage. In media enriched with spent fermentaion liquor or with cell autolyzate and purified from toxic components, considerable amounts of glycogen accumulate in the cells.

  7. Saccharomyces cerevisiae KNU5377 stress response during high-temperature ethanol fermentation.

    Science.gov (United States)

    Kim, Il-Sup; Kim, Young-Saeng; Kim, Hyun; Jin, Ingnyol; Yoon, Ho-Sung

    2013-03-01

    Fuel ethanol production is far more costly to produce than fossil fuels. There are a number of approaches to cost-effective fuel ethanol production from biomass. We characterized stress response of thermotolerant Saccharomyces cerevisiae KNU5377 during glucose-based batch fermentation at high temperature (40°C). S. cerevisiae KNU5377 (KNU5377) transcription factors (Hsf1, Msn2/4, and Yap1), metabolic enzymes (hexokinase, glyceraldehyde-3-phosphate dehydrogenase, glucose-6-phosphate dehydrogenase, isocitrate dehydrogenase, and alcohol dehydrogenase), antioxidant enzymes (thioredoxin 3, thioredoxin reductase, and porin), and molecular chaperones and its cofactors (Hsp104, Hsp82, Hsp60, Hsp42, Hsp30, Hsp26, Cpr1, Sti1, and Zpr1) are upregulated during fermentation, in comparison to S. cerevisiae S288C (S288C). Expression of glyceraldehyde-3-phosphate dehydrogenase increased significantly in KNU5377 cells. In addition, cellular hydroperoxide and protein oxidation, particularly lipid peroxidation of triosephosphate isomerase, was lower in KNU5377 than in S288C. Thus, KNU5377 activates various cell rescue proteins through transcription activators, improving tolerance and increasing alcohol yield by rapidly responding to fermentation stress through redox homeostasis and proteostasis.

  8. Characterization of rumen bacterial diversity and fermentation parameters in concentrate fed cattle with and without forage.

    Science.gov (United States)

    Petri, R M; Forster, R J; Yang, W; McKinnon, J J; McAllister, T A

    2012-06-01

    To determine the effects of the removal of forage in high-concentrate diets on rumen fermentation conditions and rumen bacterial populations using culture-independent methods. Detectable bacteria and fermentation parameters were measured in the solid and liquid fractions of digesta from cattle fed two dietary treatments, high concentrate (HC) and high concentrate without forage (HCNF). Comparison of rumen fermentation conditions showed that duration of time spent below pH 5·2 and rumen osmolality were higher in the HCNF treatment. Simpson's index of 16S PCR-DGGE images showed a greater diversity of dominant species in the HCNF treatment. Real-time qPCR showed populations of Fibrobacter succinogenes (P = 0·01) were lower in HCNF than HC diets. Ruminococcus spp., F. succinogenes and Selenomonas ruminantium were at higher (P ≤ 0·05) concentrations in the solid vs the liquid fraction of digesta regardless of diet. The detectable bacterial community structure in the rumen is highly diverse. Reducing diet complexity by removing forage increased bacterial diversity despite the associated reduction in ruminal pH being less conducive for fibrolytic bacterial populations. Quantitative PCR showed that removal of forage from the diet resulted in a decline in the density of some, but not all fibrolytic bacterial species examined. Molecular techniques such as DGGE and qPCR provide an increased understanding of the impacts of dietary changes on the nature of rumen bacterial populations, and conclusions derived using these techniques may not match those previously derived using traditional laboratory culturing techniques. © 2012 The Authors. Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

  9. Production of lactic acid from sucrose: strain selection, fermentation, and kinetic modeling.

    Science.gov (United States)

    Lunelli, Betânia H; Andrade, Rafael R; Atala, Daniel I P; Wolf Maciel, Maria Regina; Maugeri Filho, Francisco; Maciel Filho, Rubens

    2010-05-01

    Lactic acid is an important product arising from the anaerobic fermentation of sugars. It is used in the pharmaceutical, cosmetic, chemical, and food industries as well as for biodegradable polymer and green solvent production. In this work, several bacterial strains were isolated from industrial ethanol fermentation, and the most efficient strain for lactic acid production was selected. The fermentation was conducted in a batch system under anaerobic conditions for 50 h at a temperature of 34 degrees C, a pH value of 5.0, and an initial sucrose concentration of 12 g/L using diluted sugarcane molasses. Throughout the process, pulses of molasses were added in order to avoid the cell growth inhibition due to high sugar concentration as well as increased lactic acid concentrations. At the end of the fermentation, about 90% of sucrose was consumed to produce lactic acid and cells. A kinetic model has been developed to simulate the batch lactic acid fermentation results. The data obtained from the fermentation were used for determining the kinetic parameters of the model. The developed model for lactic acid production, growth cell, and sugar consumption simulates the experimental data well.

  10. Prevention of GABA reduction during dough fermentation using a baker's yeast dal81 mutant.

    Science.gov (United States)

    Ando, Akira; Nakamura, Toshihide

    2016-10-01

    γ-Aminobutyric acid (GABA) is consumed by yeasts during fermentation. To prevent GABA reduction in bread dough, a baker's yeast mutant AY77 deficient in GABA assimilation was characterized and utilized for wheat dough fermentation. An amber mutation in the DAL81 gene, which codes for a positive regulator of multiple nitrogen degradation pathways, was found in the AY77 strain. The qPCR analyses of genes involved in nitrogen utilization showed that transcriptional levels of the UGA1 and DUR3 genes encoding GABA transaminase and urea transporter, respectively, are severely decreased in the AY77 cells. The AY77 strain cultivated by fed-batch culture using cane molasses exhibited inferior gas production during dough fermentation compared to that of wild-type strain AY13. However, when fed with molasses containing 0.5% ammonium sulfate, the mutant strain exhibited gas production comparable to that of the AY13 strain. In contrast to the AY13 strain, which completely consumed GABA in dough within 5 h, the AY77 strain consumed no GABA under either culture condition. Dough fermentation with the dal81 mutant strain should be useful for suppression of GABA reduction in breads. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  11. PEI detoxification of pretreated spruce for high solids ethanol fermentation

    DEFF Research Database (Denmark)

    Cannella, David; Sveding, Per Viktor; Jørgensen, Henning

    2014-01-01

    .e. spruce) this has been difficult to reach. The main reason behind this difference is the higher recalcitrance of woody substrates which require harsher pretreatment conditions, thus generating higher amounts of inhibitory compounds, ultimately lowering fermentation performances. In this work we studied...... ethanol production from spruce performing the whole process, from pretreatment to hydrolysis and fermentation, at 30% dry matter (equivalent to similar to 20% WIS). Hydrolysis and fermentation was performed in a horizontal free fall mixing reactor enabling efficient mixing at high solids loadings....... In batch simultaneous saccharification and fermentation (SSF), up to 76% cellulose to ethanol conversion was achieved resulting in a concentration of 51 g/kg of ethanol. Key to obtaining this high ethanol yield at these conditions was the use of a detoxification technology based on applying a soluble...

  12. Stimulation of electro-fermentation in single-chamber microbial electrolysis cells driven by genetically engineered anode biofilms

    Science.gov (United States)

    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.

  13. Combined data preprocessing and multivariate statistical analysis characterizes fed-batch culture of mouse hybridoma cells for rational medium design.

    Science.gov (United States)

    Selvarasu, Suresh; Kim, Do Yun; Karimi, Iftekhar A; Lee, Dong-Yup

    2010-10-01

    We present an integrated framework for characterizing fed-batch cultures of mouse hybridoma cells producing monoclonal antibody (mAb). This framework systematically combines data preprocessing, elemental balancing and statistical analysis technique. Initially, specific rates of cell growth, glucose/amino acid consumptions and mAb/metabolite productions were calculated via curve fitting using logistic equations, with subsequent elemental balancing of the preprocessed data indicating the presence of experimental measurement errors. Multivariate statistical analysis was then employed to understand physiological characteristics of the cellular system. The results from principal component analysis (PCA) revealed three major clusters of amino acids with similar trends in their consumption profiles: (i) arginine, threonine and serine, (ii) glycine, tyrosine, phenylalanine, methionine, histidine and asparagine, and (iii) lysine, valine and isoleucine. Further analysis using partial least square (PLS) regression identified key amino acids which were positively or negatively correlated with the cell growth, mAb production and the generation of lactate and ammonia. Based on these results, the optimal concentrations of key amino acids in the feed medium can be inferred, potentially leading to an increase in cell viability and productivity, as well as a decrease in toxic waste production. The study demonstrated how the current methodological framework using multivariate statistical analysis techniques can serve as a potential tool for deriving rational medium design strategies. Copyright © 2010 Elsevier B.V. All rights reserved.

  14. Novel integration strategy for enhancing chalcopyrite bioleaching by Acidithiobacillus sp. in a 7-L fermenter.

    Science.gov (United States)

    Feng, Shoushuai; Yang, Hailin; Zhan, Xiao; Wang, Wu

    2014-06-01

    An integrated strategy (additional energy substrate-three stage pH control-fed batch) was firstly proposed for efficiently improving chalcopyrite bioleaching by Acidithiobacillus sp. in a 7-L fermenter. The strain adaptive-growing phase was greatly shortened from 8days into 4days with the supplement of additional 2g/L Fe(2+)+2g/L S(0). Jarosite passivation was effectively weakened basing on higher biomass via the three-stage pH-stat control (pH 1.3-1.0-0.7). The mineral substrate inhibition was attenuated by fed-batch fermentation. With the integrated strategy, the biochemical reaction was promoted and achieved a better balance. Meanwhile, the domination course of A. thiooxidans in the microbial community was shortened from 14days to 8days. As the results of integrated strategy, the final copper ion and productivity reached 89.1mg/L and 2.23mg/(Ld), respectively, which was improved by 52.8% compared to the uncontrolled batch bioleaching. The integrated strategy could be further exploited for industrial chalcopyrite bioleaching. Copyright © 2014 Elsevier Ltd. All rights reserved.

  15. Development of a lipase fermentation process that uses a recombinant Pseudomonas alcaligenes strain

    NARCIS (Netherlands)

    Gerritse, G; Hommes, R.W J; Quax, Wim

    Pseudomonas alcaligenes M-l secretes an alkaline lipase, which has excellent characteristics for the removal of fatty stains under modern washing conditions. A fed-batch fermentation process based on the secretion of the alkaline lipase from P. alcaligenes was developed. Due to the inability of P.

  16. A grey box model of glucose fermentation and syntrophic oxidation in microbial fuel cells.

    Science.gov (United States)

    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.

  17. Aeration-Controlled Formation of Acid in Heterolactic Fermentations

    DEFF Research Database (Denmark)

    Adler-Nissen, Jens

    1994-01-01

    fermentation processes should be analyzed as fed-batch fermentations with oxygen as the limiting substrate. Addition of fructose in limited amounts leads to the formation of one half mole of acetic acid for each mole fructose, thus offering an alternative mechanism for controlling acetic acid formation.......Controlled aeration of Leuconostoc mesenteroides was studied as a possible mechanism for control of the formation of acetic acid, a metabolite of major influence on the taste of lactic fermented foods. Fermentations were carried out in small scale in a medium in which growth was limited...... by the buffer capacity only. Ethanol and acetic acid formed during the fermentation were analyzed by rapid head space gas chromatography, and the ratio of the molar concentrations of these two volatiles quantitatively predicted the balance between the formation of acetic acid and lactic acid. The oxygen...

  18. Optimal control of switched systems arising in fermentation processes

    CERN Document Server

    Liu, Chongyang

    2014-01-01

    The book presents, in a systematic manner, the optimal controls under different mathematical models in fermentation processes. Variant mathematical models – i.e., those for multistage systems; switched autonomous systems; time-dependent and state-dependent switched systems; multistage time-delay systems and switched time-delay systems – for fed-batch fermentation processes are proposed and the theories and algorithms of their optimal control problems are studied and discussed. By putting forward novel methods and innovative tools, the book provides a state-of-the-art and comprehensive systematic treatment of optimal control problems arising in fermentation processes. It not only develops nonlinear dynamical system, optimal control theory and optimization algorithms, but can also help to increase productivity and provide valuable reference material on commercial fermentation processes.

  19. Ethanol production from biomass by repetitive solid-state fed-batch fermentation with continuous recovery of ethanol

    Energy Technology Data Exchange (ETDEWEB)

    Moukamnerd, Churairat; Kino-oka, Masahiro; Sugiyama, Minetaka; Kaneko, Yoshinobu; Harashima, Satoshi; Katakura, Yoshio [Osaka Univ. (Japan). Dept. of Biotechnology; Boonchird, Chuenchit [Mahidol Univ., Bangkok (Thailand). Dept. of Biotechnology; Noda, Hideo [Kansai Chemical Engineering Co., Ltd., Amagasaki (Japan); Ninomiya, Kazuaki [Kanazawa Univ. (Japan). Inst. of International Environment Technology; Shioya, Suteaki [Sojo Univ., Kumamoto (Japan). Dept. of Applied Life Science

    2010-09-15

    To save cost and input energy for bioethanol production, a consolidated continuous solid-state fermentation system composed of a rotating drum reactor, a humidifier, and a condenser was developed. Biomass, saccharifying enzymes, yeast, and a minimum amount of water are introduced into the system. Ethanol produced by simultaneous saccharification and fermentation is continuously recovered as vapor from the headspace of the reactor, while the humidifier compensates for the water loss. From raw corn starch as a biomass model, 95 {+-} 3, 226 {+-} 9, 458 {+-} 26, and 509 {+-} 64 g l{sup -1} of ethanol solutions were recovered continuously when the ethanol content in reactor was controlled at 10-20, 30-50, 50-70 and 75-85 g kg-mixture{sup -1}, respectively. The residue showed a lesser volume and higher solid content than that obtained by conventional liquid fermentation. The cost and energy for intensive waste water treatment are decreased, and the continuous fermentation enabled the sustainability of enzyme activity and yeast in the system. (orig.)

  20. Influence of extracellular pH on growth, viability, cell size, acidification activity, and intracellular pH of Lactococcus lactis in batch fermentations.

    Science.gov (United States)

    Hansen, Gunda; Johansen, Claus Lindvald; Marten, Gunvor; Wilmes, Jacqueline; Jespersen, Lene; Arneborg, Nils

    2016-07-01

    In this study, we investigated the influence of three extracellular pH (pHex) values (i.e., 5.5, 6.5, and 7.5) on the growth, viability, cell size, acidification activity in milk, and intracellular pH (pHi) of Lactococcus lactis subsp. lactis DGCC1212 during pH-controlled batch fermentations. A universal parameter (e.g., linked to pHi) for the description or prediction of viability, specific acidification activity, or growth behavior at a given pHex was not identified. We found viability as determined by flow cytometry to remain high during all growth phases and irrespectively of the pH set point. Furthermore, regardless of the pHex, the acidification activity per cell decreased over time which seemed to be linked to cell shrinkage. Flow cytometric pHi determination demonstrated an increase of the averaged pHi level for higher pH set points, while the pH gradient (pHi-pHex) and the extent of pHi heterogeneity decreased. Cells maintained positive pH gradients at a low pHex of 5.5 and even during substrate limitation at the more widely used pHex 6.5. Moreover, the strain proved able to grow despite small negative or even absent pH gradients at a high pHex of 7.5. The larger pHi heterogeneity at pHex 5.5 and 6.5 was associated with more stressful conditions resulting, e.g., from higher concentrations of non-dissociated lactic acid, while the low pHi heterogeneity at pHex 7.5 most probably corresponded to lower concentrations of non-dissociated lactic acid which facilitated the cells to reach the highest maximum active cell counts of the three pH set points.

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

    Science.gov (United States)

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

    2011-04-10

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

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

    Science.gov (United States)

    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.

  3. Economics of alcohol manufacture by fermentation of cane-sugar molasses

    Energy Technology Data Exchange (ETDEWEB)

    Narasimhe, I

    1964-01-01

    An improved continuous fermentation technique consists of using a special strain of yeast, GN/1942-2 (C), having high attenuating power and high alcohol tolerance which can function efficiently in the higher concentrations of sugars necessary to produce a mash containing 10 to 12% alcohol. The yeast is fed with suitable nutrients in the pre-fermentors at a regulated rate to maintain a constant cell population at all stages, thus enhancing the speed of fermentation. Thus, double the normal quantity of molasses can be fermented without sacrificing the efficiency of fermentation.

  4. Economics of alcohol manufacture by fermentation of cane-sugar molasses

    Energy Technology Data Exchange (ETDEWEB)

    Narasimha, I

    1964-01-01

    An improved continuous fermentation technique consists of using a special strain of yeast, GN/1942-2 (C), having high attenuating power and high alcohol tolerance which can function efficiently in the higher concentrations of sugars necessary to produce a mash containing 10 to 12% alcohol. The yeast is fed with suitable nutrients in the prefermentors at a regulated rate to maintain a constant cell population at all stages, thus enhancing the speed of fermentation. Thus, double the normal quantity of molasses can be fermented without sacrificing the efficiency of fermentation.

  5. EFFECTIVE FACTORS AND MODEL SYSTEMS IN THE INDUSTRIAL PRODUCTION OF NISIN

    Directory of Open Access Journals (Sweden)

    Ömer ŞİMŞEK

    2007-01-01

    Full Text Available Nisin is the first bacteriocin identified in Lactococcus lactis and belongs to type 1 lanthibiotic group. High nisin production in cultured media is related with the composition of fermentation medium, pH, produced nisin concentration and most importantly growth amount of cell. For industrial purpose, batch, fed-batch and continue fermentation systems were developed by regarding these factors. Maintaining efficient production of nisin having important potential at preservation of foods is important for both industrial production and using as starter culture. In this review the fermentation factors at nisin production were outlined and constructed model systems were compared.

  6. Fed-Batch Control and Visualization of Monomer Sequences of Individual ICAR ATRP Gradient Copolymer Chains

    Directory of Open Access Journals (Sweden)

    Dagmar R. D'hooge

    2014-04-01

    Full Text Available Based on kinetic Monte Carlo simulations of the monomer sequences of a representative number of copolymer chains (≈ 150,000, optimal synthesis procedures for linear gradient copolymers are proposed, using bulk Initiators for Continuous Activator Regeneration Atom Transfer Radical Polymerization (ICAR ATRP. Methyl methacrylate and n-butyl acrylate are considered as comonomers with CuBr2/PMDETA (N,N,N′,N′′,N′′-pentamethyldiethylenetriamine as deactivator at 80 °C. The linear gradient quality is determined in silico using the recently introduced gradient deviation ( polymer property. Careful selection or fed-batch addition of the conventional radical initiator I2 allows a reduction of the polymerization time with ca. a factor 2 compared to the corresponding batch case, while preserving control over polymer properties ( ≈ 0.30; dispersity ≈ 1.1. Fed-batch addition of not only I2, but also comonomer and deactivator (50 ppm under starved conditions yields a below 0.25 and, hence, an excellent linear gradient quality for the dormant polymer molecules, albeit at the expense of an increase of the overall polymerization time. The excellent control is confirmed by the visualization of the monomer sequences of ca. 1000 copolymer chains.

  7. Very high gravity ethanol fermentation by flocculating yeast under redox potential-controlled conditions

    Directory of Open Access Journals (Sweden)

    Liu Chen-Guang

    2012-08-01

    Full Text Available Abstract Background Very high gravity (VHG fermentation using medium in excess of 250 g/L sugars for more than 15% (v ethanol can save energy consumption, not only for ethanol distillation, but also for distillage treatment; however, stuck fermentation with prolonged fermentation time and more sugars unfermented is the biggest challenge. Controlling redox potential (ORP during VHG fermentation benefits biomass accumulation and improvement of yeast cell viability that is affected by osmotic pressure and ethanol inhibition, enhancing ethanol productivity and yield, the most important techno-economic aspect of fuel ethanol production. Results Batch fermentation was performed under different ORP conditions using the flocculating yeast and media containing glucose of 201 ± 3.1, 252 ± 2.9 and 298 ± 3.8 g/L. Compared with ethanol fermentation by non-flocculating yeast, different ORP profiles were observed with the flocculating yeast due to the morphological change associated with the flocculation of yeast cells. When ORP was controlled at −100 mV, ethanol fermentation with the high gravity (HG media containing glucose of 201 ± 3.1 and 252 ± 2.9 g/L was completed at 32 and 56 h, respectively, producing 93.0 ± 1.3 and 120.0 ± 1.8 g/L ethanol, correspondingly. In contrast, there were 24.0 ± 0.4 and 17.0 ± 0.3 g/L glucose remained unfermented without ORP control. As high as 131.0 ± 1.8 g/L ethanol was produced at 72 h when ORP was controlled at −150 mV for the VHG fermentation with medium containing 298 ± 3.8 g/L glucose, since yeast cell viability was improved more significantly. Conclusions No lag phase was observed during ethanol fermentation with the flocculating yeast, and the implementation of ORP control improved ethanol productivity and yield. When ORP was controlled at −150 mV, more reducing power was available for yeast cells to survive, which in turn improved their viability and VHG

  8. Study and management of atmospheric exhaust gas in acetic acid fermentation developing a new process for alcohol vinegar production; Etude et maitrise des rejets atmospheriques en vinaigrerie, developpement d'un procede nouveau de conduite de la fermentation acetique

    Energy Technology Data Exchange (ETDEWEB)

    Pochat Bohatier, C

    1999-12-06

    The aim of the study was to examine emissions of volatile organic compounds (VOCs) during acetic (acid) fermentation. The first part of the study presents the methodology developed to reproduce production cycles for spirit vinegar and to analyse gas effluents. The second part describes the origin and quantification of the emissions (ethanol, acetic acid, acetaldehyde and ethyl acetate). The acetic acid is produced by bacterial metabolism while the ethyl acetate is a result of the chemical reaction of esterification. By modelling the emissions during batch processing we were able to identify the various parameters involved when VOCs are carried along by the fermentation gases. The quantities of ethyl acetate depend on the length of time the diluted alcohol is stored, and on its composition. By using a fed-batch method with a continuous supply of ethanol we could reduce alcohol emissions. The third part of the study develops the kinetics studies carried out to adapt the fed-batch process to acetic acid fermentation. The influence of ethanol, either in terms of deficiency or inhibition, is minimized between 8 and 16 g.1-1. A study of the growth rate of bacteria in relation to the amount of acetic acid showed that the latter was highly inhibitive. There is a critical concentration of acetic acid at which the growth of bacteria stops, and the death rate of the culture increases rapidly. The latter depends on the composition of the culture's medium; the corresponding pH of the concentration is between 2.25 and 2.28. By limiting the formation of ethyl acetate in the diluted alcohol and by controlling the concentration of ethanol at 16 g.l -1 per fermentation, the VOC emissions are reduced by 30% and the yield increases as a result. (author)

  9. Lipid profile lowering effect of Soypro fermented with lactic acid bacteria isolated from Kimchi in high-fat diet-induced obese rats.

    Science.gov (United States)

    Kim, Na-Hyung; Moon, Phil-Dong; Kim, Su-Jin; Choi, In-Young; An, Hyo-Jin; Myung, Noh-Yil; Jeong, Hyun-Ja; Um, Jae-Young; Hong, Seung-Heon; Kim, Hyung-Min

    2008-01-01

    Lactic acid bacteria are known to exert various physiologic functions in humans. In the current study, we investigated the effects of Soypro, a new soymilk fermented with lactic acid bacteria, like Leuconostoc kimchii, Leuconostoc citreum, and Lactobacillus plantarum, isolated from Kimchi, on adipocyte differentiation in preadipocyte 3T3-L1 cell lines and weight gain or the plasma lipid profile in Sprague-Dawley rats. Adipocyte 3T3-L1 cells treated with Soypro (10 microg/ml) significantly reduced the contents of cellular triglyceride and inhibited cell differentiation by Oil red O staining. Treatment with Soypro (10 microg/ml) for an additional two days in adipocytes inhibited the expression of peroxisome proliferator-activated receptor-gamma2 and CCAAT/enhancer binding protein-alpha, transcription factors of adipocyte differentiation. Based on these in vitro studies, we examined the anti-obesity effect of Soypro in rats for six weeks. Soypro had no significant effect on high-fat diet-induced increases in body weight, food intake, or feed gain ratio. However, the administration of Soypro significantly reduced the concentration of the plasma low density lipoprotein cholesterol. Changes in the plasma levels of total cholesterol and glucose were inclined to decrease in Soypro administrated groups compared with saline treated group. Triglyceride and high density lipoprotein cholesterol values in Soypro fed groups were similar compared to those of saline fed groups. Although further research is needed, these findings suggest that Soypro decreased the levels of low density lipoprotein cholesterol in high-fat diet-induced obesity and might partially inhibit the adipocyte differentiation through the suppression of a transcription factors peroxisome proliferator-activated receptor-gamma2 and CCAAT/enhancer binding protein-alpha.

  10. Modeling of Clostridium tyrobutyricum for Butyric Acid Selectivity in Continuous Fermentation

    OpenAIRE

    Du, Jianjun; McGraw, Amy; Hestekin, Jamie

    2014-01-01

    A mathematical model was developed to describe batch and continuous fermentation of glucose to organic acids with Clostridium tyrobutyricum. A modified Monod equation was used to describe cell growth, and a Luedeking-Piret equation was used to describe the production of butyric and acetic acids. Using the batch fermentation equations, models predicting butyric acid selectivity for continuous fermentation were also developed. The model showed that butyric acid production was a strong function ...

  11. A comprehensive and quantitative review of dark fermentative biohydrogen production

    Directory of Open Access Journals (Sweden)

    Rittmann Simon

    2012-08-01

    Full Text Available Abstract Biohydrogen production (BHP can be achieved by direct or indirect biophotolysis, photo-fermentation and dark fermentation, whereof only the latter does not require the input of light energy. Our motivation to compile this review was to quantify and comprehensively report strains and process performance of dark fermentative BHP. This review summarizes the work done on pure and defined co-culture dark fermentative BHP since the year 1901. Qualitative growth characteristics and quantitative normalized results of H2 production for more than 2000 conditions are presented in a normalized and therefore comparable format to the scientific community. Statistically based evidence shows that thermophilic strains comprise high substrate conversion efficiency, but mesophilic strains achieve high volumetric productivity. Moreover, microbes of Thermoanaerobacterales (Family III have to be preferred when aiming to achieve high substrate conversion efficiency in comparison to the families Clostridiaceae and Enterobacteriaceae. The limited number of results available on dark fermentative BHP from fed-batch cultivations indicates the yet underestimated potential of this bioprocessing application. A Design of Experiments strategy should be preferred for efficient bioprocess development and optimization of BHP aiming at improving medium, cultivation conditions and revealing inhibitory effects. This will enable comparing and optimizing strains and processes independent of initial conditions and scale.

  12. Study and management of atmospheric exhaust gas in acetic acid fermentation developing a new process for alcohol vinegar production; Etude et maitrise des rejets atmospheriques en vinaigrerie, developpement d'un procede nouveau de conduite de la fermentation acetique

    Energy Technology Data Exchange (ETDEWEB)

    Pochat Bohatier, C.

    1999-12-06

    The aim of the study was to examine emissions of volatile organic compounds (VOCs) during acetic (acid) fermentation. The first part of the study presents the methodology developed to reproduce production cycles for spirit vinegar and to analyse gas effluents. The second part describes the origin and quantification of the emissions (ethanol, acetic acid, acetaldehyde and ethyl acetate). The acetic acid is produced by bacterial metabolism while the ethyl acetate is a result of the chemical reaction of esterification. By modelling the emissions during batch processing we were able to identify the various parameters involved when VOCs are carried along by the fermentation gases. The quantities of ethyl acetate depend on the length of time the diluted alcohol is stored, and on its composition. By using a fed-batch method with a continuous supply of ethanol we could reduce alcohol emissions. The third part of the study develops the kinetics studies carried out to adapt the fed-batch process to acetic acid fermentation. The influence of ethanol, either in terms of deficiency or inhibition, is minimized between 8 and 16 g.1-1. A study of the growth rate of bacteria in relation to the amount of acetic acid showed that the latter was highly inhibitive. There is a critical concentration of acetic acid at which the growth of bacteria stops, and the death rate of the culture increases rapidly. The latter depends on the composition of the culture's medium; the corresponding pH of the concentration is between 2.25 and 2.28. By limiting the formation of ethyl acetate in the diluted alcohol and by controlling the concentration of ethanol at 16 g.l -1 per fermentation, the VOC emissions are reduced by 30% and the yield increases as a result. (author)

  13. Extract of mangosteen increases high density lipoprotein levels in rats fed high lipid

    Directory of Open Access Journals (Sweden)

    Dwi Laksono Adiputro

    2013-04-01

    Full Text Available Background In cardiovascular medicine, Garcinia mangostana has been used as an antioxidant to inhibit oxidation of low density lipoproteins and as an antiobesity agent. The effect of Garcinia mangostana on hyperlipidemia is unknown. The aim of this study was to evaluate the effect of an ethanolic extract of Garcinia mangostana pericarp on lipid profile in rats fed a high lipid diet. Methods A total of 40 rats were divided into five groups control, high lipid diet, and high lipid diet + ethanolic extract of Garcinia mangostana pericarp at dosages of 200, 400, and 800 mg/kg body weight. The control group received a standard diet for 60 days. The high lipid diet group received standard diet plus egg yolk, goat fat, cholic acid, and pig fat for 60 days with or without ethanolic extract of Garcinia mangostana pericarp by the oral route. After 60 days, rats were anesthesized with ether for collection of blood by cardiac puncture. Analysis of blood lipid profile comprised colorimetric determination of cholesterol, triglyceride, low density lipoprotein (LDL, and high density lipoprotein (HDL. Results From the results of one-way ANOVA it was concluded that there were significant between-group differences in cholesterol, trygliceride, LDL, and HDL levels (p=0.000. Ethanolic extract of Garcinia mangostana pericarp significantly decreased cholesterol, trygliceride, and LDL levels, starting at 400 mg/kg body weight (p=0.000. Ethanolic extract of Garcinia mangostana pericarp significantly increased HDL level starting at 200 mg/kg body weight (p=0.000. Conclusion Ethanolic extract of Garcinia mangostana pericarp has a beneficial effect on lipid profile in rats on a high lipid diet.

  14. Extract of mangosteen increases high density lipoprotein levels in rats fed high lipid

    Directory of Open Access Journals (Sweden)

    Dwi Laksono Adiputro

    2015-12-01

    Full Text Available BACKGROUND In cardiovascular medicine, Garcinia mangostana has been used as an antioxidant to inhibit oxidation of low density lipoproteins and as an antiobesity agent. The effect of Garcinia mangostana on hyperlipidemia is unknown. The aim of this study was to evaluate the effect of an ethanolic extract of Garcinia mangostana pericarp on lipid profile in rats fed a high lipid diet. METHODS A total of 40 rats were divided into five groups control, high lipid diet, and high lipid diet + ethanolic extract of Garcinia mangostana pericarp at dosages of 200, 400, and 800 mg/kg body weight. The control group received a standard diet for 60 days. The high lipid diet group received standard diet plus egg yolk, goat fat, cholic acid, and pig fat for 60 days with or without ethanolic extract of Garcinia mangostana pericarp by the oral route. After 60 days, rats were anesthesized with ether for collection of blood by cardiac puncture. Analysis of blood lipid profile comprised colorimetric determination of cholesterol, triglyceride, low density lipoprotein (LDL, and high density lipoprotein (HDL. RESULTS From the results of one-way ANOVA it was concluded that there were significant between-group differences in cholesterol, trygliceride, LDL, and HDL levels (p=0.000. Ethanolic extract of Garcinia mangostana pericarp significantly decreased cholesterol, trygliceride, and LDL levels, starting at 400 mg/kg body weight (p=0.000. Ethanolic extract of Garcinia mangostana pericarp significantly increased HDL level starting at 200 mg/kg body weight (p=0.000. CONCLUSION Ethanolic extract of Garcinia mangostana pericarp has a beneficial effect on lipid profile in rats on a high lipid diet.

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

    Directory of Open Access Journals (Sweden)

    C. Varrone

    2015-01-01

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

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

    Science.gov (United States)

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

    2015-01-01

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

  17. The influence of total solids content and initial pH on batch biohydrogen production by solid substrate fermentation of agroindustrial wastes.

    Science.gov (United States)

    Robledo-Narváez, Paula N; Muñoz-Páez, Karla M; Poggi-Varaldo, Hector M; Ríos-Leal, Elvira; Calva-Calva, Graciano; Ortega-Clemente, L Alfredo; Rinderknecht-Seijas, Noemí; Estrada-Vázquez, Carlos; Ponce-Noyola, M Teresa; Salazar-Montoya, J Alfredo

    2013-10-15

    Hydrogen is a valuable clean energy source, and its production by biological processes is attractive and environmentally sound and friendly. In México 5 million tons/yr of agroindustrial wastes are generated; these residues are rich in fermentable organic matter that can be used for hydrogen production. On the other hand, batch, intermittently vented, solid substrate fermentation of organic waste has attracted interest in the last 10 years. Thus the objective of our work was to determine the effect of initial total solids content and initial pH on H2 production in batch fermentation of a substrate that consisted of a mixture of sugarcane bagasse, pineapple peelings, and waste activated sludge. The experiment was a response surface based on 2(2) factorial with central and axial points with initial TS (15-35%) and initial pH (6.5-7.5) as factors. Fermentation was carried out at 35 °C, with intermittent venting of minireactors and periodic flushing with inert N2 gas. Up to 5 cycles of H2 production were observed; the best treatment in our work showed cumulative H2 productions (ca. 3 mmol H2/gds) with 18% and 6.65 initial TS and pH, respectively. There was a significant effect of TS on production of hydrogen, the latter decreased with initial TS increase from 18% onwards. Cumulative H2 productions achieved in this work were higher than those reported for organic fraction of municipal solid waste (OFMSW) and mixtures of OFMSW and fruit peels waste from fruit juice industry, using the same process. Specific energetic potential due to H2 in our work was attractive and fell in the high side of the range of reported results in the open literature. Batch dark fermentation of agrowastes as practiced in our work could be useful for future biorefineries that generate biohydrogen as a first step and could influence the management of this type of agricultural wastes in México and other countries and regions as well. Copyright © 2013 Elsevier Ltd. All rights reserved.

  18. Modeling of Clostridium t yrobutyricum for Butyric Acid Selectivity in Continuous Fermentation

    OpenAIRE

    Jianjun Du; Amy McGraw; Jamie A. Hestekin

    2014-01-01

    A mathematical model was developed to describe batch and continuous fermentation of glucose to organic acids with Clostridium tyrobutyricum . A modified Monod equation was used to describe cell growth, and a Luedeking-Piret equation was used to describe the production of butyric and acetic acids. Using the batch fermentation equations, models predicting butyric acid selectivity for continuous fermentation were also developed. The model showed that butyric acid production was a strong function...

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

    Science.gov (United States)

    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

  20. Modelling of Functional States during Saccharomyces cerevisiae Fed-batch Cultivation

    Directory of Open Access Journals (Sweden)

    Stoyan Tzonkov

    2005-04-01

    Full Text Available An implementation of functional state approach for modelling of yeast fed-batch cultivation is presented in this paper. Using of functional state modelling approach aims to overcome the main disadvantage of using global process model, namely complex model structure and big number of model parameters, which complicate the model simulation and parameter estimation. This approach has computational advantages, such as the possibility to use the estimated values from the previous state as starting values for estimation of parameters of a new state. The functional state modelling approach is applied here for fedbatch cultivation of Saccharomyces cerevisiae. Four functional states are recognised and parameter estimation of local models is presented as well.

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

    Science.gov (United States)

    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.

  2. Comparison of soymilk and probiotic soymilk effects on serum high-density lipoprotein cholesterol and low-density lipoprotein cholesterol in diabetic Wistar rats

    Directory of Open Access Journals (Sweden)

    Mina Babashahi

    2015-04-01

    Full Text Available BACKGROUND: Soy milk (SM and its fermented products are identified as rich sources of bioactive compounds helping to manage and to reduce the risk of chronic disease. This study aimed to compare the effects of SM and probiotic SM (PSM consumption on serum low-density lipoprotein cholesterol (LDL-C and high-density lipoprotein cholesterol (HDL-C in diabetic Wistar rats. METHODS: Probiotic SM was prepared by fermentation of the plain SM with a native strain of Lactobacillus plantarum. 20 streptozotocin-nicotinamide-induced diabetic Wistar rats were divided into two groups based on the type of administered SM (SM group and PSM group. The animals were fed with 1 ml/day of either soy or PSM for 21 days. The serum lipoprotein levels were analyzed at baseline and the end of the intervention period. RESULTS: HDL-C increased significantly in PSM group. Furthermore, this group showed more percent of change in increased HDL-C in compression with SM group (P < 0.050. Regarding LDL-C level, rats fed with SM was not significantly different from the PSM group (P < 0.050; though, this biomarker was reduced in both group. CONCLUSION: Probiotic SM could modulate blood lipoprotein levels. Thus, it may be considered in managing diabetes complications and atherosclerotic risks. 

  3. Control of baker’s yeast fermentation : PID and fuzzy algorithms

    OpenAIRE

    Machado, Carlos; Gomes, Pedro; Soares, Rui; Pereira, Silvia; Soares, Filomena

    2001-01-01

    A MATLAB/SIMULINK-based simulator was employed for studies concerning the control of baker’s yeast fed-batch fermentation. Four control algorithms were implemented and compared: the classical PID control, two discrete versions- modified velocity and position algorithms, and a fuzzy law. The simulation package was seen to be an efficient tool for the simulation and tests of control strategies of the non-linear process.

  4. Comparison of microbial fermentation of high- and low-forage diets in Rusitec, single-flow continuous-culture fermenters and sheep rumen.

    Science.gov (United States)

    Carro, M D; Ranilla, M J; Martín-García, A I; Molina-Alcaide, E

    2009-04-01

    Eight Rusitec and eight single-flow continuous-culture fermenters (SFCCF) were used to compare the ruminal fermentation of two diets composed of alfalfa hay and concentrate in proportions of 80 : 20 (F80) and 20 : 80 (F20). Results were validated with those obtained previously in sheep fed the same diets. Rusitec fermenters were fed once daily and SFCCF twice, but liquid dilution rates were similar in both types of fermenters. Mean values of pH over the 12 h postfeeding were higher (P 0.05) were found in any in vitro system. A more precise control of pH in both types of fermenters and a reduction of concentrate retention time in Rusitec could probably improve the simulation of in vivo fermentation.

  5. High Performance Bioanode Development for Fermentable Substrates via Controlled Electroactive Biofilm Growth

    Energy Technology Data Exchange (ETDEWEB)

    Ichihashi, Osamu [ORNL; Vishnivetskaya, Tatiana A [ORNL; Borole, Abhijeet P [ORNL

    2014-11-11

    A bioanode was optimized to generate current densities reaching 38.4 4.9 A m-2, which brings bioelectrochemical systems closer to commercial consideration. Glucose and lactate were fed together in a continuous or fed-batch mode. The current density increased from 2.3 A m-2 to 38.4 A m-2 over a 33 day period and remained stable thereafter. The coulombic efficiency ranged from 50% to 80%. A change in substrate concentration from 200 mg L-1 to 5 mg L-1 decreased maximum current density from 38.4 A m-2 to 12.3 A m-2. The anode consortia included Firmicutes (55.0%), Proteobacteria (41.8%) and Bacteroidetes (2.1%) constituting two potential electrogenic genera: Geobacter (6.8%) and Aeromonas (31.9%). The current production was found to be limited by kinetics during the growth period (33 days), and mass transfer, thereafter. The results indicate the necessity of removing spent biomass for efficient long term operation and treatment of wastewater streams.

  6. Kinetics of volatile metabolites during alcoholic fermentation of cane molasses by Saccharomyces cerevisiae

    Energy Technology Data Exchange (ETDEWEB)

    Cachot, T; Mueller, M; Pons, M N [Centre National de la Recherche Scientifique, 54 - Nancy (France). Lab. des Sciences du Genie Chimique

    1991-07-01

    The kinetics of ethanol, acetaldehyde, ethyl acetate and fusel alcohols during alcoholic fermentations on cane molasses by Saccharomyces cerevisiae have been obtained via an in-situ gas membrane sensor connected to a gas chromatograph. Various operation parameters have been investigated such as inoculum rate, molasses concentration, operation mode (batch, fed-batch). The modification of fusel alcohols kinetics in response to addition of amino acids has been studied as well as the assimilation of two intermediary aldehydes (isovaleraldehyde and isobutyraldehyde) in the fusel alcohol synthesis pathway. (orig.).

  7. Wastewater recycling technology for fermentation in polyunsaturated fatty acid production.

    Science.gov (United States)

    Song, Xiaojin; Ma, Zengxin; Tan, Yanzhen; Zhang, Huidan; Cui, Qiu

    2017-07-01

    To reduce fermentation-associated wastewater discharge and the cost of wastewater treatment, which further reduces the total cost of DHA and ARA production, this study first analyzed the composition of wastewater from Aurantiochytrium (DHA) and Mortierella alpina (ARA) fermentation, after which wastewater recycling technology for these fermentation processes was developed. No negative effects of DHA and ARA production were observed when the two fermentation wastewater methods were cross-recycled. DHA and ARA yields were significantly inhibited when the wastewater from the fermentation process was directly reused. In 5-L fed-batch fermentation experiments, using this cross-recycle technology, the DHA and ARA yields were 30.4 and 5.13gL -1 , respectively, with no significant changes (P>0.05) compared to the control group, and the water consumption was reduced by half compared to the traditional process. Therefore, this technology has great potential in industrial fermentation for polyunsaturated fatty acid production. Copyright © 2017 Elsevier Ltd. All rights reserved.

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

    Science.gov (United States)

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

    2015-06-01

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

  9. Cybernetic modeling based on pathway analysis for Penicillium chrysogenum fed-batch fermentation.

    Science.gov (United States)

    Geng, Jun; Yuan, Jingqi

    2010-08-01

    A macrokinetic model employing cybernetic methodology is proposed to describe mycelium growth and penicillin production. Based on the primordial and complete metabolic network of Penicillium chrysogenum found in the literature, the modeling procedure is guided by metabolic flux analysis and cybernetic modeling framework. The abstracted cybernetic model describes the transients of the consumption rates of the substrates, the assimilation rates of intermediates, the biomass growth rate, as well as the penicillin formation rate. Combined with the bioreactor model, these reaction rates are linked with the most important state variables, i.e., mycelium, substrate and product concentrations. Simplex method is used to estimate the sensitive parameters of the model. Finally, validation of the model is carried out with 20 batches of industrial-scale penicillin cultivation.

  10. Modified ADM1 for modeling free ammonia inhibition in anaerobic acidogenic fermentation with high-solid sludge.

    Science.gov (United States)

    Bai, Jie; Liu, He; Yin, Bo; Ma, Huijun; Chen, Xinchun

    2017-02-01

    Anaerobic acidogenic fermentation with high-solid sludge is a promising method for volatile fatty acid (VFA) production to realize resource recovery. In this study, to model inhibition by free ammonia in high-solid sludge fermentation, the anaerobic digestion model No. 1 (ADM1) was modified to simulate the VFA generation in batch, semi-continuous and full scale sludge. The ADM1 was operated on the platform AQUASIM 2.0. Three kinds of inhibition forms, e.g., simple inhibition, Monod and non-inhibition forms, were integrated into the ADM1 and tested with the real experimental data for batch and semi-continuous fermentation, respectively. The improved particle swarm optimization technique was used for kinetic parameter estimation using the software MATLAB 7.0. In the modified ADM1, the K s of acetate is 0.025, the k m,ac is 12.51, and the K I_NH3 is 0.02, respectively. The results showed that the simple inhibition model could simulate the VFA generation accurately while the Monod model was the better inhibition kinetics form in semi-continuous fermentation at pH10.0. Finally, the modified ADM1 could successfully describe the VFA generation and ammonia accumulation in a 30m 3 full-scale sludge fermentation reactor, indicating that the developed model can be applicable in high-solid sludge anaerobic fermentation. Copyright © 2016. Published by Elsevier B.V.

  11. CARCASS YIELD OF BROILER CHICKENS FED BANANA (Musa paradisiaca LEAVES FERMENTED WITH Trichoderma viride

    Directory of Open Access Journals (Sweden)

    J. S. Mandey

    2016-03-01

    Full Text Available The study was conducted to evaluate the effect of level of banana (Musa paradisiaca leaves fermented with Trichoderma viride at different days on the carcass yield of broiler chickens. A hundred and eighty 3-weeks-old broiler chicks were used in this present experiment based on factorial design (3×4. The birds were randomly allocated into three experimental diets containing of 5, 10 and 15% of banana leaves fermented within 0, 5, 10 and 15 days. Each treatment was divided into three replicates of five chicks in each. The experiment was terminated after 4 weeks or when the birds were 7-weeks-old. Feed intake, body weight gain, feed efficiency and carcass yield were measured during the study. The data were subjected to the analysis of variance test followed by least significant difference (LSD test. Results showed that daily feed intake was significantly affected (P˂0.01 by the dietary treatments, in which feed intake was highest in broilers fed diet containing 10% banana leaves fermented for 10 days. The daily weight gain, feed efficiency and carcass yield were significantly affected (P˂0.01 by the treatments, in which the highest values of daily weight gain, feed efficiency, and carcass yield were observed in birds fed diet containing 10% banana leaves fermented for 10 days. It can be concluded that diet containing 10% banana leaves fermented for 10 days can be included in broiler ration without detrimental effects on the performance and carcass yield. 

  12. Analysis of L-glutamic acid fermentation by using a dynamic metabolic simulation model of Escherichia coli.

    Science.gov (United States)

    Nishio, Yousuke; Ogishima, Soichi; Ichikawa, Masao; Yamada, Yohei; Usuda, Yoshihiro; Masuda, Tadashi; Tanaka, Hiroshi

    2013-09-22

    Understanding the process of amino acid fermentation as a comprehensive system is a challenging task. Previously, we developed a literature-based dynamic simulation model, which included transcriptional regulation, transcription, translation, and enzymatic reactions related to glycolysis, the pentose phosphate pathway, the tricarboxylic acid (TCA) cycle, and the anaplerotic pathway of Escherichia coli. During simulation, cell growth was defined such as to reproduce the experimental cell growth profile of fed-batch cultivation in jar fermenters. However, to confirm the biological appropriateness of our model, sensitivity analysis and experimental validation were required. We constructed an L-glutamic acid fermentation simulation model by removing sucAB, a gene encoding α-ketoglutarate dehydrogenase. We then performed systematic sensitivity analysis for L-glutamic acid production; the results of this process corresponded with previous experimental data regarding L-glutamic acid fermentation. Furthermore, it allowed us to predicted the possibility that accumulation of 3-phosphoglycerate in the cell would regulate the carbon flux into the TCA cycle and lead to an increase in the yield of L-glutamic acid via fermentation. We validated this hypothesis through a fermentation experiment involving a model L-glutamic acid-production strain, E. coli MG1655 ΔsucA in which the phosphoglycerate kinase gene had been amplified to cause accumulation of 3-phosphoglycerate. The observed increase in L-glutamic acid production verified the biologically meaningful predictive power of our dynamic metabolic simulation model. In this study, dynamic simulation using a literature-based model was shown to be useful for elucidating the precise mechanisms involved in fermentation processes inside the cell. Further exhaustive sensitivity analysis will facilitate identification of novel factors involved in the metabolic regulation of amino acid fermentation.

  13. Single cell oil production from hydrolysate of cassava starch by marine-derived yeast Rhodotorula mucilaginosa TJY15a

    Energy Technology Data Exchange (ETDEWEB)

    Li, Mei; Liu, Guang-Lei; Chi, Zhe; Chi, Zhen-Ming [Unesco Chinese Center of Marine Biotechnology, Ocean University of China, Yushan Road, No. 5, Qingdao 266003 (China)

    2010-01-15

    Rhodotorula mucilaginosa TJY15a which was isolated from surface of marine fish could accumulate a large amount of lipid from hydrolysate of cassava starch. The cells contained 47.9% (w/w) oil during batch cultivation, whereas 52.9% (w/w) of lipid was obtained during the fed-batch cultivation. At the end of the fed-batch cultivation, all the starch were converted into reducing sugar and only 0.34 g dm{sup -3} of reducing sugar was left in the fermented medium. Therefore, the marine-derived R. mucilaginosa TJY15a was another candidate for single cell oil production. The fatty acids from R. mucilaginosa TJY15a were mainly composed of palmitic acid (C{sub 16:0}), palmitoleic acid (C{sub 16:1}), stearic acid (C{sub 18:0}), oleic acid (C{sub 18:1}) and linolenic acid (C{sub 18:2}), suggesting that the fatty acids could be used as feedstock for biodiesel production. (author)

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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.

  16. Dynamics in population heterogeneity during batch and continuous fermentation of Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Heins, Anna-Lena; Lencastre Fernandes, Rita; Lundin, L.

    2012-01-01

    Traditionally, microbial populations in optimization studies of fermentation processes have been considered homogeneous. However, research has shown that a typical microbial population in fermentation is heterogeneous. There are indications that this heterogeneity may be both beneficial...... (facilitates quick adaptation to new conditions) and harmful (reduces yields and productivities)[1,2]. Typically, gradients of e.g. dissolved oxygen, substrates, and pH are observed in industrial scale fermentation processes. Consequently, microbial cells circulating throughout a bioreactor experience rapid...... distribution during different growth stages. To further simulate which effect gradients have on population heterogeneity, glucose and ethanol perturbations during continuous cultivation were performed. Physiological changes were analyzed on single cell level by using flow cytometry followed by cell sorting...

  17. High current densities enable exoelectrogens to outcompete aerobic heterotrophs for substrate

    KAUST Repository

    Ren, Lijiao

    2014-08-05

    © 2014 Wiley Periodicals, Inc. Chemical oxygen demand (COD) removal rates could be described by first-order kinetics with respect to COD concentration at different current densities, even under open circuit conditions with no current generation. The COD concentration was reduced more quickly with current generation due to the greater consumption of substrate by exoelectrogens, and less substrate was lost to aerobic heterotrophs. Higher current densities enabled exoelectrogens to outcompete aerobic heterotrophs for substrate, allowing for increased coulombic efficiencies with current densities. © 2014 Wiley Periodicals, Inc. In mixed-culture microbial fuel cells (MFCs), exoelectrogens and other microorganisms compete for substrate. It has previously been assumed that substrate losses to other terminal electron acceptors over a fed-batch cycle, such as dissolved oxygen, are constant. However, a constant rate of substrate loss would only explain small increases in coulombic efficiencies (CEs, the fraction of substrate recovered as electrical current) with shorter cycle times, but not the large increases in CE that are usually observed with higher current densities and reduced cycle times. To better understand changes in CEs, COD concentrations were measured over time in fed-batch, single-chamber, air-cathode MFCs at different current densities (external resistances). COD degradation rates were all found to be first-order with respect to COD concentration, even under open circuit conditions with no current generation (first-order rate constant of 0.14±0.01h-1). The rate of COD removal increased when there was current generation, with the highest rate constant (0.33±0.02h-1) obtained at the lowest external resistance (100Ω). Therefore, as the substrate concentration was reduced more quickly due to current generation, the rate of loss of substrate to non-exoelectrogens decreased due to this first-order substrate-concentration dependence. As a result, coulombic

  18. High current densities enable exoelectrogens to outcompete aerobic heterotrophs for substrate

    KAUST Repository

    Ren, Lijiao; Zhang, Xiaoyuan; He, Weihua; Logan, Bruce E.

    2014-01-01

    © 2014 Wiley Periodicals, Inc. Chemical oxygen demand (COD) removal rates could be described by first-order kinetics with respect to COD concentration at different current densities, even under open circuit conditions with no current generation. The COD concentration was reduced more quickly with current generation due to the greater consumption of substrate by exoelectrogens, and less substrate was lost to aerobic heterotrophs. Higher current densities enabled exoelectrogens to outcompete aerobic heterotrophs for substrate, allowing for increased coulombic efficiencies with current densities. © 2014 Wiley Periodicals, Inc. In mixed-culture microbial fuel cells (MFCs), exoelectrogens and other microorganisms compete for substrate. It has previously been assumed that substrate losses to other terminal electron acceptors over a fed-batch cycle, such as dissolved oxygen, are constant. However, a constant rate of substrate loss would only explain small increases in coulombic efficiencies (CEs, the fraction of substrate recovered as electrical current) with shorter cycle times, but not the large increases in CE that are usually observed with higher current densities and reduced cycle times. To better understand changes in CEs, COD concentrations were measured over time in fed-batch, single-chamber, air-cathode MFCs at different current densities (external resistances). COD degradation rates were all found to be first-order with respect to COD concentration, even under open circuit conditions with no current generation (first-order rate constant of 0.14±0.01h-1). The rate of COD removal increased when there was current generation, with the highest rate constant (0.33±0.02h-1) obtained at the lowest external resistance (100Ω). Therefore, as the substrate concentration was reduced more quickly due to current generation, the rate of loss of substrate to non-exoelectrogens decreased due to this first-order substrate-concentration dependence. As a result, coulombic

  19. Enteric methane production and ruminal fermentation from forage brassica diets fed in continuous culture

    Science.gov (United States)

    Brassicas provide forage for livestock during the late fall when traditional perennial cool-season forages are not productive. However, little research exists on ruminal fermentation and methane(CH4) production of brassicas fed as forage. A continuous culture fermentor system was used to assess nutr...

  20. Process for the fermentative production of acetone, butanol and ethanol

    Science.gov (United States)

    Glassner, David A.; Jain, Mahendra K.; Datta, Rathin

    1991-01-01

    A process including multistage continuous fermentation followed by batch fermentation with carefully chosen temperatures for each fermentation step, combined with an asporogenic strain of C. acetobutylicum and a high carbohydrate substrate concentration yields extraordinarily high butanol and total solvents concentrations.

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

    Science.gov (United States)

    Petrov, Kaloyan; Popova, Luiza; Petrova, Penka

    2017-06-01

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

  2. Continuous acetone-ethanol-butanol fermentation by immobilized cells of Clostridium acetobutylicum

    Energy Technology Data Exchange (ETDEWEB)

    Badr, H.R.; Toledo, R.; Hamdy, M.K. [University of Georgia, Athens (Greece). Food Science and Technology Dept.

    2001-07-01

    Eight Clostridium acetobutylicum strains were examined for {alpha}-amylase and strains B-591, B-594 and P-262 had the highest activities. Defibered-sweet-potato-slurry (DSPS), containing 39.7 g starch l{sup -1}, supplemented with potassium phosphate (1.0 g l{sup -1}), cysteine-HCl (5.0 g l{sup -1}), the antifoam (polypropylene glycol, 0.1 mg ml{sup -1}), was used a continuous feedstock (FS) to a multistage bioreactor system for acetone-ethanol-butanol (AEB) fermentation. The system consisted on four columns (three vertical and one near horizontal) packed with beads containing immobilized cells of C. acetobutylicum P-262. When DSPS was pumped into the bioreactor system, at a flow rate of 2.36 ml min{sup -1}, the effluent has 7.73 g solvents l{sup -1} (1.56, acetone; 0.65, ethanol; 5.52 g, butanol) and no starch. Productivity of total solvents synthesized during continuous operation were 1.0 g 1{sup -1}h{sup -1} and 19.5 % yield compared to 0.12 g l{sup -1}h{sup -1} with 29% yield using the batch system. We proposed using DSPS for AEB fermentation in a continuous mode with immobilized P-262 cells that are active amylase producers which will lead to cost reduction compared to the batch system. (Author)

  3. Fermentation of hexoses to ethanol

    Energy Technology Data Exchange (ETDEWEB)

    Gustafsson, Lena [Goeteborg Univ. (Sweden). Dept. of General and Marine Microbiology]|[Chalmers Univ. of Technology, Goeteborg (Sweden). Dept of Chemical Reaction Engineering

    2000-06-01

    The Goals of the project has been: to increase the ethanol yield by reducing the by-product formation, primarily biomass and glycerol, and to prevent stuck fermentations, i.e. to maintain a high ethanol production rate simultaneously with a high ethanol yield. The studies have been performed both in defined laboratory media and in a mixture of wood- and wheat hydrolysates. The yeast strains used have been both industrial strains of bakers yeast, Saccharomyces cerevisiae, and haploid laboratory strains. The Relevance of these studies with respect to production of ethanol to be used as fuel is explained by: With the traditional process design used today, it is very difficult to reach a yield of more than 90 % of the theoretical maximal value of ethanol based on fermented hexose. During 'normal' growth and fermentation conditions in either anaerobic batch or chemostat cultures, substrate is lost as biomass and glycerol in the range of 8 to 11 % and 6 to 11 % of the substrate consumed (kg/kg). It is essential to reduce these by-products. Traditional processes are mostly batch processes, in which there is a risk that the biocatalyst, i.e. the yeast, may become inactivated. If for example yeast biomass production is avoided by use of non-growing systems, the ethanol production rate is instantaneously reduced by at least 50%. Unfortunately, even if yeast biomass production is not avoided on purpose, it is well known that stuck fermentations caused by cell death is a problem in large scale yeast processes. The main reason for stuck fermentations is nutrient imbalances. For a good process economy, it is necessary to ensure process accessibility, i.e. to maintain a high and reproducible production rate. This will both considerably reduce the necessary total volume of the fermentors (and thereby the investment costs), and moreover minimize undesirable product fall-out.

  4. Gaining electricity from in situ oxidation of hydrogen produced by fermentative cellulose degradation.

    Science.gov (United States)

    Niessen, J; Schröder, U; Harnisch, F; Scholz, F

    2005-01-01

    To exploit the fermentative hydrogen generation and direct hydrogen oxidation for the generation of electric current from the degradation of cellulose. Utilizing the metabolic activity of the mesophilic anaerobe Clostridium cellulolyticum and the thermophilic Clostridium thermocellum we show that electricity generation is possible from cellulose fermentation. The current generation is based on an in situ oxidation of microbially synthesized hydrogen at platinum-poly(tetrafluoroaniline) (Pt-PTFA) composite electrodes. Current densities of 130 mA l(-1) (with 3 g cellulose per litre medium) were achieved in poised potential experiments under batch and semi-batch conditions. The presented results show that electricity generation is possible by the in situ oxidation of hydrogen, product of the anaerobic degradation of cellulose by cellulolytic bacteria. For the first time, it is shown that an insoluble complex carbohydrate like cellulose can be used for electricity generation in a microbial fuel cell. The concept represents a first step to the utilization of macromolecular biomass components for microbial electricity generation.

  5. Glycerol production by fermenting yeast cells is essential for optimal bread dough fermentation.

    Science.gov (United States)

    Aslankoohi, Elham; Rezaei, Mohammad Naser; Vervoort, Yannick; Courtin, Christophe M; Verstrepen, Kevin J

    2015-01-01

    Glycerol is the main compatible solute in yeast Saccharomyces cerevisiae. When faced with osmotic stress, for example during semi-solid state bread dough fermentation, yeast cells produce and accumulate glycerol in order to prevent dehydration by balancing the intracellular osmolarity with that of the environment. However, increased glycerol production also results in decreased CO2 production, which may reduce dough leavening. We investigated the effect of yeast glycerol production level on bread dough fermentation capacity of a commercial bakery strain and a laboratory strain. We find that Δgpd1 mutants that show decreased glycerol production show impaired dough fermentation. In contrast, overexpression of GPD1 in the laboratory strain results in increased fermentation rates in high-sugar dough and improved gas retention in the fermenting bread dough. Together, our results reveal the crucial role of glycerol production level by fermenting yeast cells in dough fermentation efficiency as well as gas retention in dough, thereby opening up new routes for the selection of improved commercial bakery yeasts.

  6. Predictions for optimal mitigation of paracrine inhibitory signalling in haemopoietic stem cell cultures.

    Science.gov (United States)

    Berry, Joseph D; Godara, Pankaj; Liovic, Petar; Haylock, David N

    2015-04-16

    Recent studies in the literature have highlighted the critical role played by cell signalling in determining haemopoietic stem cell (HSC) fate within ex vivo culture systems. Stimulatory signals can enhance proliferation and promote differentiation, whilst inhibitory signals can significantly limit culture output. Numerical models of various mitigation strategies are presented and applied to determine effectiveness of these strategies toward mitigation of paracrine inhibitory signalling inherent in these culture systems. The strategies assessed include mixing, media-exchange, fed-batch and perfusion. The models predict that significant spatial concentration gradients exist in typical cell cultures, with important consequences for subsequent cell expansion. Media exchange is shown to be the most effective mitigation strategy, but remains labour intensive and difficult to scale-up for large culture systems. The fed-batch strategy is only effective at very small Peclet number, and its effect is diminished as the cell culture volume grows. Conversely, mixing is effective at high Peclet number, and ineffective at low Peclet number. The models predict that cell expansion in fed-batch cultures becomes independent of increasing dilution rate, consistent with experimental results previously reported in the literature. In contrast, the models predict that increasing the flow rate in perfused cultures will lead to increased cell expansion, indicating the suitability of perfusion for use as an automated, tunable strategy. The effect of initial cell seeding density is also investigated, with the model showing that perfusion outperforms dilution for all densities considered. The models predict that the impact of inhibitory signalling in HSC cultures can be mitigated against using media manipulation strategies, with the optimal strategy dependent upon the protein diffusion time-scale relative to the media manipulation time-scale. The key messages from this study can be applied to

  7. Analysis of l-glutamic acid fermentation by using a dynamic metabolic simulation model of Escherichia coli

    Science.gov (United States)

    2013-01-01

    Background Understanding the process of amino acid fermentation as a comprehensive system is a challenging task. Previously, we developed a literature-based dynamic simulation model, which included transcriptional regulation, transcription, translation, and enzymatic reactions related to glycolysis, the pentose phosphate pathway, the tricarboxylic acid (TCA) cycle, and the anaplerotic pathway of Escherichia coli. During simulation, cell growth was defined such as to reproduce the experimental cell growth profile of fed-batch cultivation in jar fermenters. However, to confirm the biological appropriateness of our model, sensitivity analysis and experimental validation were required. Results We constructed an l-glutamic acid fermentation simulation model by removing sucAB, a gene encoding α-ketoglutarate dehydrogenase. We then performed systematic sensitivity analysis for l-glutamic acid production; the results of this process corresponded with previous experimental data regarding l-glutamic acid fermentation. Furthermore, it allowed us to predicted the possibility that accumulation of 3-phosphoglycerate in the cell would regulate the carbon flux into the TCA cycle and lead to an increase in the yield of l-glutamic acid via fermentation. We validated this hypothesis through a fermentation experiment involving a model l-glutamic acid-production strain, E. coli MG1655 ΔsucA in which the phosphoglycerate kinase gene had been amplified to cause accumulation of 3-phosphoglycerate. The observed increase in l-glutamic acid production verified the biologically meaningful predictive power of our dynamic metabolic simulation model. Conclusions In this study, dynamic simulation using a literature-based model was shown to be useful for elucidating the precise mechanisms involved in fermentation processes inside the cell. Further exhaustive sensitivity analysis will facilitate identification of novel factors involved in the metabolic regulation of amino acid fermentation. PMID

  8. Paper sludge (PS) to bioethanol: Evaluation of virgin and recycle mill sludge for low enzyme, high-solids fermentation.

    Science.gov (United States)

    Boshoff, Sonja; Gottumukkala, Lalitha Devi; van Rensburg, Eugéne; Görgens, Johann

    2016-03-01

    Paper sludge (PS) from the paper and pulp industry consists primarily of cellulose and ash and has significant potential for ethanol production. Thirty-seven PS samples from 11 South African paper and pulp mills exhibited large variation in chemical composition and resulting ethanol production. Simultaneous saccharification and fermentation (SSF) of PS in fed-batch culture was investigated at high solid loadings and low enzyme dosages. Water holding capacity and viscosity of the PS influenced ethanol production at elevated solid loadings of PS. High viscosity of PS from virgin pulp mills restricted the solid loading to 18% (w/w) at an enzyme dosage of 20 FPU/gram dry PS (gdPS), whereas an optimal solid loading of 27% (w/w) was achieved with corrugated recycle mill PS at 11 FPU/gdPS. Ethanol concentration and yield of virgin pulp and corrugated recycle PS were 34.2g/L at 66.9% and 45.5 g/L at 78.2%, respectively. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. GROWTH AND COMPOSITION OF Arthrospira (Spirulina platensis IN A TUBULAR PHOTOBIOREACTOR USING AMMONIUM NITRATE AS THE NITROGEN SOURCE IN A FED-BATCH PROCESS

    Directory of Open Access Journals (Sweden)

    C. Cruz-Martínez

    2015-06-01

    Full Text Available AbstractNH4NO3 simultaneously provides a readily assimilable nitrogen source (ammonia and a reserve of nitrogen (nitrate, allowing for an increase in Arthrospira platensis biomass production while reducing the cost of the cultivation medium. In this study, a 22plus star central composite experimental design combined with response surface methodology was employed to analyze the influence of light intensity (I and the total amount of added NH4NO3 (Mt on a bench-scale tubular photobioreactor for fed-batch cultures. The maximum cell concentration (Xm, cell productivity (PX and biomass yield on nitrogen (YX/N were evaluated, as were the protein and lipid contents. Under optimized conditions (I = 148 μmol·photons·m-2·s-1 and Mt = 9.7 mM NH4NO3, Xm = 4710 ±34.4 mg·L-1, PX = 478.9 ±3.8 mg·L-1·d-1 and YX/N = 15.87 ±0.13 mg·mg-1 were obtained. The best conditions for protein content in the biomass (63.2% were not the same as those that maximized cell growth (I = 180 μmol·photons·m-2·s-1 and Mt = 22.5 mM NH4NO3. Based on these results, it is possible to conclude that ammonium nitrate is an interesting alternate nitrogen source for the cultivation of A. platensisin a fed-batch process and could be used for other photosynthetic microorganisms.

  10. Soft-sensing Modeling Based on MLS-SVM Inversion for L-lysine Fermentation Processes

    Directory of Open Access Journals (Sweden)

    Bo Wang

    2015-06-01

    Full Text Available A modeling approach 63 based on multiple output variables least squares support vector machine (MLS-SVM inversion is presented by a combination of inverse system and support vector machine theory. Firstly, a dynamic system model is developed based on material balance relation of a fed-batch fermentation process, with which it is analyzed whether an inverse system exists or not, and into which characteristic information of a fermentation process is introduced to set up an extended inversion model. Secondly, an initial extended inversion model is developed off-line by the use of the fitting capacity of MLS-SVM; on-line correction is made by the use of a differential evolution (DE algorithm on the basis of deviation information. Finally, a combined pseudo-linear system is formed by means of a serial connection of a corrected extended inversion model behind the L-lysine fermentation processes; thereby crucial biochemical parameters of a fermentation process could be predicted on-line. The simulation experiment shows that this soft-sensing modeling method features very high prediction precision and can predict crucial biochemical parameters of L-lysine fermentation process very well.

  11. Analysis of polarization methods for elimination of power overshoot in microbial fuel cells

    KAUST Repository

    Watson, Valerie J.; Logan, Bruce E.

    2011-01-01

    Polarization curves from microbial fuel cells (MFCs) often show an unexpectedly large drop in voltage with increased current densities, leading to a phenomenon in the power density curve referred to as "power overshoot". Linear sweep voltammetry (LSV, 1 mV s- 1) and variable external resistances (at fixed intervals of 20 min) over a single fed-batch cycle in an MFC both resulted in power overshoot in power density curves due to anode potentials. Increasing the anode enrichment time from 30 days to 100 days did not eliminate overshoot, suggesting that insufficient enrichment of the anode biofilm was not the primary cause. Running the reactor at a fixed resistance for a full fed-batch cycle (~ 1 to 2 days), however, completely eliminated the overshoot in the power density curve. These results show that long times at a fixed resistance are needed to stabilize current generation by bacteria in MFCs, and that even relatively slow LSV scan rates and long times between switching circuit loads during a fed-batch cycle may produce inaccurate polarization and power density results for these biological systems. © 2010 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

    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.

  13. Comparison of separate hydrolysis and fermentation and simultaneous saccharification and fermentation processes for ethanol production from wheat straw by recombinant Escherichia coli strain FBR5

    Energy Technology Data Exchange (ETDEWEB)

    Saha, Badal C.; Nichols, Nancy N.; Qureshi, Nasib; Cotta, Michael A. [U.S. Department of Agriculture, Agricultural Research Services Peoria, IL (United States). Bioenergy Reserach Unit

    2011-11-15

    Ethanol production by recombinant Escherichia coli strain FBR5 from dilute acid pretreated wheat straw (WS) by separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) was studied. The yield of total sugars from dilute acid (0.5% H2SO4) pretreated (160 C, 10 min) and enzymatically saccharified (pH 5.0, 45 C, 72 h) WS (86 g/l) was 50.0 {+-} 1.4 g/l. The hydrolyzate contained 1,184 {+-} 19 mg furfural and 161 {+-} 1 mg hydroxymethyl furfural per liter. The recombinant E. coli FBR5 could not grow at all at pH controlled at 4.5 to 6.5 in the non-abated wheat straw hydrolyzate (WSH) at 35 C. However, it produced 21.9 {+-} 0.3 g ethanol from non-abated WSH (total sugars, 44.1 {+-} 0.4 g/l) in 90 h including the lag time of 24 h at controlled pH 7.0 and 35 C. The bioabatement of WS was performed by growing Coniochaeta ligniaria NRRL 30616 in the liquid portion of the pretreated WS aerobically at pH 6.5 and 30 C for 15 h. The bacterium produced 21.6 {+-} 0.5 g ethanol per liter in 40 h from the bioabated enzymatically saccharified WSH (total sugars, 44.1 {+-} 0.4 g) at pH 6.0. It produced 24.9 {+-} 0.3 g ethanol in 96 h and 26.7 {+-} 0.0 g ethanol in 72 h per liter from bioabated WSH by batch SSF and fed-batch SSF, respectively. SSF offered a distinct advantage over SHF with respect to reducing total time required to produce ethanol from the bioabated WS. Also, fed-batch SSF performed better than the batch SSF with respect to shortening the time requirement and increase in ethanol yield. (orig.)

  14. Glycerol production by fermenting yeast cells is essential for optimal bread dough fermentation.

    Directory of Open Access Journals (Sweden)

    Elham Aslankoohi

    Full Text Available Glycerol is the main compatible solute in yeast Saccharomyces cerevisiae. When faced with osmotic stress, for example during semi-solid state bread dough fermentation, yeast cells produce and accumulate glycerol in order to prevent dehydration by balancing the intracellular osmolarity with that of the environment. However, increased glycerol production also results in decreased CO2 production, which may reduce dough leavening. We investigated the effect of yeast glycerol production level on bread dough fermentation capacity of a commercial bakery strain and a laboratory strain. We find that Δgpd1 mutants that show decreased glycerol production show impaired dough fermentation. In contrast, overexpression of GPD1 in the laboratory strain results in increased fermentation rates in high-sugar dough and improved gas retention in the fermenting bread dough. Together, our results reveal the crucial role of glycerol production level by fermenting yeast cells in dough fermentation efficiency as well as gas retention in dough, thereby opening up new routes for the selection of improved commercial bakery yeasts.

  15. Continuous fermentation of whey into alcohol using an attached film expanded bed reactor

    Energy Technology Data Exchange (ETDEWEB)

    Chen, H C; Zall, R R

    1982-01-01

    Batch and continuous processes were studied using a lactose- fermenting yeast, Saccharomyces fragi. Batch methods were used to study the effects of temperature, pH, yeast extract, and ergosterol on ethanol and cell mass production. The optimal temperature ranged from 32 to 37 degrees C. No pH control appeared necessary for whey fermentation. Supplements of yeast extract and ergosterol appeared to have pronounced effect on cell mass and ethanol production. Overall, whey reconstituted from acid whey powder containing 10% lactose could be converted to 36.5 g/l of ethanol in 44 hours when the material was supplemented with 0.7% yeast extract, 10 mg/l ergosterol, and fermented at 32 degrees C using 2.67% aerobically grown inoculum containing approximately 110 x 10 to the power of 6 CFU/ml. Ethanol productivity as high as 6.9 g/lj/h was achieved in a continuous attached film expanded bed fermentor using cellulose acetate as support media. The retention time was 1.1 hour and effluent ethanol concentration was 7.6 g/l. (Refs. 27).

  16. Ethanol fermentation of molasses by Saccharomyces cerevisiae cells immobilized onto sugar beet pulp

    Directory of Open Access Journals (Sweden)

    Vučurović Vesna M.

    2012-01-01

    Full Text Available Natural adhesion of Saccharomyces cerevisiae onto sugar beet pulp (SBP is a very simple and cheap immobilization method for retaining high cells density in the ethanol fermentation system. In the present study, yeast cells were immobilized by adhesion onto SBP suspended in the synthetic culture media under different conditions such as: glucose concentration (100, 120 and 150 g/l, inoculum concentration (5, 10 and 15 g/l dry mass and temperature (25, 30, 35 and 40°C. In order to estimate the optimal immobilization conditions the yeast cells retention (R, after each immobilization experiment was analyzed. The highest R value of 0.486 g dry mass yeast /g dry mass SBP was obtained at 30°C, glucose concentration of 150 g/l, and inoculum concentration of 15 g/l. The yeast immobilized under these conditions was used for ethanol fermentation of sugar beet molasses containing 150.2 g/l of reducing sugar. Efficient ethanol fermentation (ethanol concentration of 70.57 g/l, fermentation efficiency 93.98% of sugar beet molasses was achieved using S. cerevisiae immobilized by natural adhesion on SBP. [Projekat Ministarstva nauke Republike Srbije, br. TR-31002

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

    Science.gov (United States)

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

    2011-08-01

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

  18. Ethanol prefermentation of food waste in sequencing batch methane fermentation for improved buffering capacity and microbial community analysis.

    Science.gov (United States)

    Yu, Miao; Wu, Chuanfu; Wang, Qunhui; Sun, Xiaohong; Ren, Yuanyuan; Li, Yu-You

    2018-01-01

    This study investigates the effects of ethanol prefermentation (EP) on methane fermentation. Yeast was added to the substrate for EP in the sequencing batch methane fermentation of food waste. An Illumina MiSeq high-throughput sequencing system was used to analyze changes in the microbial community. Methane production in the EP group (254mL/g VS) was higher than in the control group (35mL/g VS) because EP not only increased the buffering capacity of the system, but also increased hydrolytic acidification. More carbon source was converted to ethanol in the EP group than in the control group, and neutral ethanol could be converted continuously to acetic acid, which promoted the growth of Methanobacterium and Methanosarcina. As a result, the relative abundance of methane-producing bacteria was significantly higher than that of the control group. Kinetic modeling indicated that the EP group had a higher hydrolysis efficiency and shorter lag phase. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Optimization and control of feb-batch fermentation processes by using artificial neural systems

    OpenAIRE

    Valencia Peroni, Catalina

    2002-01-01

    Optimización y Control de Procesos de Fermentación Fed-batch a través de Sistemas Neuronales ArtificialesLos procesos de fermentación son ampliamente utilizados en la industria química, farmacéutica y alimentaria. La producción de comida para animales, yogures, quesos, cerveza, colorantes para comida, fertilizantes, medicinas terapéuticas y penicilina, entre otros, son algunos ejemplos de procesos biotecnológicos. En una búsqueda rápida en Internet, cerca de 113000 sitios fueron encontrados, ...

  20. 2,3-Butanediol recovery from fermentation broth by alcohol precipitation and vacuum distillation.

    Science.gov (United States)

    Jeon, Sangjun; Kim, Duk-Ki; Song, Hyohak; Lee, Hee Jong; Park, Sunghoon; Seung, Doyoung; Chang, Yong Keun

    2014-04-01

    This study presents a new and effective downstream process to recover 2,3-butanediol (2,3-BD) from fermentation broth which is produced by a recombinant Klebsiella pneumoniae strain. The ldhA-deficient K. pneumoniae strain yielded about 90 g/L of 2,3-BD, along with a number of by-products, such as organic acids and alcohols, in a 65 h fed-batch fermentation. The pH-adjusted cell-free fermentation broth was firstly concentrated until 2,3-BD reached around 500 g/L by vacuum evaporation at 50°C and 50 mbar vacuum pressure. The concentrated solution was further treated using light alcohols, including methanol, ethanol, and isopropanol, for the precipitation of organic acids and inorganic salts. Isopropanol showed the highest removal efficiency, in which 92.5% and 99.8% of organic acids and inorganic salts were precipitated, respectively. At a final step, a vacuum distillation process enabled the recovery of 76.2% of the treated 2,3-BD, with 96.1% purity, indicating that fermentatively produced 2,3-BD is effectively recovered by a simple alcohol precipitation and vacuum distillation. Copyright © 2013 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  1. Development of High-Productivity Continuous Ethanol Production using PVA-Immobilized Zymomonas mobilis in an Immobilized-Cells Fermenter

    Directory of Open Access Journals (Sweden)

    Nurhayati Nurhayati

    2015-07-01

    Full Text Available Ethanol as one of renewable energy was being considered an excellent alternative clean-burning fuel to replace gasoline. Continuous ethanol fermentation systems had offered important economic advantages compared to traditional systems. Fermentation rates were significantly improved, especially when continuous fermentation was integrated with cell immobilization techniques to enrich the cells concentration in fermentor. Growing cells of Zymomonas mobilis immobilized in polyvinyl alcohol (PVA gel beads were employed in an immobilized-cells fermentor for continuous ethanol fermentation from glucose. The glucose loading, dilution rate, and cells loading were varied in order to determine which best condition employed in obtaining both high ethanol production and low residual glucose with high dilution rate. In this study, 20 g/L, 100 g/L, 125 g/L and 150 g/L of glucose concentration and 20% (w/v, 40% (w/v and 50% (w/v of cells loading were employed with range of dilution rate at 0.25 to 1 h-1. The most stable production was obtained for 25 days by employing 100 g/L of glucose loading. Meanwhile, the results also exhibited that 125 g/L of glucose loading as well as 40% (w/v of cells loading yielded high ethanol concentration, high ethanol productivity, and acceptable residual glucose at 62.97 g/L, 15.74 g/L/h and 0.16 g/L, respectively. Furthermore, the dilution rate of 4 hour with 100 g/L and 40% (w/v of glucose and cells loading was considered as the optimum condition with ethanol production, ethanol productivity and residual glucose obtained were 49.89 g/L, 12.47 g/L/h, and 2.04 g/L, respectively. This recent study investigated ethanol inhibition as well. The present research had proved that high sugar concentration was successfully converted to ethanol. These achieved results were promising for further study.

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

    Science.gov (United States)

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

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

  3. Construction of a highly efficient Bacillus subtilis 168 whole-cell biocatalyst and its application in the production of L-ornithine.

    Science.gov (United States)

    Wang, Meizhou; Xu, Meijuan; Rao, Zhiming; Yang, Taowei; Zhang, Xian

    2015-11-01

    L-Ornithine, a non-protein amino acid, is usually extracted from hydrolyzed protein as well as produced by microbial fermentation. Here, we focus on a highly efficient whole-cell biocatalyst for the production of L-ornithine. The gene argI, encoding arginase, which catalyzes the hydrolysis of L-arginine to L-ornithine and urea, was cloned from Bacillus amyloliquefaciens B10-127 and expressed in GRAS strain Bacillus subtilis 168. The recombinant strain exhibited an arginase activity of 21.9 U/mg, which is 26.7 times that of wild B. subtilis 168. The optimal pH and temperature of the purified recombinant arginase were 10.0 and 40 °C, respectively. In addition, the recombinant arginase exhibited a strong Mn(2+) preference. When using whole-cell biocatalyst-based bioconversion, a hyper L-ornithine production of 356.9 g/L was achieved with a fed-batch strategy in a 5-L reactor within 12 h. This whole-cell bioconversion study demonstrates an environmentally friendly strategy for L-ornithine production in industry.

  4. Cost-effective simultaneous saccharification and fermentation of l-lactic acid from bagasse sulfite pulp by Bacillus coagulans CC17.

    Science.gov (United States)

    Zhou, Jie; Ouyang, Jia; Xu, Qianqian; Zheng, Zhaojuan

    2016-12-01

    The main barriers to cost-effective lactic acid production from lignocellulose are the high cost of enzymes and the ineffective utilization of the xylose within the hydrolysate. In the present study, the thermophilic Bacillus coagulans strain CC17 was used for the simultaneous saccharification and fermentation (SSF) of bagasse sulfite pulp (BSP) to produce l-lactic acid. Unexpectedly, SSF by CC17 required approximately 33.33% less fungal cellulase than did separate hydrolysis and fermentation (SHF). More interestingly, CC17 can co-ferment cellobiose and xylose without any exogenous β-glucosidase in SSF. Moreover, adding xylanase could increase the concentration of lactic acid produced via SSF. Up to 110g/L of l-lactic acid was obtained using fed-batch SSF, resulting in a lactic acid yield of 0.72g/g cellulose. These results suggest that SSF using CC17 has a remarkable advantage over SHF and that a potentially low-cost and highly-efficient fermentation process can be established using this protocol. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Effect of propionic acid on citric acid fermentation in an integrated citric acid-methane fermentation process.

    Science.gov (United States)

    Xu, Jian; Bao, Jia-Wei; Su, Xian-Feng; Zhang, Hong-Jian; Zeng, Xin; Tang, Lei; Wang, Ke; Zhang, Jian-Hua; Chen, Xu-Sheng; Mao, Zhong-Gui

    2016-03-01

    In this study, an integrated citric acid-methane fermentation process was established to solve the problem of wastewater treatment in citric acid production. Citric acid wastewater was treated through anaerobic digestion and then the anaerobic digestion effluent (ADE) was further treated and recycled for the next batch citric acid fermentation. This process could eliminate wastewater discharge and reduce water resource consumption. Propionic acid was found in the ADE and its concentration continually increased in recycling. Effect of propionic acid on citric acid fermentation was investigated, and results indicated that influence of propionic acid on citric acid fermentation was contributed to the undissociated form. Citric acid fermentation was inhibited when the concentration of propionic acid was above 2, 4, and 6 mM in initial pH 4.0, 4.5 and, 5.0, respectively. However, low concentration of propionic acid could promote isomaltase activity which converted more isomaltose to available sugar, thereby increasing citric acid production. High concentration of propionic acid could influence the vitality of cell and prolong the lag phase, causing large amount of glucose still remaining in medium at the end of fermentation and decreasing citric acid production.

  6. Influence of fungal morphology on the performance of industrial fermentation processes for enzyme production

    DEFF Research Database (Denmark)

    Quintanilla Hernandez, Daniela Alejandra

    Production of industrial enzymes is usually carried out as submerged aerobic fermentations. Filamentous microorganisms are widely used as hosts in these processes due to multiple advantages. Nevertheless, they also present major drawbacks, due to the unavoidable oxygen transfer limitations...... in this work, along with its correlation to viscosity and other process variables. Considerable research work has been conducted through the years to study fungal morphology and its relation to productivity. However, the work reported in the literature lacks relevant industrial data. In this work, a platform...... was developed which was able to produce high enzyme titers in comparison with what has been reported thus far in fed-batch fermentation using a soluble inducer (lactose). Different nitrogen sources were compared, and it was found that soy meal allowed for higher enzyme titers compared to what has been reported...

  7. Versatile modeling and optimization of fed batch processes for the production of secreted heterologous proteins with Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Gasser Brigitte

    2006-12-01

    Full Text Available Abstract Background Secretion of heterologous proteins depends both on biomass concentration and on the specific product secretion rate, which in turn is not constant at varying specific growth rates. As fed batch processes usually do not maintain a steady state throughout the feed phase, it is not trivial to model and optimize such a process by mathematical means. Results We have developed a model for product accumulation in fed batch based on iterative calculation in Microsoft Excel spreadsheets, and used the Solver software to optimize the time course of the media feed in order to maximize the volumetric productivity. The optimum feed phase consisted of an exponential feed at maximum specific growth rate, followed by a phase with linearly increasing feed rate and consequently steadily decreasing specific growth rate. The latter phase could be modeled also by exact mathematical treatment by the calculus of variations, yielding the explicit shape of the growth function, however, with certain indeterminate parameters. To evaluate the latter, one needs a numerical optimum search algorithm. The explicit shape of the growth function provides additional evidence that the Excel model results in correct data. Experimental evaluation in two independent fed batch cultures resulted in a good correlation to the optimized model data, and a 2.2 fold improvement of the volumetric productivity. Conclusion The advantages of the procedure we describe here are the ease of use and the flexibility, applying software familiar to every scientist and engineer, and rapid calculation which makes predictions extremely easy, so that many options can be tested in silico quickly. Additional options like further biological and technological constraints or different functions for specific productivity and biomass yield can easily be integrated.

  8. Long-term cathode performance and the microbial communities that develop in microbial fuel cells fed different fermentation endproducts

    KAUST Repository

    Kiely, Patrick D.; Rader, Geoffrey; Regan, John M.; Logan, Bruce E.

    2011-01-01

    fermentation (acetic acid, formic acid, lactic acid, succinic acid, or ethanol). Large variations in reactor performance were primarily due to the specific substrates, with power densities ranging from 835±21 to 62±1mW/m3. Cathodes performance degraded over

  9. PEI detoxification of pretreated spruce for high solids ethanol fermentation

    International Nuclear Information System (INIS)

    Cannella, David; Sveding, Per Viktor; Jørgensen, Henning

    2014-01-01

    Highlights: • High solids (30% dry matter) pretreatment, enzymatic hydrolysis and fermentation. • Horizontal rotary reactor for hydrolysis and fermentation. • In situ hydrolysates detoxification using inhibitors adsorbing PEI polymer. • 50% of inhibitors recovered as by-product, recyclability of PEI polymer up to 5 times. • 76% of maximum theoretical ethanol was fermented at final concentration of 51 g/kg. - Abstract: Performing the bioethanol production process at high solids loading is a requirement for economic feasibility at industrial scale. So far this has successfully been achieved using wheat straw and other agricultural residues at 30% of water insoluble solids (WIS), but for softwood species (i.e. spruce) this has been difficult to reach. The main reason behind this difference is the higher recalcitrance of woody substrates which require harsher pretreatment conditions, thus generating higher amounts of inhibitory compounds, ultimately lowering fermentation performances. In this work we studied ethanol production from spruce performing the whole process, from pretreatment to hydrolysis and fermentation, at 30% dry matter (equivalent to ∼20% WIS). Hydrolysis and fermentation was performed in a horizontal free fall mixing reactor enabling efficient mixing at high solids loadings. In batch simultaneous saccharification and fermentation (SSF), up to 76% cellulose to ethanol conversion was achieved resulting in a concentration of 51 g/kg of ethanol. Key to obtaining this high ethanol yield at these conditions was the use of a detoxification technology based on applying a soluble polyelectrolyte polymer (polyethylenimine, PEI) to absorb inhibitory compounds in the material. On average 50% removal and recovery of the main inhibitors (HMF, furfural, acetic acid and formic acid) was achieved dosing 1.5% w/w of soluble PEI. The use of PEI was compatible with operating the process at high solids loadings and enabled fermentation of hydrolysates, which

  10. Sensor combination and chemometric variable selection for online monitoring of Streptomyces coelicolor fed-batch cultivations

    DEFF Research Database (Denmark)

    Ödman, Peter; Johansen, C.L.; Olsson, L.

    2010-01-01

    of biomass and substrate (casamino acids) concentrations, respectively. The effect of combination of fluorescence and gas analyzer data as well as of different variable selection methods was investigated. Improved prediction models were obtained by combination of data from the two sensors and by variable......Fed-batch cultivations of Streptomyces coelicolor, producing the antibiotic actinorhodin, were monitored online by multiwavelength fluorescence spectroscopy and off-gas analysis. Partial least squares (PLS), locally weighted regression, and multilinear PLS (N-PLS) models were built for prediction...

  11. Immobilization of Lactobacillus rhamnosus in mesoporous silica-based material: An efficiency continuous cell-recycle fermentation system for lactic acid production.

    Science.gov (United States)

    Zhao, Zijian; Xie, Xiaona; Wang, Zhi; Tao, Yanchun; Niu, Xuedun; Huang, Xuri; Liu, Li; Li, Zhengqiang

    2016-06-01

    Lactic acid bacteria immobilization methods have been widely used for lactic acid production. Until now, the most common immobilization matrix used is calcium alginate. However, Ca-alginate gel disintegrated during lactic acid fermentation. To overcome this deficiency, we developed an immobilization method in which Lactobacillus rhamnosus cells were successfully encapsulated into an ordered mesoporous silica-based material under mild conditions with a high immobilization efficiency of 78.77% by using elemental analysis. We also optimized the cultivation conditions of the immobilized L. rhamnosus and obtained a high glucose conversion yield of 92.4%. Furthermore, L. rhamnosus encapsulated in mesoporous silica-based material exhibited operational stability during repeated fermentation processes and no decrease in lactic acid production up to 8 repeated batches. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  12. Comparative study of production of Bio-Indigo by Pandoraea sp. in a two phase - fed batch and continuous bioreactor

    Directory of Open Access Journals (Sweden)

    Vaishnavi Unde

    2016-03-01

    Full Text Available Indigo, is blue of blue jeans, a synthetic dye used on large scale all over the world. Chemical production of the dye is taking a new route towards bacterial production to overcome the environmental effects that are posed by the synthetic blue powder (Indigo. In the present work a strain Pandoraea sp. isolated from the oil contaminated soil is found to produce blue pigment which is analyzed qualitatively as indigo using UV-visible scan and Thin Layer Chromatography (TLC. The strain is used for indigo production at lab scale in two different bioreactor configurations first the fed batch mode and second continuous mode using two phases. The two phases consisting of medium carrying biomass and the second phase of silicone oil carrying substrate indole. The use of second phase allows higher concentration of substrate injection reducing the inhibition effects of the substrate as well as act as a partitioning agent for removal of the product. In two phase study, the maximum indigo produced was seen to be 0.068 g/L after 22 hours of substrate injection into the Fermentor in a fed batch mode. The maximum yield obtained in this configuration was 19%. For commercial production of bio-indigo a continuous operation is required, which was studied in a bioreactor with 2.5 liter capacity under the optimized conditions. The maximum indigo produced was found to be 0.052 g/L after about 72 hours of operation. The results showed decrease in the production of indigo in continuous mode as compared to fed batch operation, which may be due to the insufficient time available for the bacteria to bio-transform indole into indigo.

  13. Characteristics of Saccharomyces cerevisiae yeasts exhibiting rough colonies and pseudohyphal morphology with respect to alcoholic fermentation.

    Science.gov (United States)

    Reis, Vanda Renata; Bassi, Ana Paula Guarnieri; da Silva, Jessica Carolina Gomes; Ceccato-Antonini, Sandra Regina

    2013-12-01

    Among the native yeasts found in alcoholic fermentation, rough colonies associated with pseudohyphal morphology belonging to the species Saccharomyces cerevisiae are very common and undesirable during the process. The aim of this work was to perform morphological and physiological characterisations of S. cerevisiae strains that exhibited rough and smooth colonies in an attempt to identify alternatives that could contribute to the management of rough colony yeasts in alcoholic fermentation. Characterisation tests for invasiveness in Agar medium, killer activity, flocculation and fermentative capacity were performed on 22 strains (11 rough and 11 smooth colonies). The effects of acid treatment at different pH values on the growth of two strains ("52"--rough and "PE-02"--smooth) as well as batch fermentation tests with cell recycling and acid treatment of the cells were also evaluated. Invasiveness in YPD Agar medium occurred at low frequency; ten of eleven rough yeasts exhibited flocculation; none of the strains showed killer activity; and the rough strains presented lower and slower fermentative capacities compared to the smooth strains in a 48-h cycle in a batch system with sugar cane juice. The growth of the rough strain was severely affected by the acid treatment at pH values of 1.0 and 1.5; however, the growth of the smooth strain was not affected. The fermentative efficiency in mixed fermentation (smooth and rough strains in the same cell mass proportion) did not differ from the efficiency obtained with the smooth strain alone, most likely because the acid treatment was conducted at pH 1.5 in a batch cell-recycle test. A fermentative efficiency as low as 60% was observed with the rough colony alone.

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

    Science.gov (United States)

    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.

  15. 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.

    Science.gov (United States)

    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.

  16. Modelling Fungal Fermentations for Enzyme Production

    DEFF Research Database (Denmark)

    Albæk, Mads Orla; Gernaey, Krist; Hansen, Morten S.

    We have developed a process model of fungal fed-batch fermentations for enzyme production. In these processes, oxygen transfer rate is limiting and controls the substrate feeding rate. The model has been shown to describe cultivations of both Aspergillus oryzae and Trichoderma reesei strains in 550......L stirred tank pilot plant reactors well. For each strain, 8 biological parameters are needed as well as a correlation of viscosity, as viscosity has a major influence on oxygen transfer. The parameters were measured averages of at least 9 batches for each strain. The model is successfully able...... to cover a wide range of process conditions (0.3-2 vvm of aeration, 0.2-10.0 kW/m3 of specific agitation power input, and 0.1-1.3 barg head space pressure). Uncertainty and sensitivity analysis have shown that the uncertainty of the model is mainly due to difficulties surrounding the estimation...

  17. Model-based optimization and scale-up of multi-feed simultaneous saccharification and co-fermentation of steam pre-treated lignocellulose enables high gravity ethanol production.

    Science.gov (United States)

    Wang, Ruifei; Unrean, Pornkamol; Franzén, Carl Johan

    2016-01-01

    High content of water-insoluble solids (WIS) is required for simultaneous saccharification and co-fermentation (SSCF) operations to reach the high ethanol concentrations that meet the techno-economic requirements of industrial-scale production. The fundamental challenges of such processes are related to the high viscosity and inhibitor contents of the medium. Poor mass transfer and inhibition of the yeast lead to decreased ethanol yield, titre and productivity. In the present work, high-solid SSCF of pre-treated wheat straw was carried out by multi-feed SSCF which is a fed-batch process with additions of substrate, enzymes and cells, integrated with yeast propagation and adaptation on the pre-treatment liquor. The combined feeding strategies were systematically compared and optimized using experiments and simulations. For high-solid SSCF process of SO2-catalyzed steam pre-treated wheat straw, the boosted solubilisation of WIS achieved by having all enzyme loaded at the beginning of the process is crucial for increased rates of both enzymatic hydrolysis and SSCF. A kinetic model was adapted to simulate the release of sugars during separate hydrolysis as well as during SSCF. Feeding of solid substrate to reach the instantaneous WIS content of 13 % (w/w) was carried out when 60 % of the cellulose was hydrolysed, according to simulation results. With this approach, accumulated WIS additions reached more than 20 % (w/w) without encountering mixing problems in a standard bioreactor. Feeding fresh cells to the SSCF reactor maintained the fermentation activity, which otherwise ceased when the ethanol concentration reached 40-45 g L(-1). In lab scale, the optimized multi-feed SSCF produced 57 g L(-1) ethanol in 72 h. The process was reproducible and resulted in 52 g L(-1) ethanol in 10 m(3) scale at the SP Biorefinery Demo Plant. SSCF of WIS content up to 22 % (w/w) is reproducible and scalable with the multi-feed SSCF configuration and model-aided process

  18. Effect of acetic acid on citric acid fermentation in an integrated citric acid-methane fermentation process.

    Science.gov (United States)

    Xu, Jian; Chen, Yang-Qiu; Zhang, Hong-Jian; Tang, Lei; Wang, Ke; Zhang, Jian-Hua; Chen, Xu-Sheng; Mao, Zhong-Gui

    2014-09-01

    An integrated citric acid-methane fermentation process was proposed to solve the problem of extraction wastewater in citric acid fermentation process. Extraction wastewater was treated by anaerobic digestion and then recycled for the next batch of citric acid fermentation to eliminate wastewater discharge and reduce water resource consumption. Acetic acid as an intermediate product of methane fermentation was present in anaerobic digestion effluent. In this study, the effect of acetic acid on citric acid fermentation was investigated and results showed that lower concentration of acetic acid could promote Aspergillus niger growth and citric acid production. 5-Cyano-2,3-ditolyl tetrazolium chloride (CTC) staining was used to quantify the activity of A. niger cells, and the results suggested that when acetic acid concentration was above 8 mM at initial pH 4.5, the morphology of A. niger became uneven and the part of the cells' activity was significantly reduced, thereby resulting in deceasing of citric acid production. Effects of acetic acid on citric acid fermentation, as influenced by initial pH and cell number in inocula, were also examined. The result indicated that inhibition by acetic acid increased as initial pH declined and was rarely influenced by cell number in inocula.

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

    Science.gov (United States)

    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.

  20. Biohydrogen production from combined dark-photo fermentation under a high ammonia content in the dark fermentation effluent

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Chun-Yen [National Cheng Kung Univ., Tainan, Taiwan (China). Dept. of Chemical Engineering; National Cheng Kung Univ., Tainan, Taiwan (China). Sustainable Environment Research Center; Lo, Yung-Chung; Yeh, Kuei-Ling [National Cheng Kung Univ., Tainan, Taiwan (China). Dept. of Chemical Engineering; Chang, Jo-Shu [National Cheng Kung Univ., Tainan, Taiwan (China). Dept. of Chemical Engineering; National Cheng Kung Univ., Tainan, Taiwan (China). Sustainable Environment Research Center; National Cheng Kung Univ., Tainan, Taiwan (China). Microalgae Biotechnology and Bioengineering Lab.

    2010-07-01

    Integrated dark and photo (two-stage) fermentation was employed to enhance the performance of H{sub 2} production. First, the continuous dark fermentation using indigenous Clostridium butyricum CGS5 was carried out at 12 h HRT and fed with sucrose at a concentration of 18750 mg/l. The overall H{sub 2} production rate and H{sub 2} yield were fairly stable with a mean value of 87.5 ml/l/h and 1.015 mol H{sub 2}/mol sucrose, respectively. In addition, a relatively high ammonia nitrogen content (574 mg/l) in the dark fermentation effluent was observed. The soluble metabolites from dark fermentation, consisting mainly of butyric, lactic and acetic acids, were directly used as the influent of continuous photo-H{sub 2} production process inoculated with Rhodopseudomonas palutris WP 3-5 under the condition of 35oC, 10000 lux irradiation, pH 7.0 and 48 h HRT. The maximum overall hydrogen production rate from photo fermentation was 16.4 ml H{sub 2}/l/h, and the utilization of the soluble metabolites could reach 90%. The maximum H{sub 2} yield dramatically increased from 1.015 mol H{sub 2}/mol sucrose (in dark fermentation only) to 6.04 mol H{sub 2}/mol sucrose in the combined dark and photo fermentation. Surprisingly, the operation strategy applied in this work was able to attain an average NH{sub 3}-N removal efficiency of 92%, implying that our photo-H{sub 2} production system has a higher NH{sub 3}-N tolerance, demonstrating its high applicability in an integrated dark-photo fermentation system. (orig.)

  1. Industrial antifoam agents impair ethanol fermentation and induce stress responses in yeast cells.

    Science.gov (United States)

    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.

  2. Comparative analysis of microbial community of novel lactic acid fermentation inoculated with different undefined mixed cultures.

    Science.gov (United States)

    Liang, Shaobo; Gliniewicz, Karol; Mendes-Soares, Helena; Settles, Matthew L; Forney, Larry J; Coats, Erik R; McDonald, Armando G

    2015-03-01

    Three undefined mixed cultures (activated sludge) from different municipal wastewater treatment plants were used as seeds in a novel lactic acid fermentation process fed with potato peel waste (PPW). Anaerobic sequencing batch fermenters were run under identical conditions to produce predominantly lactic acid. Illumina sequencing was used to examine the 16S rRNA genes of bacteria in the three seeds and fermenters. Results showed that the structure of microbial communities of three seeds were different. All three fermentation products had unique community structures that were dominated (>96%) by species of the genus Lactobacillus, while members of this genus constituted undefined mixed cultures were robust and resilient, which provided engineering prospects for the microbial utilization of carbohydrate wastes to produce lactic acid. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. Recovery of phosphorus and volatile fatty acids from wastewater and food waste with an iron-flocculation sequencing batch reactor and acidogenic co-fermentation.

    Science.gov (United States)

    Li, Ruo-Hong; Li, Xiao-Yan

    2017-12-01

    A sequencing batch reactor-based system was developed for enhanced phosphorus (P) removal and recovery from municipal wastewater. The system consists of an iron-dosing SBR for P precipitation and a side-stream anaerobic reactor for sludge co-fermentation with food waste. During co-fermentation, sludge and food waste undergo acidogenesis, releasing phosphates under acidic conditions and producing volatile fatty acids (VFAs) into the supernatant. A few types of typical food waste were investigated for their effectiveness in acidogenesis and related enzymatic activities. The results show that approximately 96.4% of total P in wastewater was retained in activated sludge. Food waste with a high starch content favoured acidogenic fermentation. Around 55.7% of P from wastewater was recovered as vivianite, and around 66% of food waste loading was converted into VFAs. The new integration formed an effective system for wastewater treatment, food waste processing and simultaneous recovery of P and VFAs. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Efficiency and rumen responses in younger and older Holstein heifers limit-fed diets of differing energy density.

    Science.gov (United States)

    Zanton, G I; Heinrichs, A J

    2016-04-01

    The objective of this study was to evaluate the effects of limit feeding diets of different predicted energy density on the efficiency of utilization of feed and nitrogen and rumen responses in younger and older Holstein heifers. Eight rumen-cannulated Holstein heifers (4 heifers beginning at 257 ± 7 d, hereafter "young," and 4 heifers beginning at 610 ± 16 d, hereafter "old") were limit-fed high [HED; 2.64 Mcal/kg of dry matter (DM), 15.31% crude protein (CP)] or low (LED; 2.42 Mcal/kg of DM, 14.15% CP) energy density diets according to a 4-period, split-plot Latin square design with 28-d periods. Diets were limit-fed to provide isonitrogenous and isoenergetic intake on a rumen empty body weight (BW) basis at a level predicted to support approximately 800 g/d of average daily gain. During the last 7d of each period, rumen contents were subsampled over a 24-h period, rumen contents were completely evacuated, and total collection of feces and urine was made over 4d. Intakes of DM and water were greater for heifers fed LED, although, by design, calculated intake of metabolizable energy did not differ between age groups or diets when expressed relative to rumen empty BW. Rumen pH was lower, ammonia (NH3-N) concentration tended to be higher, and volatile fatty acids (VFA) concentration was not different for HED compared with LED and was unaffected by age group. Rumen content mass was greater for heifers fed LED and for old heifers, so when expressing rumen fermentation responses corrected for this difference in pool size, NH3-N pool size was not different between diets and total moles of VFA in the rumen were greater for heifers fed LED, whereas these pool sizes were greater for old heifers. Total-tract digestibility of potentially digestible neutral detergent fiber (NDF) was greater in heifers fed LED and for young heifers, whereas the fractional rate of ruminal passage and digestion of NDF were both greater in heifers fed LED. Digestibility of N was greater for

  5. Continuous Ethanol Production Using Immobilized-Cell/Enzyme Biocatalysts in Fluidized-Bed Bioreactor (FBR)

    Energy Technology Data Exchange (ETDEWEB)

    Nghiem, NP

    2003-11-16

    The immobilized-cell fluidized-bed bioreactor (FBR) was developed at Oak Ridge National Laboratory (ORNL). Previous studies at ORNL using immobilized Zymomonas mobilis in FBR at both laboratory and demonstration scale (4-in-ID by 20-ft-tall) have shown that the system was more than 50 times as productive as industrial benchmarks (batch and fed-batch free cell fermentations for ethanol production from glucose). Economic analysis showed that a continuous process employing the FBR technology to produce ethanol from corn-derived glucose would offer savings of three to six cents per gallon of ethanol compared to a typical batch process. The application of the FBR technology for ethanol production was extended to investigate more complex feedstocks, which included starch and lignocellulosic-derived mixed sugars. Economic analysis and mathematical modeling of the reactor were included in the investigation. This report summarizes the results of these extensive studies.

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

    Science.gov (United States)

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

    2016-07-01

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

  7. Glycerol as a Cheaper Carbon Source in Bacterial Cellulose (BC) Production by Gluconacetobacter Xylinus DSM46604 in Batch Fermentation System

    International Nuclear Information System (INIS)

    Azila Adnan; Nair, G.R.; Roslan Umar; Roslan Umar

    2015-01-01

    Bacterial cellulose (BC) is a polymer of glucose monomers, which has unique properties including high crystallinity and high strength. It has potential to be used in biomedical applications such as making artificial blood vessel, wound dressings, and in the paper making industry. Extensive study on BC aimed to improve BC production such as by using glycerol as a cheaper carbon source. BC was produced in shake flask culture using five different concentrations of glycerol (10, 20, 30, 40 and 50 g/ L). Using concentration of glycerol above 20 g/ L inhibited culture growth and BC production. Further experiments were performed in batch culture (3-L bioreactor) using 20 g/ L glycerol. It produced yield and productivity of 0.15 g/ g and 0.29 g/ L/ day BC, respectively. This is compared with the control medium, 50 g/ L glucose, which only gave yield and productivity of 0.05 g/ g and 0.23 g/ L/ day, respectively. Twenty g/ L of glycerol enhanced BC production by Gluconacetobacter xylinus DSM46604 in batch fermentation system. (author)

  8. High power density yeast catalyzed microbial fuel cells

    Science.gov (United States)

    Ganguli, Rahul

    Microbial fuel cells leverage whole cell biocatalysis to convert the energy stored in energy-rich renewable biomolecules such as sugar, directly to electrical energy at high efficiencies. Advantages of the process include ambient temperature operation, operation in natural streams such as wastewater without the need to clean electrodes, minimal balance-of-plant requirements compared to conventional fuel cells, and environmentally friendly operation. These make the technology very attractive as portable power sources and waste-to-energy converters. The principal problem facing the technology is the low power densities compared to other conventional portable power sources such as batteries and traditional fuel cells. In this work we examined the yeast catalyzed microbial fuel cell and developed methods to increase the power density from such fuel cells. A combination of cyclic voltammetry and optical absorption measurements were used to establish significant adsorption of electron mediators by the microbes. Mediator adsorption was demonstrated to be an important limitation in achieving high power densities in yeast-catalyzed microbial fuel cells. Specifically, the power densities are low for the length of time mediator adsorption continues to occur. Once the mediator adsorption stops, the power densities increase. Rotating disk chronoamperometry was used to extract reaction rate information, and a simple kinetic expression was developed for the current observed in the anodic half-cell. Since the rate expression showed that the current was directly related to microbe concentration close to the electrode, methods to increase cell mass attached to the anode was investigated. Electrically biased electrodes were demonstrated to develop biofilm-like layers of the Baker's yeast with a high concentration of cells directly connected to the electrode. The increased cell mass did increase the power density 2 times compared to a non biofilm fuel cell, but the power density

  9. Lignocellulosic Fermentation of Wild Grass Employing Recombinant Hydrolytic Enzymes and Fermentative Microbes with Effective Bioethanol Recovery

    Directory of Open Access Journals (Sweden)

    Saprativ P. Das

    2013-01-01

    Full Text Available Simultaneous saccharification and fermentation (SSF studies of steam exploded and alkali pretreated different leafy biomass were accomplished by recombinant Clostridium thermocellum hydrolytic enzymes and fermentative microbes for bioethanol production. The recombinant C. thermocellum GH5 cellulase and GH43 hemicellulase genes expressed in Escherichia coli cells were grown in repetitive batch mode, with the aim of enhancing the cell biomass production and enzyme activity. In batch mode, the cell biomass (A600 nm of E. coli cells and enzyme activities of GH5 cellulase and GH43 hemicellulase were 1.4 and 1.6 with 2.8 and 2.2 U·mg−1, which were augmented to 2.8 and 2.9 with 5.6 and 3.8 U·mg−1 in repetitive batch mode, respectively. Steam exploded wild grass (Achnatherum hymenoides provided the best ethanol titres as compared to other biomasses. Mixed enzyme (GH5 cellulase, GH43 hemicellulase mixed culture (Saccharomyces cerevisiae, Candida shehatae system gave 2-fold higher ethanol titre than single enzyme (GH5 cellulase single culture (Saccharomyces cerevisiae system employing 1% (w/v pretreated substrate. 5% (w/v substrate gave 11.2 g·L−1 of ethanol at shake flask level which on scaling up to 2 L bioreactor resulted in 23 g·L−1 ethanol. 91.6% (v/v ethanol was recovered by rotary evaporator with 21.2% purification efficiency.

  10. Electricity generation from synthesis gas by microbial processes: CO fermentation and microbial fuel cell technology.

    Science.gov (United States)

    Kim, Daehee; Chang, In Seop

    2009-10-01

    A microbiological process was established to harvest electricity from the carbon monoxide (CO). A CO fermenter was enriched with CO as the sole carbon source. The DGGE/DNA sequencing results showed that Acetobacterium spp. were enriched from the anaerobic digester fluid. After the fermenter was operated under continuous mode, the products were then continuously fed to the microbial fuel cell (MFC) to generate electricity. Even though the conversion yield was quite low, this study proved that synthesis gas (syn-gas) can be converted to electricity with the aid of microbes that do not possess the drawbacks of metal catalysts of conventional methods.

  11. Analysis of microbial community variation during the mixed culture fermentation of agricultural peel wastes to produce lactic acid.

    Science.gov (United States)

    Liang, Shaobo; Gliniewicz, Karol; Gerritsen, Alida T; McDonald, Armando G

    2016-05-01

    Mixed cultures fermentation can be used to convert organic wastes into various chemicals and fuels. This study examined the fermentation performance of four batch reactors fed with different agricultural (orange, banana, and potato (mechanical and steam)) peel wastes using mixed cultures, and monitored the interval variation of reactor microbial communities with 16S rRNA genes using Illumina sequencing. All four reactors produced similar chemical profile with lactic acid (LA) as dominant compound. Acetic acid and ethanol were also observed with small fractions. The Illumina sequencing results revealed the diversity of microbial community decreased during fermentation and a community of largely lactic acid producing bacteria dominated by species of Lactobacillus developed. Copyright © 2016 Elsevier Ltd. All rights reserved.

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

    Science.gov (United States)

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

    2011-10-01

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

  13. Probing topological relations between high-density and low-density regions of 2MASS with hexagon cells

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Yongfeng [American Physical Society, San Diego, CA (United States); Xiao, Weike, E-mail: yongfeng.wu@maine.edu [Department of Astronautics Engineering, Harbin Institute of Technology, P.O. Box 345, Heilongjiang Province 150001 (China)

    2014-02-01

    We introduced a new two-dimensional (2D) hexagon technique for probing the topological structure of the universe in which we mapped regions of the sky with high and low galaxy densities onto a 2D lattice of hexagonal unit cells. We defined filled cells as corresponding to high-density regions and empty cells as corresponding to low-density regions. The numbers of filled cells and empty cells were kept the same by controlling the size of the cells. By analyzing the six sides of each hexagon, we could obtain and compare the statistical topological properties of high-density and low-density regions of the universe in order to have a better understanding of the evolution of the universe. We applied this hexagonal method to Two Micron All Sky Survey data and discovered significant topological differences between the high-density and low-density regions. Both regions had significant (>5σ) topological shifts from both the binomial distribution and the random distribution.

  14. L-Lactic Acid Production by Lactobacillus rhamnosus ATCC 10863

    Directory of Open Access Journals (Sweden)

    Ana Lívia Chemeli Senedese

    2015-01-01

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

  15. Improving the monitoring of methanol concentration during high cell density fermentation of Pichia pastoris.

    Science.gov (United States)

    Ramon, R; Feliu, J X; Cos, O; Montesinos, J L; Berthet, F X; Valero, F

    2004-09-01

    The Pichia pastoris expression system is widely used for the production of recombinant proteins. A simple and efficient experimental set-up allowing on-line monitoring of the methanol concentration during the fermentation of P. pastoris based on the detection of the methanol vapor concentration in the exhaust air from fermenter by a tin dioxide (SnO2) semiconductor sensor is described. An experimental procedure to allow precise calibration of the system and to reduce methanol sensor's interferences (>95% reduction) are also presented and discussed. Accuracy and measurement error were estimated about 0.05 g x l(-1) and 6%, respectively. The efficient monitoring of methanol will help to advanced control of recombinant protein production and process optimization.

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

    Science.gov (United States)

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

    2017-03-20

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

  17. [Fermentation behaviors of recombinant Pichia pastoris under inhibited methanol concentration].

    Science.gov (United States)

    Zhou, Xiang-Shan; Fan, Wei-Min; Zhang, Yuan-Xing

    2003-09-01

    Chemostat culture was performed to characterize the growth, substrate consumption and the hirudin production, and to disclose their interrelations in the fermentation of recombinant Pichia pastoris. The Andrew substrate-inhibited growth model is more suitable than Monod model to simulate the growth of Pichia pastoris on methanol. Therefore, two stationary states can be obtained in the continuous culture at a certain dilution rate because of the substrate inhibition on cell growth. The stationary state could be obtained if only the dilution rate not more than 0.048 h(-1) in the continuous fermentation. The concentrations of cell, methanol and hirudin were constant after 50 h continuous culture with dilution rate at 0.04 h(-1). However, it could not be obtained when the dilution rate more than 0.048 h(-1) because the other stationary point at S > 0.048 h(-1) is unstable. Therefore, it was found that the cell concentration declined and the methanol concentration increased from 2.9 g/L to 18.1 g/L within 18h at dilution rate 0.06 h(-1). Thus, the fed-batch culture with a constant specific growth rate was carried out to disclose the fermentation behavior at high and constant methanol concentration in aid of a methanol sensor. The theoretical maximum specific growth rate, microm = 0.0464 h(-1), was found under critical methanol concentration, Scrit = 3.1 g/L. The growth of P. pastoris was typically methanol-limited at the methanol concentration S Scrit. The maximum specific Hir65 production rate qp was obtained at 0.2 mg/(g x h) when methanol concentration and mu were 0.5 g/L and 0.02 h(-1), respectively. The specific Hir65 production rate qp increased with the increase of mu and S at mu 0.02 h(-1). The specific methanol consumption rate increased with the increase of S when S 5 g/L. At last, the high Hir65 production rate 0.2 mg/(g x h) was obtained in the fermentation conducted under methanol-limited concentration and mu controlled at 0.5 g/L and 0.02 h(-1

  18. Continuous cider fermentation with co-immobilized yeast and Leuconostoc oenos cells.

    Science.gov (United States)

    Nedovic; Durieuxb; Van Nedervelde L; Rosseels; Vandegans; Plaisant; Simon

    2000-06-01

    Ca-alginate matrix was used to co-immobilize Saccharomyces bayanus and Leuconostoc oenos in one integrated biocatalytic system in order to perform simultaneously alcoholic and malo-lactic fermentation of apple juice to produce cider, in a continuous packed bed bioreactor. The continuous process permitted much faster fermentation compared with the traditional batch process. The flavor formation was also better controlled. By adjusting the flow rate of feeding substrate through the bioreactor, i.e. its residence time, it was possible to obtain either "soft" or "dry" cider. However, the profile of volatile compounds in the final product was modified comparatively to the batch process, especially for higher alcohols, isoamylacetate, and diacetyl. This modification is due to different physiology states of yeast in two processes. Nevertheless, the taste of cider was quite acceptable.

  19. Production of ethanol from wheat straw by pretreatment and fermentation at high dry matter concentrations

    NARCIS (Netherlands)

    Groenestijn, J.W. van; Slomp, R.S.

    2011-01-01

    High concentrations of substrate and product are important for the economy of second-generation bioethanol production. By a dilute acid thermal pretreatment of large pieces of relatively dry wheat straw using a novel rapid heating method, followed by fed-batch preliquefaction with hydrolytic

  20. Microfluidic biolector-microfluidic bioprocess control in microtiter plates.

    Science.gov (United States)

    Funke, Matthias; Buchenauer, Andreas; Schnakenberg, Uwe; Mokwa, Wilfried; Diederichs, Sylvia; Mertens, Alan; Müller, Carsten; Kensy, Frank; Büchs, Jochen

    2010-10-15

    In industrial-scale biotechnological processes, the active control of the pH-value combined with the controlled feeding of substrate solutions (fed-batch) is the standard strategy to cultivate both prokaryotic and eukaryotic cells. On the contrary, for small-scale cultivations, much simpler batch experiments with no process control are performed. This lack of process control often hinders researchers to scale-up and scale-down fermentation experiments, because the microbial metabolism and thereby the growth and production kinetics drastically changes depending on the cultivation strategy applied. While small-scale batches are typically performed highly parallel and in high throughput, large-scale cultivations demand sophisticated equipment for process control which is in most cases costly and difficult to handle. Currently, there is no technical system on the market that realizes simple process control in high throughput. The novel concept of a microfermentation system described in this work combines a fiber-optic online-monitoring device for microtiter plates (MTPs)--the BioLector technology--together with microfluidic control of cultivation processes in volumes below 1 mL. In the microfluidic chip, a micropump is integrated to realize distinct substrate flow rates during fed-batch cultivation in microscale. Hence, a cultivation system with several distinct advantages could be established: (1) high information output on a microscale; (2) many experiments can be performed in parallel and be automated using MTPs; (3) this system is user-friendly and can easily be transferred to a disposable single-use system. This article elucidates this new concept and illustrates applications in fermentations of Escherichia coli under pH-controlled and fed-batch conditions in shaken MTPs. Copyright 2010 Wiley Periodicals, Inc.

  1. Cultivos de alta densidad celular por retención interna: aplicación a la fermentación continua de etanol High cell density cultures produced by internal retention: application in continuous ethanol fermentation

    Directory of Open Access Journals (Sweden)

    Godoy Rubén Darío

    2004-12-01

    . This can be achieved by using techniques leading to high cell concentration and reducing inhibition by the end-product. One of the frequently employed methods involves cell recycling. This work thus developed a membrane reactor incorporating a filtration module with 5 u,m stainless steel tubular units inside a 3L stirred jar fermenter for investigating its application in continuous ethanol production. The effects of cell concentration and transmembrane pressure difference on permeate flux were evaluated for testing the filtration module's performance. The internal cell retention system was operated in Saccharomyces cerevisiae continuous culture derived from sucrose, once fermentation conditions had been selected (30 °C, 1.25 -1.75 vvm, pH 4.5. Filter unit permeability was maintained by applying pulses of air. More than 97% of the grown cells were retained in the fermenter, reaching 51 g/L cell concentration and 8.51 g/L.h average ethanol productivity in culture with internal cell retention; this was twice that obtained in a conventional continuous culture. Key words: Membrane reactor, Saccharomyces cerevisiae, alcoholic fermentation, cell recycling.

  2. Limiting factors in Escherichia colifed-batch production of recombinant proteins

    DEFF Research Database (Denmark)

    Sanden, A.M.; Prytz, I.; Tubelekas, I.

    2003-01-01

    recombinant protein production, fed-batch, specific growth rate, feed profile, induction, mRNA, transcription, translation, acetic acid formation......recombinant protein production, fed-batch, specific growth rate, feed profile, induction, mRNA, transcription, translation, acetic acid formation...

  3. Improving simultaneous saccharification and co-fermentation of pretreated wheat straw using both enzyme and substrate feeding

    Directory of Open Access Journals (Sweden)

    Palmqvist Benny

    2010-08-01

    Full Text Available Abstract Background Simultaneous saccharification and co-fermentation (SSCF has been recognized as a feasible option for ethanol production from xylose-rich lignocellulosic materials. To reach high ethanol concentration in the broth, a high content of water-insoluble solids (WIS is needed, which creates mixing problems and, furthermore, may decrease xylose uptake. Feeding of substrate has already been proven to give a higher xylose conversion than a batch SSCF. In the current work, enzyme feeding, in addition to substrate feeding, was investigated as a means of enabling a higher WIS content with a high xylose conversion in SSCF of a xylose-rich material. A recombinant xylose-fermenting strain of Saccharomyces cerevisiae (TMB3400 was used for this purpose in fed-batch SSCF experiments of steam-pretreated wheat straw. Results By using both enzyme and substrate feeding, the xylose conversion in SSCF could be increased from 40% to 50% in comparison to substrate feeding only. In addition, by this design of the feeding strategy, it was possible to process a WIS content corresponding to 11% in SSCF and obtain an ethanol yield on fermentable sugars of 0.35 g g-1. Conclusion A combination of enzyme and substrate feeding was shown to enhance xylose uptake by yeast and increase overall ethanol yield in SSCF. This is conceptually important for the design of novel SSCF processes aiming at high-ethanol titers. Substrate feeding prevents viscosity from becoming too high and thereby allows a higher total amount of WIS to be added in the process. The enzyme feeding, furthermore, enables keeping the glucose concentration low, which kinetically favors xylose uptake and results in a higher xylose conversion.

  4. Heat-treatment, phytase and fermented liquid feeding affect the presence of inositol phosphates in ileal digesta and phosphorus digestibility in pigs fed a wheat and barley diet

    DEFF Research Database (Denmark)

    Blaabjerg, Karoline; Jørgensen, H.; Tauson, Anne-Helene

    2010-01-01

    The aim was to evaluate the effect of heat-treatment, microbial phytase addition and feeding strategy (dry feeding v. fermented liquid feeding) on degradation of phytate (myo-inositol hexakisphosphate, InsP6) and formation and further degradation of lower inositol phosphates (myo-inositol pentaki......The aim was to evaluate the effect of heat-treatment, microbial phytase addition and feeding strategy (dry feeding v. fermented liquid feeding) on degradation of phytate (myo-inositol hexakisphosphate, InsP6) and formation and further degradation of lower inositol phosphates (myo...... × 4 Latin square with four pigs fed four diets. A basal wheat/barley-based diet was fed either as non-heat-treated or heat-treated (steam-pelleted at 90°C). The heat-treatment resulted in an inactivation of plant phytase below detectable level. Diet 1 (non-heat-treated basal diet fed dry); diet 2...... (heat-treated basal diet fed dry); diet 3 (as diet 2 but with microbial phytase (750 FTU/kg as fed) fed dry); diet 4 (as diet 3 fed liquid (fermented for 17.5 h nighttime and 6.5 h daytime at 20°C with 50% residue in the tank)). Chromic oxide (Cr2O3) was included as marker and ATTD was determined both...

  5. Physiological changes of Candida tropicalis population degrading phenol in fed batch reactor

    Directory of Open Access Journals (Sweden)

    Eliska Komarkova

    2003-12-01

    Full Text Available Candida tropicalis can use phenol as the sole carbon and energy source. Experiments regarding phenol degradations from the water phase were carried out. The fermentor was operated as a fed-batch system with oxistat control. Under conditions of nutrient limitation and an excess of oxygen the respiration activity of cells was suppressed and some color metabolites (black-brown started to be formed. An accumulation of these products inhibited the cell growth under aerobic conditions. Another impact was a decrease of the phenol hydroxylase activity as the key enzyme of the phenol degradation pathway at the end of the cell respiration activity. This decrease is linked with the above mentioned product inhibition. The cell death studied by fluorescent probe proceeded very slowly after the loss of the respiration activity. The starvation stress induced an increase of the endogenous respiration rate at the expense of phenol oxidation.Candida tropicalis pode utilizar fenol como única fonte de carbono e de energia. O fermentador foi operado em um sistema ''batelada-alimentada'' e controle oxidativo. Em condições limitantes de nutrientes e excesso de oxigênio a atividade respiratória das células foi suprimida e o calor do metabolismo pode ser formado. Uma acumulação desses produtos inibiu o crescimento das células em condições aeróbicas. Outro impacto foi um decréscimo da atividade fenol hidroxilase como enzima chave da degradação do fenol no final da atividade respirométrica. Essa redução está relacionada com os fatos acima mencionados. A morte da célula estudada por sonda de fluorescência ocorreu lentamente após a perda da atividade respiratória. O ''stress'' celular induziu um aumento na taxa de respiração endógena devido à oxidação fenólica.

  6. A robotics platform for automated batch fabrication of high density, microfluidics-based DNA microarrays, with applications to single cell, multiplex assays of secreted proteins

    Science.gov (United States)

    Ahmad, Habib; Sutherland, Alex; Shin, Young Shik; Hwang, Kiwook; Qin, Lidong; Krom, Russell-John; Heath, James R.

    2011-09-01

    Microfluidics flow-patterning has been utilized for the construction of chip-scale miniaturized DNA and protein barcode arrays. Such arrays have been used for specific clinical and fundamental investigations in which many proteins are assayed from single cells or other small sample sizes. However, flow-patterned arrays are hand-prepared, and so are impractical for broad applications. We describe an integrated robotics/microfluidics platform for the automated preparation of such arrays, and we apply it to the batch fabrication of up to eighteen chips of flow-patterned DNA barcodes. The resulting substrates are comparable in quality with hand-made arrays and exhibit excellent substrate-to-substrate consistency. We demonstrate the utility and reproducibility of robotics-patterned barcodes by utilizing two flow-patterned chips for highly parallel assays of a panel of secreted proteins from single macrophage cells.

  7. A robotics platform for automated batch fabrication of high density, microfluidics-based DNA microarrays, with applications to single cell, multiplex assays of secreted proteins.

    Science.gov (United States)

    Ahmad, Habib; Sutherland, Alex; Shin, Young Shik; Hwang, Kiwook; Qin, Lidong; Krom, Russell-John; Heath, James R

    2011-09-01

    Microfluidics flow-patterning has been utilized for the construction of chip-scale miniaturized DNA and protein barcode arrays. Such arrays have been used for specific clinical and fundamental investigations in which many proteins are assayed from single cells or other small sample sizes. However, flow-patterned arrays are hand-prepared, and so are impractical for broad applications. We describe an integrated robotics/microfluidics platform for the automated preparation of such arrays, and we apply it to the batch fabrication of up to eighteen chips of flow-patterned DNA barcodes. The resulting substrates are comparable in quality with hand-made arrays and exhibit excellent substrate-to-substrate consistency. We demonstrate the utility and reproducibility of robotics-patterned barcodes by utilizing two flow-patterned chips for highly parallel assays of a panel of secreted proteins from single macrophage cells. © 2011 American Institute of Physics

  8. Multi-objective optimization of glycopeptide antibiotic production in batch and fed batch processes

    DEFF Research Database (Denmark)

    Maiti, Soumen K.; Eliasson Lantz, Anna; Bhushan, Mani

    2011-01-01

    batch operations using process model for Amycolatopsis balhimycina, a glycopeptide antibiotic producer. This resulted in a set of several pareto optimal solutions with the two objectives ranging from (0.75gl−1, 3.97g$-1) to (0.44gl−1, 5.19g$-1) for batch and from (1.5gl−1, 5.46g$-1) to (1.1gl−1, 6.34g...

  9. Effect of mixing digested slurry on the rate of biogas production from dairy manure in batch fermenter

    Energy Technology Data Exchange (ETDEWEB)

    Kalia, A.K.; Singh, S.P.

    2001-09-01

    Forty kilograms of pure cattle dung and cattle dung mixed with 10% digested slurry obtained from a field biogas plant was batch fermented in horizontal biogas digesters for 15 weeks under field conditions with mean ambient temperature 20-23{sup o}C. Compared to 821 l of biogas from digester I, containing cattle dung alone, 1457 l of biogas was obtained from digester II, containing cattle dung mixed with 10% digested slurry. Mixing of slurry not only speeded up the gas production but also enhanced its rate from 108 l/kg dry matter to 158 l/kg dry matter. It also resulted in 36.1% distraction of total volatile solid in digester II, compared to 23.93% observed in digester I. Mixing digested slurry is recommended for raising biogas production from cattle dung in dry fermenters. (author)

  10. A Shortcut to the Production of High Ethanol Concentration from Jerusalem Artichoke Tubers

    Directory of Open Access Journals (Sweden)

    Wei-Guo Zhang

    2005-01-01

    Full Text Available Aspergillus niger SL-09, a newly isolated exoinulinase-hyperproducing strain, and Saccharomyces cerevisiae Z-06, with high ethanol tolerance, were used in a fed-batch process for simultaneous saccharification and fermentation of Jerusalem artichoke tuber mash and flour. S. cerevisiae Z-06 utilized 98 % of the total sugar and produced 19.6 % of ethanol in 48 h. In this process the conversion efficiency of the fermentation of Jerusalem artichoke and the production of ethanol were 90 % of the theoretical ethanol yield and the cost of the production of flour was cut nearly into half.

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

    Directory of Open Access Journals (Sweden)

    Erdei Borbála

    2012-03-01

    Full Text Available Abstract 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

  12. Using a medium of free amino acids to produce penicillin g acylase in fed-batch cultivations of Bacillus megaterium ATCC 14945

    Directory of Open Access Journals (Sweden)

    R. G. Silva

    2006-03-01

    Full Text Available The production of penicillin G acylase (PGA, an important industrial enzyme from a wild strain of Bacillus megaterium using a pool of free amino acids as substrate was studied in a bench-scale bioreactor. Experiments carried out in shakers showed that the substitution of casein for free amino acids in the presence of cheese whey was the culture medium that provided the highest productivity. Several cultivations were carried out in a bioreactor operated in either batch or fed-batch mode. Batch runs showed that enzyme production is associated with microorganism growth. The following set of amino acids was preferentially consumed: Ala, Arg, Asp, Gly, Lys, Ser, Thr and Trp. On the other hand, the rates of consumption of His, Ile, Leu, Met, Phe, Pro, Tyr and Val were lower.

  13. Performance, carcass and ruminal fermentation characteristics of heifers fed concentrates differing in energy level and cereal type (corn vs. wheat)

    Energy Technology Data Exchange (ETDEWEB)

    Carrasco, C.; Carro, M.D.; Fuentetaja, A.; Medel, P.

    2017-07-01

    A total of 144 beef heifers (218 ± 26.4 kg body weight) were housed in 24 pens (6 animals each) and used in a 168-day feedlot study to evaluate the influence of cereal type and energy level on performance, carcass quality and ruminal fermentation. Four concentrates were formulated according to a 2×2 factorial arrangement of treatments, with two energy levels (1,452 vs. 1,700 kcal net energy/kg) and two main cereals (wheat vs. corn). Concentrate and straw were offered ad libitum. Concentrate intake and body weight were recorded on days 42, 84, 126 and 168. Ruminal fluid was obtained by ruminocentesis from 3 heifers per pen on days 1, 84 and 168; and carcass weight, classification and yield, were determined in the same animals. Heifers fed high-energy diets had lower intake (6.97 vs. 7.29 kg fresh matter/d; p=0.011), and lower concentrate to gain ratio (5.15 vs. 5.66 kg/kg; p=0.002) than those fed low energy concentrates, and tended (p=0.069) to be heavier along the time. Neither carcass yield and classification, nor ruminal pH, volatile fatty acids nor NH3-N concentrations were affected (p>0.050) by energy level. Total volatile fatty acids concentration tended (p=0.070) to be greater in heifers fed corn-based than wheat-based concentrates. No energy level x cereal type interactions were observed. These results indicate that high energy concentrates decreased feed intake and feed conversion but had minor effects on carcass performance. Cereal type had no effects on performance and ruminal fermentation and no interactions between cereal type and energy were detected.

  14. Performance, carcass and ruminal fermentation characteristics of heifers fed concentrates differing in energy level and cereal type (corn vs. wheat)

    International Nuclear Information System (INIS)

    Carrasco, C.; Carro, M.D.; Fuentetaja, A.; Medel, P.

    2017-01-01

    A total of 144 beef heifers (218 ± 26.4 kg body weight) were housed in 24 pens (6 animals each) and used in a 168-day feedlot study to evaluate the influence of cereal type and energy level on performance, carcass quality and ruminal fermentation. Four concentrates were formulated according to a 2×2 factorial arrangement of treatments, with two energy levels (1,452 vs. 1,700 kcal net energy/kg) and two main cereals (wheat vs. corn). Concentrate and straw were offered ad libitum. Concentrate intake and body weight were recorded on days 42, 84, 126 and 168. Ruminal fluid was obtained by ruminocentesis from 3 heifers per pen on days 1, 84 and 168; and carcass weight, classification and yield, were determined in the same animals. Heifers fed high-energy diets had lower intake (6.97 vs. 7.29 kg fresh matter/d; p=0.011), and lower concentrate to gain ratio (5.15 vs. 5.66 kg/kg; p=0.002) than those fed low energy concentrates, and tended (p=0.069) to be heavier along the time. Neither carcass yield and classification, nor ruminal pH, volatile fatty acids nor NH3-N concentrations were affected (p>0.050) by energy level. Total volatile fatty acids concentration tended (p=0.070) to be greater in heifers fed corn-based than wheat-based concentrates. No energy level x cereal type interactions were observed. These results indicate that high energy concentrates decreased feed intake and feed conversion but had minor effects on carcass performance. Cereal type had no effects on performance and ruminal fermentation and no interactions between cereal type and energy were detected.

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

    Science.gov (United States)

    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.

  16. Lactic acid fermentation of crude sorghum extract

    Energy Technology Data Exchange (ETDEWEB)

    Samuel, W.A.; Lee, Y.Y.; Anthony, W.B.

    1980-04-01

    Crude extract from sweet sorghum supplemented with vetch juice was utilized as the carbohydrate source for fermentative production of lactic acid. Fermentation of media containing 7% (w/v) total sugar was completed in 60-80 hours by Lactobacillus plantarum, product yield averaging 85%. Maximum acid production rates were dependent on pH, initial substrate distribution, and concentration, the rates varying from 2 to 5 g/liter per hour. Under limited medium supplementation the lactic acid yield was lowered to 67%. The fermented ammoniated product contained over eight times as much equivalent crude protein (N x 6.25) as the original medium. Unstructured kinetic models were developed for cell growth, lactic acid formation, and substrate consumption in batch fermentation. With the provision of experimentally determined kinetic parameters, the proposed models accurately described the fermentation process. 15 references.

  17. Lactic Acid Bacterial Starter Culture with Antioxidant and γ-Aminobutyric Acid Biosynthetic Activities Isolated from Flatfish-Sikhae Fermentation.

    Science.gov (United States)

    Won, Yeong Geol; Yu, Hyun-Hee; Chang, Young-Hyo; Hwang, Han-Joon

    2015-12-01

    The aim of this study is to select a lactic acid bacterial strain as a starter culture for flatfish-Sikhae fermentation and to evaluate its suitability for application in a food system. Four strains of lactic acid bacteria isolated from commercial flatfish-Sikhae were identified and selected as starter culture candidates through investigation of growth rates, salt tolerance, food safety, and functional properties such as antioxidative and antimicrobial activities. The fermentation properties of the starter candidates were also examined in food systems prepared with these strains (candidate batch) in comparison with a spontaneous fermentation process without starter culture (control batch) at 15°C. The results showed that the candidate YG331 batch had better fermentation properties such as viable cell count, pH, and acidity than the other experimental batches, including the control batch. The results are expressed according to selection criteria based on a preliminary sensory evaluation and physiochemical investigation. Also, only a small amount of histamine was detected with the candidate YG331 batch. The radical scavenging activity of the candidate batches was better compared with the control batch, and especially candidate YG331 batch showed the best radical scavenging activity. Also, we isolated another starter candidate (identified as Lactobacillus brevis PM03) with γ-aminobutyric acid (GABA)-producing activity from commercial flatfish-Sikhae products. The sensory scores of the candidate YG331 batch were better than those of the other experimental batches in terms of flavor, color, and overall acceptance. In this study, we established selection criteria for the lactic acid bacterial starter for the flatfish-Sikhae production and finally selected candidate YG331 as the most suitable starter.

  18. Fermentation conditions that affect clavulanic acid production in Streptomyces clavuligerus: a systematic review

    Directory of Open Access Journals (Sweden)

    Hooi-Leng eSer

    2016-04-01

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

  19. Mathematical model of gluconic acid fermentation by Aspergillus niger

    Energy Technology Data Exchange (ETDEWEB)

    Takamatsu, T.; Shioya, S.; Furuya, T.

    1981-11-01

    A mathematical model for the study of gluconic acid fermentation by Aspergillus niger has been developed. The model has been deduced from the basic biological concept of multicellular filamentous microorganisms, i.e. cell population balance. It can be used to explain the behaviour of both batch and continuous cultures, even when in a lag phase. A new characteristic, involving the existence of dual equilibrium stages during fermentation, has been predicted using this mathematical model. (Refs. 6).

  20. Development of high-speed and high-efficient L-lactic acid fermentation and P (3HB) fermentative production for realizing Lactate Industry as Post petrochemistry; Posuto petorokemisutori to shiteno Lactate Industry wo jitsugensuru tameno kosoku kokoritsu L-nyusan kakko to P(3HB) kakko seisan no kaihatsu

    Energy Technology Data Exchange (ETDEWEB)

    Ishizaki, Fumiaki; Kobayashi, Genta; Vonktaveesuk, P.; Tsuge, Takeharu; Tanaka, Kenji [Kyushu University, Fukuoka (Japan)

    1999-04-05

    We are streptococci For the purpose of maintaining thing, it is high in the culture of Lactococcus lactis IO-1 in respect of perfect use of the substrate and metabolism activity of bacterial cell, and it pHs pH Substrate feed control method (pH-dependent substrate feed system) as an index was developed. In addition, advanced and integrated continuous culture system which enabled stabilized culture system and high lactic acid production speed was constructed by using furnace concentrator and on-line laser turbidity controller. And, Lactococcus lactis IO-1 By going through the organic acid such as the lactic acid from the xylose, it seems to be possible to carry out bio conversion of which it is efficient, since it is also very much excellent in the utilization-ness of the xylose which is a major component of tree biomass of the glucose otherwise. Then, culture medium supply method which enabled new batch culture system and substrate concentration automatic control for making this lactic bacteria and fermented milk acid liquid produced using culture system and organic acid liquid to be a substrate, and for producing polyhydroxy butyric acid [P (3HB)] which is the biodegradable plastic material by Alcaligenes eutrophus at high speed high-density was developed. (translated by NEDO)

  1. Comparison of fermentation of diets of variable composition and microbial populations in the rumen of sheep and Rusitec fermenters. II. Protozoa population and diversity of bacterial communities.

    Science.gov (United States)

    Martínez, M E; Ranilla, M J; Tejido, M L; Saro, C; Carro, M D

    2010-08-01

    Four ruminally and duodenally cannulated sheep and 8 Rusitec fermenters were used to determine the effects of dietary characteristics on microbial populations and bacterial diversity. The purpose of the study was to assess how closely fermenters can mimic the differences between diets found in vivo. The 4 experimental diets contained forage to concentrate (F:C) ratios of 70:30 (high forage; HF) or 30:70 (high concentrate; HC) with either alfalfa hay (A) or grass hay (G) as the forage. Total bacterial numbers were greater in the rumen of sheep fed HF diets compared with those fed HC diets, whereas the opposite was found in fermenters. The numbers of cellulolytic bacteria were not affected by F:C ratio in any fermentation system, but cellulolytic numbers were 2.7 and 1.8 times greater in sheep than in fermenters for HF and HC diets, respectively. Neither total bacterial nor cellulolytic numbers were affected by the type of forage in sheep or fermenters. Decreasing F:C ratio increased total protozoa and Entodiniae numbers in sheep by about 29 and 25%, respectively, but it had no effect in fermenters. Isotrichidae and Ophryoscolecinae numbers in sheep were not affected by changing F:C ratio, but both disappeared completely from fermenters fed HC diets. Total protozoa and Entodiniae numbers were greater in sheep fed A diets than in those fed G diets, whereas the opposite was found in fermenters. Results indicate that under the conditions of the present study, protozoa population in Rusitec fermenters was not representative of that in the rumen of sheep fed the same diets. In addition, protozoa numbers in fermenters were 121 and 226 times lower than those in the sheep rumen for HF and HC diets, respectively. The automated ribosomal intergenic spacer analysis of the 16S ribosomal DNA was used to analyze the diversity of liquid- and solid-associated bacteria in both systems. A total of 170 peaks were detected in the automated ribosomal intergenic spacer analysis

  2. High volumetric power density, non-enzymatic, glucose fuel cells.

    Science.gov (United States)

    Oncescu, Vlad; Erickson, David

    2013-01-01

    The development of new implantable medical devices has been limited in the past by slow advances in lithium battery technology. Non-enzymatic glucose fuel cells are promising replacement candidates for lithium batteries because of good long-term stability and adequate power density. The devices developed to date however use an "oxygen depletion design" whereby the electrodes are stacked on top of each other leading to low volumetric power density and complicated fabrication protocols. Here we have developed a novel single-layer fuel cell with good performance (2 μW cm⁻²) and stability that can be integrated directly as a coating layer on large implantable devices, or stacked to obtain a high volumetric power density (over 16 μW cm⁻³). This represents the first demonstration of a low volume non-enzymatic fuel cell stack with high power density, greatly increasing the range of applications for non-enzymatic glucose fuel cells.

  3. [Optimization of cultural condition of genetic engineering strain for antibiotic peptide adenoregulin and research on its fed-batch cultivation].

    Science.gov (United States)

    Zhou, Yu-Xun; Cao, Wei; Wei, Dong-Zhi; Luo, Qing-Ping; Wang, Jin-Zhi

    2005-07-01

    33 amino acid antibiotic peptide adenoregulin (ADR), which were firstly isolated from the skin of South America arboreal frog Phyllomedusa bicolor, forms alpha-helix amphipathic structure in apolar medium and has a wide spectrum of antimicrobial activity and high potency of lytic ability. Adr gene was cloned in pET32a and transformed into Escherichia coli BL21(DE3) . The cultural and inductive conditions of E. coli BL21(DE3)/pET32a-adr have been optimized. The effect of three factors which were time point of induction, concentration of IPTG in the culture and time of induction on the expression level of Trx-ADR was investigated. The results indicated that the expression level was affected by the time point of induction most predominantly. 9 veriaties of media in which BL21 (DE3)/pET32a-adr was cultured and induced were tested to achieve high expression level of target protein. It was found that glucose in the medium played an important role in keeping stable and high expression level of Trx-ADR. The optimal inductive condition is as follows: the culture medium is 2 x YT + 0.5% glucose, the time point of induction is OD600 = 0.9, the final concentration of IPTG in the culture is 0.1 mmol/L and the induction time is 4 h. BL21 (DE3)/pET32a-adr was cultivated according to the strategy of constant pH at early stage and exponential feeding at later stage to obtain high cell density. During the entire fed-batch phase, by controlling the feeding of glucose, the specific growth rate of the culture was controlled at about 0.15 h(-1), the accumulation of acetic acid was controlled at low level (<2 g/L), but the plasmid stability could not be maintained well. At the end of the cultivation, 40% of the bacteria in the culture lost their plasmids. As a result, the expression level of the target protein declined dramatically, but 90% of Trx-ADR was in soluble form. The expressed fusion protein showed no antibacterial activity, while the native form of ADR lysed from Trx-ADR showed

  4. Efficient Simultaneous Saccharification and Fermentation of Inulin to 2,3-Butanediol by Thermophilic Bacillus licheniformis ATCC 14580

    Science.gov (United States)

    Li, Lixiang; Chen, Chao; Li, Kun; Wang, Yu; Gao, Chao; Ma, Cuiqing

    2014-01-01

    2,3-Butanediol (2,3-BD) is an important starting material for the manufacture of bulk chemicals. For efficient and large-scale production of 2,3-BD through fermentation, low-cost substrates are required. One such substrate, inulin, is a polydisperse fructan found in a wide variety of plants. In this study, a levanase with high inulinase activity and high pH and temperature stability was identified in Bacillus licheniformis strain ATCC 14580. B. licheniformis strain ATCC 14580 was found to efficiently produce 2,3-BD from fructose at 50°C. Then, the levanase was used for simultaneous saccharification and fermentation (SSF) of inulin to 2,3-BD. A fed-batch SSF yielded 103.0 g/liter 2,3-BD in 30 h, with a high productivity of 3.4 g/liter · h. The results suggest that the SSF process developed with the thermophilic B. licheniformis strain used might be a promising alternative for efficient 2,3-BD production from the favorable substrate inulin. PMID:25107977

  5. Fermentative hydrogen production from Jerusalem artichoke by Clostridium tyrobutyricum expressing exo-inulinase gene.

    Science.gov (United States)

    Jiang, Ling; Wu, Qian; Xu, Qing; Zhu, Liying; Huang, He

    2017-08-11

    Clostridium tyrobutyricum ATCC25755 has been reported as being able to produce significant quantities of hydrogen. In this study, the exo-inulinase encoding gene cloned from Paenibacillus polymyxa SC-2 was into the expression plasmid pSY6 and expressed in the cells of C. tyrobutyricum. The engineered C. tyrobutyricum strain efficiently fermented the inulin-type carbohydrates from Jerusalem artichoke, without any pretreatment being necessary for the production of hydrogen. A comparatively high hydrogen yield (3.7 mol/mol inulin-type sugar) was achieved after 96 h in a batch process with simultaneous saccharification and fermentation (SSF), with an overall volumetric productivity rate of 620 ± 60 mL/h/L when the initial total sugar concentration of the inulin extract was increased to 100 g/L. Synthesis of inulinase in the batch SSF culture was closely associated with strain growth until the end of the exponential phase, reaching a maximum activity of 28.4 ± 0.26 U/mL. The overall results show that the highly productive and abundant biomass crop Jerusalem artichoke can be a good substrate for hydrogen production, and that the application of batch SSF for its conversion has the potential to become a cost-effective process in the near future.

  6. Production of ethanol at high temperatures in the fermentation of Jerusalem artichoke juice and a simple medium by Kluyveromyces marxianus

    Energy Technology Data Exchange (ETDEWEB)

    Rosa, M.F.; Correia, I.S.; Novais, J.M.

    1987-01-01

    Temperatures as high as 36 degrees C and 40 degrees C did not negatively affect the ethanol productivity of Jerusalem artichoke (J.a.) juice batch fermentation and the final concentrations of ethanol were close to those produced at lower temperatures. At higher process temperatures (36-40 degrees C), ethanol toxicity in Kluyveromyces marxianus was less important during the fermentation of J.a. juice as compared with a simple medium. In simple medium, the heat-sticking of fermentation was observed and the percentage of unfermented sugars steeply increased from 28 degrees C up to 40 degrees C. (Refs. 13).

  7. Comparative technoeconomic analysis of a softwood ethanol process featuring posthydrolysis sugars concentration operations and continuous fermentation with cell recycle.

    Science.gov (United States)

    Schneiderman, Steven J; Gurram, Raghu N; Menkhaus, Todd J; Gilcrease, Patrick C

    2015-01-01

    Economical production of second generation ethanol from Ponderosa pine is of interest due to widespread mountain pine beetle infestation in the western United States and Canada. The conversion process is limited by low glucose and high inhibitor concentrations resulting from conventional low-solids dilute acid pretreatment and enzymatic hydrolysis. Inhibited fermentations require larger fermentors (due to reduced volumetric productivity) and low sugars lead to low ethanol titers, increasing distillation costs. In this work, multiple effect evaporation (MEE) and nanofiltration (NF) were evaluated to concentrate the hydrolysate from 30 g/l to 100, 150, or 200 g/l glucose. To ferment this high gravity, inhibitor containing stream, traditional batch fermentation was compared with continuous stirred tank fermentation (CSTF) and continuous fermentation with cell recycle (CSTF-CR). Equivalent annual operating cost (EAOC = amortized capital + yearly operating expenses) was used to compare these potential improvements for a local-scale 5 MGY ethanol production facility. Hydrolysate concentration via evaporation increased EAOC over the base process due to the capital and energy intensive nature of evaporating a very dilute sugar stream; however, concentration via NF decreased EAOC for several of the cases (by 2 to 15%). NF concentration to 100 g/l glucose with a CSTF-CR was the most economical option, reducing EAOC by $0.15 per gallon ethanol produced. Sensitivity analyses on NF options showed that EAOC improvement over the base case could still be realized for even higher solids removal requirements (up to two times higher centrifuge requirement for the best case) or decreased NF performance. © 2015 American Institute of Chemical Engineers.

  8. Digestibility, Milk Production, and Udder Health of Etawah Goats Fed with Fermented Coffee Husk

    Directory of Open Access Journals (Sweden)

    I. Badarina

    2015-04-01

    Full Text Available This study was carried out to assess the utilization of coffee husk fermented by Pleurotus ostreatus as feed supplement by measuring the digestibility, milk production and udder health of Etawah goats suffered from subclinical mastitis (+1. There were three experimental diets consisted of T0 (control diet/basal diet without fermented coffee husk, T1 (basal diet with 6% fermented coffee husk and T2 (basal diet with 6% fermented coffee husk soaked in crude palm oil for an hour before using. Basal diet consisted of napier grass (60% and concentrate (40%. The results showed that supplementation of lactating Etawah does with fermented coffee husk did not affect the palatability of the diets, but increased the protein and crude fiber consumption (P<0.05. There was no significant effect on nutrient digestibility and milk production while milk composition (protein, fat, total solid increased in supplemented groups (P<0.05. The persistency of milk production and the somatic cells count were not different. There was an improvement of somatic cells count on supplemented groups. In conclusion, fermented coffee husk could be used as feed supplement without any negative effects on digestibility and milk production. The positive effects to udder health could be expected from including fermented coffee husk in diets.

  9. Passively operated vapor-fed direct methanol fuel cells for portable applications

    Energy Technology Data Exchange (ETDEWEB)

    Eccarius, Steffen; Krause, Falko; Agert, Carsten [Fraunhofer Institute for Solar Energy Systems ISE, Department of Energy Systems, Heidenhofstrasse 2, 79110 Freiburg (Germany); Beard, Kevin [Department of Chemical Engineering, University of South Carolina, Columbia (United States)

    2008-08-01

    The impact of structural parameters and operating conditions has not been researched yet for vapor-fed operation of a DMFC at near-ambient conditions. Thus, a detailed parameter study that included reference cell measurements to assess anode and cathode losses separately was performed. Among other parameters like temperature or air stoichiometry, different opening ratios that controlled evaporation of methanol into the vapor chamber were examined. Water management was found to be a critical parameter for a vapor-fed DMFC. Depletion of water inside the anode catalyst layer, especially at higher current densities, decreased performance of the fuel cell substantially. Back diffusion of water from the cathode to the anode was examined. A micro-structured cathode electrode that increased water back diffusion due to a reduced mass transfer resistance was developed and investigated. Finally, efficiencies and heat losses of a vapor-fed DMFC were determined. (author)

  10. Differential effects of two fermentable carbohydrates on central appetite regulation and body composition.

    Directory of Open Access Journals (Sweden)

    Tulika Arora

    Full Text Available Obesity is rising at an alarming rate globally. Different fermentable carbohydrates have been shown to reduce obesity. The aim of the present study was to investigate if two different fermentable carbohydrates (inulin and β-glucan exert similar effects on body composition and central appetite regulation in high fat fed mice.Thirty six C57BL/6 male mice were randomized and maintained for 8 weeks on a high fat diet containing 0% (w/w fermentable carbohydrate, 10% (w/w inulin or 10% (w/w β-glucan individually. Fecal and cecal microbial changes were measured using fluorescent in situ hybridization, fecal metabolic profiling was obtained by proton nuclear magnetic resonance ((1H NMR, colonic short chain fatty acids were measured by gas chromatography, body composition and hypothalamic neuronal activation were measured using magnetic resonance imaging (MRI and manganese enhanced MRI (MEMRI, respectively, PYY (peptide YY concentration was determined by radioimmunoassay, adipocyte cell size and number were also measured. Both inulin and β-glucan fed groups revealed significantly lower cumulative body weight gain compared with high fat controls. Energy intake was significantly lower in β-glucan than inulin fed mice, with the latter having the greatest effect on total adipose tissue content. Both groups also showed an increase in the numbers of Bifidobacterium and Lactobacillus-Enterococcus in cecal contents as well as feces. β-Glucan appeared to have marked effects on suppressing MEMRI associated neuronal signals in the arcuate nucleus, ventromedial hypothalamus, paraventricular nucleus, periventricular nucleus and the nucleus of the tractus solitarius, suggesting a satiated state.Although both fermentable carbohydrates are protective against increased body weight gain, the lower body fat content induced by inulin may be metabolically advantageous. β-Glucan appears to suppress neuronal activity in the hypothalamic appetite centers. Differential

  11. Fermentation of lignocellulosic hydrolysates: Inhibition and detoxification

    Energy Technology Data Exchange (ETDEWEB)

    Palmqvist, E.

    1998-02-01

    The ethanol yield and productivity obtained during fermentation of lignocellulosic hydrolysates is decreased due to the presence of inhibiting compounds, such as weak acids, furans and phenolic compounds produced during hydrolysis. Evaluation of the effect of various biological, physical and chemical detoxification treatments by fermentation assays using Saccharomyces cerevisiae was used to characterise inhibitors. Inhibition of fermentation was decreased after removal of the non-volatile compounds, pre-fermentation by the filamentous fungus Trichoderma reesei, treatment with the lignolytic enzyme laccase, extraction with ether, and treatment with alkali. Yeast growth in lignocellulosic hydrolysates was inhibited below a certain fermentation pH, most likely due to high concentrations of undissociated weak acids. The effect of individual compounds were studied in model fermentations. Furfural is reduced to furfuryl alcohol by yeast dehydrogenases, thereby affecting the intracellular redox balance. As a result, acetaldehyde accumulated during furfural reduction, which most likely contributed to inhibition of growth. Acetic acid (10 g 1{sup -1}) and furfural (3 g 1{sup -1}) interacted antagonistically causing decreased specific growth rate, whereas no significant individual or interaction effects were detected by the lignin-derived compound 4-hydroxybenzoic acid (2 g 1{sup -1}). By maintaining a high cell mass density in the fermentor, the process was less sensitive to inhibitors affecting growth and to fluctuations in fermentation pH, and in addition the depletion rate of bioconvertible inhibitors was increased. A theoretical ethanol yield and high productivity was obtained in continuous fermentation of spruce hydrolysate when the cell mass concentration was maintained at a high level by applying cell recirculation 164 refs, 16 figs, 5 tabs

  12. High Production of 2,3-Butanediol (2,3-BD by Raoultella ornithinolytica B6 via Optimizing Fermentation Conditions and Overexpressing 2,3-BD Synthesis Genes.

    Directory of Open Access Journals (Sweden)

    Taeyeon Kim

    Full Text Available Biological production of 2,3-butandiol (2,3-BD has received great attention as an alternative to the petroleum-based 2,3-BD production. In this study, a high production of 2,3-BD in fed-batch fermentation was investigated with a newly isolated bacterium designated as Raoultella ornithinolytica B6. The isolate produced 2,3-BD as the main product using hexoses (glucose, galactose, and fructose, pentose (xylose and disaccharide (sucrose. The effects of temperature, pH-control schemes, and agitation speeds on 2,3-BD production were explored to optimize the fermentation conditions. Notably, cell growth and 2,3-BD production by R. ornithinolytica B6 were higher at 25°C than at 30°C. When three pH control schemes (no pH control, pH control at 7, and pH control at 5.5 after the pH was decreased to 5.5 during fermentation were tested, the best 2,3-BD titer and productivity along with reduced by-product formation were achieved with pH control at 5.5. Among different agitation speeds (300, 400, and 500 rpm, the optimum agitation speed was 400 rpm with 2,3-BD titer of 68.27 g/L, but acetic acid was accumulated up to 23.32 g/L. Further enhancement of the 2,3-BD titer (112.19 g/L, yield (0.38 g/g, and productivity (1.35 g/L/h as well as a significant reduction of acetic acid accumulation (9.71 g/L was achieved by the overexpression of homologous budABC genes, the 2,3-BD-synthesis genes involved in the conversion of pyruvate to 2,3-BD. This is the first report presenting a high 2,3-BD production by R.ornithinolytica which has attracted little attention with respect to 2,3-BD production, extending the microbial spectrum of 2,3-BD producers.

  13. Enhanced lipid production with undetoxified corncob hydrolysate by Rhodotorula glutinis using a high cell density culture strategy.

    Science.gov (United States)

    Liu, Yating; Wang, Yanping; Liu, Hongjuan; Zhang, Jian'an

    2015-03-01

    In recent years, energy crisis and environmental issues such as greenhouse effect, global warming, etc. has roused peoples' concern. Biodiesel, as renewable energy, has attracted much attention to deal with such problems. This work studied the lipid production by Rhodotorula glutinis with undetoxified corncob hydrolysate. The results indicated that R. glutinis had high tolerance to the inhibitors in corncob hydrolysate and it could utilize undetoxified corncob hydrolysate directly for lipid production. The cell grew well with undetoxified hydrolysate in the batch culture of 5L fermentor with the optimized C/N ratio of 75, lipid titer and lipid content reached 5.5g/L and 36.4%, respectively. High cell density culture with two-stage nitrogen feeding strategy was studied to enhance the lipid production, biomass, lipid concentration and lipid content of 70.8, 33.5g/L and 47.2% were obtained. The results indicated the potential application for lipid production by R. glutinis with corncob hydrolysate directly. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. On-line monitoring of monoclonal antibody formation in high density perfusion culture using FIA.

    Science.gov (United States)

    Fenge, C; Fraune, E; Freitag, R; Scheper, T; Schügerl, K

    1991-05-01

    An automated flow injection system for on-line analysis of proteins in real fermentation fluids was developed by combining the principles of stopped-flow, merging zones flow injection analysis (FIA) with antigen-antibody reactions. IgG in the sample reacted with its corresponding antibody (a-IgG) in the reagent solution. Formation of insoluble immunocomplexes resulted in an increase of the turbidity which was determined photometrically. This system was used to monitor monoclonal antibody production in high cell density perfusion culture of hybridoma cells. Perfusion was performed with a newly developed static filtration unit equipped with hydrophilic microporous tubular membranes. Different sampling devices were tested to obtain a cell-free sample stream for on-line product analysis of high molecular weight (e.g., monoclonal antibodies) and low molecular weight (e.g., glucose, lactate) medium components. In fermentation fluids a good correlation (coefficient: 0.996) between the FIA method and an ELISA test was demonstrated. In a high density perfusion cultivation process mAb formation was successfully monitored on-line over a period of 400 h using a reliable sampling system. Glucose and lactate were measured over the same period of time using a commercially available automatic analyser based on immobilized enzyme technology.

  15. Kinetic Modeling of Corn Fermentation with S. cerevisiae Using a Variable Temperature Strategy

    Directory of Open Access Journals (Sweden)

    Augusto C. M. Souza

    2018-04-01

    Full Text Available While fermentation is usually done at a fixed temperature, in this study, the effect of having a controlled variable temperature was analyzed. A nonlinear system was used to model batch ethanol fermentation, using corn as substrate and the yeast Saccharomyces cerevisiae, at five different fixed and controlled variable temperatures. The lower temperatures presented higher ethanol yields but took a longer time to reach equilibrium. Higher temperatures had higher initial growth rates, but the decay of yeast cells was faster compared to the lower temperatures. However, in a controlled variable temperature model, the temperature decreased with time with the initial value of 40 ∘ C. When analyzing a time window of 60 h, the ethanol production increased 20% compared to the batch with the highest temperature; however, the yield was still 12% lower compared to the 20 ∘ C batch. When the 24 h’ simulation was analyzed, the controlled model had a higher ethanol concentration compared to both fixed temperature batches.

  16. Kinetic Modeling of Corn Fermentation with S. cerevisiae Using a Variable Temperature Strategy.

    Science.gov (United States)

    Souza, Augusto C M; Mousaviraad, Mohammad; Mapoka, Kenneth O M; Rosentrater, Kurt A

    2018-04-24

    While fermentation is usually done at a fixed temperature, in this study, the effect of having a controlled variable temperature was analyzed. A nonlinear system was used to model batch ethanol fermentation, using corn as substrate and the yeast Saccharomyces cerevisiae , at five different fixed and controlled variable temperatures. The lower temperatures presented higher ethanol yields but took a longer time to reach equilibrium. Higher temperatures had higher initial growth rates, but the decay of yeast cells was faster compared to the lower temperatures. However, in a controlled variable temperature model, the temperature decreased with time with the initial value of 40 ∘ C. When analyzing a time window of 60 h, the ethanol production increased 20% compared to the batch with the highest temperature; however, the yield was still 12% lower compared to the 20 ∘ C batch. When the 24 h’ simulation was analyzed, the controlled model had a higher ethanol concentration compared to both fixed temperature batches.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

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

    2018-01-02

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

  19. Re-fermentation of washed spent solids from batch hydrogenogenic fermentation for additional production of biohydrogen from the organic fraction of municipal solid waste.

    Science.gov (United States)

    Muñoz-Páez, Karla M; Ríos-Leal, Elvira; Valdez-Vazquez, Idania; Rinderknecht-Seijas, Noemí; Poggi-Varaldo, Héctor M

    2012-03-01

    In the first batch solid substrate anaerobic hydrogenogenic fermentation with intermittent venting (SSAHF-IV) of the organic fraction of municipal solid waste (OFMSW), a cumulative production of 16.6 mmol H(2)/reactor was obtained. Releases of hydrogen partial pressure first by intermittent venting and afterward by flushing headspace of reactors with inert gas N(2) allowed for further hydrogen production in a second to fourth incubation cycle, with no new inoculum nor substrate nor inhibitor added. After the fourth cycle, no more H(2) could be harvested. Interestingly, accumulated hydrogen in 4 cycles was 100% higher than that produced in the first cycle alone. At the end of incubation, partial pressure of H(2) was near zero whereas high concentrations of organic acids and solvents remained in the spent solids. So, since approximate mass balances indicated that there was still a moderate amount of biodegradable matter in the spent solids we hypothesized that the organic metabolites imposed some kind of inhibition on further fermentation of digestates. Spent solids were washed to eliminate organic metabolites and they were used in a second SSAHF-IV. Two more cycles of H(2) production were obtained, with a cumulative production of ca. 2.4 mmol H(2)/mini-reactor. As a conclusion, washing of spent solids of a previous SSAHF-IV allowed for an increase of hydrogen production by 15% in a second run of SSAHF-IV, leading to the validation of our hypothesis. Copyright © 2011 Elsevier Ltd. All rights reserved.

  20. Spontaneous large-scale autolysis in Clostridium acetobutylicum contributes to generation of more spores

    Directory of Open Access Journals (Sweden)

    Zhen eLiu

    2015-09-01

    Full Text Available Autolysis is a widespread phenomenon in bacteria. In batch fermentation of Clostridium acetobutylicum ATCC 824, there is a spontaneous large-scale autolysis phenomenon with significant decrease of cell density immediately after exponential phase. To unravel the role of autolysis, an autolysin-coding gene, CA_C0554, was disrupted by using ClosTron system to obtain the mutant C. acetobutylicum lyc::int(72. The lower final cell density and faster cell density decrease rate of C. acetobutylicum ATCC 824 than those of C. acetobutylicum lyc::int(72 indicates that CA_C0554 was an important but not the sole autolysin-coding gene responding for the large-scale autolysis. Similar glucose utilization and solvents production but obvious lower cell density of C. acetobutylicum ATCC 824 comparing to C. acetobutylicum lyc::int(72 suggests that lysed C. acetobutylicum ATCC 824 cells were metabolic inactive. On the contrary, the spore density of C. acetobutylicum ATCC 824 is 26.1% higher than that of C. acetobutylicum lyc::int(72 in the final culture broth of batch fermentation. We speculated that spontaneous autolysis of metabolic-inactive cells provided nutrients for the sporulating cells. The present study suggests that one important biological role of spontaneous large-scale autolysis in C. acetobutylicum ATCC 824 batch fermentation is contributing to generation of more spores during sporulation.

  1. Spontaneous large-scale autolysis in Clostridium acetobutylicum contributes to generation of more spores.

    Science.gov (United States)

    Liu, Zhen; Qiao, Kai; Tian, Lei; Zhang, Quan; Liu, Zi-Yong; Li, Fu-Li

    2015-01-01

    Autolysis is a widespread phenomenon in bacteria. In batch fermentation of Clostridium acetobutylicum ATCC 824, there is a spontaneous large-scale autolysis phenomenon with significant decrease of cell density immediately after exponential phase. To unravel the role of autolysis, an autolysin-coding gene, CA_C0554, was disrupted by using ClosTron system to obtain the mutant C. acetobutylicum lyc::int(72). The lower final cell density and faster cell density decrease rate of C. acetobutylicum ATCC 824 than those of C. acetobutylicum lyc::int(72) indicates that CA_C0554 was an important but not the sole autolysin-coding gene responding for the large-scale autolysis. Similar glucose utilization and solvents production but obvious lower cell density of C. acetobutylicum ATCC 824 comparing to C. acetobutylicum lyc::int(72) suggests that lysed C. acetobutylicum ATCC 824 cells were metabolic inactive. On the contrary, the spore density of C. acetobutylicum ATCC 824 is 26.1% higher than that of C. acetobutylicum lyc::int(72) in the final culture broth of batch fermentation. We speculated that spontaneous autolysis of metabolic-inactive cells provided nutrients for the sporulating cells. The present study suggests that one important biological role of spontaneous large-scale autolysis in C. acetobutylicum ATCC 824 batch fermentation is contributing to generation of more spores during sporulation.

  2. Community proteomics provides functional insight into polyhydroxyalkanoate production by a mixed microbial culture cultivated on fermented dairy manure.

    Science.gov (United States)

    Hanson, Andrea J; Guho, Nicholas M; Paszczynski, Andrzej J; Coats, Erik R

    2016-09-01

    Polyhydroxyalkanoates (PHAs) are bio-based, biodegradable polyesters that can be produced from organic-rich waste streams using mixed microbial cultures (MMCs). To maximize PHA production, MMCs are enriched for bacteria with a high polymer storage capacity through the application of aerobic dynamic feeding (ADF) in a sequencing batch reactor (SBR), which consequently induces a feast-famine metabolic response. Though the feast-famine response is generally understood empirically at a macro-level, the molecular level is less refined. The objective of this study was to investigate the microbial community composition and proteome profile of an enriched MMC cultivated on fermented dairy manure. The enriched MMC exhibited a feast-famine response and was capable of producing up to 40 % (wt. basis) PHA in a fed-batch reactor. High-throughput 16S rRNA gene sequencing revealed a microbial community dominated by Meganema, a known PHA-producing genus not often observed in high abundance in enrichment SBRs. The application of the proteomic methods two-dimensional electrophoresis and LC-MS/MS revealed PHA synthesis, energy generation, and protein synthesis prominently occurring during the feast phase, corroborating bulk solution variable observations and theoretical expectations. During the famine phase, nutrient transport, acyl-CoA metabolism, additional energy generation, and housekeeping functions were more pronounced, informing previously under-determined MMC functionality under famine conditions. During fed-batch PHA production, acetyl-CoA acetyltransferase and PHA granule-bound phasin proteins were in increased abundance relative to the SBR, supporting the higher PHA content observed. Collectively, the results provide unique microbial community structural and functional insight into feast-famine PHA production from waste feedstocks using MMCs.

  3. Ethanol fermentation by immobilized cells of Zymomonas mobilis

    Energy Technology Data Exchange (ETDEWEB)

    Grote, W.

    1985-01-01

    Previous studies have shown that immobilized yeast cell cultures have commercial potential for fuel ethanol production. In this study the suitability of strains of Z. mobilis for whole cell immobilization was investigated. Experiments revealed that immobilization in Ca-alginate or K-carrageenan gel or use of flocculating strains was effective for ethanol production at relatively high productivities. Two laboratory size reactors were designed and constructed. These were a compartmented multiple discshaft column and a tower fermentor. Results of this work supported other studies that established that growth and fermentation could be uncoupled. The data indicated that specific metabolic rates were dependent on the nature of the fermentation media. The addition of lactobacilli to Z. mobilis continuous fermentations had only a transient effect, and was unlikely to affect an immobilized Z. mobilis process. With 150 gl/sup -1/ glucose media and a Z. mobilis ZM4 immobilized cell reactor, a maximum volumetric ethanol productivity of 55 gl/sup -1/h/sup -1/ was obtained. The fermentation of sucrose media or sucrose-based raw materials (molasses, cane juice, synthetic mill liquor) by immobilized Z. mobilis ZM4 revealed a pattern of rapid sucrose hydrolysis, preferential glucose utilization and the conversion of fructose to the undesirable by-products levan and sorbitol.

  4. Change in microbial communities in acetate- and glucose-fed microbial fuel cells in the presence of light

    KAUST Repository

    Xing, Defeng

    2009-09-01

    Power densities produced by microbial fuel cells (MFCs) in natural systems are changed by exposure to light through the enrichment of photosynthetic microorganisms. When MFCs with brush anodes were exposed to light (4000 lx), power densities increased by 8-10% for glucose-fed reactors, and 34% for acetate-fed reactors. Denaturing gradient gel electrophoresis (DGGE) profiles based on the 16S rRNA gene showed that exposure to high light levels changed the microbial communities on the anodes. Based on 16S rRNA gene clone libraries of light-exposed systems the anode communities using glucose were also significantly different than those fed acetate. Dominant bacteria that are known exoelectrogens were identified in the anode biofilm, including a purple nonsulfur (PNS) photosynthetic bacterium, Rhodopseudomonas palustris, and a dissimilatory iron-reducing bacterium, Geobacter sulfurreducens. Pure culture tests confirmed that PNS photosynthetic bacteria increased power production when exposed to high light intensities (4000 lx). These results demonstrate that power production and community composition are affected by light conditions as well as electron donors in single-chamber air-cathode MFCs. © 2009 Elsevier B.V. All rights reserved.

  5. Change in microbial communities in acetate- and glucose-fed microbial fuel cells in the presence of light

    KAUST Repository

    Xing, Defeng; Cheng, Shaoan; Regan, John M.; Logan, Bruce E.

    2009-01-01

    Power densities produced by microbial fuel cells (MFCs) in natural systems are changed by exposure to light through the enrichment of photosynthetic microorganisms. When MFCs with brush anodes were exposed to light (4000 lx), power densities increased by 8-10% for glucose-fed reactors, and 34% for acetate-fed reactors. Denaturing gradient gel electrophoresis (DGGE) profiles based on the 16S rRNA gene showed that exposure to high light levels changed the microbial communities on the anodes. Based on 16S rRNA gene clone libraries of light-exposed systems the anode communities using glucose were also significantly different than those fed acetate. Dominant bacteria that are known exoelectrogens were identified in the anode biofilm, including a purple nonsulfur (PNS) photosynthetic bacterium, Rhodopseudomonas palustris, and a dissimilatory iron-reducing bacterium, Geobacter sulfurreducens. Pure culture tests confirmed that PNS photosynthetic bacteria increased power production when exposed to high light intensities (4000 lx). These results demonstrate that power production and community composition are affected by light conditions as well as electron donors in single-chamber air-cathode MFCs. © 2009 Elsevier B.V. All rights reserved.

  6. Molecular diversity of lactic acid bacteria on ileum broiler chicken fed by bran and bran fermentation

    Science.gov (United States)

    Baniyah, Laelatul; Nur Jannah, Siti; Rukmi, Isworo; Sugiharto

    2018-05-01

    Lactic Acid Bacteria (LAB) is a digestive tract microflora that have a positive role in poultry health. The number and diversity of LAB in the digestive tract affected by several factors, among them was the kind of feed. The purpose of this research was to know diversity of Lactic Acid Bacteria (LAB) ileum broiler’s after feeding with prebiotic bran and Rhizopus oryzae fermented bran which was added to commercial feed. As much as 15 broilers were used to determine the diversity of LAB. All broilers were fed using commercial feed. The control used commercial feed no addition of bran or fermented bran, and commercial feed with fermented bran and nonfermented bran were as a treatment. To determine the diversity of LAB, T-RFLP method was applied. The Hae III and Msp I were used as restriction enzymes. The number of phylotype, relative abundance, Shannon diversity index (H '), evenness (E), and Dominance (D) were examined. The results indicated that the addition of prebiotic bran on commercial feed showed a higher diversity of lactic acid bacteria on broiler’s ileum, compared with control and addition of Rhizopus oryzae fermented bran. LAB group that dominates in the ileum is Lactobacillus sp. and L. delbruecii subs bulgaricus.

  7. Exergy Analysis of an Intermediate Temperature Solid Oxide Fuel Cell-Gas Turbine Hybrid System Fed with Ethanol

    Directory of Open Access Journals (Sweden)

    Fotini Tzorbatzoglou

    2012-10-01

    Full Text Available In the present work, an ethanol fed Solid Oxide Fuel Cell-Gas Turbine (SOFC-GT system has been parametrically analyzed in terms of exergy and compared with a single SOFC system. The solid oxide fuel cell was fed with hydrogen produced from ethanol steam reforming. The hydrogen utilization factor values were kept between 0.7 and 1. The SOFC’s Current-Volt performance was considered in the range of 0.1–3 A/cm2 at 0.9–0.3 V, respectively, and at the intermediate operating temperatures of 550 and 600 °C, respectively. The curves used represent experimental results obtained from the available bibliography. Results indicated that for low current density values the single SOFC system prevails over the SOFC-GT hybrid system in terms of exergy efficiency, while at higher current density values the latter is more efficient. It was found that as the value of the utilization factor increases the SOFC system becomes more efficient than the SOFC-GT system over a wider range of current density values. It was also revealed that at high current density values the increase of SOFC operation temperature leads in both cases to higher system efficiency values.

  8. Effects of a specific blend of essential oils on apparent nutrient digestion, rumen fermentation and rumen microbial populations in sheep fed a 50:50 alfalfa hay:concentrate diet

    Directory of Open Access Journals (Sweden)

    N. Khateri

    2017-03-01

    Full Text Available Objective An experiment was conducted to investigate the effects of a specific mixture of essential oils (MEO, containing thyme, clove and cinnamon EO, on rumen microbial fermentation, nutrient apparent digestibility and blood metabolites in fistulated sheep. Methods Six sheep fitted with ruminal fistulas were used in a repeated measurement design with two 24-d periods to investigate the effect of adding MEO at 0 (control, 0.8, and 1.6 mL/d on apparent nutrient digestibility, rumen fermentation characteristics, rumen microbial population and blood chemical metabolites. Animals were fed with a 50:50 alfalfa hay:concentrate diet. Results Ruminal pH, total volatile fatty acids (VFA concentration, molar proportion of individual VFA, acetate: propionate ratio and methane production were not affected with MEO. Relative to the control, Small peptides plus amino acid nitrogen and large peptides nitrogen concentration in rumen fluid were not affected with MEO supplementation; while, rumen fluid ammonia nitrogen concentration at 0 and 6 h after morning feeding in sheep fed with 1.6 mL/d of MEO was lower (p<0.05 compared to the control and 0.8 mL/d of MEO. At 0 h after morning feeding, ammonia nitrogen concentration was higher (p<0.05 in sheep fed 0.8 mL/d of MEO relative to 1.6 mL/d and control diet. Ruminal protozoa and hyper ammonia producing (HAP bacteria counts were not affected by addition of MEO in the diet. Relative to the control, no changes were observed in the red and white blood cells, hemoglobin, hematocrit, glucose, beta-hydroxybutyric acid, cholesterol, total protein, albumin, blood urea nitrogen and aspartate aminotransferase and alanine aminotransferase concentration. Apparent total tract digestibility of dry matter, crude proten, organic matter, and neutral detergent fiber were not influenced by MEO supplementation. Conclusion The results of the present study suggested that supplementation of MEO may have limited effects on apparent

  9. Single-cell analysis of S. cerevisiae growth recovery after a sublethal heat-stress applied during an alcoholic fermentation.

    Science.gov (United States)

    Tibayrenc, Pierre; Preziosi-Belloy, Laurence; Ghommidh, Charles

    2011-06-01

    Interest in bioethanol production has experienced a resurgence in the last few years. Poor temperature control in industrial fermentation tanks exposes the yeast cells used for this production to intermittent heat stress which impairs fermentation efficiency. Therefore, there is a need for yeast strains with improved tolerance, able to recover from such temperature variations. Accordingly, this paper reports the development of methods for the characterization of Saccharomyces cerevisiae growth recovery after a sublethal heat stress. Single-cell measurements were carried out in order to detect cell-to-cell variability. Alcoholic batch fermentations were performed on a defined medium in a 2 l instrumented bioreactor. A rapid temperature shift from 33 to 43 °C was applied when ethanol concentration reached 50 g l⁻¹. Samples were collected at different times after the temperature shift. Single cell growth capability, lag-time and initial growth rate were determined by monitoring the growth of a statistically significant number of cells after agar medium plating. The rapid temperature shift resulted in an immediate arrest of growth and triggered a progressive loss of cultivability from 100 to 0.0001% within 8 h. Heat-injured cells were able to recover their growth capability on agar medium after a lag phase. Lag-time was longer and more widely distributed as the time of heat exposure increased. Thus, lag-time distribution gives an insight into strain sensitivity to heat-stress, and could be helpful for the selection of yeast strains of technological interest.

  10. Lignocellulosic sugar management for xylitol and ethanol fermentation with multiple cell recycling by Kluyveromyces marxianus IIPE453.

    Science.gov (United States)

    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.

  11. Effects of oxygen and ethanol on recombinant yeast fermentation for hepatitis B virus surface antigen production: modeling and simulation studies.

    Science.gov (United States)

    Shi, Y; Ryu, D D; Yuan, W K

    1993-01-05

    A model was formulated to examine the competitive growth of two phenotypes (Leu(+) and Leu(-)) and the product formation with recombinant Saccharomyces cerevisiae strain DBY-745, which contains the shuttle vector pYGH3-16-s with the foreign gene HBsAg (hepatitis B virus surface antigen) as well as experimental fedbatch fermentation data. The important state variables and the process parameters evaluated include (1) the ratio of the plasmid-free cell concentration to the plasmid-containing cell concentration (rho = X(-)X(+)), (2) the expression of human hepatitis B surface antigen g (CH), (3) the glucose consumption (S), (4) the ethanol production (/), (5) the change of working volume (V) in the fermentor, (6) the different specific growth rates of two phenotype cells, and (7) the plasmid loss frequency coefficient (alpha ). These variables and other parameters were carefully defined, their correlations were studied, and a mathematical model using a set of nonlinear ordinary differential equations (ODEs) for fed-batch fermentation was then obtained based on the theoretical considerations and the experimental results. The extended Kalman filter (EKF) methods was applied for the best estimate of these variables based on the experimentally observable variables: rhoV, and g (CH). Each of these variable was affected by random measuring errors under the different operating conditions. Simulation results presented for verification of the model agreed with our observations and provided useful information relevant to the operation and the control of the fedbatch recombinant yeast fermentation. The method of predicting an optimal profile of the cell growth was also demonstrated under the different dissolved oxygen concentrations.

  12. Breeding of Coenzyme Q10 Produced Strain by Low-Energy Ion Implantation and Optimization of Coenzyme Q10 Fermentation

    International Nuclear Information System (INIS)

    Xu Dejun; Zheng Zhiming; Wang Peng; Wang Li; Yuan Hang; Yu Zengliang

    2008-01-01

    In order to increase the production efficiency of coenzyme Q 10 , the original strain Agrobacterium tumefaciens ATCC 4452 was mutated by means of Nitrogen ions implantation. A mutant strain, ATX 12, with high contents of coenzyme Q 10 was selected. Subsequently, the conditions such as carbohydrate concentration, nitrogen source concentration, inoculum's size, seed age, aeration and temperature which might affect the production of CoQ 10 were investigated in detail. Under optimal conditions, the maximum concentration of the intracellular CoQ 10 reached 200.3 mg/L after 80 h fed-batch fermentation, about 245% increasing in CoQ 10 production after ion implantation, compared to the original strain. (ion beam bioengineering)

  13. [Construction and high-density fermentation of alkaline pectate lyase high-yield yeast].

    Science.gov (United States)

    Wang, Xiaowen; Xiang, La; Xu, Ting; Lu, Zhenghui; Zhang, Guimin

    2017-12-25

    Pectate lyase is widely applied in ramie degumming and fabric bioscouring in the textile industry. Compared to conventional processes that involve high alkaline and high temperature treatment, enzyme based treatments have significant advantages in fibers protectiveness, improved efficiency of refining, reduced energy consumption and pollution. Hence, it would be highly desirable to construct high-yield alkaline pectate lyase engineered strains and reduce the pectate lyase production cost. In the previous study, pectate lyase gene pel from Bacillus subtilis168 was expressed in Pichia pastoris GS115 after codon usage optimization based on the vector pHBM905A. To improve the expression level, the vector pHBM905BDM with optimized promoter and signal peptide was used to express the optimized gene pels in GS115. The transformant had increased activity from 68 U/mL to 100 U/mL with the improvement in the transcription level by 27% measured by qPCR. The transformants were further screened on pectin plates, where higher halo forming strains were picked for shake-flask fermentation and strain GS115-pHBM905BDM-pels4 showed the highest activity of 536 U/mL. Then plasmid pPIC9K-pels was constructed and electroporated into the GS115-pHBM905BDM-pels4 cells. Subsequently, high-copy transformant was screened by using the medium containing antibiotics G418, strain GS115-pHBM905BDMpPIC9K- pels1 was identified with increased activity of 770 U/mL and the copy number of pels was 7 confirmed by qPCR. Finally, the activity of pectate lyase produced by GS115-pHBM905BDM-pPIC9K-pels1reached to 2 271 U/mL in a 5-L fermentor. The activity of pectate lyase in our study reached the highest level of expression in P. pastoris, showing good application potential in the textile industry.

  14. Two-stage pretreatment of excess sludge for electricity generation in microbial fuel cell.

    Science.gov (United States)

    Zhang, Yi; Zhao, Yang-Guo; Guo, Liang; Gao, Mengchun

    2018-01-12

    Thermophiles hydrolysis and acidogens fermentation were sequentially adopted to pretreat excess sludge for microbial fuel cell (MFC) electricity production. The results indicated that MFC fed with the thermophiles-acidogens pretreated sludge (MFC AB), reached a higher removal of ammonia nitrogen than the MFC fed with the heating hydrolysis and acidogens fermentation pretreated sludge (MFC NB). However, compared with the MFC AB, MFC NB presented a better performance for removal of soluble chemical oxygen demand (SCOD) (90.08%) and protein (82.42%). As for the electricity production, MFC NB obtained higher voltage of 0.632 V and maximum power density with 1.05 W/m 3 while MFC AB reached maximum voltage of 0.373 V and maximum power density of 0.58 W/m 3 . Bacterial 16S rRNA-based molecular microbial techniques showed that microbial communities on both MFC anode biofilms was diverse and different. The cooperation of fermentation bacteria and electricigen Shewanella baltica in the MFC NB may have contributed towards the improvement of electricity generation.

  15. Application of gain scheduling to the control of batch bioreactors

    Science.gov (United States)

    Cardello, Ralph; San, Ka-Yiu

    1987-01-01

    The implementation of control algorithms to batch bioreactors is often complicated by the inherent variations in process dynamics during the course of fermentation. Such a wide operating range may render the performance of fixed gain PID controllers unsatisfactory. In this work, a detailed study on the control of batch fermentation is performed. Furthermore, a simple batch controller design is proposed which incorporates the concept of gain-scheduling, a subclass of adaptive control, with oxygen uptake rate as an auxiliary variable. The control of oxygen tension in the biorector is used as a vehicle to convey the proposed idea, analysis and results. Simulation experiments indicate significant improvement in controller performance can be achieved by the proposed approach even in the presence of measurement noise.

  16. Polyhexamethyl biguanide can eliminate contaminant yeasts from fuel-ethanol fermentation process.

    Science.gov (United States)

    Elsztein, Carolina; de Menezes, João Assis Scavuzzi; de Morais, Marcos Antonio

    2008-09-01

    Industrial ethanol fermentation is a non-sterile process and contaminant microorganisms can lead to a decrease in industrial productivity and significant economic loss. Nowadays, some distilleries in Northeastern Brazil deal with bacterial contamination by decreasing must pH and adding bactericides. Alternatively, contamination can be challenged by adding a pure batch of Saccharomyces cerevisiae-a time-consuming and costly process. A better strategy might involve the development of a fungicide that kills contaminant yeasts while preserving S. cerevisiae cells. Here, we show that polyhexamethyl biguanide (PHMB) inhibits and kills the most important contaminant yeasts detected in the distilleries of Northeastern Brazil without affecting the cell viability and fermentation capacity of S. cerevisiae. Moreover, some physiological data suggest that PHMB acts through interaction with the yeast membrane. These results support the development of a new strategy for controlling contaminant yeast population whilst keeping industrial yields high.

  17. Lactic acid production from potato peel waste by anaerobic sequencing batch fermentation using undefined mixed culture.

    Science.gov (United States)

    Liang, Shaobo; McDonald, Armando G; Coats, Erik R

    2015-11-01

    Lactic acid (LA) is a necessary industrial feedstock for producing the bioplastic, polylactic acid (PLA), which is currently produced by pure culture fermentation of food carbohydrates. This work presents an alternative to produce LA from potato peel waste (PPW) by anaerobic fermentation in a sequencing batch reactor (SBR) inoculated with undefined mixed culture from a municipal wastewater treatment plant. A statistical design of experiments approach was employed using set of 0.8L SBRs using gelatinized PPW at a solids content range from 30 to 50 g L(-1), solids retention time of 2-4 days for yield and productivity optimization. The maximum LA production yield of 0.25 g g(-1) PPW and highest productivity of 125 mg g(-1) d(-1) were achieved. A scale-up SBR trial using neat gelatinized PPW (at 80 g L(-1) solids content) at the 3 L scale was employed and the highest LA yield of 0.14 g g(-1) PPW and a productivity of 138 mg g(-1) d(-1) were achieved with a 1 d SRT. Copyright © 2015 Elsevier Ltd. All rights reserved.

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

    Science.gov (United States)

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

    2014-01-01

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

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

    Science.gov (United States)

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

    2018-01-01

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

  20. Thermodynamic and exergoeconomic analysis of biogas fed solid oxide fuel cell power plants emphasizing on anode and cathode recycling: A comparative study

    International Nuclear Information System (INIS)

    Mehr, A.S.; Mahmoudi, S.M.S.; Yari, M.; Chitsaz, A.

    2015-01-01

    Highlights: • Four biogas-fed solid oxide fuel cell power plants are proposed. • Performance of systems is compared with each other economically. • Efficiency of biogas fed fuel cell with anode–cathode recycling is the highest. • For current density of 6000 A/m"2 the optimum anode recycle ratio is around 0.25. • Unit product cost of biogas fed fuel cell with anode–cathode recycling is 19.07$/GJ. - Abstract: Four different configurations of natural gas and biogas fed solid oxide fuel cell are proposed and analyzed thermoeconomically, focusing on the influence of anode and/or cathode gas recycling. It is observed that the net output power is maximized at an optimum current density the value of which is lowered as the methane concentration in the biogas is decreased. Results indicate that when the current density is low, there is an optimum anode recycling ratio at which the thermal efficiency is maximized. In addition, an increase in the anode recycling ratio increases the unit product cost of the system while an increase in the cathode recycling ratio has a revers effect. For the same working conditions, the solid oxide fuel cell with anode and cathode recycling is superior to the other configurations and its thermal efficiency is calculated as 46.09% being 6.81% higher than that of the simple solid oxide fuel cell fed by natural gas. The unit product cost of the solid oxide fuel cell-anode and cathode recycling system is calculated as 19.07$/GJ which is about 35% lower than the corresponding value for the simple natural gas fed solid oxide fuel cell system.

  1. Microbiology and optimization of hydrogen fermentation and bioelectricity production

    Energy Technology Data Exchange (ETDEWEB)

    Makinen, A.

    2013-11-01

    This work investigated dark fermentative hydrogen (H{sub 2}) and bioelectricity production from carbohydrates. Meso- and thermophilic fermentative and mesophilic exoelectrogenic bacteria were enriched from different natural sources. The H{sub 2} production from different hexoses and pentoses, them main constituents of lignocellulose, was studied in batch assays. H{sub 2} production from xylose was examined in continuous stirred tank reactor (CSTR). Operational parameters for H{sub 2} production were optimized. Bioelectricity production was studied in microbial fuel cells and process parameters were optimized. Dynamics of microbial communities in H{sub 2} and bioelectricity production processes were determined. A novel thermophilic dark fermentative H{sub 2} producing bacterium, Thermovorax subterraneus, was enriched and isolated from geothermal underground mine. T. subterraneus had the optimum growth temperature of 72 deg C and the maximum H{sub 2} yield of 1.4 mol/mol glucose in batch assay. The main soluble fermentative end products of T. subterraneus were acetate and ethanol. Thermophilic dark fermentative mixed culture enriched from hot spring (Hisarlan, Turkey) had the maximum H{sub 2} yield of 1.7 mol/mol glucose. The optimal environmental parameters to maximize H{sub 2} yield were temperature 52 deg C, initial pH 6.5, 40 mg/L Fe{sup 2+}, 4.5 g/L yeast extract and glucose concentration of 4 g/L. Increasing the glucose concentration to 18 g/L increased the maximum H{sub 2} production rate to 56.2 mmol H{sub 2}/h/L. Environmental parameters had a significant effect on metabolic pathways of fermentation. Another hot spring (Hisarkoy, Turkey) enrichment culture was able to ferment different sugars to H{sub 2} favoring pentoses over hexoses. The best H{sub 2} yields in batch assays were obtained from pentoses: xylose, arabinose and ribose yielded 21, 15 and 8 % of the theoretical yield, respectively; whilst on glucose the yield was only 2 % of the theoretical

  2. Thermodynamics of metabolic pathways for penicillin production: Analysis of thermodynamic feasibility and free energy changes during fed-batch cultivation

    DEFF Research Database (Denmark)

    Pissarra, P.D.; Nielsen, Jens Bredal

    1997-01-01

    This paper describes the thermodynamic analysis of pathways related to penicillin production in Penicillium chrysogenum. First a thermodynamic feasibility analysis is performed of the L-lysine pathway of which one of the precursors for penicillin biosynthesis (alpha-aminoadipic acid......) is an intermediate. It is found that the L-lysine pathway in P. chrysogenum is thermodynamically feasible and that the calculated standard Gibbs free energy values of the two enzymes controlling the pathway flux indicate that they operate far from equilibrium. It is therefore proposed that the regulation of alpha......-aminoadipate reductase by lysine is important to maintain a high concentration of alpha-aminoadipate in order to direct the carbon flux to penicillin production. Secondly the changes in Gibbs free energy in the penicillin biosynthetic pathway during fed-batch cultivation were studied. The analysis showed that all...

  3. Coupling limonene formation and oxyfunctionalization by mixed-culture resting cell fermentation.

    Science.gov (United States)

    Willrodt, Christian; Hoschek, Anna; Bühler, Bruno; Schmid, Andreas; Julsing, Mattijs K

    2015-09-01

    Metabolic engineering strategies mark a milestone for the fermentative production of bulk and fine chemicals. Yet, toxic products and volatile reaction intermediates with low solubilities remain challenging. Prominent examples are artificial multistep pathways like the production of perillyl acetate (POHAc) from glucose via limonene. For POHAc, these limitations can be overcome by mixed-culture fermentations. A limonene biosynthesis pathway and cytochrome P450 153A6 (CYP153A6) as regioselective hydroxylase are used in two distinct recombinant E. coli. POHAc formation from glucose in one recombinant cell was hindered by ineffective coupling of limonene synthesis and low rates of oxyfunctionalization. The optimization of P450 gene expression led to the formation of 6.20 ± 0.06 mg gcdw (-1) POHAc in a biphasic batch cultivation with glucose as sole carbon and energy source. Increasing the spatial proximity between limonene synthase and CYP153A6 by a genetic fusion of both enzymes changed the molar limonene/POHAc ratio from 3.2 to 1.6. Spatial separation of limonene biosynthesis from its oxyfunctionalization improved POHAc concentration 3.3-fold to 21.7 mg L(-1) as compared to a biphasic fermentation. Mixed-cultures of E. coli BL21 (DE3) containing the limonene biosynthesis pathway and E. coli MG1655 harboring either CYP153A6, or alternatively a cymene monooxygenase, showed POHAc formation rates of 0.06 or 0.11 U gcdw (-1) , respectively. This concept provides a novel framework for fermentative syntheses involving toxic, volatile, or barely soluble compounds or pathway intermediates. © 2015 Wiley Periodicals, Inc.

  4. Simulated Batch Production of Penicillin

    Science.gov (United States)

    Whitaker, A.; Walker, J. D.

    1973-01-01

    Describes a program in applied biology in which the simulation of the production of penicillin in a batch fermentor is used as a teaching technique to give students experience before handling a genuine industrial fermentation process. Details are given for the calculation of minimum production cost. (JR)

  5. Development of a high temperature microbial fermentation process for butanol

    Energy Technology Data Exchange (ETDEWEB)

    Jeor, Jeffery D. St. [Idaho National Lab. (INL), Idaho Falls, ID (United States); Reed, David W. [Idaho National Lab. (INL), Idaho Falls, ID (United States); Daubaras, Dayna L. [Idaho National Lab. (INL), Idaho Falls, ID (United States); Thompson, Vicki S. [Idaho National Lab. (INL), Idaho Falls, ID (United States)

    2015-08-01

    Transforming renewable biomass into cost-competitive high-performance biofuels and bioproducts is key to the U.S. future energy and chemical needs. Butanol production by microbial fermentation for chemical conversion to polyolefins, elastomers, drop-in jet or diesel fuel, and other chemicals is a promising solution. A high temperature fermentation process could decrease energy costs, capital cost, give higher butanol production, and allow for continuous fermentation. In this paper, we describe our approach to genetically transform Geobacillus caldoxylosiliticus, using a pUCG18 plasmid, for potential insertion of a butanol production pathway. Transformation methods tested were electroporation of electrocompetent cells, ternary conjugation with E. coli donor and helper strains, and protoplast fusion. These methods have not been successful using the current plasmid. Growth controls show cells survive the various methods tested, suggesting the possibility of transformation inhibition from a DNA restriction modification system in G. caldoxylosiliticus, as reported in the literature.

  6. Characterization of a microfluidic microbial fuel cell as a power generator based on a nickel electrode.

    Science.gov (United States)

    Mardanpour, Mohammad Mahdi; Yaghmaei, Soheila

    2016-05-15

    This study reports the fabrication of a microfluidic microbial fuel cell (MFC) using nickel as a novel alternative for conventional electrodes and a non-phatogenic strain of Escherichia coli as the biocatalyst. The feasibility of a microfluidic MFC as an efficient power generator for production of bioelectricity from glucose and urea as organic substrates in human blood and urine for implantable medical devices (IMDs) was investigated. A maximum open circuit potential of 459 mV was achieved for the batch-fed microfluidic MFC. During continuous mode operation, a maximum power density of 104 Wm(-3) was obtained with nutrient broth. For the glucose-fed microfluidic MFC, the maximum power density of 5.2 μW cm(-2) obtained in this study is significantly greater than the power densities reported previously for microsized MFCs and glucose fuel cells. The maximum power density of 14 Wm(-3) obtained using urea indicates the successful performance of a microfluidic MFC using human excreta. It features high power density, self-regeneration, waste management and a low production cost (microfluidic MFC as a power supply was characterized based on polarization behavior and cell potential in different substrates, operational modes, and concentrations. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. Development of a rapid high-efficiency scalable process for acetylated Sus scrofa cationic trypsin production from Escherichia coli inclusion bodies.

    Science.gov (United States)

    Zhao, Mingzhi; Wu, Feilin; Xu, Ping

    2015-12-01

    Trypsin is one of the most important enzymatic tools in proteomics and biopharmaceutical studies. Here, we describe the complete recombinant expression and purification from a trypsinogen expression vector construct. The Sus scrofa cationic trypsin gene with a propeptide sequence was optimized according to Escherichia coli codon-usage bias and chemically synthesized. The gene was inserted into pET-11c plasmid to yield an expression vector. Using high-density E. coli fed-batch fermentation, trypsinogen was expressed in inclusion bodies at 1.47 g/L. The inclusion body was refolded with a high yield of 36%. The purified trypsinogen was then activated to produce trypsin. To address stability problems, the trypsin thus produced was acetylated. The final product was generated upon gel filtration. The final yield of acetylated trypsin was 182 mg/L from a 5-L fermenter. Our acetylated trypsin product demonstrated higher BAEE activity (30,100 BAEE unit/mg) than a commercial product (9500 BAEE unit/mg, Promega). It also demonstrated resistance to autolysis. This is the first report of production of acetylated recombinant trypsin that is stable and suitable for scale-up. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. On-line prediction of the feeding phase in high-cell density cultivation of rE. coli using constructive neural networks.

    Science.gov (United States)

    Nicoletti, M C; Bertini, J R; Tanizaki, M M; Zangirolami, T C; Gonçalves, V M; Horta, A C L; Giordano, R C

    2013-07-01

    Streptococcus pneumoniae (pneumococcus) is a bacterium responsible for a wide spectrum of illnesses. The surface of the bacterium consists of three distinctive membranes: plasmatic, cellular and the polysaccharide (PS) capsule. PS capsules may mediate several biological processes, particularly invasive infections of human beings. Prevention against pneumococcal related illnesses can be provided by vaccines. There is a sound investment worldwide in the investigation of a proteic antigen as a possible alternative to pneumococcal vaccines based exclusively on PS. A few proteins which are part of the membrane of the pneumococcus seem to have antigen potential to be part of a vaccine, particularly the PspA. A vital aspect in the production of the intended conjugate pneumococcal vaccine is the efficient production (in industrial scale) of both, the chosen PS serotypes as well as the PspA protein. Growing recombinant Escherichia coli (rE. coli) in high-cell density cultures (HCDC) under a fed-batch regime requires a refined continuous control over various process variables where the on-line prediction of the feeding phase is of particular relevance and one of the focuses of this paper. The viability of an on-line monitoring software system, based on constructive neural networks (CoNN), for automatically detecting the time to start the fed-phase of a HCDC of rE. coli that contains a plasmid used for PspA expression is investigated. The paper describes the data and methodology used for training five different types of CoNNs, four of them suitable for classification tasks and one suitable for regression tasks, aiming at comparatively investigate both approaches. Results of software simulations implementing five CoNN algorithms as well as conventional neural networks (FFNN), decision trees (DT) and support vector machines (SVM) are also presented and discussed. A modified CasCor algorithm, implementing a data softening process, has shown to be an efficient candidate to be

  9. Bioaggregate of photo-fermentative bacteria for enhancing continuous hydrogen production in a sequencing batch photobioreactor.

    Science.gov (United States)

    Xie, Guo-Jun; Liu, Bing-Feng; Wang, Rui-Qing; Ding, Jie; Ren, Hong-Yu; Zhou, Xu; Ren, Nan-Qi

    2015-11-05

    Hydrogen recovery through solar-driven biomass conversion by photo-fermentative bacteria (PFB) has been regarded as a promising way for sustainable energy production. However, a considerable fraction of organic substrate was consumed for the growth of PFB as biocatalysts, furthermore, these PFB were continuously washed out from the photobioreactor in continuous operation because of their poor flocculation. In this work, PFB bioaggregate induced by L-cysteine was applied in a sequencing batch photobioreactor to enhance continuous hydrogen production and reduce biomass washout. The effects of the hydraulic retention time (HRT), influent concentration and light intensity on hydrogen production of the photobioreactor were investigated. The maximum hydrogen yield (3.35 mol H2/mol acetate) and production rate (1044 ml/l/d) were obtained at the HRT of 96 h, influent concentration of 3.84 g COD/l, and light intensity of 200 W/m(2). With excellent settling ability, biomass accumulated in the photobioreactor and reached 2.15 g/l under the optimum conditions. Structural analysis of bioaggregate showed that bacterial cells were covered and tightly linked together by extracellular polymeric substances, and formed a stable structure. Therefore, PFB bioaggregate induced by L-cysteine is an efficient strategy to improve biomass retention capacity of the photobioreactor and enhance hydrogen recovery efficiency from organic wastes.

  10. Bioaggregate of photo-fermentative bacteria for enhancing continuous hydrogen production in a sequencing batch photobioreactor

    Science.gov (United States)

    Xie, Guo-Jun; Liu, Bing-Feng; Wang, Rui-Qing; Ding, Jie; Ren, Hong-Yu; Zhou, Xu; Ren, Nan-Qi

    2015-11-01

    Hydrogen recovery through solar-driven biomass conversion by photo-fermentative bacteria (PFB) has been regarded as a promising way for sustainable energy production. However, a considerable fraction of organic substrate was consumed for the growth of PFB as biocatalysts, furthermore, these PFB were continuously washed out from the photobioreactor in continuous operation because of their poor flocculation. In this work, PFB bioaggregate induced by L-cysteine was applied in a sequencing batch photobioreactor to enhance continuous hydrogen production and reduce biomass washout. The effects of the hydraulic retention time (HRT), influent concentration and light intensity on hydrogen production of the photobioreactor were investigated. The maximum hydrogen yield (3.35 mol H2/mol acetate) and production rate (1044 ml/l/d) were obtained at the HRT of 96 h, influent concentration of 3.84 g COD/l, and light intensity of 200 W/m2. With excellent settling ability, biomass accumulated in the photobioreactor and reached 2.15 g/l under the optimum conditions. Structural analysis of bioaggregate showed that bacterial cells were covered and tightly linked together by extracellular polymeric substances, and formed a stable structure. Therefore, PFB bioaggregate induced by L-cysteine is an efficient strategy to improve biomass retention capacity of the photobioreactor and enhance hydrogen recovery efficiency from organic wastes.

  11. Differential Effects of Two Fermentable Carbohydrates on Central Appetite Regulation and Body Composition

    Science.gov (United States)

    Gibson, Glenn R.; Tuohy, Kieran M.; Sharma, Raj Kumar; Swann, Jonathan R.; Deaville, Eddie R.; Sleeth, Michele L.; Thomas, E. Louise; Holmes, Elaine; Bell, Jimmy D.; Frost, Gary

    2012-01-01

    Background Obesity is rising at an alarming rate globally. Different fermentable carbohydrates have been shown to reduce obesity. The aim of the present study was to investigate if two different fermentable carbohydrates (inulin and β-glucan) exert similar effects on body composition and central appetite regulation in high fat fed mice. Methodology/Principal Findings Thirty six C57BL/6 male mice were randomized and maintained for 8 weeks on a high fat diet containing 0% (w/w) fermentable carbohydrate, 10% (w/w) inulin or 10% (w/w) β-glucan individually. Fecal and cecal microbial changes were measured using fluorescent in situ hybridization, fecal metabolic profiling was obtained by proton nuclear magnetic resonance (1H NMR), colonic short chain fatty acids were measured by gas chromatography, body composition and hypothalamic neuronal activation were measured using magnetic resonance imaging (MRI) and manganese enhanced MRI (MEMRI), respectively, PYY (peptide YY) concentration was determined by radioimmunoassay, adipocyte cell size and number were also measured. Both inulin and β-glucan fed groups revealed significantly lower cumulative body weight gain compared with high fat controls. Energy intake was significantly lower in β-glucan than inulin fed mice, with the latter having the greatest effect on total adipose tissue content. Both groups also showed an increase in the numbers of Bifidobacterium and Lactobacillus-Enterococcus in cecal contents as well as feces. β- glucan appeared to have marked effects on suppressing MEMRI associated neuronal signals in the arcuate nucleus, ventromedial hypothalamus, paraventricular nucleus, periventricular nucleus and the nucleus of the tractus solitarius, suggesting a satiated state. Conclusions/Significance Although both fermentable carbohydrates are protective against increased body weight gain, the lower body fat content induced by inulin may be metabolically advantageous. β-glucan appears to suppress neuronal

  12. Adaptive evolution of Saccharomyces cerevisiae with enhanced ethanol tolerance for Chinese rice wine fermentation.

    Science.gov (United States)

    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.

  13. Fermentative hydrogen production from liquid swine manure with glucose supplement using an anaerobic sequencing batch reactor

    Science.gov (United States)

    Wu, Xiao

    2009-12-01

    The idea of coupling renewable energy production and agricultural waste management inspired this thesis. The production of an important future fuel---hydrogen gas---from high strength waste stream-liquid swine manure---using anaerobic treatment processes makes the most sustainable sense for both wastewater utilization and energy generation. The objectives of this thesis were to develop a fermentation process for converting liquid swine manure to hydrogen and to maximize hydrogen productivity. Anaerobic sequencing batch reactor (ASBR) systems were constructed to carry out this fermentation process, and seed sludge obtained from a dairy manure anaerobic digester and pretreated by nutrient acclimation, heat and pH treatment was used as inoculum. High system stability was indicated by a short startup period of 12 days followed by stable hydrogen production, and successful sludge granulation occurred within 23 days of startup at a hydraulic retention time (HRT) of 24 hours. Operation at a progressively decreasing HRT from 24 to 8h gave rise to an increasing biogas production rate from 15.2-34.4L/d, while good linear relationships were observed between both total biogas and hydrogen production rates correlated to HRT, with R2 values of 0.993 and 0.997, respectively. The maximum hydrogen yield of 1.63 mol-H 2/mol-hexose-feed occurred at HRT of 16h, while the HRT of 12h was highly suggested to achieve both high production rate and efficient yield. Hexose utilization efficiencies over 98%, considerable hydrogen production rate up to 14.3 L/d and hydrogen percentage of off-gas up to 43% (i.e., a CO 2/H2 ratio of 1.2) with the absence of CH4 production throughout the whole course of experiment at a pH of 5.0 strongly validated the feasibility of the fermentative H2 production from liquid swine manure using an ASBR system. Ethanol as well as acetic, butyric and valeric acids were produced in the system accompanying the hydrogen production, with acetic acid being the dominant

  14. High power density carbonate fuel cell

    Energy Technology Data Exchange (ETDEWEB)

    Yuh, C.; Johnsen, R.; Doyon, J.; Allen, J. [Energy Research Corp., Danbury, CT (United States)

    1996-12-31

    Carbonate fuel cell is a highly efficient and environmentally clean source of power generation. Many organizations worldwide are actively pursuing the development of the technology. Field demonstration of multi-MW size power plant has been initiated in 1996, a step toward commercialization before the turn of the century, Energy Research Corporation (ERC) is planning to introduce a 2.85MW commercial fuel cell power plant with an efficiency of 58%, which is quite attractive for distributed power generation. However, to further expand competitive edge over alternative systems and to achieve wider market penetration, ERC is exploring advanced carbonate fuel cells having significantly higher power densities. A more compact power plant would also stimulate interest in new markets such as ships and submarines where space limitations exist. The activities focused on reducing cell polarization and internal resistance as well as on advanced thin cell components.

  15. Hypolipidemic effect of fruit fibers in rats fed with high dietary fat.

    Science.gov (United States)

    Esmael, O A; Sonbul, S N; Kumosani, T A; Moselhy, S S

    2015-03-01

    The hypolipidemic effect of 10% fruit fibers in rats fed with high-fat diet (HFD) was evaluated. This study was conducted on a total of 50 male Albino rats divided into 10 equal groups fed with different types of dietary fruits. The feeding period lasted for 24 weeks. Fasting blood samples were collected and sera separated and subjected to lipid profile assay and atherogenic index. In addition, total antioxidant activity of different fruits was determined. The results obtained showed that pomegranate had higher content of antioxidants followed by apple, strawberry and guava compared with other fruits. Rats fed with 20% coconut oil showed a highly significant elevation in the levels of serum total cholesterol, low-density lipoprotein cholesterol and atherogenic factor while the level of high-density lipoprotein cholesterol was significantly decreased when compared with control rats. Histological examination revealed that there was a large lipid and cholesterol deposition in the livers of rats fed with HFD. The potential in lowering the levels of plasma total cholesterol and triglyceride is in the following order: pomegranate > apple > strawberry > guava > papaya > mandarin and orange. Accumulation of hepatic lipid droplets was diminished when compared with the HFD group. Also, antiatherogenic is better than the untreated groups. Accordingly these hypolipidemic effects may be due to high-fiber content and antioxidant activity of these fruits. © The Author(s) 2012.

  16. Effects of potato fiber and potato-resistant starch on biomarkers of colonic health in rats fed diets containing red meat.

    Science.gov (United States)

    Paturi, Gunaranjan; Nyanhanda, Tafadzwa; Butts, Christine A; Herath, Thanuja D; Monro, John A; Ansell, Juliet

    2012-10-01

    The effects of red meat consumption with and without fermentable carbohydrates on indices of large bowel health in rats were examined. Sprague-Dawley rats were fed cellulose, potato fiber, or potato-resistant starch diets containing 12% casein for 2 wk, then similar diets containing 25% cooked beef for 6 wk. After week 8, cecal and colonic microbiota composition, fermentation end-products, colon structure, and colonocyte DNA damage were analyzed. Rats fed potato fiber had lower Bacteroides-Prevotella-Porphyromonas group compared to other diet groups. Colonic Bifidobacterium spp. and/or Lactobacillus spp. were higher in potato fiber and potato-resistant starch diets than in the cellulose diet. Beneficial changes were observed in short-chain fatty acid concentrations (acetic, butyric, and propionic acids) in rats fed potato fiber compared with rats fed cellulose. Phenol and p-cresol concentrations were lower in the cecum and colon of rats fed potato fiber. An increase in goblet cells per crypt and longer crypts were found in the colon of rats fed potato fiber and potato-resistant starch diets. Fermentable carbohydrates had no effect on colonic DNA damage. Dietary combinations of red meat with potato fiber or potato-resistant starch have distinctive effects in the large bowel. Future studies are essential to examine the efficacy of different types of nondigestible carbohydrates in maintaining colonic health during long-term consumption of high-protein diets. Improved understanding of interactions between the food consumed and gut microbiota provides knowledge needed to make healthier food choices for large bowel health. The impact of red meat on large bowel health may be ameliorated by consuming with fermentable dietary fiber, a colonic energy source that produces less harmful by-products than the microbial breakdown of colonic protein for energy. Developing functional red meat products with fermentable dietary fiber could be one way to promote a healthy and balanced

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

    Science.gov (United States)

    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.

  18. Direct dimethyl ether high temperature polymer electrolyte membrane fuel cells

    DEFF Research Database (Denmark)

    Vassiliev, Anton; Jensen, Jens Oluf; Li, Qingfeng

    and suffers from low DME solubility in water. When the DME - water mixture is fed as vapour miscibility is no longer a problem. The increased temperature is more beneficial for the kinetics of the direct oxidation of DME than of methanol. The Open Circuit Voltage (OCV) with DME operation was 50 to 100 m......A high temperature polybenzimidazole (PBI) polymer fuel cell was fed with dimethyl ether (DME) and water vapour mixture on the anode at ambient pressure with air as oxidant. A peak power density of 79 mW/cm2 was achieved at 200°C. A conventional polymer based direct DME fuel cell is liquid fed......V higher than that of methanol, indicating less fuel crossover....

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

    Science.gov (United States)

    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.

  20. Microfluidic engineered high cell density three-dimensional neural cultures

    Science.gov (United States)

    Cullen, D. Kacy; Vukasinovic, Jelena; Glezer, Ari; La Placa, Michelle C.

    2007-06-01

    Three-dimensional (3D) neural cultures with cells distributed throughout a thick, bioactive protein scaffold may better represent neurobiological phenomena than planar correlates lacking matrix support. Neural cells in vivo interact within a complex, multicellular environment with tightly coupled 3D cell-cell/cell-matrix interactions; however, thick 3D neural cultures at cell densities approaching that of brain rapidly decay, presumably due to diffusion limited interstitial mass transport. To address this issue, we have developed a novel perfusion platform that utilizes forced intercellular convection to enhance mass transport. First, we demonstrated that in thick (>500 µm) 3D neural cultures supported by passive diffusion, cell densities =104 cells mm-3), continuous medium perfusion at 2.0-11.0 µL min-1 improved viability compared to non-perfused cultures (p death and matrix degradation. In perfused cultures, survival was dependent on proximity to the perfusion source at 2.00-6.25 µL min-1 (p 90% viability in both neuronal cultures and neuronal-astrocytic co-cultures. This work demonstrates the utility of forced interstitial convection in improving the survival of high cell density 3D engineered neural constructs and may aid in the development of novel tissue-engineered systems reconstituting 3D cell-cell/cell-matrix interactions.