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.

Xylitol production by Candida parapsilosis under fed-batch culture

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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.

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.

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

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Wiebe Marilyn G

2012-05-01

carbohydrates than from glucose or xylose. Although high biomass and lipid production were achieved in both batch and fed-batch cultures with glucose as carbon source, for lipid production from mixtures of carbohydrates fed-batch cultivation was preferable. Constant feeding was better than intermittent feeding. The feeding strategy did not affect the relative proportion of different fatty acids in the lipid, but the presence of C5 sugars did.

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

ENHANCED PRODUCTION OF POLYHYDROXYBUTYRATE (PHB FROM AGRO-INDUSTRIAL WASTES; FED-BATCH CULTIVATION AND STATISTICAL MEDIA OPTIMIZATION

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Mahmoud M. Berekaa

2016-06-01

Full Text Available Bacillus megaterium SW1-2 showed enhanced growth and polyhydroxybutyrate (PHB production during cultivation on date palm syrup (DEPS or sugar cane molasses. FT-IR and NMR spectroscopic analyses of the polymer accumulated during growth on DEPS revealed specific absorption peaks characteristic for PHB. 1.65 g/L of PHB (56.9% CDW was produced during growth on medium supplemented with 2 g/L of DEPS. Approximately, 36.1% CDW of PHB were recorded during growth on sugar cane molasses. Six runs of different fed-batch cultivation strategies were tested, the optimal run showed approximately 6.87-fold increase. Modified E2 medium was prefered recording 10.11 and 11.34 g/L of total PHB produced for runs 1 and 2, at the end of 96 h incubation period, respectively. Decrease in PHB was recorded during growth on complex medium (run 3 and run 4. In another independent optimization strategy, ten variables were concurrently examined for their significance on PHB production by Plackett-Burman statistical design for the first time. Among variables, DEPS-II and inoculum concentration followed by KH2PO4 and (NH42SO4 were found to be the most significant variables encourage PHB production. Indeed, DEPS-II or Fresh syrup is more significant than commercial syrup DEPS-I (p-value= 0.05. RPM, incubation period have highly negative effect on PHB production. Role of ago-industrial wastes, especially DEPS, in enhancement of PHB production was closely discussed.

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...

Functional State Modelling of Cultivation Processes: Dissolved Oxygen Limitation State

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Olympia Roeva

2015-04-01

Full Text Available A new functional state, namely dissolved oxygen limitation state for both bacteria Escherichia coli and yeast Saccharomyces cerevisiae fed-batch cultivation processes is presented in this study. Functional state modelling approach is applied to cultivation processes in order to overcome the main disadvantages of using global process model, namely complex model structure and a big number of model parameters. Alongwith the newly introduced dissolved oxygen limitation state, second acetate production state and first acetate production state are recognized during the fed-batch cultivation of E. coli, while mixed oxidative state and first ethanol production state are recognized during the fed-batch cultivation of S. cerevisiae. For all mentioned above functional states both structural and parameter identification is here performed based on experimental data of E. coli and S. cerevisiae fed-batch cultivations.

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

was found to be limited by UDP-Gal biosynthesis, which was observed to be both cell line and cultivation condition-dependent. Extracellular glucose and glutamine concentrations and uptake rates were positively correlated with intracellular UDP-Gal availability. All these findings are important for optimization of fed-batch culture for improving IgG production and directing glycosylation quality. © 2014 Wiley Periodicals, Inc.

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.

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.

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....

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

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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.

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

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

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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.

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.

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

Science.gov (United States)

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

2013-01-01

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

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.

CONVERSION OF PINEAPPLE JUICE WASTE INTO LACTIC ACID IN BATCH AND FED – BATCH FERMENTATION SYSTEMS

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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

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....

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.

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...

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.).

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

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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.

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.

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).

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.

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.

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

Following an Optimal Batch Bioreactor Operations Model

DEFF Research Database (Denmark)

Ibarra-Junquera, V.; Jørgensen, Sten Bay; Virgen-Ortíz, J.J.

2012-01-01

The problem of following an optimal batch operation model for a bioreactor in the presence of uncertainties is studied. The optimal batch bioreactor operation model (OBBOM) refers to the bioreactor trajectory for nominal cultivation to be optimal. A multiple-variable dynamic optimization of fed...... as the master system which includes the optimal cultivation trajectory for the feed flow rate and the substrate concentration. The “real” bioreactor, the one with unknown dynamics and perturbations, is considered as the slave system. Finally, the controller is designed such that the real bioreactor...

HPLC-ESI-MS(n) Analysis, Fed-Batch Cultivation Enhances Bioactive Compound Biosynthesis and Immune-Regulative Effect of Adventitious Roots in Pseudostellaria heterophylla.

Science.gov (United States)

Wang, Juan; Li, Jing; Li, Hongfa; Wu, Xiaolei; Gao, Wenyuan

2015-09-01

A electrospray ionization tandem mass spectrometry (ESI-MS(n)) analysis was performed in order to identify the active composition in Pseudostellaria heterophylla adventitious roots. Pseudostellarin A, C, D, and G were identified from P. heterophylla adventitious roots on the basis of LC-MS(n) analysis. The culture conditions of adventitious roots were optimized, and datasets were subjected to a partial least squares discriminant analysis (PLS-DA), in which the growth ratio and some compounds showed a positive correlation with an aeration volume of 0.3 vvm and inoculum density of 0.15 %. Fed-batch cultivation enhanced the contents of total saponin, polysaccharides, and specific oxygen uptaker rate (SOUR). The maximum dry root weight (4.728 g l(-1)) was achieved in the 3/4 Murashige and Skoog (MS) medium group. PLS-DA showed that polysaccharides contributed significantly to the clustering of different groups and showed a positive correlation in the MS medium group. The delayed-type hypersensitivity (DTH) reaction on the mice induced by 2,4-dinitrofluorobenzene (DNFB) was applied to compare the immunocompetence effects of adventitious roots (AR) with field native roots (NR) of P. heterophylla. As a result, AR possessed a similar immunoregulation function as NR.

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

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....

Different cultivation methods to acclimatise ammonia-tolerant methanogenic consortia

DEFF Research Database (Denmark)

Tian, Hailin; Fotidis, Ioannis; Mancini, Enrico

2017-01-01

reactors (i.e. batch, fed-batch and continuous stirred-tank reactors (CSTR)) operated at mesophilic (37°C) and thermophilic (55°C) conditions were assessed, based on methane production efficiency, incubation time, TAN/FAN (total ammonium nitrogen/free ammonia nitrogen) levels and maximum methanogenic...... activity. Overall, fed-batch cultivation was clearly the most efficient method compared to batch and CSTR. Specifically, by saving incubation time up to 150%, fed-batch reactors were acclimatised to nearly 2-fold higher FAN levels with a 37%-153% methanogenic activity improvement, compared to batch method....... Meanwhile, CSTR reactors were inhibited at lower ammonia levels. Finally, specific methanogenic activity test showed that hydrogenotrophic methanogens were more active than aceticlastic methanogens in all FAN levels above 540 mg NH3-N L-1....

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.

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

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

[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

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.

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.

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 ...

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.

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.

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.

Functional State Modelling of Saccharomyces cerevisiae Cultivations

Directory of Open Access Journals (Sweden)

Iasen Hristozov

2004-10-01

Full Text Available The implementation of functional state approach for modelling of yeast cultivation is considered in this paper. This concept helps in monitoring and control of complex processes such as bioprocesses. Using of functional state modelling approach for fermentation processes aims to overcome the main disadvantage of using global process model, namely complex model structure and big number of model parameters. The main advantage of functional state modelling is that the parameters of each local model can be separately estimated from other local models parameters. The results achieved from batch, as well as from fed-batch, cultivations are presented.

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....

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.

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.

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

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.

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.

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

Science.gov (United States)

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

2018-07-01

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

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.

Different cultivation methods to acclimatise ammonia-tolerant methanogenic consortia.

Science.gov (United States)

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

2017-05-01

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

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.

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.

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.

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.

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...

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

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...

GROWTH AND COMPOSITION OF Arthrospira (Spirulina platensis IN A TUBULAR PHOTOBIOREACTOR USING AMMONIUM NITRATE AS THE NITROGEN SOURCE IN A FED-BATCH PROCESS

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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.

Chemometric analysis of in-line multi-wavelength fluorescence measurements obtained during cultivations with a lipase producing Aspergillus oryzae strain

DEFF Research Database (Denmark)

Haack, Martin Brian; Eliasson Lantz, Anna; Mortensen, P.P.

2007-01-01

The filamentous fungus, Aspergillus oryzae, was cultivated in batch and fed-batch cultivations in order to investigate the use of multi-wavelength fluorescence for monitoring course of events during filamentous fungi cultivations. The A. oryzae strain applied expressed a fungal lipase from...... Thermomyces lanuginosus. Spectra of multi-wavelength fluorescence were collected every 5 min with the BioView system (DELTA, Denmark) and both explorative and predictive models, correlating the fluorescence data with cell mass and lipase activity, were built. During the cultivations, A. oryzae displayed...

Growth physiology and dimorphism of Mucor circinelloides (syn. racemosus) during submerged batch cultivation

DEFF Research Database (Denmark)

Mcintyre, Mhairi; Breum, J.; Arnau, J.

2002-01-01

Mucor circinelloides is being investigated as a possible host for the production of heterologous proteins. Thus, the environmental conditions defining the physiology and morphology of this dimorphic fungus have been investigated in submerged batch cultivation. The optimal conditions for growth...

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

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

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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.

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.

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.

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 ...

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

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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.

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

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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.

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

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.

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

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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.

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.

Effect of culture medium, host strain and oxygen transfer on recombinant Fab antibody fragment yield and leakage to medium in shaken E. coli cultures

Science.gov (United States)

2013-01-01

Background Fab antibody fragments in E. coli are usually directed to the oxidizing periplasmic space for correct folding. From periplasm Fab fragments may further leak into extracellular medium. Information on the cultivation parameters affecting this leakage is scarce, and the unpredictable nature of Fab leakage is problematic regarding consistent product recovery. To elucidate the effects of cultivation conditions, we investigated Fab expression and accumulation into either periplasm or medium in E. coli K-12 and E. coli BL21 when grown in different types of media and under different aeration conditions. Results Small-scale Fab expression demonstrated significant differences in yield and ratio of periplasmic to extracellular Fab between different culture media and host strains. Expression in a medium with fed-batch-like glucose feeding provided highest total and extracellular yields in both strains. Unexpectedly, cultivation in baffled shake flasks at 150 rpm shaking speed resulted in higher yield and accumulation of Fabs into culture medium as compared to cultivation at 250 rpm. In the fed-batch medium, extracellular fraction in E. coli K-12 increased from 2-17% of total Fab at 250 rpm up to 75% at 150 rpm. This was partly due to increased lysis, but also leakage from intact cells increased at the lower shaking speed. Total Fab yield in E. coli BL21 in glycerol-based autoinduction medium was 5 to 9-fold higher at the lower shaking speed, and the extracellular fraction increased from ≤ 10% to 20-90%. The effect of aeration on Fab localization was reproduced in multiwell plate by variation of culture volume. Conclusions Yield and leakage of Fab fragments are dependent on expression strain, culture medium, aeration rate, and the combination of these parameters. Maximum productivity in fed-batch-like conditions and in autoinduction medium is achieved under sufficiently oxygen-limited conditions, and lower aeration also promotes increased Fab accumulation into

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.

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.

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.

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.

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.

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

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.

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).

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

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.

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...

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.

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.

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.

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).

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.

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.

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.

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.).

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.

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

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.

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)

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

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...

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.

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.

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.

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%.

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.

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.

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.

To What Extent Do Improved Practices Increase Productivity of Small-Scale Rice Cultivation in A Rain-fed Area? : Evidence from Tanzania

OpenAIRE

Yuko Nakano; Yuki Tanaka; Keijiro Otsuka

2014-01-01

This paper investigates the impact of training provided by a large-scale private farm on the performance of surrounding small-scale rice farmers in a rain-fed area in Tanzania. We found that the training effectively enhances the adoption of improved rice cultivation practices, paddy yield, and profit of rice cultivation by small-holder farmers. In fact, the trainees achieve paddy yield of 5 tons per hectare on average, which is remarkably high for rain-fed rice cultivation. Our results sugges...

Diversity of both the cultivable protease-producing bacteria and bacterial extracellular proteases in the coastal sediments of King George Island, Antarctica.

Directory of Open Access Journals (Sweden)

Ming-Yang Zhou

Full Text Available Protease-producing bacteria play a vital role in degrading sedimentary organic nitrogen. However, the diversity of these bacteria and their extracellular proteases in most regions remain unknown. In this paper, the diversity of the cultivable protease-producing bacteria and of bacterial extracellular proteases in the sediments of Maxwell Bay, King George Island, Antarctica was investigated. The cultivable protease-producing bacteria reached 10(5 cells/g in all 8 sediment samples. The cultivated protease-producing bacteria were mainly affiliated with the phyla Actinobacteria, Firmicutes, Bacteroidetes, and Proteobacteria, and the predominant genera were Bacillus (22.9%, Flavobacterium (21.0% and Lacinutrix (16.2%. Among these strains, Pseudoalteromonas and Flavobacteria showed relatively high protease production. Inhibitor analysis showed that nearly all the extracellular proteases from the bacteria were serine proteases or metalloproteases. These results begin to address the diversity of protease-producing bacteria and bacterial extracellular proteases in the sediments of the Antarctic Sea.

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.

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...

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

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

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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.

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

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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.

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.

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.

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

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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.

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.

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.

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.

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.

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

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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.

Versatile modeling and optimization of fed batch processes for the production of secreted heterologous proteins with Pichia pastoris

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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.

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

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

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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.

Comparative study of production of Bio-Indigo by Pandoraea sp. in a two phase - fed batch and continuous bioreactor

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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.

Characterization of Co-Cultivation of Cyanobacteria on Growth, Productions of Polysaccharides and Extracellular Proteins, Nitrogenase Activity, and Photosynthetic Activity.

Science.gov (United States)

Xue, Chuizhao; Wang, Libo; Wu, Tong; Zhang, Shiping; Tang, Tao; Wang, Liang; Zhao, Quanyu; Sun, Yuhan

2017-01-01

Cyanobacteria as biofertilizers are benefit to reduce the use of chemical fertilizers and reestablish the ecological system in soil. In general, several strains of cyanobacteria were involved in the biofertilizers. The co-cultivation of cyanobacteria were characterized on growth profile, production of polysaccharides and extracellular proteins, nitrogenase activity, and photosynthetic activity for three selected N 2 -fixing cyanobacteria, Anabaena cylindrica (B1611 and F243) and Nostoc sp. (F280). After eight-day culture, the highest dry weights were obtained in F280 pure culture and co-cultivation of B1611 and F280. Higher production of extracellular proteins and cell-bonding polysaccharides (CPS) were observed in co-cultivations compared with pure culture. The highest released polysaccharides (RPS) contents were obtained in pure culture of F280 and co-cultivation of F280 and F243. Galactose and glucose were major components of CPS and RPS in all samples. Trehalose was a specific component of RPS in F280 pure culture. Based on the monosaccharide contents of CPS and RPS, F280 was the dominant species in the related treatments of co-cultivation. The nitrogenase activities in all treatments exhibited a sharp rise at the late stage while a significant decrease existed when three cyanobacteria strains were mixed. Photosynthetic activities for all treatments were determined with rapid light curve, and the related parameters were estimated.

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

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

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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.

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.

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...

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)

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.

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

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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.

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...

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...

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

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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.

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...

Fed batch fermentation scale up in the production of recombinant streptokinase

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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.

Morphology and physiology of an alpha Amylase producing strain of Aspergillus oryzae during batch cultivations

DEFF Research Database (Denmark)

Carlsen, Morten; Spohr, Anders Bendsen; Nielsen, Jens Bredal

1996-01-01

, whereas the alpha-amylase production has a sharper maximum at about pH 6. During batch cultivation with pellets the growth is described well by the cube-root law when pellet fragmentation can be neglected. The kinetic parameter k in the cube-root law is derived from the growth kinetics with no mass......, suggesting that ethanol is produced in the oxygen limited part of the biomass. A constitutive, low alpha-amylase production was observed at high glucose concentration. The specific alpha-amylase production was significantly higher for filamentous growth than for pellets and oxygen appears to be necessary...... for production of alpha-amylase. (C) 1996 John Wiley & Sons, Inc....

Physiological studies in aerobic batch cultivations of Saccharomyces cerevisiae strains harboring the MEL1 gene

DEFF Research Database (Denmark)

Østergaard, Simon; Roca, Christophe Francois Aime; Ronnow, B.

2000-01-01

Physiological studies of Saccharomyces cerevisiae strains harboring the MEL1 gene were carried out in aerobic batch cultivations on glucose-galactose mixtures and on the disaccharide melibiose, which is hydrolyzed by the enzyme melibiase (Mel1, EC 3.2.1.22) into a glucose and a galactose moiety...... rates were 2.5-3.3-fold higher on glucose than on galactose for all the strains examined, and hence, ethanol production was pronounced on glucose due to respiro-fermentative metabolism. The T256 strain and the T200 strain having the MEL1 gene inserted in the HXK2 locus and the LEU2 locus, respectively...

Production and partial characterization of alkaline feruloyl esterases by Fusarium oxysporum during submerged batch cultivation

DEFF Research Database (Denmark)

Topakas, E.; Christakopoulos, Paul

2004-01-01

Production of feruloyl esterases (FAEs) by Fusarium oxysporum was enhanced by optimization of initial pH of the culture medium, the type and concentration of nitrogen and carbon source. Submerged batch cultivation in a laboratory bioreactor (17 1) produced activity at 82 nkat g(-1) dry substrate....... Production of FAE does not therefore, require FA, however, production is diminished by the removal of esterified FA from the growth substrate. Optimal FAE activity was observed at pH 7 and 50 degreesC with 68 and 55% activity at pH 8 and pH 9, respectively. The esterase was fully stable at pH 5-8 and up...

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

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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.

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

Hydrolytic activities of extracellular enzymes in thermophilic and mesophilic anaerobic sequencing-batch reactors treating organic fractions of municipal solid wastes.

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Kim, Hyun-Woo; Nam, Joo-Youn; Kang, Seok-Tae; Kim, Dong-Hoon; Jung, Kyung-Won; Shin, Hang-Sik

2012-04-01

Extracellular enzymes offer active catalysis for hydrolysis of organic solid wastes in anaerobic digestion. To evidence the quantitative significance of hydrolytic enzyme activities for major waste components, track studies of thermophilic and mesophilic anaerobic sequencing-batch reactors (TASBR and MASBR) were conducted using a co-substrate of real organic wastes. During 1day batch cycle, TASBR showed higher amylase activity for carbohydrate (46%), protease activity for proteins (270%), and lipase activity for lipids (19%) than MASBR. In particular, the track study of protease identified that thermophilic anaerobes degraded protein polymers much more rapidly. Results revealed that differences in enzyme activities eventually affected acidogenic and methanogenic performances. It was demonstrated that the superior nature of enzymatic capability at thermophilic condition led to successive high-rate acidogenesis and 32% higher CH(4) recovery. Consequently, these results evidence that the coupling thermophilic digestion with sequencing-batch operation is a viable option to promote enzymatic hydrolysis of organic particulates. Copyright © 2012 Elsevier Ltd. All rights reserved.

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

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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

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

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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.

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

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.

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.

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

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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.

Nitrite survival and nitrous oxide production of denitrifying phosphorus removal sludges in long-term nitrite/nitrate-fed sequencing batch reactors.

Science.gov (United States)

Wang, Yayi; Zhou, Shuai; Ye, Liu; Wang, Hong; Stephenson, Tom; Jiang, Xuxin

2014-12-15

Nitrite-based phosphorus (P) removal could be useful for innovative biological P removal systems where energy and carbon savings are a priority. However, using nitrite for denitrification may cause nitrous oxide (N2O) accumulation and emissions. A denitrifying nitrite-fed P removal system [Formula: see text] was successfully set up in a sequencing batch reactor (SBR) and was run for 210 days. The maximum pulse addition of nitrite to [Formula: see text] was 11 mg NO2(-)-N/L in the bulk, and a total of 34 mg NO2(-)-N/L of nitrite was added over three additions. Fluorescent in situ hybridization results indicated that the P-accumulating organisms (PAOs) abundance was 75 ± 1.1% in [Formula: see text] , approximately 13.6% higher than that in a parallel P removal SBR using nitrate [Formula: see text] . Type II Accumulibacter (PAOII) (unable to use nitrate as an electron acceptor) was the main PAOs species in [Formula: see text] , contributing 72% to total PAOs. Compared with [Formula: see text] , [Formula: see text] biomass had enhanced nitrite/free nitrous acid (FNA) endurance, as demonstrated by its higher nitrite denitrification and P uptake rates. N2O accumulated temporarily in [Formula: see text] after each pulse of nitrite. Peak N2O concentrations in the bulk for [Formula: see text] were generally 6-11 times higher than that in [Formula: see text] ; these accumulations were rapidly denitrified to nitrogen gases. N2O concentration increased rapidly in nitrate-cultivated biomass when 5 or 10 mg NO2(-)-N/L per pulse was added. Whereas, N2O accumulation did not occur in nitrite-cultivated biomass until up to 30 mg NO2(-)-N/L per pulse was added. Long-term acclimation to nitrite and pulse addition of nitrite in [Formula: see text] reduced the risk of nitrite accumulation, and mitigated N2O accumulation and emissions from denitrifying P removal by nitrite. Copyright © 2014 Elsevier Ltd. All rights reserved.

Determination of volumetric gas-liquid mass transfer coefficient of carbon monoxide in a batch cultivation system using kinetic simulations.

Science.gov (United States)

Jang, Nulee; Yasin, Muhammad; Park, Shinyoung; Lovitt, Robert W; Chang, In Seop

2017-09-01

A mathematical model of microbial kinetics was introduced to predict the overall volumetric gas-liquid mass transfer coefficient (k L a) of carbon monoxide (CO) in a batch cultivation system. The cell concentration (X), acetate concentration (C ace ), headspace gas (N co and [Formula: see text] ), dissolved CO concentration in the fermentation medium (C co ), and mass transfer rate (R) were simulated using a variety of k L a values. The simulated results showed excellent agreement with the experimental data for a k L a of 13/hr. The C co values decreased with increase in cultivation times, whereas the maximum mass transfer rate was achieved at the mid-log phase due to vigorous microbial CO consumption rate higher than R. The model suggested in this study may be applied to a variety of microbial systems involving gaseous substrates. Copyright © 2017 Elsevier Ltd. All rights reserved.

Repeated batch cultivation of the hydrocarbon-degrading, micro-algal strain Prototheca zopfii RND16 immobilized in polyurethane foam.

Science.gov (United States)

Ueno, Ryohei; Wada, Shun; Urano, Naoto

2008-01-01

This study reports on the stability of the cells of a heterotrophic green micro-algal strain Prototheca zopfii RND16 immobilized in polyurethane foam (PUF) cubes during degradation of mixed hydrocarbon substrate, which was composed of n-alkanes and polycyclic aromatic hydrocarbons (PAHs), in 5 successive cycles of repeated batch cultivation at 30 degrees C. Both RND16 cells and mixed hydrocarbon substrate components had been entrapped in PUF cubes through cultivation. PUF-immobilized RND16 degraded n-alkanes almost completely, whereas the strain hardly degraded PAHs in PUFs, rather they accumulated in the matrices. It is noteworthy that this result is strikingly different from that of the free-living cell culture, where RND16 reduced concentrations of both n-alkanes and PAHs. However, PAHs accumulation in the PUFs did not impair the performance of the immobilized alga to utilize n-alkanes. These results suggest that the PUFs harboring RND16 cells could be used repeatedly for selective retrieval of PAHs from oil-polluted waters after preferential biodegradation of n-alkanes by algae.

Long-term effect of inoculum pretreatment on fermentative hydrogen production by repeated batch cultivations: homoacetogenesis and methanogenesis as competitors to hydrogen production

DEFF Research Database (Denmark)

Luo, Gang; Karakashev, Dimitar Borisov; Xie, Li

2011-01-01

Long-term effects of inoculum pretreatments(heat, acid, loading-shock) on hydrogen production from glucose under different temperatures (378C, 558C) and initial pH (7 and 5.5) were studied by repeated batch cultivations. Results obtained showed that it was necessary to investigate the long......-term effect of inoculum pretreatment on hydrogen production since pretreatments may just temporarily inhibit the hydrogen consuming processes. After long-term cultivation, pretreated inocula did not enhance hydrogen production compared to untreated inocula under mesophilic conditions (initial pH 7 and pH 5.......5) and thermophilic conditions (initial pH 7). However, pretreatment could inhibit lactate production and lead to higher hydrogen yield under thermophilic conditions at initial pH 5.5. The results further demonstrated that inoculum pretreatment could not permanently inhibit either methanogenesis or homoacetogenesis...

Cultivation of the bacterium Azotobacter chroococcum for ...

African Journals Online (AJOL)

... increase of bioprocess efficiency parameters (yield coefficient and productivity) were observed compared with the batch cultivation. On the basis of the obtained results, repeated batch technique appeared to be the most suitable for the bacterial biomass production at industrial scale. Key words: Azotobacter chroococcum, ...

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.

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.

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.

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.

Robotic platform for parallelized cultivation and monitoring of microbial growth parameters in microwell plates.

Science.gov (United States)

Knepper, Andreas; Heiser, Michael; Glauche, Florian; Neubauer, Peter

2014-12-01

The enormous variation possibilities of bioprocesses challenge process development to fix a commercial process with respect to costs and time. Although some cultivation systems and some devices for unit operations combine the latest technology on miniaturization, parallelization, and sensing, the degree of automation in upstream and downstream bioprocess development is still limited to single steps. We aim to face this challenge by an interdisciplinary approach to significantly shorten development times and costs. As a first step, we scaled down analytical assays to the microliter scale and created automated procedures for starting the cultivation and monitoring the optical density (OD), pH, concentrations of glucose and acetate in the culture medium, and product formation in fed-batch cultures in the 96-well format. Then, the separate measurements of pH, OD, and concentrations of acetate and glucose were combined to one method. This method enables automated process monitoring at dedicated intervals (e.g., also during the night). By this approach, we managed to increase the information content of cultivations in 96-microwell plates, thus turning them into a suitable tool for high-throughput bioprocess development. Here, we present the flowcharts as well as cultivation data of our automation approach. © 2014 Society for Laboratory Automation and Screening.

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.

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.

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.

Water-saving analysis on an effective water reuse system in biodiesel feedstock production based on Chlorella zofingiensis fed-batch cultivation.

Science.gov (United States)

Yang, Kang; Qin, Lei; Wang, Zhongming; Feng, Wei; Feng, Pingzhong; Zhu, Shunni; Xu, Jingliang; Yuan, Zhenhong

2015-01-01

The micralgae-based biofuel obtained from dairy wastewater (DWW) is considered a promising source of energy. However, this process consumes water due to the concentration of wastewater being normally too high for some micoralgae cultivation, and dilution is always needed. In this work, the cultivation of microalgae has been examined in non-recirculated water (NR) and recirculated water systems (R). The growth of Chlorella zofingiensis and the nutrient removal of DWW have been recorded. The comparison indicates the R had a little more advantage in biomass and lipid output (1.55, 0.22 g, respectively) than the NR (1.51, 0.20 g, respectively). However, the total chemical oxygen demand (COD), total Kjeldahl nitrogen (TKN), and total phosphorus (TP) removals of the R were lower than those of the NR system during the culture. The highest removal of total COD, TKN, and TP were 85.05%, 93.64%, and 98.45%, respectively. Furthermore, no significant difference has been observed in the higher heating value and lipid content of the biomass of the R and NR. The results show the R can save 30% of the total water input during the culture. All above results indicate the R system has great potential in industry.

Production of extracellular fatty acid using engineered Escherichia coli

Directory of Open Access Journals (Sweden)

Liu Hui

2012-04-01

Full Text Available Abstract Background As an alternative for economic biodiesel production, the microbial production of extracellular fatty acid from renewable resources is receiving more concerns recently, since the separation of fatty acid from microorganism cells is normally involved in a series of energy-intensive steps. Many attempts have been made to construct fatty acid producing strains by targeting genes in the fatty acid biosynthetic pathway, while few studies focused on the cultivation process and the mass transfer kinetics. Results In this study, both strain improvements and cultivation process strategies were applied to increase extracellular fatty acid production by engineered Escherichia coli. Our results showed overexpressing ‘TesA and the deletion of fadL in E. coli BL21 (DE3 improved extracellular fatty acid production, while deletion of fadD didn’t strengthen the extracellular fatty acid production for an undetermined mechanism. Moreover, the cultivation process controls contributed greatly to extracellular fatty acid production with respect to titer, cell growth and productivity by adjusting the temperature, adding ampicillin and employing on-line extraction. Under optimal conditions, the E. coli strain (pACY-‘tesA-ΔfadL produced 4.8 g L−1 extracellular fatty acid, with the specific productivity of 0.02 g h−1 g−1dry cell mass, and the yield of 4.4% on glucose, while the ratios of cell-associated fatty acid versus extracellular fatty acid were kept below 0.5 after 15 h of cultivation. The fatty acids included C12:1, C12:0, C14:1, C14:0, C16:1, C16:0, C18:1, C18:0. The composition was dominated by C14 and C16 saturated and unsaturated fatty acids. Using the strain pACY-‘tesA, similar results appeared under the same culture conditions and the titer was also much higher than that ever reported previously, which suggested that the supposedly superior strain did not necessarily perform best for the efficient production of desired

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.

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.

Biohydrogen production from glucose in upflow biofilm reactors with plastic carriers under extreme thermophilic conditions (70(degree)C)

DEFF Research Database (Denmark)

Zheng, H.; Zeng, Raymond Jianxiong; Angelidaki, Irini

2008-01-01

Biohydrogen could efficiently be produced in glucose-fed biofilm reactors filled with plastic carriers and operated at 70°C. Batch experiments were, in addition, conducted to enrich and cultivate glucose-fed extremethermophilic hydrogen producing microorganisms from a biohydrogen CSTR reactor fed...

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

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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

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.

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.

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

Science.gov (United States)

Gahlawat, Geeta; Srivastava, Ashok K

2017-10-01

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

Sludge Batch 5 Slurry Fed Melt Rate Furnace Test with Frits 418 and 550

International Nuclear Information System (INIS)

Miller, Donald; Pickenheim, Bradley

2009-01-01

Based on Melt Rate Furnace (MRF) testing for the Sludge Batch 5 (SB5) projected composition and assessments of the potential frits with reasonable operating windows, the Savannah River National Laboratory (SRNL) recommended Slurry Fed Melt Rate Furnace (SMRF) testing with Frits 418 and 550. DWPF is currently using Frit 418 with SB5 based on SRNL's recommendation due to its ability to accommodate significant sodium variation in the sludge composition. However, experience with high boron containing frits in DWPF indicated a potential advantage for Frit 550 might exist. Therefore, SRNL performed SMRF testing to assess Frit 550's potential advantages. The results of SMRF testing with SB5 simulant indicate that there is no appreciable difference in melt rate between Frit 418 and Frit 550 at a targeted 34 weight % waste loading. Both batches exhibited comparable behavior when delivered through the feed tube by the peristaltic pump. Limited observation of the cold cap during both runs showed no indication of major cold cap mounding. MRF testing, performed after the SMRF runs due to time constraints, with the same two Slurry Mix Evaporator (SME) dried products led to the same conclusion. Although visual observations of the cross-sectioned MRF beakers indicated differences in the appearance of the two systems, the measured melt rates were both ∼0.6 in/hr. Therefore, SRNL does not recommend a change from Frit 418 for the initial SB5 processing in DWPF. Once the actual SB5 composition is known and revised projections of SB5 after the neptunium stream addition and any decants is provided, SRNL will perform an additional compositional window assessment with Frit 418. If requested, SRNL can also include other potential frits in this assessment should processing of SB5 with Frit 418 result in less than desirable melter throughput in DWPF. The frits would then be subjected to melt rate testing at SRNL to determine any potential advantages

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

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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.

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.

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

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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.

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)

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.

Capacity Planning for Batch and Perfusion Bioprocesses Across Multiple Biopharmaceutical Facilities

OpenAIRE

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

2014-01-01

Production planning for biopharmaceutical portfolios becomes more complex when products switch between fed-batch and continuous perfusion culture processes. This article describes the development of a discrete-time mixed integer linear programming (MILP) model to optimize capacity plans for multiple biopharmaceutical products, with either batch or perfusion bioprocesses, across multiple facilities to meet quarterly demands. The model comprised specific features to account for products with fe...

Experimental and analytical study on removal of strontium from cultivated soil

International Nuclear Information System (INIS)

Fukutani, Satoshi; Takahashi, Tomoyuki

2003-01-01

Experimental and analytical study was done to estimate the removal of strontium from cultivated soil. The continuous batch tests were made and uneasy desorption form or immobility form was proved to exist. 2-Component Model, which considers easy desorption and uneasy desorption form fraction, was constructed and it showed good explanation of the continuous batch test results. (author)

Neural Network-Based State Estimation for a Closed-Loop Control Strategy Applied to a Fed-Batch Bioreactor

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Santiago Rómoli

2017-01-01

Full Text Available The lack of online information on some bioprocess variables and the presence of model and parametric uncertainties pose significant challenges to the design of efficient closed-loop control strategies. To address this issue, this work proposes an online state estimator based on a Radial Basis Function (RBF neural network that operates in closed loop together with a control law derived on a linear algebra-based design strategy. The proposed methodology is applied to a class of nonlinear systems with three types of uncertainties: (i time-varying parameters, (ii uncertain nonlinearities, and (iii unmodeled dynamics. To reduce the effect of uncertainties on the bioreactor, some integrators of the tracking error are introduced, which in turn allow the derivation of the proper control actions. This new control scheme guarantees that all signals are uniformly and ultimately bounded, and the tracking error converges to small values. The effectiveness of the proposed approach is illustrated on the basis of simulated experiments on a fed-batch bioreactor, and its performance is compared with two controllers available in the literature.

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.

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...

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.

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.

Reducing the variability of antibiotic production in Streptomyces by cultivation in 24-square deepwell plates

DEFF Research Database (Denmark)

Siebenberg, S.; Bapat, Prashant Madhusudhan; Eliasson Lantz, Anna

2010-01-01

Highly reproducible production values of the aminocoumarin antibiotic novobiocin were achieved by cultivation of a heterologous Streptomyces producer strain in commercially available square deepwell plates consisting of 24 wells of 3 ml culture volume each. Between parallel cultivation batches...

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

International Nuclear Information System (INIS)

Smith, M; Timothy Jones, T; Donald02 Miller, D

2007-01-01

Several Slurry-Fed Melt Rate Furnace (SMRF) tests with earlier projections of the Sludge Batch 4 (SB4) composition have been performed.1,2 The first SB4 SMRF test used Frits 418 and 320, however it was found after the test that the REDuction/OXidation (REDOX) correlation at that time did not have the proper oxidation state for manganese. Because the manganese level in the SB4 sludge was higher than previous sludge batches tested, the impact of the higher manganese oxidation state was greater. The glasses were highly oxidized and very foamy, and therefore the results were inconclusive. After resolving this REDOX issue, Frits 418, 425, and 503 were tested in the SMRF with the updated baseline SB4 projection. Based on dry-fed Melt Rate Furnace (MRF) tests and the above mentioned SMRF tests, two previous frit recommendations were made by the Savannah River National Laboratory (SRNL) for processing of SB4 in the Defense Waste Processing Facility (DWPF). The first was Frit 503 based on the June 2006 composition projections.3 The recommendation was changed to Frit 418 as a result of the October 2006 composition projections (after the Tank 40 decant was implemented as part of the preparation plan). However, the start of SB4 processing was delayed due to the control room consolidation outage and the repair of the valve box in the Tank 51 to Tank 40 transfer line. These delays resulted in changes to the projected SB4 composition. Due to the slight change in composition and based on preliminary dry-fed MRF testing, SRNL believed that Frit 510 would increase throughput in processing SB4 in DWPF. Frit 418, which was used in processing Sludge Batch 3 (SB3), was a viable candidate and available in DWPF. Therefore, it was used during the initial SB4 processing. Due to the potential for higher melt rates with Frit 510, SMRF tests with the latest SB4 composition (1298 canisters) and Frits 510 and 418 were performed at a targeted waste loading (WL) of 35%. The '1298 canisters

Recovery of Extracellular Lipolytic Enzymes from Macrophomina phaseolina by Foam Fractionation with Air

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Claudia Schinke

2013-01-01

Full Text Available Macrophomina phaseolina was cultivated in complex and simple media for the production of extracellular lipolytic enzymes. Culture supernatants were batch foam fractionated for the recovery of these enzymes, and column design and operation included the use of P 2 frit (porosity 40 to 100 μm, air as sparging gas at variable flow rates, and Triton X-100 added at the beginning or gradually in aliquots. Samples taken at intervals showed the progress of the kinetic and the efficiency parameters. Best results were obtained with the simple medium supernatant by combining the stepwise addition of small amounts of the surfactant with the variation of the air flow rates along the separation. Inert proteins were foamed out first, and the subsequent foamate was enriched in the enzymes, showing estimated activity recovery (R, enrichment ratio (E, and purification factor (P of 45%, 34.7, and 2.9, respectively. Lipases were present in the enriched foamate.

Biomass production of Lactobacillus plantarum LP02 isolated from ...

African Journals Online (AJOL)

The potentially hypocholesterolemic strain, designated PL02, of Lactobacillus plantarum, was isolated from infant feces. The aim of this study was to characterize and to cultivate this isolate for biomass production in a 5 L fermentor by batch or fed-batch fermentation. A modified medium composition without peptone was ...

Batch cultivation of kluyveromyces fragilis in cheese whey

Energy Technology Data Exchange (ETDEWEB)

Beausejour, D; Leduy, A; Ramalho, R S

1981-01-01

Kluyveromyces fragilis was cultivated batchwise in an open pond rectangular bioreactor at 30 degrees Celcius with aeration, under non-sterile conditions and uncontrolled pH. The culture medium contained 7% cheese whey powder, 0.25% KH2PO4, and 0.5% (NH4)2SO4 and was adjusted to an initial pH of 4.0 with H3PO4. The lactose was almost completely consumed after 16 hours and COD reduction attained 80% after 64 hours. The maximum suspended solids concentration obtained was 11.7 g/L. The cheese whey which had initially low protein and high lactose contents was converted by this system into a high protein and low lactose carbohydrate product.

Cultivation characteristics of immobilized Aspergillus oryzae for kojic acid production.

Science.gov (United States)

Kwak, M Y; Rhee, J S

1992-04-15

Aspergillus oryzae in situ grown from spores entrapped in calcium alginate gel beads was used for the production of kojic acid. The immobilized cells in flask cultures produced kojic acid in a linear proportion while maintaining the stable metabolic activity for a prolonged production period. Kojic acid was accumulated up to a high concentration of 83 g/L, at which the kojic acid began to crystallize, and, thus, the culture had to be replaced with fresh media for the next batch culture. The overall productivities of two consecutive cultivations were higher than that of free mycelial fermentation. However, the production rate of kojic acid by the immobilized cells was suddenly decreased with the appearance of central cavernae inside the immobilized gel beads after 12 days of the third batch cultivation.

Fed-Batch Feeding Strategies for Enzymatic Biodiesel Production

DEFF Research Database (Denmark)

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

2014-01-01

of the differences in the interfacial and bulk concentrations of the enzyme. The model is then used to evaluate various feeding strategies to improve the enzymatic biodiesel production. The feeding strategies investigated, gave insight into how the methanol should be fed to potentially mitigate enzyme deactivation...... while improving the biodiesel yield. The best experimental results gave a yield of 703 .76 g FAME L-1 and a reactor productivity of 28.12 g FAME L-1 h-1. In comparison, to reach the same yield, the optimised two step feeding strategy took 6.25 hours less, which equates to an increase the reactor...

Modelling of a Batch Whey Cultivation of Kluyveromyces marxianus var. lactis MC 5 with Investigation of Mass Transfer Processes in the Bioreactor

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Mitko Petrov

2015-04-01

Full Text Available This study presents a mathematical model of a batch fermentation of lactose oxidation from a natural substratum in a cultivation by the strain Kluyweromyces marxianus var. lactis MC 5. In the model of the process, the mass transfer in the bioreactor for oxygen concentration in the gas phase (GP and in the liquid phase (LP is based on the dispersion model of the GP. In addition, perfect mixing in LP is included. Nine models were investigated for specific growth rate and specific oxygen consumptions rate: Monod, Mink, Tessier, Aiba, Andrews, Haldane, Luong, Edward and Han-Levenspiel. In regard to the parameter estimation, the worst observed error was used for all experiments as an objective function. This approach is a special case of multi objective parameter estimation problems allowing the parameter estimation problem to become a min-max problem. The results obtained (values of criteria, relative error and statistics λ for the specific growth rate showed that the best fit to experimental data is achieved when applying the Mink model. In a combination a Mink, and Monod, Mink, Luong, Haldane, and Han-Levenspiel are used for specific oxygen consumptions rate. Based on the investigation, it was discovered that the best fit belonged to the models of Mink and Haldane, Mink and Luong and Mink and Han-Levenspiel. Therefore, these particular models are used for modeling the batch processes.

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.

Batch cultivation of kluyveromyces fragilis in cheese whey

Energy Technology Data Exchange (ETDEWEB)

Beausejour, D; Leduy, A; Ramalho, R S

1981-08-01

Kluyveromyces fragilis was cultivated batchwise in an open pond rectangular bioreactor at 30 degrees Centigrade, 2vvm of aeration, under non-sterile conditions and uncontrolled pH. The culture medium contained 7% cheese whey powder, 0.25% KH/sub 2/PO/sub 4/, 0.5% (NH/sub 4/)/sub 2/SO/sub 4/ and was adjusted to an initial pH of 4.0 with phosphoric acid. The lactose was almost completely consumed after 16 hours and COD reduction attained 80% after 64 hours. The maximum suspended solids concentration obtained was 11.7 g/L. The cheese whey which had initially low protein and high lactose contents was converted by this system into a high protein and low lactose carbohydrate product. (Refs. 26).

Antioxidant properties of mushroom mycelia obtained by batch cultivation and tocopherol content affected by extraction procedures.

Science.gov (United States)

Vamanu, Emanuel

2014-01-01

The determination of the antioxidant potential of lyophilized mushroom mycelia from 5 strains of the species Pleurotus ostreatus and Coprinus comatus (obtained by submerged cultivation in batch system) was analyzed as ethanolic extracts by evaluating ABTS and the hydroxyl scavenging activity, FRAP method, the chelating capacity, the inhibition of human erythrocyte hemolysis, and the inhibition of xanthine oxidase activity. The main compounds present in all extracts were determined by HPLC chromatography. Overall, results demonstrated that the biologically active substances content is modulated by the extraction method used. The most beneficial extract, characterized by determining the EC50 value, was that of C. comatus M8102, followed by P. ostreatus PQMZ91109. Significant amount of α-tocopherol (179.51 ± 1.51 mg/100 g extract) was determined as well as flavones such as rutin and apigenin. In the P. ostreatus PQMZ91109 extract, 4.8 ± 0.05 mg/100 g extract of tocopherol acetate known to play a significant role as an antioxidant in skin protection against oxidative stress generated by UV rays was determined. The various correlations (r (2) = 0.7665-0.9426 for tocopherol content) assessed and the composition of extracts in fluidized bed from the mycelia of the tested species depicted a significant pharmacological potential as well as the possibility of usage in the development of new functional products.

Volatile fatty acid formation and utilization in anaerobic sulphidogenic batch reactors

CSIR Research Space (South Africa)

Greben, HA

2006-05-01

Full Text Available four stirred batch-test reactors (2 l) were operated, fed with artificial SO4 rich (1700 mg/l) feed water and tap water (controls). The reactors received sulphate reducing bacteria, compost bacteria and grass cuttings. The experimental period was 25...

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

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...

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).

The association between measurements of antimicrobial use and resistance in the faeces microbiota of finisher batches.

Science.gov (United States)

Andersen, V D; DE Knegt, L V; Munk, P; Jensen, M S; Agersø, Y; Aarestrup, F M; Vigre, H

2017-10-01

The objectives were to present three approaches for calculating antimicrobial (AM) use in pigs that take into account the rearing period and rearing site, and to study the association between these measurements and phenotypical resistance and abundance of resistance genes in faeces samples from 10 finisher batches. The AM use was calculated relative to the rearing period of the batches as (i) 'Finisher Unit Exposure' at unit level, (ii) 'Lifetime Exposure' at batch level and (iii) 'Herd Exposure' at herd level. A significant effect on the occurrence of tetracycline resistance measured by cultivation was identified for Lifetime Exposure for the AM class: tetracycline. Furthermore, for Lifetime Exposure for the AM classes: macrolide, broad-spectrum penicillin, sulfonamide and tetracycline use as well as Herd Unit Exposure for the AM classes: aminoglycoside, lincosamide and tetracycline use, a significant effect was observed on the occurrence of genes coding for the AM resistance classes: aminoglycoside, lincosamide, macrolide, β-lactam, sulfonamide and tetracycline. No effect was observed for Finisher Unit Exposure. Overall, the study shows that Lifetime Exposure is an efficient measurement of AM use in finisher batches, and has a significant effect on the occurrence of resistance, measured either by cultivation or metagenomics.

Ethanol Production from Different Intermediates of Sugar Beet Processing

OpenAIRE

Mladen Pavlečić; Ivna Vrana; Kristijan Vibovec; Mirela Ivančić Šantek; Predrag Horvat; Božidar Šantek

2010-01-01

In this investigation, the production of ethanol from the raw sugar beet juice and raw sugar beet cossettes has been studied. For ethanol production from the raw sugar beet juice, batch and fed-batch cultivation techniques in the stirred tank bioreactor were used, while batch ethanol production from the raw sugar beet cossettes was carried out in horizontal rotating tubular bioreactor (HRTB). In both cases, Saccharomyces cerevisiae was used as a production microorganism. During batch ethanol ...

Immobilization of anaerobic thermophilic bacteria for the production of cell-free thermostable. alpha. -amylases and pullulanases

Energy Technology Data Exchange (ETDEWEB)

Klingeberg, M [Goettingen Univ. (Germany, F.R.). Inst. fuer Mikrobiologie; Vorlop, K D [Technische Univ. Braunschweig (Germany, F.R.). Inst. fuer Technische Chemie; Antranikian, G [Technische Univ. Hamburg-Harburg, Hamburg (Germany, F. R.). Arbeitsbereich Biotechnologie 1

1990-08-01

For the production of cell-free thermostable {alpha}-amylases and pullulanases various anaerobic thermophilic bacteria that belong to the genera Clostridium and Thermoanaerobacter were immobilized in calcium alginate gel beads. The entrapment of bacteria was performed in full was well as in hollow spheres. An optimal limited medium, which avoided bacterial outgrowth, was developed for the cultivation of immobilized organisms at 60deg C using 0.4% starch as substrate. Compared to non-immobilized cells these techniques allowed a significant increase (up to 5.6-fold) in the specific activities of the extracellular enzymes formed. An increase in the productivity of extracellular enzymes was observed after immobilization of bacteria in full spheres. In the case of C. thermosaccharolyticum, for instance, the productivity was raised from 90 units (U)/10{sup 12} cells up to 700 U/10{sup 12} cells. Electrophoretic analysis of the secreted proteins showed that in all cases most of the amylolytic enzymes formed were released into the culture medium. Proteins that had a molecular mass of less than 450 000 daltons could easily diffuse through the gel matrix. Cultivation of immobilized bacteria in semi-continuous and fed-batch cultures was also accompanied by an elevation in the concentration of cell-free enzymes. (orig.).

Enhancing the Production of Polyhydroxyalkanoate Biopolymer by Azohydromonas Australica Using a Simple Empty and Fill Bioreactor Cultivation Strategy

Directory of Open Access Journals (Sweden)

G. Gahlawat

2018-01-01

Full Text Available Polyhydroxyalkanoates (PHAs are biodegradable polymers which are considered as an effective alternative for conventional plastics due to their mechanical properties similar to the latter. However, widespread use of these polymers is still hampered due to their high cost of production. This shortcoming could partly be resolved by obtaining high yields and productivity. In the present study, a drain-and-fill strategy of repeated-batch cultivation was adopted for the enhanced production of polyhydroxybutyrate PHB using Azohydromonas australica. In this strategy, 20 % (v/v of the culture broth was removed from the reactor and supplemented with an equal volume of fresh medium. This strategy demonstrated a 3.3 fold and 1.8 fold increase in PHB concentration and productivity, respectively, as compared to batch cultivation. Repeated cultivation had also the benefit of avoiding non-productive time required for cleaning, refilling and sterilization of bioreactor during batch, thereby increasing the overall volumetric productivity and industrial importance of the process.

Novel pH control strategy for glutathione overproduction in batch ...

African Journals Online (AJOL)

The effects of pH values on cell growth and glutathione (GSH) production were studied in batch cultivation of Candida utilis. According to the fact that lower pH value favors cells growth but retards GSH production and higher pH value promotes GSH production while inhibits cells growth, a pH-shift strategy, optimized via ...

Physiological changes of Candida tropicalis population degrading phenol in fed batch reactor

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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.

Extracellular Glycoproteins in Embryogenic Culture of Pumpkin (Cucurbita pepo L.

Directory of Open Access Journals (Sweden)

Hana Čipčić Paljetak

2011-01-01

Full Text Available The extracellular proteins in three distinctly induced embryogenic lines of pumpkin (Cucurbita pepo L. cultivated in four MS media modified regarding the nitrogen composition or auxin presence/absence have been analyzed. Extracellular glycoproteins containing α-D-mannose were specifically detected by the lectine concavalin A. During the cultivation of embryogenic tissue in the medium supplemented with reduced nitrogen, the embryos were mostly arrested at preglobular and globular developmental stages, which coincide with the absence of protein secretion. Secreted glycoproteins of 76, 68, 37 and 34 kDa were detected only if any of the three lines were cultivated in the medium that stimulates embryo development, irrespectively of the addition of 2,4-dichlorophenoxyacetic acid or tunicamycin. The glycoprotein of 64 kDa was detected in all lines cultivated in hormone-free MS medium with conventional nitrogen sources and it appears to be associated with embryo maturation. Tunicamycin treatment did not influence embryogenesis, although it specifically affected glycosylation of proteins in the investigated lines. Our results show that besides auxin, the source of nitrate is of great importance for proper protein glycosylation, excretion and developmental transition of pumpkin somatic embryos.

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

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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.

Reducing conditions are the key for efficient production of active ribonuclease inhibitor in Escherichia coli

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Neubauer Peter

2011-05-01

Full Text Available Abstract Background The eukaryotic RNase ribonuclease/angiogenin inhibitors (RI are a protein group distinguished by a unique structure - they are composed of hydrophobic leucine-rich repeat motifs (LRR and contain a high amount of reduced cysteine residues. The members of this group are difficult to produce in E. coli and other recombinant hosts due to their high aggregation tendency. Results In this work dithiothreitol (DTT was successfully applied for improving the yield of correctly folded ribonuclease/angiogenin inhibitor in E. coli K12 periplasmic and cytoplasmic compartments. The feasibility of the in vivo folding concepts for cytoplasmic and periplasmic production were demonstrated at batch and fed-batch cultivation modes in shake flasks and at the bioreactor scale. Firstly, the best secretion conditions of RI in the periplasmic space were evaluated by using a high throughput multifactorial screening approach of a vector library, directly with the Enbase fed-batch production mode in 96-well plates. Secondly, the effect of the redox environment was evaluated in isogenic dsbA+ and dsbA- strains at the various cultivation conditions with reducing agents in the cultivation medium. Despite the fusion to the signal peptide, highest activities were found in the cytoplasmic fraction. Thus by removing the signal peptide the positive effect of the reducing agent DTT was clearly proven also for the cytoplasmic compartment. Finally, optimal periplasmic and cytoplasmic RI fed-batch production processes involving externally added DTT were developed in shake flasks and scaled up to the bioreactor scale. Conclusions DTT highly improved both, periplasmic and cytoplasmic accumulation and activity of RI at low synthesis rate, i.e. in constructs harbouring weak recombinant synthesis rate stipulating genetic elements together with cultivation at low temperature. In a stirred bioreactor environment RI folding was strongly improved by repeated pulse addition

Cultivation of oleaginous Rhodotorula mucilaginosa in airlift bioreactor by using seawater.

Science.gov (United States)

Yen, Hong-Wei; Liao, Yu-Ting; Liu, Yi Xian

2016-02-01

The enormous water resource consumption is a concern to the scale-up fermentation process, especially for those cheap fermentation commodities, such as microbial oils as the feedstock for biodiesel production. The direct cultivation of oleaginous Rhodotorula mucilaginosa in a 5-L airlift bioreactor using seawater instead of pure water led to a slightly lower biomass being achieved, at 17.2 compared to 18.1 g/L, respectively. Nevertheless, a higher lipid content of 65 ± 5% was measured in the batch using seawater as compared to the pure water batch. Both the salinity and osmotic pressure decreased as the cultivation time increased in the seawater batch, and these effects may contribute to the high tolerance for salinity. No effects were observed for the seawater on the fatty acid profiles. The major components for both batches using seawater and pure water were C16:0 (palmitic acid), C18:1 (oleic acid) and C18:2 (linoleic acid), which together accounted for over 85% of total lipids. The results of this study indicated that seawater could be a suitable option for scaling up the growth of oleaginous R. mucilaginosa, especially from the perspective of water resource utilization. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Greenalgae as a substrate for biogas production - cultivation and biogas potentials

OpenAIRE

Liu, Yang

2010-01-01

Algae is regarded as a good potential substrate for biogas production, due to high cells productivity, low cellulose and zero lignin content. Two parts were included in this study: first, cultivations of micro-algae (Chlorella sorokiniana and Tetraselmis suecica) at two different nitrate concentrations, also the effect of addition of CO2 on algae grow was investigated in this first part. Second, batch fermentations of the cultivated micro-algae as well as a powder Chlorella (obtained from Raw...

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.

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.

Progressing batch hydrolysis process

Science.gov (United States)

Wright, J.D.

1985-01-10

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

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

African Journals Online (AJOL)

2010-12-16

Dec 16, 2010 ... water is fed to the biological treatment unit either intermittently or ... the same culture in a binary mixture and in ternary mixture of 2,4,6TCP; 2,3,5 .... Solving the balance equation (Eq. (2)) with the initial condi- tions of the ...

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.

Mini review: Recombinant production of tailored bio-pharmaceuticals in different Bacillus strains and future perspectives.

Science.gov (United States)

Lakowitz, Antonia; Godard, Thibault; Biedendieck, Rebekka; Krull, Rainer

2018-05-01

Bio-pharmaceuticals like antibodies, hormones and growth factors represent about one-fifth of commercial pharmaceuticals. Host candidates of growing interest for recombinant production of these proteins are strains of the genus Bacillus, long being established for biotechnological production of homologous and heterologous proteins. Bacillus strains benefit from development of efficient expression systems in the last decades and emerge as major industrial workhorses for recombinant proteins due to easy cultivation, non-pathogenicity and their ability to secrete recombinant proteins directly into extracellular medium allowing cost-effective downstream processing. Their broad product portfolio of pharmaceutically relevant recombinant proteins described in research include antibody fragments, growth factors, interferons and interleukins, insulin, penicillin G acylase, streptavidin and different kinases produced in various cultivation systems like microtiter plates, shake flasks and bioreactor systems in batch, fed-batch and continuous mode. To further improve production and secretion performance of Bacillus, bottlenecks and limiting factors concerning proteases, chaperones, secretion machinery or feedback mechanisms can be identified on different cell levels from genomics and transcriptomics via proteomics to metabolomics and fluxomics. For systematical identification of recurring patterns characteristic of given regulatory systems and key genetic targets, systems biology and omics-technology provide suitable and promising approaches, pushing Bacillus further towards industrial application for recombinant pharmaceutical protein production. Copyright © 2017. Published by Elsevier B.V.

Enhanced cellulase production by Trichoderma harzianum by cultivation on glycerol followed by induction on cellulosic substrates.

Science.gov (United States)

Delabona, Priscila da Silva; Lima, Deise Juliana; Robl, Diogo; Rabelo, Sarita Cândida; Farinas, Cristiane Sanchez; Pradella, José Geraldo da Cruz

2016-05-01

The use of glycerol obtained as an intermediate of the biodiesel manufacturing process as carbon source for microbial growth is a potential alternative strategy for the production of enzymes and other high-value bioproducts. This work evaluates the production of cellulase enzymes using glycerol for high cell density growth of Trichoderma harzianum followed by induction with a cellulosic material. Firstly, the influence of the carbon source used in the pre-culture step was investigated in terms of total protein secretion and fungal morphology. Enzymatic productivity was then determined for cultivation strategies using different types and concentrations of carbon source, as well as different feeding procedures (batch and fed-batch). The best strategy for cellulase production was then further studied on a larger scale using a stirred tank bioreactor. The proposed strategy for cellulase production, using glycerol to achieve high cell density growth followed by induction with pretreated sugarcane bagasse, achieved enzymatic activities up to 2.27 ± 0.37 FPU/mL, 106.40 ± 8.87 IU/mL, and 9.04 ± 0.39 IU/mL of cellulase, xylanase, and β-glucosidase, respectively. These values were 2 times higher when compared to the control experiments using glucose instead of glycerol. This novel strategy proved to be a promising approach for improving cellulolytic enzymes production, and could potentially contribute to adding value to biomass within the biofuels sector.

Increased riboflavin production by knockout of 6-phosphofructokinase I and blocking the Entner-Doudoroff pathway in Escherichia coli.

Science.gov (United States)

Liu, Shuang; Kang, Pei; Cui, Zhenzhen; Wang, Zhiwen; Chen, Tao

2016-08-01

To construct an Escherichia coli strain capable of producing riboflavin with high titer and yield. A low copy number plasmid pLS01 containing a riboflavin operon under the control of a constitutive promoter was constructed and introduced into Escherichia coli MG1655. Subsequently, the pfkA, edd and ead genes were disrupted, and the resulting strain LS02T produced 667 mg riboflavin/l in MSY medium supplied with 10 g glucose/l in flask cultivation. In a fed-batch process, riboflavin production of the strain reached 10.4 g/l with a yield of 56.8 mg riboflavin/g glucose. To our knowledge, this is the first report of engineered E. coli strains that can produce more than 10 g riboflavin/l in fed-batch cultivation, indicating that E. coli has potential for riboflavin production.

Extracellular Enzymes Produced by the Cultivated Mushroom Lentinus edodes during Degradation of a Lignocellulosic Medium

Science.gov (United States)

Leatham, Gary F.

1985-01-01

Although the commercially important mushroom Lentinus (= Lentinula) edodes (Berk.) Sing. can be rapidly cultivated on supplemented wood particles, fruiting is not reliable. This study addressed the problem by developing more information about growth and development on a practical oakwood-oatmeal medium. The study determined (i) the components degraded during a 150-day incubation at 22°C, (ii) the apparent vegetative growth pattern, (iii) the likely growth-limiting nutrient, and (iv) assays that can be used to study key extracellular enzymes. All major components of the medium were degraded, lignin selectively so. The vegetative growth rate was most rapid during the initial 90 days, during which weight loss correlated with glucosamine accumulation (assayed after acid hydrolysis). The rate then slowed; in apparent preparation for fruiting, the cultures rapidly accumulated glucosamine (or its oligomer or polymer). Nitrogen was growth limiting. Certain enzyme activities were associated with the pattern of medium degradation, with growth, or with development. They included cellulolytic system enzymes, hemicellulases, the ligninolytic system, (gluco-)amylase, pectinase, acid protease, cell wall lytic enzymes (laminarinase, 1,4-β-d-glucosidase, β-N-acetyl-d-glucosaminidase, α-d-galactosidase, β-d-mannosidase), acid phosphatase, and laccase. Enzyme activities over the 150-day incubation period with and without a fruiting stimulus are reported. These results provide a basis for future investigations into the physiology and biochemistry of growth and fruiting. PMID:16346918

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.

Are agrochemicals present in high fructose corn syrup fed to honey bees (Apis mellifera L.)?

Science.gov (United States)

Honey bee colonies are commonly fed high fructose corn syrup (HFCS) as a nectar substitute. Many agrochemicals are applied to corn during cultivation including systemic neonicotinoids. Whether agrochemicals are present in HFCS fed to bees is unknown. Samples from the major manufacturers and distri...

Transformation of ferulic acid to vanillin using a fed-batch solid-liquid two-phase partitioning bioreactor.

Science.gov (United States)

Ma, Xiao-kui; Daugulis, Andrew J

2014-01-01

Amycolatopsis sp. ATCC 39116 (formerly Streptomyces setonii) has shown promising results in converting ferulic acid (trans-4-hydroxy-3-methoxycinnamic acid; substrate), which can be derived from natural plant wastes, to vanillin (4-hydroxy-3-methoxybenzaldehyde). After exploring the influence of adding vanillin at different times during the growth cycle on cell growth and transformation performance of this strain and demonstrating the inhibitory effect of vanillin, a solid-liquid two-phase partitioning bioreactor (TPPB) system was used as an in situ product removal technique to enhance transformation productivity by this strain. The thermoplastic polymer Hytrel(®) G4078W was found to have superior partitioning capacity for vanillin with a partition coefficient of 12 and a low affinity for the substrate. A 3-L working volume solid-liquid fed-batch TPPB mode, using 300 g Hytrel G4078W as the sequestering phase, produced a final vanillin concentration of 19.5 g/L. The overall productivity of this reactor system was 450 mg/L. h, among the highest reported in literature. Vanillin was easily and quantitatively recovered from the polymers mostly by single stage extraction into methanol or other organic solvents used in food industry, simultaneously regenerating polymer beads for reuse. A polymer-liquid two phase bioreactor was again confirmed to easily outperform single phase systems that feature inhibitory or easily further degraded substrates/products. This enhancement strategy might reasonably be expected in the production of other flavor and fragrance compounds obtained by biotransformations. © 2013 American Institute of Chemical Engineers.

Classification system for rain fed wheat grain cultivars using artificial ...

African Journals Online (AJOL)

Artificial neural network (ANN) models have found wide applications, including ... of grains is essential for various applications as wheat grain industry and cultivation. In order to classify the rain fed wheat cultivars using artificial neural network ...

High level extracellular production of a recombinant alkaline catalase in E. coli BL21 under ethanol stress and its application in hydrogen peroxide removal after cotton fabrics bleaching.

Science.gov (United States)

Yu, Zhenxiao; Zheng, Hongchen; Zhao, Xingya; Li, Shufang; Xu, Jianyong; Song, Hui

2016-08-01

The effects of induction parameters, osmolytes and ethanol stress on the productivity of the recombinant alkaline catalase (KatA) in Escherichia coli BL21 (pET26b-KatA) were investigated. The yield of soluble KatA was significantly enhanced by 2% ethanol stress. And a certain amount of Triton X-100 supplementation could markedly improved extracellular ratio of KatA. A total soluble catalase activity of 78,762U/mL with the extracellular ratio of 92.5% was achieved by fed-batch fermentation in a 10L fermentor, which was the highest yield so far. The purified KatA showed high stability at 50°C and pH 6-10. Application of KatA for elimination of H2O2 after cotton fabrics bleaching led to less consumption of water, steam and electric power by 25%, 12% and 16.7% respectively without productivity and quality losing of cotton fabrics. Thus, the recombinant KatA is a promising candidate for industrial production and applications. Copyright © 2016 Elsevier Ltd. All rights reserved.

Increasing the Photoautotrophic Growth Rate of Synechocystis sp. PCC 6803 by Identifying the Limitations of Its Cultivation.

Science.gov (United States)

van Alphen, Pascal; Abedini Najafabadi, Hamed; Branco Dos Santos, Filipe; Hellingwerf, Klaas J

2018-03-25

Many conditions have to be optimized in order to be able to grow the cyanobacterium Synechocystis sp. PCC 6803 (Synechocystis) for an extended period of time under physiologically well-defined and constant conditions. It is still poorly understood what limits growth of this organism in batch and continuous cultures in BG-11, the standard medium used to grow Synechocystis. Through a series of batch experiments in flasks and continuous mode experiments in advanced photobioreactors, it is shown that the limiting nutrient during batch cultivation is sulfate, the depletion of which leads to ROS formation and rapid bleaching of pigments after entry into stationary phase. In continuous mode, however, the limiting nutrient is iron. Optimizing these growth conditions resulted in a so far highest growth rate of 0.16 h -1 (4.3 h doubling time), which is significantly higher than the textbook value of 0.09 h -1 (8 h doubling time). An improved medium, BG-11 for prolonged cultivation (BG-11-PC) is introduced, that allows for controlled, extended cultivation of Synechocystis, under well-defined physiological conditions. The data present here have implications for mass-culturing of cyanobacteria. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Comparison of laser diffraction and image analysis for measurement of Streptomyces coelicolor cell clumps and pellets

DEFF Research Database (Denmark)

Rønnest, Nanna Petersen; Stocks, Stuart M; Eliasson Lantz, Anna

2012-01-01

and pellets of Streptomyces coelicolor compare to image analysis. Samples, taken five times during fed-batch cultivation, were analyzed by image analysis and laser diffraction. The volume-weighted size distribution was calculated for each sample. Laser diffraction and image analysis yielded similar size...

Stable isotope probing of acetate fed anaerobic batch incubations shows a partial resistance of acetoclastic methanogenesis catalyzed by Methanosarcina to sudden increase of ammonia level.

Science.gov (United States)

Hao, Liping; Lü, Fan; Mazéas, Laurent; Desmond-Le Quéméner, Elie; Madigou, Céline; Guenne, Angéline; Shao, Liming; Bouchez, Théodore; He, Pinjing

2015-02-01

Ammonia inhibition represents a major operational issue for anaerobic digestion. In order to refine our understanding of the terminal catabolic steps in thermophilic anaerobic digestion under ammonia stress, we studied batch thermophilic acetate fed experiments at low (0.26 g L(-1)) and high (7.00 g L(-1)) Total Ammonia Nitrogen concentrations (TAN). Although methane production started immediately for all incubations and resulted in methane yields close to stoichiometric expectations, a 62-72% decrease of methanogenic rate was observed throughout the incubation at 7.00 g L(-1) of TAN compared to 0.26 g L(-1). Stable Isotope Probing analysis of active microbial communities in (13)C-acetate fed experiments coupled to automated ribosomal intergenic spacer analysis and 16S rDNA pyrotag sequencing confirmed that microbial communities were similar for both TAN conditions. At both TAN levels, the (13)C-labeled bacterial community was mainly affiliated to Clostridia-relatives, with OPB54 bacteria being the most abundant sequence in the heavy DNA 16S rDNA pyrotag library. Sequences closely related to Methanosarcina thermophila were also abundantly retrieved in the heavy DNA fractions, showing that this methanogen was still actively assimilating labeled carbon from acetate at free ammonia nitrogen concentrations up to 916 mg L(-1). Stable isotopic signature analysis of biogas, measured in unlabeled acetate fed experiments that were conducted in parallel, confirmed that acetoclastic methanogenic pathway was dominant at both ammonia concentrations. Our work demonstrates that, besides the syntrophic acetate oxidation pathway, acetoclastic methanogenesis catalyzed by Methanosarcina can also play a major role in methane production at high ammonia levels. Copyright © 2014 Elsevier Ltd. All rights reserved.

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.

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.

Heuristics for batching and sequencing in batch processing machines

Directory of Open Access Journals (Sweden)

Chuda Basnet

2016-12-01

Full Text Available In this paper, we discuss the “batch processing” problem, where there are multiple jobs to be processed in flow shops. These jobs can however be formed into batches and the number of jobs in a batch is limited by the capacity of the processing machines to accommodate the jobs. The processing time required by a batch in a machine is determined by the greatest processing time of the jobs included in the batch. Thus, the batch processing problem is a mix of batching and sequencing – the jobs need to be grouped into distinct batches, the batches then need to be sequenced through the flow shop. We apply certain newly developed heuristics to the problem and present computational results. The contributions of this paper are deriving a lower bound, and the heuristics developed and tested in this paper.

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.

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.

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.

Establishment and Characterization of an Anaerobic Thermophilic (55 degrees C) Enrichment Culture Degrading Long-Chain Fatty Acids

DEFF Research Database (Denmark)

Angelidaki, Irini; Ahring, Birgitte Kiær

1995-01-01

A thermophilic, long-chain fatty acid-oxidizing culture was enriched. Stearate was used as the substrate, and methane and carbon dioxide were the sole end products. Cultivation was possible only when a fed-batch system was used or with addition of activated carbon or bentonite. The enrichment...

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.

In vitro azadirachtin production by hairy root cultivation of Azadirachta indica in nutrient mist bioreactor.

Science.gov (United States)

Srivastava, Smita; Srivastava, A K

2012-01-01

Azadirachtin, a well-known biopesticide is a secondary metabolite conventionally extracted from the seeds of Azadirachta indica. The present study involved in vitro azadirachtin production by developing hairy roots of A. indica via Agrobacterium rhizogenes-mediated transformation of A. indica explants. Liquid culture of hairy roots was established in shake flask to study the kinetics of growth and azadirachtin production. A biomass production of 13.3 g/L dry weight (specific growth rate of 0.7 day(-1)) was obtained after 25 days of cultivation period with an azadirachtin yield of 3.3 mg/g root biomass. To overcome the mass transfer limitation in conventionally used liquid-phase reactors, batch cultivation of hairy roots was carried out in gas-phase reactors (nutrient spray and nutrient mist bioreactor) to investigate the possible scale-up of A. indica hairy root culture. The nano-size nutrient mist particles generated from the nozzle of the nutrient mist bioreactor could penetrate till the inner core of the inoculated root matrix, facilitating uniform growth during high-density cultivation of hairy roots. A biomass production of 9.8 g/L dry weight with azadirachtin accumulation of 2.8 mg/g biomass (27.4 mg/L) could be achieved in 25 days of batch cultivation period, which was equivalent to a volumetric productivity of 1.09 mg/L per day of azadirachtin.

High-level extracellular production and characterization of Candida antarctica lipase B in Pichia pastoris.

Science.gov (United States)

Eom, Gyeong Tae; Lee, Seung Hwan; Song, Bong Keun; Chung, Keun-Wo; Kim, Young-Wun; Song, Jae Kwang

2013-08-01

The gene encoding lipase B from Candida antarctica (CalB) was expressed in Pichia pastoris after it was synthesized by the recursive PCR and cloned into the Pichia expression plasmid, pPICZαA. The CalB was successfully secreted in the recombinant P. pastoris strain X-33 with an apparent molecular weight of 34 kDa. For 140 h flask culture, the dry cell weight and the extracellular lipase activity reached at 5.4 g/l and 57.9 U/l toward p-nitrophenyl palmitate, respectively. When we performed the fed-batch fermentation using a methanol feeding strategy for 110 h, the dry cell weight and the extracellular lipase activity were increased to 135.7 g/l and 11,900 U/l; the CalB protein concentration was 1.18 g/l of culture supernatant. The characteristics of CalB recovered from the P. pastoris culture were compared with the commercial form of CalB produced in Aspergillus oryzae. The kinetic constants and specific activity, the effects of activity and stability on temperature and pH, the glycosylation extent, the degree of immobilization on macroporous resin and the yield of esterification reaction between oleic acid and n-butanol were almost identical to each other. Therefore, we successfully proved that the Pichia-based expression system for CalB in this study was industrially promising compared with one of the most efficient production systems. Copyright © 2013 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

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

Science.gov (United States)

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

2014-01-01

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

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.

Biohydrogen production from desugared molasses (DM) using thermophilic mixed cultures immobilized on heat treated anaerobic sludge granules

DEFF Research Database (Denmark)

Kongjan, Prawit; O-Thong, Sompong; Angelidaki, Irini

2011-01-01

Hydrogen production from desugared molasses (DM) was investigated in both batch and continuous reactors using thermophilic mixed cultures enriched from digested manure by load shock (loading with DM concentration of 50.1 g-sugar/L) to suppress methanogens. H2 gas, free of methane, was produced......) and Thermoanaerobacterium thermosaccharolyticum with a relative abundance of 36%, 27%, and 10% of total microorganisms, respectively. This study shows that hydrogen production could be efficiently facilitated by using anaerobic granules as a carrier, where microbes from mixed culture enriched in the DM batch cultivation....... The enriched hydrogen producing mixed culture achieved from the 16.7 g-sugars/L DM batch cultivation was immobilized on heat treated anaerobic sludge granules in an up-flow anaerobic sludge blanket (UASB) reactor. The UASB reactor, operated at a hydraulic retention time (HRT) of 24 h fed with 16.7 g...

Isolation and characterization of extracellular polysaccharides from the epipelic diatoms Cylindrotheca closterium and Navicula salinarum

NARCIS (Netherlands)

Staats, N.; de Winder, B.; Stal, L.J.; Mur, L.R.

1999-01-01

The production and composition of extracellular polymeric substances (EPS) in axenic batch cultures of the benthic marine epipelic diatoms Navicula salinarum and Cylindrotheca closterium were investigated. EPS was secreted into the medium and the bulk was loosely associated with the cells. Neither

Comparison of some characteristics of aerobic granules and sludge flocs from sequencing batch reactors.

Science.gov (United States)

Li, J; Garny, K; Neu, T; He, M; Lindenblatt, C; Horn, H

2007-01-01

Physical, chemical and biological characteristics were investigated for aerobic granules and sludge flocs from three laboratory-scale sequencing batch reactors (SBRs). One reactor was operated as normal SBR (N-SBR) and two reactors were operated as granular SBRs (G-SBR1 and G-SBR2). G-SBR1 was inoculated with activated sludge and G-SBR2 with granules from the municipal wastewater plant in Garching (Germany). The following major parameters and functions were measured and compared between the three reactors: morphology, settling velocity, specific gravity (SG), sludge volume index (SVI), specific oxygen uptake rate (SOUR), distribution of the volume fraction of extracellular polymeric substances (EPS) and bacteria, organic carbon and nitrogen removal. Compared with sludge flocs, granular sludge had excellent settling properties, good solid-liquid separation, high biomass concentration, simultaneous nitrification and denitrification. Aerobic granular sludge does not have a higher microbial activity and there are some problems including higher effluent suspended solids, lower ratio of VSS/SS and no nitrification at the beginning of cultivation. Measurement with CLSM and additional image analysis showed that EPS glycoconjugates build one main fraction inside the granules. The aerobic granules from G-SBR1 prove to be heavier, smaller and have a higher microbial activity compared with G-SBR2. Furthermore, the granules were more compact, with lower SVI and less filamentous bacteria.

Cassava processing wastewater as a platform for third generation biodiesel production

Directory of Open Access Journals (Sweden)

Cristina Neves

Full Text Available ABSTRACT This study aimed to evaluate third generation biodiesel production by microalgae Phormidium autumnale using cassava processing wastewater as a platform. Experiments were performed in a heterotrophic bubble column bioreactor. The study focused on the evaluation of the bioreactor (batch and fed-batch of different operational modes and the analysis of biofuel quality. Results indicate that fed-batch cultivations improved system performance, elevating biomass and oil productions to 12.0 g L−1 and 1.19 g L−1, respectively. The composition of this oil is predominantly saturated (60 % and monounsaturated (39 %, resulting in a biodiesel that complys with U.S., European and Brazilian standards. The technological route developed indicates potential for sustainable production of bulk oil and biodiesel, through the minimization of water and chemical demands required to support such a process.

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

Science.gov (United States)

Ogawa, Morimasa

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

Pro Spring Batch

CERN Document Server

Minella, Michael T

2011-01-01

Since its release, Spring Framework has transformed virtually every aspect of Java development including web applications, security, aspect-oriented programming, persistence, and messaging. Spring Batch, one of its newer additions, now brings the same familiar Spring idioms to batch processing. Spring Batch addresses the needs of any batch process, from the complex calculations performed in the biggest financial institutions to simple data migrations that occur with many software development projects. Pro Spring Batch is intended to answer three questions: *What? What is batch processing? What

Acid protease and formation of multiple forms of glucoamylase in batch and continuous cultures of Aspergillus niger

DEFF Research Database (Denmark)

Aalbæk, Thomas; Reeslev, Morten; Jensen, Bo

2002-01-01

with molecular weights of approx. 91 (GAI), 73 (GAII), and 59 kDa (GAIII). Data from batch fermentations with constant pH 3.0 and 5.0 showed a uniform distribution of extracellular GA forms throughout the fermentations and independent of culture growth phases. Furthermore, steady-state data from chemostat...

Ethanol Production from Different Intermediates of Sugar Beet Processing

Directory of Open Access Journals (Sweden)

Mladen Pavlečić

2010-01-01

Full Text Available In this investigation, the production of ethanol from the raw sugar beet juice and raw sugar beet cossettes has been studied. For ethanol production from the raw sugar beet juice, batch and fed-batch cultivation techniques in the stirred tank bioreactor were used, while batch ethanol production from the raw sugar beet cossettes was carried out in horizontal rotating tubular bioreactor (HRTB. In both cases, Saccharomyces cerevisiae was used as a production microorganism. During batch ethanol production from the raw sugar beet juice, ethanol yield was 59.89 g/L and production efficiency 78.8 %, and in fed-batch process the yield was 92.78 g/L and efficiency 93.4 %. At the same time, ethanol production in HRTB from the raw sugar beet cossettes with inoculum of 16.7 % V/m (raw sugar beet cossettes resulted in the highest ethanol yield of 54.53 g/L and production efficiency of 79.5 %. The obtained results clearly show that both intermediates of sugar beet processing can be successfully used for ethanol production.

21 CFR 573.920 - Selenium.

Science.gov (United States)

2010-04-01

... section, or as selenium yeast, as provided in paragraph (h) of this section. (c) It is added to feed as... months.” (h) Selenium yeast is a dried, non-viable yeast (Saccharomyces cerevisiae) cultivated in a fed-batch fermentation which provides incremental amounts of cane molasses and selenium salts in a manner...

Substrate Channelling and Energetics of Saccharomyces cerevisiae ...

African Journals Online (AJOL)

Data collected during the high-cell-density cultivation of Saccharomyces cerevisiae DSM 2155 on glucose in a simulated five-phase feeding strategy of fed-batch process, executed on the Universal BIoprocess CONtrol (UBICON) system using 150L bioreactor over a period of 24h have been analysed. The consistency of the ...

Capacity Planning for Batch and Perfusion Bioprocesses Across Multiple Biopharmaceutical Facilities

Science.gov (United States)

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

2014-01-01

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

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...

Contrasting evolution patterns between glacier-fed and non-glacier-fed lakes in the central Tibetan Plateau and driving force analysis

Science.gov (United States)

Song, C.; Sheng, Y.

2015-12-01

High-altitude lakes in the Tibetan Plateau (TP) showed strong spatio-temporal variability during past decades. The lake dynamics can be associated with several key factors including lake type, supply of glacial meltwater, local climate variations. It is important to differentiate these factors when analyzing the driving force of lakes dynamics. With a focus on lakes over the Tanggula Mountains of the central TP, this study investigates the temporal evolution patterns of lake area and water level of different types: glacier-fed closed lake, non-glacier-fed closed lake and upstream lake (draining into closed lakes). We collected all available Landsat archive data and quantified the inter-annual variability of lake extents. Results show accelerated expansions of both glacier-fed and non-glacier-fed lakes during 1970s-2013, and different temporal patterns of the two types of lakes: the non-glacier-fed lakes displayed a batch-wise growth pattern, with obvious growth in 2002, 2005 and 2011 and slight changes in other years, while glacier-fed lakes showed steady expanding tendency. The contrasting patterns are confirmed by the distinction of lake level change between the two groups derived from satellite altimetry during 2003-2009. The upstream lakes remained largely stable due to natural drainage regulation. The intermittent expansions for non-glacier-fed lakes were found to be related to excessive precipitation events and positive "precipitation-evaporation". In contrast, glacier-fed lake changes showed weak correlations with precipitation variations, which imply a joint contribution from glacial meltwater to water budgets. A simple estimation reveals that the increased water storage for all of examined lakes contributed from precipitation/evaporation (0.31±0.09 Gt/yr) slightly overweighed the glacial meltwater supply (0.26±0.08 Gt/yr).

Semicontinuous cultivation of the cyanobacterium Spirulina platensis in a closed photobioreactor

Directory of Open Access Journals (Sweden)

C. C. Reichert

2006-03-01

Full Text Available The cultivation of photosynthetic microorganisms such as the cyanobacterium Spirulina platensis has been studied by researchers in many countries because these organisms can produce products with industrial potential. We studied the specific growth rate (µx, day-1 and productivity (Px, in mg/L/day of Spirulina platensis biomass, dry weight basis of two S. platensis strains (LEB-52 and Paracas growing in aerated semicontinuous culture in two-liter Erlenmeyer flasks for 90 days (2160 h at 30°C under 2500 lux of illumination in a 12 h photoperiod. Independent of the S. platensis strain used we found that low biomass concentrations (0.50 g/L and high renewal rates (50% v/v resulted in a high specific growth rate (µx = 0.111 day-1 and high productivity (Px = 42.3 mg/L/day. These values are two to four times higher than those obtained in simple batch cultivation and indicate that the semicontinuous cultivation of S. platensis is viable.

The Kinetic of Biogas Production Rate from Cattle Manure in Batch Mode

OpenAIRE

Budiyono; I N. Widiasa; S. Johari; Sunarso

2010-01-01

In this study, the kinetic of biogas production was studied by performing a series laboratory experiment using rumen fluid of animal ruminant as inoculums. Cattle manure as substrate was inoculated by rumen fluid to the anaerobic biodigester. Laboratory experiments using 400 ml biodigester were performed in batch operation mode. Given 100 grams of fresh cattle manure was fed to each biodigester and mixed with rumen fluid by manure : rumen weight ratio of 1:1 (MR11). The operating temperatures...

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

The Cultivation and SeleniumEnrichment of SeleniumEnriched Earthworm

Directory of Open Access Journals (Sweden)

SUN Xiao-fei

2014-12-01

Full Text Available As a bio-carrier, Eisenia fetida was fed with cow dung that added with sodium selenite in order to transfer inorganic selenium(Se into organic selenium. Targeting on survival rate and selenium content, the effects of five Se concentrations(0, 20, 40, 60, 80, 100 mg·kg-1 and four cultivation periods(15, 30, 45, 60 don earthworm growth and Se contents were investigated. The cultivation method with high survival rate, high Se content of earthworm and short breeding time would be screened out. The experimental results showed that the earthworm survival rate decreased and the Se content in earthworm increased with the increase of Se application and the extension of breeding time. The most optimummethod was screened out when the Se concentration was 80 mg·kg-1 and the cultivation period was 45 days, Se content in earthworm was up to 33.25 mg·kg-1.

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.

Cultivation Of Deep Subsurface Microbial Communities

Science.gov (United States)

Obrzut, Natalia; Casar, Caitlin; Osburn, Magdalena R.

2018-01-01

The potential habitability of surface environments on other planets in our solar system is limited by exposure to extreme radiation and desiccation. In contrast, subsurface environments may offer protection from these stressors and are potential reservoirs for liquid water and energy that support microbial life (Michalski et al., 2013) and are thus of interest to the astrobiology community. The samples used in this project were extracted from the Deep Mine Microbial Observatory (DeMMO) in the former Homestake Mine at depths of 800 to 2000 feet underground (Osburn et al., 2014). Phylogenetic data from these sites indicates the lack of cultured representatives within the community. We used geochemical data to guide media design to cultivate and isolate organisms from the DeMMO communities. Media used for cultivation varied from heterotrophic with oxygen, nitrate or sulfate to autotrophic media with ammonia or ferrous iron. Environmental fluid was used as inoculum in batch cultivation and strains were isolated via serial transfers or dilution to extinction. These methods resulted in isolating aerobic heterotrophs, nitrate reducers, sulfate reducers, ammonia oxidizers, and ferric iron reducers. DNA sequencing of these strains is underway to confirm which species they belong to. This project is part of the NASA Astrobiology Institute Life Underground initiative to detect and characterize subsurface microbial life; by characterizing the intraterrestrials, the life living deep within Earth’s crust, we aim to understand the controls on how and where life survives in subsurface settings. Cultivation of terrestrial deep subsurface microbes will provide insight into the survival mechanisms of intraterrestrials guiding the search for these life forms on other planets.

Extracellular Metabolites from Industrial Microalgae and Their Biotechnological Potential

Directory of Open Access Journals (Sweden)

Lu Liu

2016-10-01

Full Text Available Industrial microalgae, as a big family of promising producers of renewable biomass feedstock, have been commercially exploited for functional food, living feed and feed additives, high-value chemicals in nutraceuticals, cosmeceuticals, and chemical reagents. Recently, microalgae have also been considered as a group that might play an important role in biofuel development and environmental protection. Almost all current products of industrial microalgae are derived from their biomass; however, large amounts of spent cell-free media are available from mass cultivation that is mostly unexploited. In this contribution we discuss that these media, which may contain a remarkable diversity of bioactive substances are worthy to be recovered for further use. Obviously, the extracellular metabolites from industrial microalgae have long been neglected in the development of production methods for valuable metabolites. With the advances in the last ten years, more and more structures and properties from extracellular metabolites have been identified, and the potential utilization over wide fields is attracting attention. Some of these extracellular metabolites can be potentially used as drugs, antioxidants, growth regulators or metal chelators. The purpose of this review is to provide an overview of the known extracellular metabolites from industrial microalgae which might be of commercial interest. The attention mainly focuses on the reports of extracellular bioactive metabolites and their potential application in biotechnology.

Energy efficiency of batch and semi-batch (CCRO) reverse osmosis desalination.

Science.gov (United States)

Warsinger, David M; Tow, Emily W; Nayar, Kishor G; Maswadeh, Laith A; Lienhard V, John H

2016-12-01

As reverse osmosis (RO) desalination capacity increases worldwide, the need to reduce its specific energy consumption becomes more urgent. In addition to the incremental changes attainable with improved components such as membranes and pumps, more significant reduction of energy consumption can be achieved through time-varying RO processes including semi-batch processes such as closed-circuit reverse osmosis (CCRO) and fully-batch processes that have not yet been commercialized or modelled in detail. In this study, numerical models of the energy consumption of batch RO (BRO), CCRO, and the standard continuous RO process are detailed. Two new energy-efficient configurations of batch RO are analyzed. Batch systems use significantly less energy than continuous RO over a wide range of recovery ratios and source water salinities. Relative to continuous RO, models predict that CCRO and batch RO demonstrate up to 37% and 64% energy savings, respectively, for brackish water desalination at high water recovery. For batch RO and CCRO, the primary reductions in energy use stem from atmospheric pressure brine discharge and reduced streamwise variation in driving pressure. Fully-batch systems further reduce energy consumption by not mixing streams of different concentrations, which CCRO does. These results demonstrate that time-varying processes can significantly raise RO energy efficiency. Copyright © 2016 Elsevier Ltd. All rights reserved.

Differential proteomic analysis highlights metabolic strategies associated with balhimycin production in Amycolatopsis balhimycina chemostat cultivations

DEFF Research Database (Denmark)

Gallo, Giuseppe; Alduina, Rosa; Renzone, Giovanni

2010-01-01

Background Proteomics was recently used to reveal enzymes whose expression is associated with the production of the glycopeptide antibiotic balhimycin in Amycolatopsis balhimycina batch cultivations. Combining chemostat fermentation technology, where cells proliferate with constant parameters...... in a highly reproducible steady-state, and differential proteomics, the relationships between physiological status and metabolic pathways during antibiotic producing and non-producing conditions could be highlighted. Results Two minimal defined media, one with low Pi (0.6 mM; LP) and proficient glucose (12 g....../l) concentrations and the other one with high Pi (1.8 mM) and limiting (6 g/l; LG) glucose concentrations, were developed to promote and repress antibiotic production, respectively, in A. balhimycina chemostat cultivations. Applying the same dilution rate (0.03 h-1), both LG and LP chemostat cultivations showed...

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.

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

Science.gov (United States)

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

2018-04-24

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

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

Science.gov (United States)

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

2014-07-04

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

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.

Azadirachtin production by hairy root cultivation of Azadirachta indica in a modified stirred tank reactor.

Science.gov (United States)

Srivastava, Smita; Srivastava, A K

2012-11-01

Present investigation involves hairy root cultivation of Azadirachta indica in a modified stirred tank reactor under optimized culture conditions for maximum volumetric productivity of azadirachtin. The selected hairy root line (Az-35) was induced via Agrobacterium rhizogenes LBA 920-mediated transformation of A. indica leaf explants (Coimbatore variety, India). Liquid culture of the hairy roots was developed in a modified Murashige and Skoog medium (MM2). To further enhance the productivity of azadirachtin, selected growth regulators (1.0 mg/l IAA and 0.025 mg/l GA(3)), permeabilizing agent (0.5 % v/v DNBP), a biotic elicitor (1 % v/v Curvularia (culture filtrate)) and an indirectly linked biosynthetic precursor (50 mg/l cholesterol) were added in the growth medium on 15th day of the hairy root cultivation period in shake flask. Highest azadirachtin production (113 mg/l) was obtained on 25th day of the growth cycle with a biomass of 21 g/l DW. Further, batch cultivation of hairy roots was carried out in a novel liquid-phase bioreactor configuration (modified stirred tank reactor with polyurethane foam as root support) to investigate the possible scale-up of the established A. indica hairy root culture. A biomass production of 15.2 g/l with azadirachtin accumulation in the hairy roots of 6.4 mg/g (97.28 mg/l) could be achieved after 25 days of the batch cultivation period, which was ~27 and ~14 % less biomass and azadirachtin concentration obtained respectively, in shake flasks. An overall volumetric productivity of 3.89 mg/(l day) of azadirachtin was obtained in the bioreactor.

Semicontinuous cultivation of the Cyanobacterium Spirulina platensis in a closed photobioreactor

Energy Technology Data Exchange (ETDEWEB)

Reichert, C.C.; Costa, J.A.V. [Fundacao Universidade Federal do Rio Grande (FURG), Rio Grande, RS (Brazil). Dept. de Quimica], Email: dqmjorge@furg.br; Reinehr, C.O. [Universidade de Passo Fundo, RS (Brazil). Centro de Pesquisa em Alimentacao], Email: reinehr@upf.br

2006-01-15

The cultivation of photosynthetic microorganisms such as the cyanobacterium Spirulina platensis has been studied by researchers in many countries because these organisms can produce products with industrial potential. We studied the specific growth rate ({mu}{sub x}, day{sup -1}) and productivity (P{sub x}, in mg/L/day of Spirulina platensis biomass, dry weight basis) of two S. platensis strains (LEB-52 and Paracas) growing in aerated semicontinuous culture in two-liter Erlenmeyer flasks for 90 days (2160 h) at 30 deg C under 2500 lux of illumination in a 12 h photoperiod. Independent of the S. platensis strain used we found that low biomass concentrations (0.50 g/L) and high renewal rates (50% v/v) resulted in a high specific growth rate ({mu}{sub x} = 0.111 day{sup -1}) and high productivity (P{sub x} = 42.3 mg/L/day). These values are two to four times higher than those obtained in simple batch cultivation and indicate that the semicontinuous cultivation of S. platensis is viable. (author)

Cultivating the Deep Subsurface Microbiome

Science.gov (United States)

Casar, C. P.; Osburn, M. R.; Flynn, T. M.; Masterson, A.; Kruger, B.

2017-12-01

Subterranean ecosystems are poorly understood because many microbes detected in metagenomic surveys are only distantly related to characterized isolates. Cultivating microorganisms from the deep subsurface is challenging due to its inaccessibility and potential for contamination. The Deep Mine Microbial Observatory (DeMMO) in Lead, SD however, offers access to deep microbial life via pristine fracture fluids in bedrock to a depth of 1478 m. The metabolic landscape of DeMMO was previously characterized via thermodynamic modeling coupled with genomic data, illustrating the potential for microbial inhabitants of DeMMO to utilize mineral substrates as energy sources. Here, we employ field and lab based cultivation approaches with pure minerals to link phylogeny to metabolism at DeMMO. Fracture fluids were directed through reactors filled with Fe3O4, Fe2O3, FeS2, MnO2, and FeCO3 at two sites (610 m and 1478 m) for 2 months prior to harvesting for subsequent analyses. We examined mineralogical, geochemical, and microbiological composition of the reactors via DNA sequencing, microscopy, lipid biomarker characterization, and bulk C and N isotope ratios to determine the influence of mineralogy on biofilm community development. Pre-characterized mineral chips were imaged via SEM to assay microbial growth; preliminary results suggest MnO2, Fe3O4, and Fe2O3 were most conducive to colonization. Solid materials from reactors were used as inoculum for batch cultivation experiments. Media designed to mimic fracture fluid chemistry was supplemented with mineral substrates targeting metal reducers. DNA sequences and microscopy of iron oxide-rich biofilms and fracture fluids suggest iron oxidation is a major energy source at redox transition zones where anaerobic fluids meet more oxidizing conditions. We utilized these biofilms and fluids as inoculum in gradient cultivation experiments targeting microaerophilic iron oxidizers. Cultivation of microbes endemic to DeMMO, a system

Inhibitory effect of self-generated extracellular dissolved organic carbon on carbon dioxide fixation in sulfur-oxidizing bacteria during a chemoautotrophic cultivation process and its elimination.

Science.gov (United States)

Wang, Ya-Nan; Tsang, Yiu Fai; Wang, Lei; Fu, Xiaohua; Hu, Jiajun; Li, Huan; Le, Yiquan

2018-03-01

The features of extracellular dissolved organic carbon (EDOC) generation in two typical aerobic sulfur-oxidizing bacteria (Thiobacillus thioparus DSM 505 and Halothiobacillus neapolitanus DSM 15147) and its impact on CO 2 fixation during chemoautotrophic cultivation process were investigated. The results showed that EDOC accumulated in both strains during CO 2 fixation process. Large molecular weight (MW) EDOC derived from cell lysis and decay was dominant during the entire process in DSM 505, whereas small MW EDOC accounted for a large proportion during initial and middle stages of DSM 15147 as its cytoskeleton synthesis rate did not keep up with CO 2 assimilation rate. The self-generated EDOC feedback repressed cbb gene transcription and thus decreased total bacterial cell number and CO 2 fixation yield in both strains, but DSM 505 was more sensitive to this inhibition effect. Moreover, the membrane bioreactor effectively decreased the EDOC/TOC ratio and improved carbon fixation yield of DSM 505. Copyright © 2017 Elsevier Ltd. All rights reserved.

Evaluation of Nitrification Inhibition Using Sequencing Batch Reactors and BioWin Modeling, and the Effect of Aqueous Film Forming Foam on Biological Nutrient Removal

OpenAIRE

Hingley, Daniel McCabe

2011-01-01

To evaluate continuous and sporadic nitrification inhibition at the HRSD Nansemond Wastewater Treatment Plant, which has a history of nitrification upsets, continuous sequencing batch reactors (SBRs) were operated to simulate the full-scale plant. Four reactors were operated in this study. One reactor was fed with raw influent (RWI) from the Nansemond Wastewater Treatment Plant (NP). Another was fed with NP primary clarifier influent (PCI), which includes the raw influent, as well as plant re...

Carbohydrates in Ankistrodesmus braunii biomass cultivated in tubular photobioreactors

Directory of Open Access Journals (Sweden)

Ana Lucía Morocho-Jácome

2017-12-01

Full Text Available The great need for microalgae biomass production in tubular photobioreactors has increased for use in biofuels, pharmaceuticals and even cosmetic applications. In order to better understand the potential applications of this material, it is imperative to know in detail its composition. Ankistrodesmus braunii was cultivated in 3.5 L tubular air-lift photobioreactors using 10 mM sodium nitrate as nitrogen source in batch mode at 60 µmol photons m-2 s-1. The maximum biomass concentration (Xm and the biomass productivity (PX reached at 6th day of cultivation was 1249 ± 72 mg L-1 and 165 ± 13 mg L-1 d-1, respectively. Carbohydrates productivity expressed in terms of glucose, galactose and glucose+galactose (1:1 were 2.57 ± 0.04, 4.12 ± 0.06 and 3.22 ± 0.05 mg L-1 d-1, respectively. Results show a statistical difference that was found between carbohydrate productivity values expressed as glucose, galactose and glucose+galactose (1:1.

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

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.

Anaerobic digestion of solid waste in RAS: Effect of reactor type on the biochemical acidogenic potential (BAP) and assessment of the biochemical methane potential (BMP) by a batch assay

DEFF Research Database (Denmark)

Suhr, Karin Isabel; Letelier-Gordo, Carlos Octavio; Lund, Ivar

2015-01-01

the biochemical acidogenic potential of solid waste from juvenile rainbow trout was evaluated by measuring the yield of volatile fatty acids (VFA) during anaerobic digestion by batch or fed-batch reactor operation at hydrolysis time (HT) / hydraulic retention time (HRT) of 1, 5, or 10 days (and for batch......Anaerobic digestion is a way to utilize the potential energy contained in solid waste produced in recirculating aquaculture systems (RASs), either by providing acidogenic products for driving heterotrophic denitrification on site or by directly producing combustive methane. In this study...

Use of diluted urine for cultivation of Chlorella vulgaris.

Science.gov (United States)

Jaatinen, Sanna; Lakaniemi, Aino-Maija; Rintala, Jukka

2016-01-01

Our aim was to study the biomass growth of microalga Chlorella vulgaris using diluted human urine as a sole nutrient source. Batch cultivations (21 days) were conducted in five different urine dilutions (1:25-1:300), in 1:100-diluted urine as such and with added trace elements, and as a reference, in artificial growth medium. The highest biomass density was obtained in 1:100-diluted urine with and without additional trace elements (0.73 and 0.60 g L(-1), respectively). Similar biomass growth trends and densities were obtained with 1:25- and 1:300-diluted urine (0.52 vs. 0.48 gVSS L(-1)) indicating that urine at dilution 1:25 can be used to cultivate microalgal based biomass. Interestingly, even 1:300-diluted urine contained sufficiently nutrients and trace elements to support biomass growth. Biomass production was similar despite pH-variation from < 5 to 9 in different incubations indicating robustness of the biomass growth. Ammonium formation did not inhibit overall biomass growth. At the beginning of cultivation, the majority of the biomass consisted of living algal cells, while towards the end, their share decreased and the estimated share of bacteria and cell debris increased.

Production of oleic acid ethyl ester catalyzed by crude rice bran (Oryza sativa lipase in a modified fed-batch system: problem and its solution

Directory of Open Access Journals (Sweden)

Indro Prastowo

2015-01-01

Full Text Available A fed-batch system was modified for the enzymatic production of Oleic Acid Ethyl Ester (OAEE using rice bran (Oryza sativa lipase by retaining the substrate molar ratio (ethanol/oleic acid at 2.05: 1 during the reaction. It resulted in an increase in the ester conversion up to 76.8% in the first 6 h of the reaction, and then followed by a decrease from 76.8% to 22.9% in 6 h later. Meanwhile, the production of water in the reaction system also showed a similar trend to the trend of ester production. The water was hypothesized to lead lipase to reverse the reaction which resulted in a decrease in both (water and esters in the last 6 h of the reaction. In order to overcome the problem, zeolite powders (25 and 50 mg/ml were added into the reaction system at 5 h of the reaction. As the result, final ester conversions increased drastically up to 90 - 95.7% (1.17 – 1.24 times. The addition also proved a hypothesis that the water was involved in reducing the ester conversion in the last 6 h of the reaction. Thus, the combination was effective to produce the high final ester conversion.

Bicarbonate-based cultivation of Dunaliella salina for enhancing carbon utilization efficiency.

Science.gov (United States)

Kim, Ga-Yeong; Heo, Jina; Kim, Hee-Sik; Han, Jong-In

2017-08-01

In this study, bicarbonate was proposed as an alternative carbon source to overcome exceedingly low CO 2 fixation efficiency of conventional microalgae cultivation system. 5gL -1 of sodium bicarbonate was found to well support the growth of Dunaliella salina, showing 2.84-fold higher specific growth rate than a bicarbonate-free control. This bicarbonate-fed cultivation also could yield biomass productivity similar to that of CO 2 -based system as long as pH was controlled. While the supplied CO 2 , because of its being a gas, was mostly lost and only 3.59% of it was used for biomass synthesis, bicarbonate was effectively incorporated into the biomass with 91.40% of carbon utilization efficiency. This study showed that the bicarbonate-based microalgae cultivation is indeed possible, and can even become a truly environment-friendly and workable approach, provided that a CO 2 mineralization technology is concomitantly established. Copyright © 2017 Elsevier Ltd. All rights reserved.

Cashew apple juice as microbial cultivation medium for non-immunogenic hyaluronic acid production

OpenAIRE

Oliveira, Adriano H.; Ogrodowski, Cristiane C.; de Macedo, André C.; Santana, Maria Helena A.; Gonçalves, Luciana R.B.

2014-01-01

In this work, natural cashew apple juice was used as cultivation medium as an alternative to substitute brain heart infusion medium. The effect of aeration and juice supplementation with yeast extract on the production of hyaluronic acid in batch fermentation was also investigated. Similar levels of cell mass were obtained in inoculum using cashew apple juice supplemented with yeast extract or the conventional brain heart infusion medium. Fermentation in Erlenmeyer flasks produced low biomass...

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, ...

Extracellular Membrane Vesicles and Phytopathogenicity of Acholeplasma laidlawii PG8

Directory of Open Access Journals (Sweden)

Vladislav M. Chernov

2012-01-01

Full Text Available For the first time, the phytopathogenicity of extracellular vesicles of Acholeplasma laidlawii PG8 (a ubiquitous mycoplasma that is one of the five common species of cell culture contaminants and is a causative agent for phytomycoplasmoses in Oryza sativa L. plants was studied. Data on the ability of extracellular vesicles of Acholeplasma laidlawii PG8 to penetrate from the nutrient medium into overground parts of Oryza sativa L. through the root system and to cause alterations in ultrastructural organization of the plants were presented. As a result of the analysis of ultrathin leaf sections of plants grown in medium with A. laidlawii PG8 vesicles, we detected significant changes in tissue ultrastructure characteristic to oxidative stress in plants as well as their cultivation along with bacterial cells. The presence of nucleotide sequences of some mycoplasma genes within extracellular vesicles of Acholeplasma laidlawii PG8 allowed a possibility to use PCR (with the following sequencing to perform differential detection of cells and bacterial vesicles in samples under study. The obtained data may suggest the ability of extracellular vesicles of the mycoplasma to display in plants the features of infection from the viewpoint of virulence criteria—invasivity, infectivity—and toxigenicity—and to favor to bacterial phytopathogenicity.

Biological treatment of PAH-contaminated sediments in a Sequencing Batch Reactor

International Nuclear Information System (INIS)

Chiavola, Agostina; Baciocchi, Renato; Gavasci, Renato

2010-01-01

The technical feasibility of a sequential batch process for the biological treatment of sediments contaminated by polycyclic aromatic hydrocarbons (PAHs) was evaluated through an experimental study. A bench-scale Sediment Slurry Sequencing Batch Reactor (SS-SBR) was fed with river sediments contaminated by a PAH mixture made by fluorene, anthracene, pyrene and crysene. The process performance was evaluated under different operating conditions, obtained by modifying the influent organic load, the feed composition and the hydraulic residence time. Measurements of the Oxygen Uptake Rates (OURs) provided useful insights on the biological kinetics occurring in the SS-SBR, suggesting the minimum applied cycle time-length of 7 days could be eventually halved, as also confirmed by the trend observed in the volatile solid and total organic carbon data. The removal efficiencies gradually improved during the SS-SBR operation, achieving at the end of the study rather constant removal rates above 80% for both 3-rings PAHs (fluorene and anthracene) and 4-ring PAHs (pyrene and crysene) for an inlet total PAH concentration of 70 mg/kg as dry weight (dw).

Sunki mandarin and Swingle citrumelo as rootstocks for rain-fed cultivation of late-season sweet orange selections in northern São Paulo state, Brazil

Directory of Open Access Journals (Sweden)

Eduardo Augusto Girardi

2017-08-01

Full Text Available ABSTRACT In Brazilian regions affected by the citrus sudden death disease, sweet orange cultivation depends on the use of resistant rootstocks. Rangpur lime was mainly replaced by Swingle citrumelo and Sunki mandarin rootstocks, more drought-sensitive ones. The diversification of scion selections is also desirable aiming at the increasing demand for not from concentrate orange juice (NFC that requires high-quality fruits. In this work, we evaluated the performance of 6 selections of Valencia (IAC, Dom João, Late Burjasot IVIA 35-2, Rhode Red SRA 360, Temprana IVIA 25 and Campbell and Natal IAC sweet oranges grafted onto Swingle citrumelo and Sunki mandarin. The planting occurred in 2001 under rain-fed cultivation in Bebedouro, northern São Paulo state, Brazil. The outline was made through randomized blocks in a 7 × 2 factorial design (selections × rootstock, with 4 replications and 2 trees in unit. Both rootstocks performed well in the region. Sunki mandarin rootstock induced greater tree size and production per plant to the scion selections, 38 and 21%, respectively, plus higher precocity of production compared to Swingle citrumelo. The later determined a greater productive efficiency, as well as a greater percentage of juice in general, albeit with lower concentrations of soluble solids and acidity. Natal IAC, Valencia IAC and Rhode Red Valencia selections presented a higher accumulated production, on average, 218.6 kg∙plant−1 (2004 – 2008, and a higher productive efficiency (kg fruit∙m−3 of canopy due to their smaller tree size. All assessed selections produced fruits with high soluble solids content that were suitable for juice processing.

pH-oscillations in the bromate-sulfite reaction in semibatch and in gel-fed batch reactors

Science.gov (United States)

Poros, Eszter; Kurin-Csörgei, Krisztina; Szalai, István; Rábai, Gyula; Orbán, Miklós

2015-06-01

The simplest bromate oxidation based pH-oscillator, the two component BrO3--SO32- flow system was transformed to operate under semibatch and closed arrangements. The experimental preconditions of the pH-oscillations in semibatch configuration were predicted by model calculations. Using this information as guideline large amplitude (ΔpH˜3), long lasting (11-24 h) pH-oscillations accompanied with only a 20% increase of the volume in the reactor were measured when a mixture of Na2SO3 and H2SO4 was pumped into the solution of BrO3- with a very low rate. Batch-like pH-oscillations, similar in amplitude and period time appeared when the sulfite supply was substituted by its dissolution from a gel layer prepared previously in the reactor in presence of high concentration of Na2SO3. The dissolution vs time curve and the pH-oscillations in the semibatch and closed systems were successfully simulated. Due to the simplicity in composition and in experimental technique, the semibatch and batch-like BrO3--SO32- pH-oscillators may become superior to their CSTR (continuous flow stirred tank reactor) version in some present and future applications.

Production of extracellular protease and glucose uptake in Bacillus clausii in steady-state and transient continuous cultures

DEFF Research Database (Denmark)

Christiansen, Torben; Nielsen, Jens

2002-01-01

The production of the extracellular alkaline protease Savinase(R) (EC 3.4.21.62) and glucose uptake in a non-sporulating strain of Bacillus clausii were investigated by analysing steady-state and transients during continuous cultivations. The specific production rate was found to have an optimum...

SPS batch spacing optimisation

CERN Document Server

Velotti, F M; Carlier, E; Goddard, B; Kain, V; Kotzian, G

2017-01-01

Until 2015, the LHC filling schemes used the batch spac-ing as specified in the LHC design report. The maximumnumber of bunches injectable in the LHC directly dependson the batch spacing at injection in the SPS and hence onthe MKP rise time.As part of the LHC Injectors Upgrade project for LHCheavy ions, a reduction of the batch spacing is needed. In thisdirection, studies to approach the MKP design rise time of150ns(2-98%) have been carried out. These measurementsgave clear indications that such optimisation, and beyond,could be done also for higher injection momentum beams,where the additional slower MKP (MKP-L) is needed.After the successful results from 2015 SPS batch spacingoptimisation for the Pb-Pb run [1], the same concept wasthought to be used also for proton beams. In fact, thanksto the SPS transverse feed back, it was already observedthat lower batch spacing than the design one (225ns) couldbe achieved. For the 2016 p-Pb run, a batch spacing of200nsfor the proton beam with100nsbunch spacing wasreque...

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.

pH-oscillations in the bromate–sulfite reaction in semibatch and in gel-fed batch reactors

Energy Technology Data Exchange (ETDEWEB)

Poros, Eszter; Kurin-Csörgei, Krisztina; Szalai, István; Orbán, Miklós, E-mail: orbanm@chem.elte.hu [Department of Analytical Chemistry, Institute of Chemistry, L. Eötvös University, P.O. Box 32, H-1518 Budapest 112 (Hungary); Rábai, Gyula [Institute of Physical Chemistry, University of Debrecen, P.O. Box 7, H-4010 Debrecen (Hungary)

2015-06-15

The simplest bromate oxidation based pH-oscillator, the two component BrO{sub 3}{sup −}–SO{sub 3}{sup 2–} flow system was transformed to operate under semibatch and closed arrangements. The experimental preconditions of the pH-oscillations in semibatch configuration were predicted by model calculations. Using this information as guideline large amplitude (ΔpH∼3), long lasting (11–24 h) pH-oscillations accompanied with only a 20% increase of the volume in the reactor were measured when a mixture of Na{sub 2}SO{sub 3} and H{sub 2}SO{sub 4} was pumped into the solution of BrO{sub 3}{sup −} with a very low rate. Batch-like pH-oscillations, similar in amplitude and period time appeared when the sulfite supply was substituted by its dissolution from a gel layer prepared previously in the reactor in presence of high concentration of Na{sub 2}SO{sub 3}. The dissolution vs time curve and the pH-oscillations in the semibatch and closed systems were successfully simulated. Due to the simplicity in composition and in experimental technique, the semibatch and batch-like BrO{sub 3}{sup −}–SO{sub 3}{sup 2–} pH-oscillators may become superior to their CSTR (continuous flow stirred tank reactor) version in some present and future applications.

Fibronectin distribution in the extracellular matrix in the cells grown in deuterated media

International Nuclear Information System (INIS)

Buzgariu, Wanda; Caloianu, Maria; Moldovan, Lucia; Stefanescu, I.; Titescu, Gh.

2003-01-01

The aim of this work is the study of the influence of deuterated water upon the synthesis and organization of fibronectin (FN) in extracellular matrices. Changes were evidenced at the level of extracellular matrix in case of embryo fibroblast cultivation in media with different concentrations of heavy water (20%, 40% and 65%). FN was identified in the extracellular matrix by means of indirect immunocytochemical technique, using a secondary antibody coupled with peroxydase. In the presence of heavy water in culture medium, the arrangement and localization of cellular FN showed changes depending on the exposure time, D 2 O concentration in the medium and the FN polymerization step in the extra cellular matrix in correlation with the culture stage of the monolayer. The heavy water determined a strong reduction of the FN amount released by the cells. This reduction was most evident in the 65% D 2 O medium following a 5 day exposure. The FN distribution after 2 day exposure in an early stage with regards to the FN network formation in a the deuterated medium presented a FN pericellular distribution arranged in aggregates. The heavy water can act upon formation of FN fibrils immediately due to solvent role in the FN polymerization process but also indirectly through metabolic processes and so upon the protein synthesis and FN cellular secretion.The FN network arrangement in the cells cultivated in deuterated media as aggregates might be the effect of solvent role played by D 2 O while the quantitative reduction of FN results from perturbation of protein synthesis as well from biochemical synthesis reactions

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.

Denitrifying capability and community dynamics of glycogen accumulating organisms during sludge granulation in an anaerobic-aerobic sequencing batch reactor

Science.gov (United States)

Bin, Zhang; Bin, Xue; Zhigang, Qiu; Zhiqiang, Chen; Junwen, Li; Taishi, Gong; Wenci, Zou; Jingfeng, Wang

2015-08-01

Denitrifying capability of glycogen accumulating organisms (GAOs) has received great attention in environmental science and microbial ecology. Combining this ability with granule processes would be an interesting attempt. Here, a laboratory-scale sequencing batch reactor (SBR) was operated to enrich GAOs and enable sludge granulation. The results showed that the GAO granules were cultivated successfully and the granules had denitrifying capability. The batch experiments demonstrated that all NO3--N could be removed or reduced, some amount of NO2--N were accumulated in the reactor, and N2 was the main gaseous product. SEM analysis suggested that the granules were tightly packed with a large amount of tetrad-forming organisms (TFOs); filamentous bacteria served as the supporting structures for the granules. The microbial community structure of GAO granules was differed substantially from the inoculant conventional activated sludge. Most of the bacteria in the seed sludge grouped with members of Proteobacterium. FISH analysis confirmed that GAOs were the predominant members in the granules and were distributed evenly throughout the granular space. In contrast, PAOs were severely inhibited. Overall, cultivation of the GAO granules and utilizing their denitrifying capability can provide us with a new approach of nitrogen removal and saving more energy.

Production of the {sup 14}C-labeled insecticidal protein Cry1Ab for soil metabolic studies using a recombinant Escherichia coli in small-scale batch fermentations

Energy Technology Data Exchange (ETDEWEB)

Valldor, Petra; Miethling-Graff, Rona; Dockhorn, Susanne; Martens, Rainer; Tebbe, Christoph C. [Federal Research Institute for Rural Areas, Forestry and Fisheries, Braunschweig (Germany). Thuenen Institute (vTI) for Biodiversity

2012-10-15

Insecticidal Cry proteins naturally produced by Bacillus thuringiensis are a major recombinant trait expressed by genetically modified crops. They are released into the soil during and after cropping. The objective of this study was to produce {sup 14}C-labeled Cry1Ab proteins for soil metabolic studies in scope of their environmental risk assessment. Cry1Ab was synthesized as a protoxin by Escherichia coli HB101 pMP in 200-mL liquid batch culture fermentations and purified from inclusion bodies after trypsin digestion. For cultivation, U-{sup 14}C-glycerol was the main carbon source. Inclusion bodies were smaller and Cry1Ab yield was lower when the initial amount of total organic carbon in the cultivation broth was below 6.4 mg C L{sup -1}. Concentrations of 12.6 g {sup 14}C-labeled glycerol L{sup -1} (1 % v/v) resulted in the production of 17.1 mg {sup 14}C-Cry1Ab L{sup -1} cultivation medium. {sup 14}C mass balances showed that approx. 50 % of the label was lost by respiration and 20 % remained in the growth media, while the residual activity was associated with biomass. Depending on the production batch, 0.01 to 0.05 % of the total {sup 14}C originated from Cry1Ab. In the presence of 2.04 MBq {sup 14}C-labeled carbon sources, a specific activity of up to 268 Bq mg{sup -1} {sup 14}C-Cry1Ab was obtained. A more than threefold higher specific activity was achieved with 4.63 MBq and an extended cultivation period of 144 h. This study demonstrates that {sup 14}C-labeled Cry1Ab can be obtained from batch fermentations with E. coli in the presence of a simple {sup 14}C-labeled carbon source. It also provides a general strategy to produce {sup 14}C-labeled proteins useful for soil metabolic studies. (orig.)

Immobilization of yeast inulinase on chitosan beads for the hydrolysis of inulin in a batch system.

Science.gov (United States)

Singh, R S; Singh, R P; Kennedy, J F

2017-02-01

An extracellular inulinase was partially purified by ethanol precipitation and gel exclusion chromatography from a cell free extract of Kluyveromyces marxianus. Partially purified inulinase exhibited 420 IU/mg specific activity and it was immobilized on chitosan beads. Activity yield of immobilized inulinase was optimized with glutaraldehyde concentration (1-5%), glutaraldehyde treatment time (30-240min), enzyme coupling-time (2-16h) and enzyme loading (5-30 IU) as functions. Under the optimized conditions maximum yield 65.5% of immobilized inulinase was obtained. Maximum hydrolysis of inulin 84.5% and 78.2% was observed at 125rpm after 4h by immobilized and free enzyme, respectively. A retention-time of 4h and 5h was found optimal for the hydrolysis of inulin under agitation (125rpm) by free and immobilized enzyme, respectively. The recycling of the developed immobilized biocatalyst was carried out after 5h of inulin hydrolysis in a batch system. The developed immobilized biocatalyst was successfully used for the hydrolysis of inulin for 14 batches. This is the first report on the immobilization of yeast inulinase on chitosan beads for the hydrolysis of inulin in a batch system. Copyright © 2016 Elsevier B.V. All rights reserved.

Development and application of a milliliter-scale bioreactor for continuous microbial cultivations

DEFF Research Database (Denmark)

Bolic, Andrijana

measurementwhere light was sent through the MSBR bottom and sample to a mirror-like surface in the MSBR and returned back to a fiber bundle. Aerobic and anaerobic batch cultivations were performed with Saccharomyces cerevisiae and Lactobacillus paracasei, respectively. A high evaporation rate was experienced...... bioreactor functionality usually comes in regular lab size, which then transforms a smallscale bioreactor platform to a regular size experimental set up. To address this issue, effort was placed in developing 2 push/pull pumps that were able to deliver gas and medium ina controlled manner as a part...

High-level extracellular protein production in Bacillus subtilis using an optimized dual-promoter expression system.

Science.gov (United States)

Zhang, Kang; Su, Lingqia; Duan, Xuguo; Liu, Lina; Wu, Jing

2017-02-20

We recently constructed a Bacillus subtilis strain (CCTCC M 2016536) from which we had deleted the srfC, spoIIAC, nprE, aprE and amyE genes. This strain is capable of robust recombinant protein production and amenable to high-cell-density fermentation. Because the promoter is among the factors that influence the production of target proteins, optimization of the initial promoter, P amyQ from Bacillus amyloliquefaciens, should improve protein expression using this strain. This study was undertaken to develop a new, high-level expression system in B. subtilis CCTCC M 2016536. Using the enzyme β-cyclodextrin glycosyltransferase (β-CGTase) as a reporter protein and B. subtilis CCTCC M 2016536 as the host, nine plasmids equipped with single promoters were screened using shake-flask cultivation. The plasmid containing the P amyQ' promoter produced the greatest extracellular β-CGTase activity; 24.1 U/mL. Subsequently, six plasmids equipped with dual promoters were constructed and evaluated using this same method. The plasmid containing the dual promoter P HpaII -P amyQ' produced the highest extracellular β-CGTase activity (30.5 U/mL) and was relatively glucose repressed. The dual promoter P HpaII -P amyQ' also mediated substantial extracellular pullulanase (90.7 U/mL) and α-CGTase expression (9.5 U/mL) during shake-flask cultivation, demonstrating the general applicability of this system. Finally, the production of β-CGTase using the dual-promoter P HpaII -P amyQ' system was investigated in a 3-L fermenter. Extracellular expression of β-CGTase reached 571.2 U/mL (2.5 mg/mL), demonstrating the potential of this system for use in industrial applications. The dual-promoter P HpaII -P amyQ' system was found to support superior expression of extracellular proteins in B. subtilis CCTCC M 2016536. This system appears generally applicable and is amenable to scale-up.

Statistical process control applied to the liquid-fed ceramic melter process

International Nuclear Information System (INIS)

Pulsipher, B.A.; Kuhn, W.L.

1987-09-01

In this report, an application of control charts to the apparent feed composition of a Liquid-Fed Ceramic Melter (LFCM) is demonstrated by using results from a simulation of the LFCM system. Usual applications of control charts require the assumption of uncorrelated observations over time. This assumption is violated in the LFCM system because of the heels left in tanks from previous batches. Methods for dealing with this problem have been developed to create control charts for individual batches sent to the feed preparation tank (FPT). These control charts are capable of detecting changes in the process average as well as changes in the process variation. All numbers reported in this document were derived from a simulated demonstration of a plausible LFCM system. In practice, site-specific data must be used as input to a simulation tailored to that site. These data directly affect all variance estimates used to develop control charts. 64 refs., 3 figs., 2 tabs

Modelling of Batch Process Operations

DEFF Research Database (Denmark)

Abdul Samad, Noor Asma Fazli; Cameron, Ian; Gani, Rafiqul

2011-01-01

Here a batch cooling crystalliser is modelled and simulated as is a batch distillation system. In the batch crystalliser four operational modes of the crystalliser are considered, namely: initial cooling, nucleation, crystal growth and product removal. A model generation procedure is shown that s...

Extracellular Degradative Enzymes from Pleurotus pulmonarius Cultivated on Various Solid Cellulose- Radioactive Waste Simulates

International Nuclear Information System (INIS)

Abd El-Aziz, S.M.; El-Sayad, H.; Abu El- Soud, S.M.; Awad Alah, O.A.; Eskander, S.B.

2008-01-01

The present work was devoted to search the behavior of some extracellular enzymes secreted by P. pulmonarius during the bioremediation process of some cellulose based solid radioactive waste simulates. Four categories of this group, namely contaminated protective clothes, spent paper, and ruined cotton and mixture of them were subject to the fungal biodegradation and the variations in P. pulmonarius cellulase, xylanase and laccase enzymes activates were followed during three microbial growing stages. In addition, the changes in reducing sugars and total protein as end products of the degradation process were determined. Also the variations in both the secreted enzymes and the metabolism end products were measured as function of exposing the inoculated P. pulmonarius spawns to increasing doses of gamma irradiation(0.0,0.1,0.25,0.5,0.75,1.0,2.0 kGy). Based on the data so far obtained, it could be stated that the extracellular cellulase enzyme and total protein in the degraded substrate were increased throughout the whole incubation period for all types of cellulose based waste. In addition, it have been concluded that the enzymatic activities and consequently the biodegradation of the cellulose based solid radioactive simulates is enhanced by the gamma irradiation up to the dose 0.75 kGy

Spring batch essentials

CERN Document Server

Rao, P Raja Malleswara

2015-01-01

If you are a Java developer with basic knowledge of Spring and some experience in the development of enterprise applications, and want to learn about batch application development in detail, then this book is ideal for you. This book will be perfect as your next step towards building simple yet powerful batch applications on a Java-based platform.

Cultivation of Scenedesmus obliquus in liquid hydrolysate from flash hydrolysis for nutrient recycling

Science.gov (United States)

Barbera, Elena; Sforza, Eleonora; Kumar, Sandeep; Morosinotto, Tomas; Bertucco, Alberto

2016-01-01

The production of biofuels from microalgae is associated with high demands of nutrients (nitrogen and phosphorus) required for growth. Recycling nutrients from the residual biomass is essential to obtain a sustainable production. In this work, the aqueous phase obtained from flash hydrolysis of Scenedesmus sp. was used as cultivation medium for a microalga of the same genus, to assess the feasibility of this technique for nutrient recycling purposes. Batch and continuous cultivations were carried out, to determine growth performances in this substrate compared to standard media, and verify if a stable biomass production could be obtained. In continuous experiments, the effect of hydrolysate inlet concentration and of residence time were assessed to optimize nutrient supply in relation to productivity. Results obtained show that nutrient recycling is feasible by treating biomass with flash hydrolysis, and Scenedesmus is capable of recycling large amounts of recovered nutrients. PMID:26868157

Monte Carlo simulation on kinetics of batch and semi-batch free radical polymerization

KAUST Repository

Shao, Jing; Tang, Wei; Xia, Ru; Feng, Xiaoshuang; Chen, Peng; Qian, Jiasheng; Song, Changjiang

2015-01-01

experimental and simulation studies, we showed the capability of our Monte Carlo scheme on representing polymerization kinetics in batch and semi-batch processes. Various kinetics information, such as instant monomer conversion, molecular weight

Evaluation of biomass production in unleaded gasoline and BTEX-fed batch reactors.

Science.gov (United States)

Acuna-Askar, K; Englande, A J; Ramirez-Medrano, A; Coronado-Guardiola, J E; Chavez-Gomez, B

2003-01-01

BTEX removal under aerobic conditions by unleaded gasoline acclimated biomass and BTEX acclimated biomass, and the effect of surfactant on BTEX biodegradation were evaluated. The effect of BTEX concentration as the sole source of carbon for biomass acclimation and the effect of yeast extract on cell growth in unleaded gasoline-fed reactors were also evaluated. For the unleaded gasoline acclimated biomass, benzene was shown the most recalcitrant among all BTEX, followed by o-xylene and toluene with 16-23%, 35-41% and 57-69% biodegradation, respectively. Ethylbenzene was consistently the fastest BTEX chemical removed with 99% biodegradation for the four bioreactor acclimated biomasses tested. For the 1,200 ppm BTEX acclimated biomass, benzene showed the highest removal efficiency (99%) among the four biomass environmental conditions tested, along with 99% toluene and 99% ethylbenzene biodegradation. O-xylene showed 92-94% removal. In all bioassays tested Tergitol NP-10 was fully removed, and did not have a substantial effect on BTEX biodegradation at the end of a 10-day evaluation.

Antibacterial Activity of Extracellular Protease Isolated From an Algicolous Fungus Xylaria psidii KT30 Against Gram-Positive Bacteria

Directory of Open Access Journals (Sweden)

Taufik Indarmawan

2016-04-01

Full Text Available Infectious diseases became more serious problem for public health in recent years. Although existing antibacterial drugs have been relatively effective, they do not rule out the emergence of resistance to the drug. Therefore, the intensive exploration of new bioactive compounds from natural, especially peptide compounds began in recent decades in order-handling infection. This study aimed to isolate, purify and test the potential application of Xylaria psidii KT30 extracellular protease as antibacterial agent against Gram-positive bacteria. X. psidii KT30, a marine fungus isolated from red seaweed Kappaphycus alvarezii showed antibacterial activity against Bacillus subtilis and Staphylococcus aureus. Antibacterial compounds of this fungus were predicted as a group of proteases. Extracellular protease exhibited an optimum activity when potato dextrose broth was used as cultivation medium. Furthermore, the highest activity of these proteases was found on fungal extract after day 15 of cultivation with value of 2.33 ± 0.19 U/mL. The partial purification of proteases using G-75 column chromatography resulted in 2 groups of fractions and showed protease activity based on zymogram assay. The extracellular proteases obtained from those fractions have 3 patterns of molecular mass based on sodium dodecyl sulfate–polyacrylamide gel electrophoresis which are 56.62, 89.12, 162.18 kDa.

Acid protease and formation of multiple forms of glycoamylase in batch and continuous cultures of Aspergillus niger

DEFF Research Database (Denmark)

Aalbæk, Thomas; Reeslev, Morten; Jensen, Bo

2002-01-01

In order to identify factors responsible for production of multiple forms of glucoamylase (GA) by Aspergillus niger Bo-1, the fungus was cultured in both complex and defined media in pH-controlled batch fermenters and chemostats. At all culture conditions three forms of GA were produced...... degradation of the GA forms at low pH. It was concluded that the observed modifications of the extracellular profile of GA isoforms in A. niger Bo-1 are due to changes in pH and medium composition....

Monte Carlo simulation on kinetics of batch and semi-batch free radical polymerization

KAUST Repository

Shao, Jing

2015-10-27

Based on Monte Carlo simulation technology, we proposed a hybrid routine which combines reaction mechanism together with coarse-grained molecular simulation to study the kinetics of free radical polymerization. By comparing with previous experimental and simulation studies, we showed the capability of our Monte Carlo scheme on representing polymerization kinetics in batch and semi-batch processes. Various kinetics information, such as instant monomer conversion, molecular weight, and polydispersity etc. are readily calculated from Monte Carlo simulation. The kinetic constants such as polymerization rate k p is determined in the simulation without of “steady-state” hypothesis. We explored the mechanism for the variation of polymerization kinetics those observed in previous studies, as well as polymerization-induced phase separation. Our Monte Carlo simulation scheme is versatile on studying polymerization kinetics in batch and semi-batch processes.

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.

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...

Data-driven batch schuduling

Energy Technology Data Exchange (ETDEWEB)

Bent, John [Los Alamos National Laboratory; Denehy, Tim [GOOGLE; Arpaci - Dusseau, Remzi [UNIV OF WISCONSIN; Livny, Miron [UNIV OF WISCONSIN; Arpaci - Dusseau, Andrea C [NON LANL

2009-01-01

In this paper, we develop data-driven strategies for batch computing schedulers. Current CPU-centric batch schedulers ignore the data needs within workloads and execute them by linking them transparently and directly to their needed data. When scheduled on remote computational resources, this elegant solution of direct data access can incur an order of magnitude performance penalty for data-intensive workloads. Adding data-awareness to batch schedulers allows a careful coordination of data and CPU allocation thereby reducing the cost of remote execution. We offer here new techniques by which batch schedulers can become data-driven. Such systems can use our analytical predictive models to select one of the four data-driven scheduling policies that we have created. Through simulation, we demonstrate the accuracy of our predictive models and show how they can reduce time to completion for some workloads by as much as 80%.

Highly active promoters and native secretion signals for protein production during extremely low growth rates in Aspergillus niger.

Science.gov (United States)

Wanka, Franziska; Arentshorst, Mark; Cairns, Timothy C; Jørgensen, Thomas; Ram, Arthur F J; Meyer, Vera

2016-08-20

The filamentous ascomycete Aspergillus niger is used in many industrial processes for the production of enzymes and organic acids by batch and fed-batch cultivation. An alternative technique is continuous cultivation, which promises improved yield and optimized pipeline efficiency. In this work, we have used perfusion (retentostat) cultivation to validate two promoters that are suitable for A. niger continuous cultivation of industrially relevant products. Firstly, promoters of genes encoding either an antifungal protein (Panafp) or putative hydrophobin (PhfbD) were confirmed as active throughout retentostat culture by assessing mRNA and protein levels using a luciferase (mluc) reporter system. This demonstrated the anafp promoter mediates a high but temporally variable expression profile, whereas the hfbD promoter mediates a semi-constant, moderate-to-high protein expression during retentostat culture. In order to assess whether these promoters were suitable to produce heterologous proteins during retentostat cultivation, the secreted antifungal protein (AFP) from Aspergillus giganteus, which has many potential biotechnological applications, was expressed in A. niger during retentostat cultivation. Additionally, this assay was used to concomitantly validate that native secretion signals encoded in anafp and hfbD genes can be harnessed for secretion of heterologous proteins. Afp mRNA and protein abundance were comparable to luciferase measurements throughout retentostat cultivation, validating the use of Panafp and PhfbD for perfusion cultivation. Finally, a gene encoding the highly commercially relevant thermal hysteresis protein (THP) was expressed in this system, which did not yield detectable protein. Both hfbD and anafp promoters are suitable for production of useful products in A. niger during perfusion cultivation. These findings provide a platform for further optimisations for high production of heterologous proteins with industrial relevance.

Genetic Algorithm Tuning of PID Controller in Smith Predictor for Glucose Concentration Control

OpenAIRE

Tsonyo Slavov; Olympia Roeva

2011-01-01

This paper focuses on design of a glucose concentration control system based on nonlinear model plant of E. coli MC4110 fed-batch cultivation process. Due to significant time delay in real time glucose concentration measurement, a correction is proposed in glucose concentration measurement and a Smith predictor (SP) control structure based on universal PID controller is designed. To reduce the influence of model error in SP structure the estimate of measured glucose concentration is used. For...

Production of FucoPol by Enterobacter A47 using waste tomato paste by-product as sole carbon source.

Science.gov (United States)

Antunes, Sílvia; Freitas, Filomena; Sevrin, Chantal; Grandfils, Christian; Reis, Maria A M

2017-03-01

Out-of-specification tomato paste, a by-product from the tomato processing industry, was used as the sole substrate for cultivation of the bacterium Enterobacter A47 and production of FucoPol, a value-added fucose-rich extracellular polysaccharide. Among the different tested fed-batch strategies, pH-stat, DO-stat and continuous substrate feeding, the highest production (8.77gL -1 ) and overall volumetric productivity (2.92gL -1 d -1 ) were obtained with continuous substrate feeding at a constant flow rate of 11gh -1 . The polymer produced had the typical FucoPol composition (37mol% fucose, 27mol% galactose, 23mol% glucose and 12mol% glucuronic acid, with an acyl groups content of 13wt%). The average molecular weight was 4.4×10 6 Da and the polydispersity index was 1.2. This study demonstrated that out-of-specification tomato paste is a suitable low-cost substrate for the production of FucoPol, thus providing a route for the valorization of this by-product into a high-value microbial product. Copyright © 2016 Elsevier Ltd. All rights reserved.

Phenotypical analysis of the Lactobacillus rhamnosus GG fimbrial spaFED operon: surface expression and functional characterization of recombinant SpaFED pili in Lactococcus lactis.

Directory of Open Access Journals (Sweden)

Johanna Rintahaka

Full Text Available A noticeable genomic feature of many piliated Gram-positive bacterial species is the presence of more than one pilus-encoding operon. Paradigmatically, the gut-adapted Lactobacillus rhamnosus GG strain contains two different fimbrial operons in its genome. However, whereas one of these operons (called spaCBA is encoding for the functionally mucus-/collagen-binding SpaCBA pilus, for the other operon (called spaFED any native expression of the SpaFED-called pili is still the subject of some uncertainty. Irrespective of such considerations, we decided it would be of relevance or interest to decipher the gross structure of this pilus type, and as well assess its functional capabilities for cellular adhesion and immunostimulation. For this, and by following the approach we had used previously to explicate the immuno-properties of SpaCBA pili, we constructed nisin-inducible expression clones producing either wild-type or SpaF pilin-deleted surface-assembled L. rhamnosus GG SpaFED pili on Lactococcus lactis cells. Using these piliated lactococcal constructs, we found that the pilin-polymerized architecture of a recombinant-produced SpaFED pilus coincides with sequence-based functional predictions of the related pilins, and in fact is prototypical of those other sortase-dependent pilus-like structures thus far characterized for piliated Gram-positive bacteria. Moreover, we confirmed that among the different pilin subunits encompassing spaFED operon-encoded pili, the SpaF pilin is a main adhesion determinant, and when present in the assembled structure can mediate pilus binding to mucus, certain extracellular matrix proteins, and different gut epithelial cell lines. However, somewhat unexpectedly, when recombinant SpaFED pili are surface-attached, we found that they could not potentiate the existing lactococcal cell-induced immune responses so elicited from intestinal- and immune-related cells, but rather instead, they could dampen them. Accordingly, we

Phenotypical analysis of the Lactobacillus rhamnosus GG fimbrial spaFED operon: surface expression and functional characterization of recombinant SpaFED pili in Lactococcus lactis.

Science.gov (United States)

Rintahaka, Johanna; Yu, Xia; Kant, Ravi; Palva, Airi; von Ossowski, Ingemar

2014-01-01

A noticeable genomic feature of many piliated Gram-positive bacterial species is the presence of more than one pilus-encoding operon. Paradigmatically, the gut-adapted Lactobacillus rhamnosus GG strain contains two different fimbrial operons in its genome. However, whereas one of these operons (called spaCBA) is encoding for the functionally mucus-/collagen-binding SpaCBA pilus, for the other operon (called spaFED) any native expression of the SpaFED-called pili is still the subject of some uncertainty. Irrespective of such considerations, we decided it would be of relevance or interest to decipher the gross structure of this pilus type, and as well assess its functional capabilities for cellular adhesion and immunostimulation. For this, and by following the approach we had used previously to explicate the immuno-properties of SpaCBA pili, we constructed nisin-inducible expression clones producing either wild-type or SpaF pilin-deleted surface-assembled L. rhamnosus GG SpaFED pili on Lactococcus lactis cells. Using these piliated lactococcal constructs, we found that the pilin-polymerized architecture of a recombinant-produced SpaFED pilus coincides with sequence-based functional predictions of the related pilins, and in fact is prototypical of those other sortase-dependent pilus-like structures thus far characterized for piliated Gram-positive bacteria. Moreover, we confirmed that among the different pilin subunits encompassing spaFED operon-encoded pili, the SpaF pilin is a main adhesion determinant, and when present in the assembled structure can mediate pilus binding to mucus, certain extracellular matrix proteins, and different gut epithelial cell lines. However, somewhat unexpectedly, when recombinant SpaFED pili are surface-attached, we found that they could not potentiate the existing lactococcal cell-induced immune responses so elicited from intestinal- and immune-related cells, but rather instead, they could dampen them. Accordingly, we have now provided

Characterization of Extracellular Penicilin G Acylase Produced by A New Local Strain of Bacillus subtilis BAC4

Directory of Open Access Journals (Sweden)

SUPARTONO

2008-06-01

Full Text Available Penicillin G acylase (PGA which catalyses penicillin G hydrolysis reaction is a key enzyme for the industrial production of penicilin G derivatives used in therapeutics. A new local strain of Bacillus subtilis BAC4 was found capable of producing extracellular PGA. However, characteristics of this extracellular PGA are not known. The goal of this research was to characterize the extracellular PGA produced by B. subtilis BAC4. Enzyme production was carried out by batch fermentation, followed by enzyme purification and characterization of the PGA. The PGA activity was determined by the Kornfeld method, with optimal activity for hydrolysing penicillin G observed at 43 °C and pH 8.5. The activation energy of penicillin G hydrolysis by the PGA of B. subtilis BAC4 was determined as 4.9 kcal.mol−1 and Vmax and Km values were found to be 0.7 μmole.min−1.mg−1 and 3.5 mM respectively. PGA catalytic activity was competitively inhibited by phenylacetic acid with an inhibition constant, Ki(PAA, of 347.2 mM. It was concluded that the extracellular PGA of B. subtilis BAC4 can hydrolyse penicillin G efficiently.

Characterization of Extracellular Penicilin G Acylase Produced by A New Local Strain of Bacillus subtilis BAC4

Directory of Open Access Journals (Sweden)

SUPARTONO

2008-06-01

Full Text Available Penicillin G acylase (PGA which catalyses penicillin G hydrolysis reaction is a key enzyme for the industrial production of penicilin G derivatives used in therapeutics. A new local strain of Bacillus subtilis BAC4 was found capable of producing extracellular PGA. However, characteristics of this extracellular PGA are not known. The goal of this research was to characterize the extracellular PGA produced by B. subtilis BAC4. Enzyme production was carried out by batch fermentation, followed by enzyme purification and characterization of the PGA. The PGA activity was determined by the Kornfeld method, with optimal activity for hydrolysing penicillin G observed at 43 oC and pH 8.5. The activation energy of penicillin G hydrolysis by the PGA of B. subtilis BAC4 was determined as 4.9 kcal.mol-1 and Vmax and Km values were found to be 0.7 µmole.min-1.mg-1 and 3.5 mM respectively. PGA catalytic activity was competitively inhibited by phenylacetic acid with an inhibition constant, Ki(PAA, of 347.2 mM. It was concluded that the extracellular PGA of B. subtilis BAC4 can hydrolyse penicillin G efficiently.

The effect of toxic carbon source on the reaction of activated sludge in the batch reactor.

Science.gov (United States)

Wu, Changyong; Zhou, Yuexi; Zhang, Siyu; Xu, Min; Song, Jiamei

2018-03-01

The toxic carbon source can cause higher residual effluent dissolved organic carbon than easily biodegraded carbon source in activated sludge process. In this study, an integrated activated sludge model is developed as the tool to understand the mechanism of toxic carbon source (phenol) on the reaction, regarding the carbon flows during the aeration period in the batch reactor. To estimate the toxic function of phenol, the microbial cells death rate (k death ) is introduced into the model. The integrated model was calibrated and validated by the experimental data and it was found the model simulations matched the all experimental measurements. In the steady state, the toxicity of phenol can result in higher microbial cells death rate (0.1637 h -1 vs 0.0028 h -1 ) and decay rate coefficient of biomass (0.0115 h -1 vs 0.0107 h -1 ) than acetate. In addition, the utilization-associated products (UAP) and extracellular polymeric substances (EPS) formation coefficients of phenol are higher than that of acetate, indicating that more carbon flows into the extracellular components, such as soluble microbial products (SMP), when degrading toxic organics. In the non-steady state of feeding phenol, the yield coefficient for growth and maximum specific growth rate are very low in the first few days (1-10 d), while the decay rate coefficient of biomass and microbial cells death rate are relatively high. The model provides insights into the difference of the dynamic reaction with different carbon sources in the batch reactor. Copyright © 2017 Elsevier Ltd. All rights reserved.

Enhancing the Production of Polyhydroxyalkanoate Biopolymer by Azohydromonas Australica Using a Simple Empty and Fill Bioreactor Cultivation Strategy

OpenAIRE

G. Gahlawat; A. K. Srivastava

2018-01-01

Polyhydroxyalkanoates (PHAs) are biodegradable polymers which are considered as an effective alternative for conventional plastics due to their mechanical properties similar to the latter. However, widespread use of these polymers is still hampered due to their high cost of production. This shortcoming could partly be resolved by obtaining high yields and productivity. In the present study, a drain-and-fill strategy of repeated-batch cultivation was adopted for the enhanced production of p...

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)

Production of extracellular lipase by the phytopathogenic fungus Fusarium solani FS1

OpenAIRE

Maia, Maria de Mascena Diniz; Morais, Marcia Maria Camargo de; Morais Jr., Marcos Antonio de; Melo, Eduardo Henrique Magalhães; Lima Filho, José Luiz de

1999-01-01

A Brazilian strain of Fusarium solani was tested for extracellular lipase production in peptone-olive oil medium. The fungus produced 10,500 U.l-1 of lipase after 72 hours of cultivation at 25oC in shake-flask at 120 rpm in a medium containing 3% (w/v) peptone plus 0.5% (v/v) olive oil. Glucose (1% w/v) was found to inhibit the inductive effect of olive oil. Peptone concentrations below 3% (w/v) resulted in a reduced lipase production while increased olive oil concentration (above 0.5%) did n...

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.

Cellulase enzyme production during continuous culture growth of Sporotrichum (Chrysosporium) thermophile

Energy Technology Data Exchange (ETDEWEB)

Cossar, D; Canevascini, G

1986-07-01

The cellulolytic fungus Sporotrichum (Chrysosporium) thermophile produces an extracellular cellobiose dehydrogenase during batch culture on cellulose or cellobiose. In chemostat culture at pH 5.6 on cellobiose this enzyme was produced in parallel with endo-cellulase. At pH 5.0 in continuous or fed-batch culture such a pattern was not evident. At constant growth rate in a chemostat with varying pH, activity of these enzymes was found to be poorly correlated. Thus the induction of cellobiose dehydrogenase shows a dependence on pH and cellobiose concentration which is different to that for endo-cellulase. The natural inducer of these enzymes and the role of cellubiose dehydrogenase remain to be elucidated.

Economic Evaluation for Integrated Use of Glyphosate Herbicide and Tillage Combinations Applied before Sowing of Rain-Fed Wheat (Triticum Aestivum L.)

International Nuclear Information System (INIS)

Ali, S.; Malik, M. A.; Ansar, M.; Qureshi, R.

2016-01-01

Low average yield, scarce soil moisture and less soil fertility are major problems of rain-fed wheat. Economic feasibility of different tillage systems integrated with glyphosate herbicide and wheat crop productivity was determined through field experiments conducted at the University Research Farm of Pir Mehr Ali Shah Arid Agriculture University Rawalpindi, Pakistan during summer and winter seasons of 2012-13 and 2013-14. Different combinations of tillage and glyphosate herbicide were used in the fallow period (summer season) that were consisted of following treatments viz. T1 = 1 Mould board Plowing + 8 Cultivations, T2 = No-Till + Glyphosate, T3 = 1 Mould board Plowing + Glyphosate, T4 = 1 MB Plowing + 4 Cultivations, T5 = 1 Disc Harrowing + Glyphosate, T6 = 1 Disc Harrowing + 4 Cultivations and T7 = 1 Chiseling + Glyphosate. Results showed that the highest yield viz. 3.5132, 3.1242 t ha-1 were obtained in the case of conventional tillage (T1) and reduced tillage (T4), respectively with a net profit of 888.92 and 839.35 $ ha-1. The yield was positively affected by tillage intensity. In conclusion, T1 is recommended for getting maximum net return from wheat grown in rain-fed areas of Pakistan. (author)

BATCH-GE: Batch analysis of Next-Generation Sequencing data for genome editing assessment

Science.gov (United States)

Boel, Annekatrien; Steyaert, Woutert; De Rocker, Nina; Menten, Björn; Callewaert, Bert; De Paepe, Anne; Coucke, Paul; Willaert, Andy

2016-01-01

Targeted mutagenesis by the CRISPR/Cas9 system is currently revolutionizing genetics. The ease of this technique has enabled genome engineering in-vitro and in a range of model organisms and has pushed experimental dimensions to unprecedented proportions. Due to its tremendous progress in terms of speed, read length, throughput and cost, Next-Generation Sequencing (NGS) has been increasingly used for the analysis of CRISPR/Cas9 genome editing experiments. However, the current tools for genome editing assessment lack flexibility and fall short in the analysis of large amounts of NGS data. Therefore, we designed BATCH-GE, an easy-to-use bioinformatics tool for batch analysis of NGS-generated genome editing data, available from https://github.com/WouterSteyaert/BATCH-GE.git. BATCH-GE detects and reports indel mutations and other precise genome editing events and calculates the corresponding mutagenesis efficiencies for a large number of samples in parallel. Furthermore, this new tool provides flexibility by allowing the user to adapt a number of input variables. The performance of BATCH-GE was evaluated in two genome editing experiments, aiming to generate knock-out and knock-in zebrafish mutants. This tool will not only contribute to the evaluation of CRISPR/Cas9-based experiments, but will be of use in any genome editing experiment and has the ability to analyze data from every organism with a sequenced genome. PMID:27461955

Effects of chlortetracycline amended feed on anaerobic sequencing batch reactor performance of swine manure digestion.

Science.gov (United States)

Dreher, Teal M; Mott, Henry V; Lupo, Christopher D; Oswald, Aaron S; Clay, Sharon A; Stone, James J

2012-12-01

The effects of antimicrobial chlortetracycline (CTC) on the anaerobic digestion (AD) of swine manure slurry using anaerobic sequencing batch reactors (ASBRs) was investigated. Reactors were loaded with manure collected from pigs receiving CTC and no-antimicrobial amended diets at 2.5 g/L/d. The slurry was intermittently fed to four 9.5L lab-scale anaerobic sequencing batch reactors, two with no-antimicrobial manure, and two with CTC-amended manure, and four 28 day ASBR cycles were completed. The CTC concentration within the manure was 2 8 mg/L immediately after collection and 1.02 mg/L after dilution and 250 days of storage. CTC did not inhibit ASBR biogas production extent, however the volumetric composition of methane was significantly less (approximately 13% and 15% for cycles 1 and 2, respectively) than the no-antimicrobial through 56 d. CTC decreased soluble chemical oxygen demand and acetic acid utilization through 56 d, after which acclimation to CTC was apparent for the duration of the experiment. Copyright © 2012 Elsevier Ltd. All rights reserved.

Production of sup 5 sup 5 Co via the sup 5 sup 4 Fe(d, n)-process and excitation functions of sup 5 sup 4 Fe(d, t) sup 5 sup 3 Fe and sup 5 sup 4 Fe(d, alpha) sup 5 sup 2 sup m Mn reactions from threshold up to 13.8 MeV

CERN Document Server

Zaman, M R; Qaim, S M

2003-01-01

For production of the medically interesting beta; sup + -emitter sup 5 sup 5 Co (T sup 1 sup / sup 2 = 17.6 h) via the sup 5 sup 4 Fe(d, n)-reaction, 91.6% enriched sup 5 sup 4 Fe sub 2 O sub 3 , mixed with Al powder, was pressed to a pellet which could be irradiated with 14 MeV deuterons at 4 mu A in a water-cooled target system. A separation method was developed which led to > 99.9% pure sup 5 sup 5 Co and allowed a recovery of the enriched target material. For a target thickness of E sub d = 12.6 -> 5 MeV, the experimental thick target yield of sup 5 sup 5 Co after chemical separation amounted to about 13 MBq/mu A.h, which is about 60% of the theoretical value. In a 3 h irradiation at 4 mu A, the batch yield of sup 5 sup 5 Co achieved was 160 MBq (4.3 mCi). An 8 h irradiation could lead to a batch yield of about 400 MBq. The sup 5 sup 4 Fe(d, n) reaction leads to the highest purity sup 5 sup 5 Co but it is essential that the isotopic enrichment of the target is not less than 90%. In addition to the product...

Cultivo da levedura Phaffia rhodozyma (Xanthophyllomyces dendrorhous em processo descontínuo alimentado para produção de astaxantina Cultivation of Phaffia rhodozyma (Xanthophyllomyces dendrorhous yeast in discontinuous system to obtain astaxanthin

Directory of Open Access Journals (Sweden)

Miriam Blümel Chociai

2002-12-01

Full Text Available A levedura Phaffia rhodozyma, produtora de astaxantina, pigmento carotenóide largamente empregado na aqüicultura de peixes e crustáceos, pode ser eficientemente cultivada num meio de cultura de baixo custo, à base de caldo de cana diluído 1:10 e uréia a 1 g/L. No entanto, a produção de biomassa e a formação do carotenóide sofrem a inibição pelo substrato (efeito "Crabtree", limitando desta forma a utilização do caldo de cana com concentrações da fonte de carbono superiores a 20 g/L, importante consideração na produção industrial de astaxantina. No presente trabalho, o cultivo da levedura P. rhodozyma foi realizado em processo descontínuo alimentado, no qual se obteve produtividade volumétrica de 0,024 mg astaxantina/L.h. em relação aos 0,013 mg astaxantina/L.h. obtidos no cultivo controle, que não sofreu alimentação da fonte de carbono.The yeast Phaffia rhodozyma produces astaxanthin, a carotenoid pigment widely applied in fish and crustaceous cultivation. This yeast can be efficiently cultured in a low cost medium, sugar cane broth diluted 1:10 and supplemented with 1 g/L urea. However, the biomass and astaxanthin production undergo inhibition by the substrate (Crabtree effect, limiting the utilization of sugar cane broth up to 20 g/L total sugar concentration. Therefore, this effect must be considered during the industrial production of astaxanthin. In the present work, using fed batch system to cultivate P. rhodozyma we were able to obtain 0.024 mg astaxanthin/l.h compared to 0.013 mg astaxanthin/l.h obtained by the discontinuous cultivation system.

Medication waste reduction in pediatric pharmacy batch processes.

Science.gov (United States)

Toerper, Matthew F; Veltri, Michael A; Hamrock, Eric; Mollenkopf, Nicole L; Holt, Kristen; Levin, Scott

2014-04-01

To inform pediatric cart-fill batch scheduling for reductions in pharmaceutical waste using a case study and simulation analysis. A pre and post intervention and simulation analysis was conducted during 3 months at a 205-bed children's center. An algorithm was developed to detect wasted medication based on time-stamped computerized provider order entry information. The algorithm was used to quantify pharmaceutical waste and associated costs for both preintervention (1 batch per day) and postintervention (3 batches per day) schedules. Further, simulation was used to systematically test 108 batch schedules outlining general characteristics that have an impact on the likelihood for waste. Switching from a 1-batch-per-day to a 3-batch-per-day schedule resulted in a 31.3% decrease in pharmaceutical waste (28.7% to 19.7%) and annual cost savings of $183,380. Simulation results demonstrate how increasing batch frequency facilitates a more just-in-time process that reduces waste. The most substantial gains are realized by shifting from a schedule of 1 batch per day to at least 2 batches per day. The simulation exhibits how waste reduction is also achievable by avoiding batch preparation during daily time periods where medication administration or medication discontinuations are frequent. Last, the simulation was used to show how reducing batch preparation time per batch provides some, albeit minimal, opportunity to decrease waste. The case study and simulation analysis demonstrate characteristics of batch scheduling that may support pediatric pharmacy managers in redesign toward minimizing pharmaceutical waste.

Batch-to-batch quality consistency evaluation of botanical drug products using multivariate statistical analysis of the chromatographic fingerprint.

Science.gov (United States)

Xiong, Haoshu; Yu, Lawrence X; Qu, Haibin

2013-06-01

Botanical drug products have batch-to-batch quality variability due to botanical raw materials and the current manufacturing process. The rational evaluation and control of product quality consistency are essential to ensure the efficacy and safety. Chromatographic fingerprinting is an important and widely used tool to characterize the chemical composition of botanical drug products. Multivariate statistical analysis has showed its efficacy and applicability in the quality evaluation of many kinds of industrial products. In this paper, the combined use of multivariate statistical analysis and chromatographic fingerprinting is presented here to evaluate batch-to-batch quality consistency of botanical drug products. A typical botanical drug product in China, Shenmai injection, was selected as the example to demonstrate the feasibility of this approach. The high-performance liquid chromatographic fingerprint data of historical batches were collected from a traditional Chinese medicine manufacturing factory. Characteristic peaks were weighted by their variability among production batches. A principal component analysis model was established after outliers were modified or removed. Multivariate (Hotelling T(2) and DModX) control charts were finally successfully applied to evaluate the quality consistency. The results suggest useful applications for a combination of multivariate statistical analysis with chromatographic fingerprinting in batch-to-batch quality consistency evaluation for the manufacture of botanical drug products.

Cadmium removal using Cladophora in batch, semi-batch and flow reactors.

Science.gov (United States)

Sternberg, Steven P K; Dorn, Ryan W

2002-02-01

This study presents the results of using viable algae to remove cadmium from a synthetic wastewater. In batch and semi-batch tests, a local strain of Cladophora algae removed 80-94% of the cadmium introduced. The flow experiments that followed were conducted using non-local Cladophora parriaudii. Results showed that the alga removed only 12.7(+/-6.4)% of the cadmium introduced into the reactor. Limited removal was the result of insufficient algal quantities and poor contact between the algae and cadmium solution.

Noise Reduction of Measurement Data using Linear Digital Filters

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Hitzmann B.

2007-12-01

Full Text Available In this paper Butterworth, Chebyshev (Type I and II and Elliptic digital filters are designed for signal noise reduction. On-line data measurements of substrate concentration from E. coli fed-batch cultivation process are used. Application of the designed filters leads to a successful noise reduction of on-line glucose measurements. The digital filters presented here are simple, easy to implement and effective - the used filters allow for a smart compromise between signal information and noise corruption.

Kubernetes as a batch scheduler

OpenAIRE

Souza, Clenimar; Brito Da Rocha, Ricardo

2017-01-01

This project aims at executing a CERN batch use case using Kubernetes, in order to figure out what are the advantages and disadvantages, as well as the functionality that can be replicated or is missing. The reference for the batch system is the CERN Batch System, which uses HTCondor. Another goal of this project is to evaluate the current status of federated resources in Kubernetes, in comparison to the single-cluster API resources. Finally, the last goal of this project is to implement buil...

Extracellular laccase production and phenolic degradation by an olive mill wastewater isolate

Directory of Open Access Journals (Sweden)

R. Kumar

2018-03-01

Full Text Available Olive mill wastewater (OMWW presents a challenge to the control of effluents due to the presence of a high organic load, antimicrobial agents (monomeric-polymeric phenols, volatile acids, polyalcohols, and tannins, salinity and acidity. In this study, the production of extracellular laccase, monomeric or polymeric phenol, from an OMWW isolate based on its ability to biodegrade phenols and gallic acid as a model of phenolic compounds in OMWW was investigated. Phylogenetic analysis of the 16S RNA gene sequences identified the bacterial isolate (Acinetobacter REY as being closest to Acinetobacter pittii. This isolate exhibited a constitutive production of extracellular laccase with an activity of 1.5 and 1.3 U ml/L when supplemented with the inducers CuSO4 and CuSO4+phenols, respectively. Batch experiments containing minimal media supplemented with phenols or gallic acid as the sole carbon and energy source were performed in order to characterize their phenolic biodegradability. Acinetobacter REY was capable of biodegrading up to 200 mg/L of phenols and gallic acid both after 10 h and 72 h, respectively.

High-Temperature Induced Changes of Extracellular Metabolites in Pleurotus ostreatus and Their Positive Effects on the Growth of Trichoderma asperellum.

Science.gov (United States)

Qiu, Zhiheng; Wu, Xiangli; Zhang, Jinxia; Huang, Chenyang

2018-01-01

Pleurotus ostreatus is a widely cultivated edible fungus in China. Green mold disease of P. ostreatus which can seriously affect yield is a common disease during cultivation. It occurs mostly after P. ostreatus mycelia have been subjected to high temperatures. However, little information is available on the relationship between high temperature and green mold disease. The aim of this study is to prove that extracellular metabolites of P. ostreatus affected by high temperature can promote the growth of Trichoderma asperellum . After P. ostreatus mycelia was subjected to high temperature, the extracellular fluid of P. ostreatus showed a higher promoting effect on mycelial growth and conidial germination of T. asperellum . The thiobarbituric acid reactive substance (TBARS) content reached the maximum after 48 h at 36°C. A comprehensive metabolite profiling strategy involving gas chromatography-mass spectrometry (GC/MS) combined with liquid chromatography-mass spectrometry (LC/MS) was used to analyze the changes of extracellular metabolites in response to high temperature. A total of 141 differential metabolites were identified, including 84.4% up-regulated and 15.6% down-regulated. Exogenous metabolites whose concentrations were increased after high temperature were randomly selected, and nearly all of them were able to promote the mycelial growth and conidial germination of T. asperellum . The combination of all selected exogenous metabolites also has the promotion effects on the mycelial growth and conidial germination of T. asperellum in a given concentration range in vitro . Overall, these results provide a first view that high temperature affects the extracellular metabolites of P. ostreatus , and the extensive change in metabolites promotes T. asperellum growth.

BatchJS: Implementing Batches in JavaScript

NARCIS (Netherlands)

D. Kasemier

2014-01-01

htmlabstractNone of our popular programming languages know how to handle distribution well. Yet our programs interact more and more with each other and our data resorts in databases and web services. Batches are a new addition to languages that can finally bring native support for distribution to

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.

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.

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

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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

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

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.

Improving Former Shifted Cultivation Land Using Wetland Cultivation in Kapuas District, Central Kalimantan

Directory of Open Access Journals (Sweden)

Wahyudi Wahyudi

2016-06-01

Full Text Available Degraded forest area in Kalimantan could be caused by shifted cultivation activity that be conducted by local peoples in the surrounding forest areas. Efforts to improve the former shifted cultivation area (non productive land is developing the settled cultivation by use of irrigation system, better paddy seed, land processing, fertilizing, spraying pesticide, weeding, and better acces to the market.  Local peoples, especially in Kalimantan, has been depended their food on the shifted cultivation pattern since the long time ago.  This tradition could cause forest damage, forest fire, forest degradation, deforestation, and lose out of children education because they were following shifted cultivation activity although itsspace is very far from their home.  This research was aimed to improve former shifted cultivation lands using wetland cultivation in order to improve land productivity and to support food securityin the local community. This research was administratively located in Tanjung Rendan Village, Kapuas Hulu Sub-Ddistrict, Kapuas District, Central Kalimantan Province, Indonesia.  Data of rice yield from settled cultivation and shifted cultivation were got from 15 households that was taking by random at 2010 to 2011. Homogeneity test, analysis of variants, and least significant different (LSD test using SPSS 15.0 for Windows. Result of this research showed that     paddy yield at settled cultivation was significantly differentand better than shifted cultivation at 0.05 level. LSD test also indicated that all paddy yields from settled cultivation were significantly different compare to shifted cultivation at the 0.05 level.  The community in Tanjung Rendan Villages preferred settled cultivation than shifted cultivation, especially due to higher paddy production. Profit for settled cultivation was IDR10.95 million ha-1, meanwhile profit for shifted cultivation was just IDR 2.81 million ha-1 only.  Settled cultivation pattern could

In Vitro Expression of the Extracellular Matrix Components Aggrecan, Collagen Types I and II by Articular Cartilage-Derived Chondrocytes.

Science.gov (United States)

Schneevoigt, J; Fabian, C; Leovsky, C; Seeger, J; Bahramsoltani, M

2017-02-01

The extracellular matrix (ECM) of hyaline cartilage is perfectly suited to transmit articular pressure load to the subchondral bone. Pressure is transferred by a high amount of aggrecan-based proteoglycans and collagen type II fibres in particular. After any injury, the hyaline cartilage is replaced by fibrocartilage, which is low in proteoglycans and contains collagen type I predominantly. Until now, long-term results of therapeutic procedures including cell-based therapies like autologous chondrocyte transplantation (ACT) lead to a replacement tissue meeting the composition of fibrocartilage. Therefore, it is of particular interest to discover how and to what extent isolation and in vitro cultivation of chondrocytes affect the cells and their expression of ECM components. Hyaline cartilage-derived chondrocytes were cultivated in vitro and observed microscopically over a time period of 35 days. The expression of collagen type I, collagen type II and aggrecan was analysed using RT-qPCR and Western blot at several days of cultivation. Chondrocytes presented a longitudinal shape for the entire cultivation period. While expression of collagen type I prevailed within the first days, only prolonged cultivation led to an increase in collagen type II and aggrecan expression. The results indicate that chondrocyte isolation and in vitro cultivation lead to a dedifferentiation at least to the stage of chondroprogenitor cells. © 2016 Blackwell Verlag GmbH.

Lactobacillus plantarum BL011 cultivation in industrial isolated soybean protein acid residue

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Chaline Caren Coghetto

Full Text Available Abstract In this study, physiological aspects of Lactobacillus plantarum BL011 growing in a new, all-animal free medium in bioreactors were evaluated aiming at the production of this important lactic acid bacterium. Cultivations were performed in submerged batch bioreactors using the Plackett-Burman methodology to evaluate the influence of temperature, aeration rate and stirring speed as well as the concentrations of liquid acid protein residue of soybean, soy peptone, corn steep liquor, and raw yeast extract. The results showed that all variables, except for corn steep liquor, significantly influenced biomass production. The best condition was applied to bioreactor cultures, which produced a maximal biomass of 17.87 g L-1, whereas lactic acid, the most important lactic acid bacteria metabolite, peaked at 37.59 g L-1, corresponding to a productivity of 1.46 g L-1 h-1. This is the first report on the use of liquid acid protein residue of soybean medium for L. plantarum growth. These results support the industrial use of this system as an alternative to produce probiotics without animal-derived ingredients to obtain high biomass concentrations in batch bioreactors.

Energy and economic analysis of traditional versus introduced crops cultivation in the mountains of the Indian Himalayas: A case study

Energy Technology Data Exchange (ETDEWEB)

Nautiyal, Sunil; Kaechele, H. [Leibniz-Centre for Agricultural Landscape Research (ZALF), Institute of Socioeconomics, Eberswalder Str. 84, 15374 Muencheberg (Germany); Rao, K.S. [Centre for Inter-disciplinary Studies of Mountain and Hill Environment, Academic Research Center, University of Delhi, Delhi 110007 (India); Maikhuri, R.K. [G.B. Pant Institute of Himalayan Environment and Development, Garhwal Unit, P.O. Box 92, Srinagar (Garhwal) 246174 (India); Saxena, K.G. [School of Environmental Sciences, Jawaharlal Nehru University, New Delhi 110067 (India)

2007-12-15

This study analyzed the energy and economics associated with cultivation of traditional and introduced crops in the mountains of the Central Himalaya, India. The production cost in terms of energy for introduced crops such as tomato (Lycopersicon esculentum) and bell pepper (Capsicum annuum) cultivation was 90,358-320,516 MJ ha{sup -1} as compared to between 19,814 and 42,380 MJ ha{sup -1} for traditional crops within Himalayan agroecosystems. For the introduced crops, high energy and monetary input was associated with human labor, forest resources, chemical fertilizer and pesticides. However, energy threshold/projection for farmyard manure in traditional crop cultivation was 80-90% of the total energy cost, thus traditional crop cultivation was more efficient in energy and economics. During the study, the farm productivity of introduced crops cultivation declined with increasing years of cultivation. Consequently, the energy output from the system has been declining at the rate of -y20,598 to y20,748 MJ ha{sup -1} yr{sup -1} for tomato and y12,072 to y15,056 MJ ha{sup -1} yr{sup -1} for bell pepper under irrigated and rain-fed land use in the mountains, respectively. The comparative analysis on this paradigm shift indicates that more research is needed to support sustainable crop cultivation in the fragile Himalayan environment. (author)

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

Science.gov (United States)

Persson, Oliver; Andersson, Niklas; Nilsson, Bernt

2018-01-05

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

Optimization of the production of extracellular α-amylase by Kluyveromyces marxianus IF0 0288 by response surface methodology

Directory of Open Access Journals (Sweden)

Panagiota-Yiolanda Stergiou

2014-06-01

Full Text Available The aim of this work was to study the production of extracellular α-amylase by Kluyveromyces marxianus IF0 0288 using optimized nutritional and cultural conditions in a complex yeast medium under aerobic batch fermentation. By applying the conventional "one-variable-at-a-time" approach and the response surface methodology, the effect of four fermentation parameters (type of carbon source, initial culture pH, temperature, and incubation time on the growth and α-amylase production was evaluated. The production of α-amylase during 60 h of fermentation increased 13-fold under optimized conditions (1% starch, pH 6.0, 30ºC in comparison to the conventional optimization method. The initial pH value of 6.13 and temperature of 30.3ºC were optimal conditions by the response surface methodology, leading to further improvement (up to 13-fold in the production of extracellular α-amylase. These results constituted first evidence that K. marxianus could be potentially used as an effective source of extracellular α-amylase.

Biokinetic model-based multi-objective optimization of Dunaliella tertiolecta cultivation using elitist non-dominated sorting genetic algorithm with inheritance.

Science.gov (United States)

Sinha, Snehal K; Kumar, Mithilesh; Guria, Chandan; Kumar, Anup; Banerjee, Chiranjib

2017-10-01

Algal model based multi-objective optimization using elitist non-dominated sorting genetic algorithm with inheritance was carried out for batch cultivation of Dunaliella tertiolecta using NPK-fertilizer. Optimization problems involving two- and three-objective functions were solved simultaneously. The objective functions are: maximization of algae-biomass and lipid productivity with minimization of cultivation time and cost. Time variant light intensity and temperature including NPK-fertilizer, NaCl and NaHCO 3 loadings are the important decision variables. Algal model involving Monod/Andrews adsorption kinetics and Droop model with internal nutrient cell quota was used for optimization studies. Sets of non-dominated (equally good) Pareto optimal solutions were obtained for the problems studied. It was observed that time variant optimal light intensity and temperature trajectories, including optimum NPK fertilizer, NaCl and NaHCO 3 concentration has significant influence to improve biomass and lipid productivity under minimum cultivation time and cost. Proposed optimization studies may be helpful to implement the control strategy in scale-up operation. Copyright © 2017 Elsevier Ltd. All rights reserved.

7 CFR 58.728 - Cooking the batch.

Science.gov (United States)

2010-01-01

... 7 Agriculture 3 2010-01-01 2010-01-01 false Cooking the batch. 58.728 Section 58.728 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Standards... Procedures § 58.728 Cooking the batch. Each batch of cheese within the cooker, including the optional...

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...

Bioaugmentation of an acetate-oxidising anaerobic consortium in up-flow sludge blanket reactor subjected to high ammonia loads

DEFF Research Database (Denmark)

Fotidis, Ioannis; Karakashev, Dimitar Borisov; Angelidaki, Irini

Ammonia is the major inhibitor of anaerobic digestion (AD) process leading to suboptimal utilisation of the biogas potential of the feedstocks and causing economical losses to the biogas plants. However, ammonia is mainly inhibiting the aceticlastic methanogens, while the hydrogenotrophic...... was tested. The co-cultivation in fed-batch of a fast-growing hydrogenotrophic methanogen (i.e. Methanoculleus bourgensis) with the SAO culture was also investigated. Results obtained clearly demonstrated that bioaugmentation of SAO culture in a UASB reactor was not possible most probably due to the slow...

NGBAuth - Next Generation Batch Authentication for long running batch jobs.

CERN Document Server

Juto, Zakarias

2015-01-01

This document describes the prototyping of a new solution for the CERN batch authentication of long running jobs. While the job submission requires valid user credentials, these have to be renewed due to long queuing and execution times. Described within is a new system which will guarantee a similar level of security as the old LSFAuth while simplifying the implementation and the overall architecture. The new system is being built on solid, streamlined and tested components (notably OpenSSL) and a priority has been to make it more generic in order to facilitate the evolution of the current system such as for the expected migration from LSF to Condor as backend batch system.

Optimization of Cultural Conditions for Production of Extracellular Polymeric Substances (EPS) by Serpentine Rhizobacterium Cupriavidus pauculus KPS 201

OpenAIRE

Arundhati Pal; A. K. Paul

2013-01-01

Extracellular polymeric substances (EPS) are complex biopolymers produced by a wide array of microorganisms for protection against dessication, aggregation, adhesion, and expression of virulence. Growth associated production of EPS by Ni-resistant Cupriavidus pauculus KPS 201 was determined in batch culture using sodium gluconate as the sole carbon source. The optimum pH and temperature for EPS production were 6.5 and 25°C, respectively. Optimal EPS yield (118 μg/mL) was attained at 0.35% Na-...

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

Science.gov (United States)

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

2015-01-01

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

Strategies for Automated Control of the Bioproduction of Mcl-PHA Biopolymers

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P. Hrnčiřík

2017-10-01

Full Text Available Medium-chain-length polyhydroxyalkanoates (mcl-PHAs are polyesters synthesized by numerous bacteria as storage material. Despite being promising candidates for biodegradable materials of industrial interest and environmental value, their usage is still rather limited because of high production costs. One of the areas with considerable potential for further improvements is control of the production process. This paper deals with the experimental work related to the design of control strategies for mcl-PHA biopolymer production process (Pseudomonas putida KT2442 fed-batch cultivations. For this bioprocess, a set of five control strategies (two main and three auxiliary strategies have been proposed, together with the proper sequence of their switching during the fed-batch part of the production process. The application of these strategies with octanoic acid as a sole carbon source resulted in intracellular PHA content (max. mass fraction 65 % of mcl-PHA in cell dry mass (g g–1 and PHA productivity (max. 0.89 g L–1 h–1 comparable to the best results reported in the literature for this type of strain and carbon substrate.

The extracellular redox state modulates mitochondrial function, gluconeogenesis, and glycogen synthesis in murine hepatocytes.

Science.gov (United States)

Nocito, Laura; Kleckner, Amber S; Yoo, Elsia J; Jones Iv, Albert R; Liesa, Marc; Corkey, Barbara E

2015-01-01

Circulating redox state changes, determined by the ratio of reduced/oxidized pairs of different metabolites, have been associated with metabolic diseases. However, the pathogenic contribution of these changes and whether they modulate normal tissue function is unclear. As alterations in hepatic gluconeogenesis and glycogen metabolism are hallmarks that characterize insulin resistance and type 2 diabetes, we tested whether imposed changes in the extracellular redox state could modulate these processes. Thus, primary hepatocytes were treated with different ratios of the following physiological extracellular redox couples: β-hydroxybutyrate (βOHB)/acetoacetate (Acoc), reduced glutathione (GSH)/oxidized glutathione (GSSG), and cysteine/cystine. Exposure to a more oxidized ratio via extracellular βOHB/Acoc, GSH/GSSG, and cysteine/cystine in hepatocytes from fed mice increased intracellular hydrogen peroxide without causing oxidative damage. On the other hand, addition of more reduced ratios of extracellular βOHB/Acoc led to increased NAD(P)H and maximal mitochondrial respiratory capacity in hepatocytes. Greater βOHB/Acoc ratios were also associated with decreased β-oxidation, as expected with enhanced lipogenesis. In hepatocytes from fasted mice, a more extracellular reduced state of βOHB/Acoc led to increased alanine-stimulated gluconeogenesis and enhanced glycogen synthesis capacity from added glucose. Thus, we demonstrated for the first time that the extracellular redox state regulates the major metabolic functions of the liver and involves changes in intracellular NADH, hydrogen peroxide, and mitochondrial respiration. Because redox state in the blood can be communicated to all metabolically sensitive tissues, this work confirms the hypothesis that circulating redox state may be an important regulator of whole body metabolism and contribute to alterations associated with metabolic diseases.

The extracellular redox state modulates mitochondrial function, gluconeogenesis, and glycogen synthesis in murine hepatocytes.

Directory of Open Access Journals (Sweden)

Laura Nocito

Full Text Available Circulating redox state changes, determined by the ratio of reduced/oxidized pairs of different metabolites, have been associated with metabolic diseases. However, the pathogenic contribution of these changes and whether they modulate normal tissue function is unclear. As alterations in hepatic gluconeogenesis and glycogen metabolism are hallmarks that characterize insulin resistance and type 2 diabetes, we tested whether imposed changes in the extracellular redox state could modulate these processes. Thus, primary hepatocytes were treated with different ratios of the following physiological extracellular redox couples: β-hydroxybutyrate (βOHB/acetoacetate (Acoc, reduced glutathione (GSH/oxidized glutathione (GSSG, and cysteine/cystine. Exposure to a more oxidized ratio via extracellular βOHB/Acoc, GSH/GSSG, and cysteine/cystine in hepatocytes from fed mice increased intracellular hydrogen peroxide without causing oxidative damage. On the other hand, addition of more reduced ratios of extracellular βOHB/Acoc led to increased NAD(PH and maximal mitochondrial respiratory capacity in hepatocytes. Greater βOHB/Acoc ratios were also associated with decreased β-oxidation, as expected with enhanced lipogenesis. In hepatocytes from fasted mice, a more extracellular reduced state of βOHB/Acoc led to increased alanine-stimulated gluconeogenesis and enhanced glycogen synthesis capacity from added glucose. Thus, we demonstrated for the first time that the extracellular redox state regulates the major metabolic functions of the liver and involves changes in intracellular NADH, hydrogen peroxide, and mitochondrial respiration. Because redox state in the blood can be communicated to all metabolically sensitive tissues, this work confirms the hypothesis that circulating redox state may be an important regulator of whole body metabolism and contribute to alterations associated with metabolic diseases.

Nitrate to ammonia and ceramic (NAC) process during batch and continuous operation

International Nuclear Information System (INIS)

Muguercia, I.; Solomon, S.; Ebadian, M.A.

1996-01-01

The nitrate to ammonia and ceramic (NAC) process is an innovative technology for the denitration of radioactive sodium nitrate-based liquid waste found throughout Department of Energy (DOE) facilities in the United States. In the present investigation, two reaction systems were studied. The first utilized only sodium nitrate as the substrate for the aluminum. The second consisted of the multication composition of waste forms located at the Hanford facility. Studies were carried out on the batch reaction at three different starting nitrate ion concentrations, each at three different temperatures. For each of these conditions, the rate of nitrate depletion was determined, and rate constants were calculated. The reaction did not demonstrate simple kinetics; rather, it appeared to involve two zero order reactions. Certain generalities were obtained in both the batch reaction and in the continuous process, nonetheless. It was found that the conversion of nitrate to ammonia seemed to be most efficient at the lowest temperature studied, 50 degrees C. This behavior was more obvious in the case of the unadulterated nitrate solution than with the Hanford simulant. To elaborate a practical, marketable product, it was necessary to develop a process that could be carried out in a continuous matter, whereby reactants were continuously fed into a reactor while the products of the reaction were simultaneously removed. Thus, the objective has been to develop the prototype procedures for carrying out this continuous reaction. As a corollary of this research, it was first necessary to define the characteristics of the reaction with respect to rate, conversion efficiency, and safety. To achieve this end, reactions were run under various batch conditions, and an attempt was made to measure the rates of the depletion of nitrate and the production of ammonia and hydrogen as well as pH and temperature changes

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

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

Importance of Thickness in Human Cardiomyocyte Network for Effective Electrophysiological Stimulation Using On-Chip Extracellular Microelectrodes

Science.gov (United States)

Hamada, Tomoyo; Nomura, Fumimasa; Kaneko, Tomoyuki; Yasuda, Kenji

2012-06-01

We have developed a three-dimensionally controlled in vitro human cardiomyocyte network assay for the measurements of drug-induced conductivity changes and the appearance of fatal arrhythmia such as ventricular tachycardia/fibrillation for more precise in vitro predictive cardiotoxicity. To construct an artificial conductance propagation model of a human cardiomyocyte network, first, we examined the cell concentration dependence of the cell network heights and found the existence of a height limit of cell networks, which was double-layer height, whereas the cardiomyocytes were effectively and homogeneously cultivated within the microchamber maintaining their spatial distribution constant and their electrophysiological conductance and propagation were successfully recorded using a microelectrode array set on the bottom of the microchamber. The pacing ability of a cardiomyocyte's electrophysiological response has been evaluated using microelectrode extracellular stimulation, and the stimulation for pacing also successfully regulated the beating frequencies of two-layered cardiomyocyte networks, whereas monolayered cardiomyocyte networks were hardly stimulated by the external electrodes using the two-layered cardiomyocyte stimulation condition. The stability of the lined-up shape of human cardiomyocytes within the rectangularly arranged agarose microchambers was limited for a two-layered cardiomyocyte network because their stronger force generation shrunk those cells after peeling off the substrate. The results indicate the importance of fabrication technology of thickness control of cellular networks for effective extracellular stimulation and the potential concerning thick cardiomyocyte networks for long-term cultivation.

Enhanced extracellular production of L-asparaginase from Bacillus subtilis 168 by B. subtilis WB600 through a combined strategy.

Science.gov (United States)

Feng, Yue; Liu, Song; Jiao, Yun; Gao, Hui; Wang, Miao; Du, Guocheng; Chen, Jian

2017-02-01

L-asparaginase (EC 3.5.1.1, ASN) exhibits great commercial value due to its uses in the food and medicine industry. In this study, we reported the enhanced expression of type II ASN from Bacillus subtilis 168 in B. subtilis WB600 through a combined strategy. First, eight signal peptides (the signal peptide of the ASN, ywbN, yvgO, amyE, oppA, vpr, lipA, and wapA) were used for ASN secretion in B. subtilis by using Hpa II promoter, respectively. The signal peptide wapA achieved the highest extracellular ASN activity (28.91 U/mL). Second, Hpa II promoter was replaced by a strong promoter, P43 promoter, resulting in 38.1 % enhanced ASN activity. By two rounds of error-prone PCR mutation, the P43 promoter variants with remarkably enhanced strength (D7, E2, H6, B2, and F3) were identified. B2 (-28: A → G, -13: A → G) achieved ASN activity up to 51.13 U/mL. Third, after deletion of the N-terminal 25-residues, ASN activity reached 102.41 U/mL, which was 100 % higher than that of the intact ASN. At last, the extracellular ASN of the B. subtilis arrived at 407.6 U/mL (2.5 g/L of ASN protein) in a 3-L bioreactor by using a fed-batch strategy. The purified ASN showed maximal activity at 65 °C and its half-life at 65 °C was 61 min. The K m and k cat of the ASN were 5.29 mM and 54.4 s -1 , respectively. To the best of our knowledge, we obtained the highest yield of ASN in a food-grade host ever reported, which may benefit the industrial production and application of ASN.

Heat-treated (in single aliquot or batch) colostrum outperforms non-heat-treated colostrum in terms of quality and transfer of immunoglobulin G in neonatal Jersey calves.

Science.gov (United States)

Kryzer, A A; Godden, S M; Schell, R

2015-03-01

The objective of this randomized clinical trial was to describe the effect on colostrum characteristics and passive transfer of IgG in neonatal calves when using the Perfect Udder colostrum management system (single-aliquot treatment; Dairy Tech Inc., Greeley, CO) compared with a negative control (fresh refrigerated or fresh frozen colostrum) and a positive control (batch heat-treated colostrum). First-milking Jersey colostrum was pooled to achieve 31 unique batches with a minimum of 22.8 L per batch. The batch was then divided into 4 with 3.8 L allocated to each treatment group: (1) heat-treated in Perfect Udder bag at 60°C for 60 min and then stored at -20°C (PU); (2) heat-treated in a batch pasteurizer (Dairy Tech Inc.) at 60°C for 60 min and then stored at -20°C in Perfect Udder bag (DTB; positive control); (3) fresh frozen colostrum stored at -20°C in Perfect Udder bag (FF; negative control); and (4) fresh refrigerated colostrum stored at 4°C in Perfect Udder bag (FR; negative control). Colostrum from all treatments was sampled for analysis of IgG concentration and bacterial culture immediately after batch assembly, after processing, and before feeding. Newborn Jersey calves were randomly assigned to be fed 3.8 L of colostrum from 1 of the 4 treatment groups. A prefeeding, 0-h blood sample was collected, calves were fed by esophageal tube within 2 h of birth, and then a 24-h postfeeding blood sample was collected. Paired serum samples from 0- and 24-h blood samples were analyzed for IgG concentration (mg/mL) using radial immunodiffusion analysis. The overall mean IgG concentration in colostrum was 77.9 g/L and was not affected by treatment. Prefeeding total plate counts (log10 cfu/mL) were significantly different for all 4 treatments and were lower for heat-treated colostrum (PU=4.23, DTB=3.63) compared with fresh colostrum (FF=5.68, FR=6.53). Total coliform counts (log10 cfu/mL) were also significantly different for all 4 treatments and were lower for

Oral microbial profile discriminates breast-fed from formula-fed infants.

Science.gov (United States)

Holgerson, Pernilla L; Vestman, Nelly R; Claesson, Rolf; Ohman, Carina; Domellöf, Magnus; Tanner, Anne C R; Hernell, Olle; Johansson, Ingegerd

2013-02-01

Little is known about the effect of diet on the oral microbiota of infants, although diet is known to affect the gut microbiota. The aims of the present study were to compare the oral microbiota in breast-fed and formula-fed infants, and investigate growth inhibition of streptococci by infant-isolated lactobacilli. A total of 207 mothers consented to participation of their 3-month-old infants. A total of 146 (70.5%) infants were exclusively and 38 (18.4%) partially breast-fed, and 23 (11.1%) were exclusively formula-fed. Saliva from all of their infants was cultured for Lactobacillus species, with isolate identifications from 21 infants. Lactobacillus isolates were tested for their ability to suppress Streptococcus mutans and S sanguinis. Oral swabs from 73 infants were analysed by the Human Oral Microbe Identification Microarray (HOMIM) and by quantitative polymerase chain reaction for Lactobacillus gasseri. Lactobacilli were cultured from 27.8% of exclusively and partially breast-fed infants, but not from formula-fed infants. The prevalence of 14 HOMIM-detected taxa, and total salivary lactobacilli counts differed by feeding method. Multivariate modelling of HOMIM-detected bacteria and possible confounders clustered samples from breast-fed infants separately from formula-fed infants. The microbiota of breast-fed infants differed based on vaginal or C-section delivery. Isolates of L plantarum, L gasseri, and L vaginalis inhibited growth of the cariogenic S mutans and the commensal S sanguinis: L plantarum >L gasseri >L vaginalis. The microbiota of the mouth differs between 3-month-old breast-fed and formula-fed infants. Possible mechanisms for microbial differences observed include species suppression by lactobacilli indigenous to breast milk.

NDA BATCH 2002-02

Energy Technology Data Exchange (ETDEWEB)

Lawrence Livermore National Laboratory

2009-12-09

QC sample results (daily background checks, 20-gram and 100-gram SGS drum checks) were within acceptable criteria established by WIPP's Quality Assurance Objectives for TRU Waste Characterization. Replicate runs were performed on 5 drums with IDs LL85101099TRU, LL85801147TRU, LL85801109TRU, LL85300999TRU and LL85500979TRU. All replicate measurement results are identical at the 95% confidence level as established by WIPP criteria. Note that the batch covered 5 weeks of SGS measurements from 23-Jan-2002 through 22-Feb-2002. Data packet for SGS Batch 2002-02 generated using gamma spectroscopy with the Pu Facility SGS unit is technically reasonable. All QC samples are in compliance with established control limits. The batch data packet has been reviewed for correctness, completeness, consistency and compliance with WIPP's Quality Assurance Objectives and determined to be acceptable. An Expert Review was performed on the data packet between 28-Feb-02 and 09-Jul-02 to check for potential U-235, Np-237 and Am-241 interferences and address drum cases where specific scan segments showed Se gamma ray transmissions for the 136-keV gamma to be below 0.1 %. Two drums in the batch showed Pu-238 at a relative mass ratio more than 2% of all the Pu isotopes.

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.

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.

Continuous cultivation of a thermophilic bacterium Aeribacillus pallidus 418 for production of an exopolysaccharide applicable in cosmetic creams.

Science.gov (United States)

Radchenkova, N; Panchev, I; Vassilev, S; Kuncheva, M; Dobreva, S; Kambourova, M

2015-11-01

The aim of this study was to evaluate the effectiveness of continuous cultivation approach for exopolysaccharide (EPS) production by a thermophilic micro-organism and the potential of the synthesized EPS for application in cosmetic industry. Study on the ability of Aeribacillus pallidus 418, isolated as a good EPS producer, to synthesize the polymer in continuous cultures showed higher production in comparison with batch cultures. The degree of the EPS in the precipitate after continuous cultivation significantly increased. Non-Newtonian pseudoplastic and thixotropic behaviour of EPS determines the ability of the received cream to become more fluid after increasing time of application on the skin. This study demonstrates a highly efficient way for production of EPS from a continuous growth culture of A. pallidus 418 that have many advantages and can outperform batch culture by eliminating time for cleaning and sterilization of the vessel and the comparatively long lag phases before the organisms enter a brief period of high productivity. The valuable physico-chemical properties of the synthesized EPS influenced positively the properties of a commercial cream. EPSs from thermophilic micro-organisms are of special interest due to the advantages of the thermophilic processes and nonpathogenic nature of the polymer molecules. However, their industrial application is hindered by the comparatively low biomass and correspondingly EPS yield. Suggested continuous approach for EPS could have an enormous economic potential for an industrial scale production of thermophilic EPSs. © 2015 The Society for Applied Microbiology.

Dynamic cultivation of human mesenchymal stem cells in a rotating bed bioreactor system based on the Z RP platform.

Science.gov (United States)

Diederichs, Solvig; Röker, Stefanie; Marten, Dana; Peterbauer, Anja; Scheper, Thomas; van Griensven, Martijn; Kasper, Cornelia

2009-01-01

Because the regeneration of large bone defects is limited by quantitative restrictions and risks of infections, the development of bioartificial bone substitutes is of great importance. To obtain a three-dimensional functional tissue-like graft, static cultivation is inexpedient due to limitations in cell density, nutrition and oxygen support. Dynamic cultivation in a bioreactor system can overcome these restrictions and furthermore provide the possibility to control the environment with regard to pH, oxygen content, and temperature. In this study, a three-dimensional bone construct was engineered by the use of dynamic bioreactor technology. Human adipose tissue derived mesenchymal stem cells were cultivated on a macroporous zirconium dioxide based ceramic disc called Sponceram. Furthermore, hydroxyapatite coated Sponceram was used. The cells were cultivated under dynamic conditions and compared with statically cultivated cells. The differentiation into osteoblasts was initiated by osteogenic supplements. Cellular proliferation during static and dynamic cultivation was compared measuring glucose and lactate concentration. The differentiation process was analysed determining AP-expression and using different specific staining methods. Our results demonstrate much higher proliferation rates during dynamic conditions in the bioreactor system compared to static cultivation measured by glucose consumption and lactate production. Cell densities on the scaffolds indicated higher proliferation on native Sponceram compared to hydroxyapatite coated Sponceram. With this study, we present an excellent method to enhance cellular proliferation and bone lineage specific growth of tissue like structures comprising fibrous (collagen) and globular (mineral) extracellular components. (c) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009.

Bioprocess iterative batch-to-batch optimization based on hybrid parametric/nonparametric models.

Science.gov (United States)

Teixeira, Ana P; Clemente, João J; Cunha, António E; Carrondo, Manuel J T; Oliveira, Rui

2006-01-01

This paper presents a novel method for iterative batch-to-batch dynamic optimization of bioprocesses. The relationship between process performance and control inputs is established by means of hybrid grey-box models combining parametric and nonparametric structures. The bioreactor dynamics are defined by material balance equations, whereas the cell population subsystem is represented by an adjustable mixture of nonparametric and parametric models. Thus optimizations are possible without detailed mechanistic knowledge concerning the biological system. A clustering technique is used to supervise the reliability of the nonparametric subsystem during the optimization. Whenever the nonparametric outputs are unreliable, the objective function is penalized. The technique was evaluated with three simulation case studies. The overall results suggest that the convergence to the optimal process performance may be achieved after a small number of batches. The model unreliability risk constraint along with sampling scheduling are crucial to minimize the experimental effort required to attain a given process performance. In general terms, it may be concluded that the proposed method broadens the application of the hybrid parametric/nonparametric modeling technique to "newer" processes with higher potential for optimization.

A kinetic model for the penicillin biosynthetic pathway in

DEFF Research Database (Denmark)

Nielsen, Jens; Jørgensen, Henrik

1996-01-01

A kinetic model for the first two steps in the penicillin biosynthetic pathway, i.e. the ACV synthetase (ACVS) and the isopenicillin N synthetase (IPNS) is proposed. The model is based on Michaelis-Menten type kinetics with non-competitive inhibition of the ACVS by ACV, and competitive inhibition...... of the IPNS by glutathione. The model predicted flux through the pathway corresponds well with the measured rate of penicillin biosynthesis. From the kinetic model the elasticity coefficients and the flux control coefficients are calculated throughout a fed-batch cultivation, and it is found...

Impact of carbon and nitrogen feeding strategy on high production of biomass and docosahexaenoic acid (DHA) by Schizochytrium sp. LU310.

Science.gov (United States)

Ling, Xueping; Guo, Jing; Liu, Xiaoting; Zhang, Xia; Wang, Nan; Lu, Yinghua; Ng, I-Son

2015-05-01

A new isolated Schizochytrium sp. LU310 from the mangrove forest of Wenzhou, China, was found as a high producing microalga of docosahexaenoic acid (DHA). In this study, the significant improvements for DHA fermentation by the batch mode in the baffled flasks (i.e. higher oxygen supply) were achieved. By applied the nitrogen-feeding strategy in 1000 mL baffled flasks, the biomass, DHA concentration and DHA productivity were increased by 110.4%, 117.9% and 110.4%, respectively. Moreover, DHA concentration of 21.06 g/L was obtained by feeding 15 g/L of glucose intermittently, which was an increase of 41.25% over that of the batch mode. Finally, an innovative strategy was carried out by intermittent feeding carbon and simultaneously feeding nitrogen. The maximum DHA concentration and DHA productivity in the fed-batch cultivation reached to 24.74 g/L and 241.5 mg/L/h, respectively. Copyright © 2014 Elsevier Ltd. All rights reserved.

Kinetic analysis and modeling of daptomycin batch fermentation by Streptomyces roseosporus.

Science.gov (United States)

Lu, Wenyu; Fan, Jinghua; Wen, Jianping; Xia, Zhendong; Caiyin, Qinggele

2011-02-01

In this study, Streptomyces roseosporus was subjected to helium-neon (He-Ne) laser (632.8 nm) irradiation to improve the production ability of extracellular antibiotic daptomycin. Under the optimum irradiation dosage of 18 mW for 22 min, a stable positive mutant strain S. roseosporus LC-54 was obtained. The maximum A21978C (daptomycin is a semisynthetic antimicrobial substance derived from the A21978C complex) yield of this mutant strain was 296 mg/l, which was 146% higher than that of the wild strain. The mutant strain grew more quickly and utilized carbohydrate sources more efficiently than the wild strain. The batch culture kinetics was investigated in a 7 l bioreactor. The logistic equation for growth, the Luedeking-Piret equation for daptomycin production, and Luedeking-Piret-like equations for carbon substrate consumption were established. This model appeared to provide a reasonable description for each parameter during the growth phase and fitted fairly well with the experiment data.

Passing in Command Line Arguments and Parallel Cluster/Multicore Batching in R with batch.

Science.gov (United States)

Hoffmann, Thomas J

2011-03-01

It is often useful to rerun a command line R script with some slight change in the parameters used to run it - a new set of parameters for a simulation, a different dataset to process, etc. The R package batch provides a means to pass in multiple command line options, including vectors of values in the usual R format, easily into R. The same script can be setup to run things in parallel via different command line arguments. The R package batch also provides a means to simplify this parallel batching by allowing one to use R and an R-like syntax for arguments to spread a script across a cluster or local multicore/multiprocessor computer, with automated syntax for several popular cluster types. Finally it provides a means to aggregate the results together of multiple processes run on a cluster.

Cultivo mixotrófico da microalga Spirulina platensis em batelada alimentada Mixotrophic growth of Spirulina platensis in fed-batch mode

Directory of Open Access Journals (Sweden)

Adriana Muliterno

2005-12-01

Full Text Available A Spirulina platensis tem sido estudada devido a seu alto valor protéico, digestibilidade e por apresentar quantidades significativas de ácidos graxos poliinsaturados, vitaminas, fenólicos e ficocianina, podendo ser utilizada na alimentação humana. A utilização de nutrientes de baixo custo é um fator importante na produção da cianobactéria por possibilitar a redução de custos de processo. Objetivou-se com este trabalho estudar o cultivo mixotrófico da S. platensis por meio da adição de uma fonte orgânica de carbono (glicose em modo bateladaalimentada. Foi utilizado um Planejamento Fatorial Completo 2³ para o cultivo e as variáveis de estudo foram a concentração de glicose (0,5 gL-1 e 1,0 gL-1, a diluição do meio Zarrouk (50% e 75% e a iluminância (1800 lux e 3000 lux. A concentração celular máxima obtida foi de 5,38 gL-1 com uma velocidade específica máxima de crescimento de 0,0063 h-1, nas condições de 0,5 gL-1 de glicose, diluição do meio de 75% e iluminância de 3000 lux.The cyanobacterium Spirulina platensis has been studied due to its high content (~65% of highly digestible protein as well as significant amounts of polyunsaturated fatty acids, phenolics, vitamins, minerals and phycocyanin which could be useful in the human nutrition. The use of nutrients of low costs in the cyanobacterium growth could reduce the costs of production. We studied the fed-batch mixotrophic growth of the S. platensis in Zarrouk's medium with glucose (0.5 gL-1 and 1.0 gL-1 as carbon source and also investigated the effects of dilution (50% and 75%, with water and illumination (1,800 lux and 3,000 lux using a 2³ factorial design. The maximum celular concentration of 5.38 gL-1 and maximum specific growth rate of 0.0063 h-1 were obtained with a glucose concentration of 0.5 gL-1, 50% dilution and 1800 lux of illuminance.

Lactobacillus plantarum BL011 cultivation in industrial isolated soybean protein acid residue.

Science.gov (United States)

Coghetto, Chaline Caren; Vasconcelos, Carolina Bettker; Brinques, Graziela Brusch; Ayub, Marco Antônio Záchia

In this study, physiological aspects of Lactobacillus plantarum BL011 growing in a new, all-animal free medium in bioreactors were evaluated aiming at the production of this important lactic acid bacterium. Cultivations were performed in submerged batch bioreactors using the Plackett-Burman methodology to evaluate the influence of temperature, aeration rate and stirring speed as well as the concentrations of liquid acid protein residue of soybean, soy peptone, corn steep liquor, and raw yeast extract. The results showed that all variables, except for corn steep liquor, significantly influenced biomass production. The best condition was applied to bioreactor cultures, which produced a maximal biomass of 17.87gL -1 , whereas lactic acid, the most important lactic acid bacteria metabolite, peaked at 37.59gL -1 , corresponding to a productivity of 1.46gL -1 h -1 . This is the first report on the use of liquid acid protein residue of soybean medium for L. plantarum growth. These results support the industrial use of this system as an alternative to produce probiotics without animal-derived ingredients to obtain high biomass concentrations in batch bioreactors. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

On-line Scheduling Of Multi-Server Batch Operations

NARCIS (Netherlands)

van der Zee, D.J.; van Harten, A.; Schuur, P.C.

1999-01-01

Batching jobs in a manufacturing system is a very common policy in most industries. Main reasons for batching are avoidance of setups and/or facilitation of material handling. Good examples of batch-wise production systems are ovens found in aircraft industry and in semiconductor manufacturing.

[Batch release of immunoglobulin and monoclonal antibody products].

Science.gov (United States)

Gross, S

2014-10-01

The Paul-Ehrlich Institute (PEI) is an independent institution of the Federal Republic of Germany responsible for performing official experimental batch testing of sera. The institute decides about the release of each batch and performs experimental research in the field. The experimental quality control ensures the potency of the product and also the absence of harmful impurities. For release of an immunoglobulin batch the marketing authorization holder has to submit the documentation of the manufacture and the results of quality control measures together with samples of the batch to the PEI. Experimental testing is performed according to the approved specifications regarding the efficacy and safety. Since implementation of the 15th German drug law amendment, the source of antibody is not defined anymore. According to § 32 German drug law, all batches of sera need to be released by an official control laboratory. Sera are medicinal products, which contain antibodies, antibody fragments or fusion proteins with a functional antibody portion. Therefore, all batches of monoclonal antibodies and derivatives must also be released by the PEI and the marketing authorization holder has to submit a batch release application. Under certain circumstances a waiver for certain products can be issued with regard to batch release. The conditions for such a waiver apply to the majority of monoclonal antibodies.

Fate of H2S during the cultivation of Chlorella sp. deployed for biogas upgrading.

Science.gov (United States)

González-Sánchez, Armando; Posten, Clemens

2017-04-15

The H 2 S may play a key role in the sulfur cycle among the biogas production by the anaerobic digestion of wastes and the biogas upgrading by a microalgae based technology. The biogas is upgraded by contacting with slightly alkaline aqueous microalgae culture, then CO 2 and H 2 S are absorbed. The dissolved H 2 S could limit or inhibit the microalgae growth. This paper evaluated the role of dissolved H 2 S and other sulfured byproducts under prevailing biogas upgrading conditions using a microalgal technology. At initial stages of batch cultivation the growth of Chlorella sp. was presumably inhibited by dissolved H 2 S. After 2 days, the sulfides were oxidized mainly by oxic chemical reactions to sulfate, which was later rapidly assimilated by Chlorella sp., allowing high growing rates. The fate of H 2 S during the microalgae cultivation at pH > 8.5 was assessed by a mathematical model where the pentasulfide, thiosulfate and sulfite were firstly produced and converted finally to sulfate for posterior assimilation. Copyright © 2017 Elsevier Ltd. All rights reserved.

A practical approach for exploration and modeling of the design space of a bacterial vaccine cultivation process.

Science.gov (United States)

Streefland, M; Van Herpen, P F G; Van de Waterbeemd, B; Van der Pol, L A; Beuvery, E C; Tramper, J; Martens, D E; Toft, M

2009-10-15

A licensed pharmaceutical process is required to be executed within the validated ranges throughout the lifetime of product manufacturing. Changes to the process, especially for processes involving biological products, usually require the manufacturer to demonstrate that the safety and efficacy of the product remains unchanged by new or additional clinical testing. Recent changes in the regulations for pharmaceutical processing allow broader ranges of process settings to be submitted for regulatory approval, the so-called process design space, which means that a manufacturer can optimize his process within the submitted ranges after the product has entered the market, which allows flexible processes. In this article, the applicability of this concept of the process design space is investigated for the cultivation process step for a vaccine against whooping cough disease. An experimental design (DoE) is applied to investigate the ranges of critical process parameters that still result in a product that meets specifications. The on-line process data, including near infrared spectroscopy, are used to build a descriptive model of the processes used in the experimental design. Finally, the data of all processes are integrated in a multivariate batch monitoring model that represents the investigated process design space. This article demonstrates how the general principles of PAT and process design space can be applied for an undefined biological product such as a whole cell vaccine. The approach chosen for model development described here, allows on line monitoring and control of cultivation batches in order to assure in real time that a process is running within the process design space.

LSF usage for batch at CERN

CERN Multimedia

Schwickerath, Ulrich

2007-01-01

Contributed poster to the CHEP07. Original abstract: LSF 7, the latest version of Platform's batch workload management system, addresses many issues which limited the ability of LSF 6.1 to support large scale batch farms, such as the lxbatch service at CERN. In this paper we will present the status of the evaluation and deployment of LSF 7 at CERN, including issues concerning the integration of LSF 7 with the gLite grid middleware suite and, in particular, the steps taken to endure an efficient reporting of the local batch system status and usage to the Grid Information System

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

efficiency of C. salexigens can be improved by careful design of culture feeding schemes. Metabolic shifts observed at low ammonium concentrations were explained by a shift in the energy required for nitrogen assimilation. Carbon-limited fed-batch cultures with reduced ammonium supply were the best conditions for cultivation of C. salexigens, supporting high density growth and maintaining high ectoines production.

[Dendrobium officinale stereoscopic cultivation method].

Science.gov (United States)

Si, Jin-Ping; Dong, Hong-Xiu; Liao, Xin-Yan; Zhu, Yu-Qiu; Li, Hui

2014-12-01

The study is aimed to make the most of available space of Dendrobium officinale cultivation facility, reveal the yield and functional components variation of stereoscopic cultivated D. officinale, and improve quality, yield and efficiency. The agronomic traits and yield variation of stereoscopic cultivated D. officinale were studied by operating field experiment. The content of polysaccharide and extractum were determined by using phenol-sulfuric acid method and 2010 edition of "Chinese Pharmacopoeia" Appendix X A. The results showed that the land utilization of stereoscopic cultivated D. officinale increased 2.74 times, the stems, leaves and their total fresh or dry weight in unit area of stereoscopic cultivated D. officinale were all heavier than those of the ground cultivated ones. There was no significant difference in polysaccharide content between stereoscopic cultivation and ground cultivation. But the extractum content and total content of polysaccharide and extractum were significantly higher than those of the ground cultivated ones. In additional, the polysaccharide content and total content of polysaccharide and extractum from the top two levels of stereoscopic culture matrix were significantly higher than that of the ones from the other levels and ground cultivation. Steroscopic cultivation can effectively improves the utilization of space and yield, while the total content of polysaccharides and extractum were significantly higher than that of the ground cultivated ones. The significant difference in Dendrobium polysaccharides among the plants from different height of stereo- scopic culture matrix may be associated with light factor.

Family based dispatching with batch availability

NARCIS (Netherlands)

van der Zee, D.J.

2013-01-01

Family based dispatching rules seek to lower set-up frequencies by grouping (batching) similar types of jobs for joint processing. Hence shop flow times may be improved, as less time is spent on set-ups. Motivated by an industrial project we study the control of machines with batch availability,

On-line scheduling of multi-server batch operations

NARCIS (Netherlands)

Zee, Durk Jouke van der; Harten, Aart van; Schuur, Peter

The batching of jobs in a manufacturing system is a very common policy in many industries. The main reasons for batching are the avoidance of setups and/or facilitation of material handling. Good examples of batch-wise production systems are the ovens that are found in the aircraft industry and in

Batching System for Superior Service

Science.gov (United States)

2001-01-01

Veridian's Portable Batch System (PBS) was the recipient of the 1997 NASA Space Act Award for outstanding software. A batch system is a set of processes for managing queues and jobs. Without a batch system, it is difficult to manage the workload of a computer system. By bundling the enterprise's computing resources, the PBS technology offers users a single coherent interface, resulting in efficient management of the batch services. Users choose which information to package into "containers" for system-wide use. PBS also provides detailed system usage data, a procedure not easily executed without this software. PBS operates on networked, multi-platform UNIX environments. Veridian's new version, PBS Pro,TM has additional features and enhancements, including support for additional operating systems. Veridian distributes the original version of PBS as Open Source software via the PBS website. Customers can register and download the software at no cost. PBS Pro is also available via the web and offers additional features such as increased stability, reliability, and fault tolerance.A company using PBS can expect a significant increase in the effective management of its computing resources. Tangible benefits include increased utilization of costly resources and enhanced understanding of computational requirements and user needs.

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.

Batch-To-Batch Rational Feedforward Control : From Iterative Learning to Identification Approaches, with Application to a Wafer Stage

NARCIS (Netherlands)

Blanken, L.; Boeren, F.A.J.; Bruijnen, D.J.H.; Oomen, T.A.E.

2017-01-01

Feedforward control enables high performance for industrial motion systems that perform nonrepeating motion tasks. Recently, learning techniques have been proposed that improve both performance and flexibility to nonrepeating tasks in a batch-To-batch fashion by using a rational parameterization in

Evaluation of vitrification factors from DWPF's macro-batch 1

International Nuclear Information System (INIS)

Edwards, T.B.

2000-01-01

The Defense Waste Processing Facility (DWPF) is evaluating new sampling and analytical methods that may be used to support future Slurry Mix Evaporator (SME) batch acceptability decisions. This report uses data acquired during DWPF's processing of macro-batch 1 to determine a set of vitrification factors covering several SME and Melter Feed Tank (MFT) batches. Such values are needed for converting the cation measurements derived from the new methods to a ''glass'' basis. The available data from macro-batch 1 were used to examine the stability of these vitrification factors, to estimate their uncertainty over the course of a macro-batch, and to provide a recommendation on the use of a single factor for an entire macro-batch. The report is in response to Technical Task Request HLW/DWPF/TTR-980015

Sojourn time distributions in a Markovian G-queue with batch arrival and batch removal

Directory of Open Access Journals (Sweden)

Yang Woo Shin

1999-01-01

Full Text Available We consider a single server Markovian queue with two types of customers; positive and negative, where positive customers arrive in batches and arrivals of negative customers remove positive customers in batches. Only positive customers form a queue and negative customers just reduce the system congestion by removing positive ones upon their arrivals. We derive the LSTs of sojourn time distributions for a single server Markovian queue with positive customers and negative customers by using the first passage time arguments for Markov chains.

Optimization of Growth Medium for Efficient Cultivation of Lactobacillus salivarius i 24 using Response Surface Method

Directory of Open Access Journals (Sweden)

Lim, C. H.

2007-01-01

Full Text Available Production of Lactobacillus salivarius i 24, a probiotic strain for chicken, was studied in batch fermentation using 500 mL Erlenmeyer flask. Response surface method (RSM was used to optimize the medium for efficient cultivation of the bacterium. The factors investigated were yeast extract, glucose and initial culture pH. A polynomial regression model with cubic and quartic terms was used for the analysis of the experimental data. Estimated optimal conditions of the factors for growth of L. salivarius i 24 were; 3.32 % (w/v glucose, 4.31 % (w/v yeast extract and initial culture pH of 6.10.

Extracellular enzymatic activities and physiological profiles of yeasts colonizing fruit trees.

Science.gov (United States)

Molnárová, Jana; Vadkertiová, Renáta; Stratilová, Eva

2014-07-01

Yeasts form a significant and diverse part of the phyllosphere microbiota. Some yeasts that inhabit plants have been found to exhibit extracellular enzymatic activities. The aim of the present study was to investigate the ability of yeasts isolated from leaves, fruits, and blossoms of fruit trees cultivated in Southwest Slovakia to produce extracellular enzymes, and to discover whether the yeasts originating from these plant organs differ from each other in their physiological properties. In total, 92 strains belonging to 29 different species were tested for: extracellular protease, β-glucosidase, lipase, and polygalacturonase activities; fermentation abilities; the assimilation of xylose, saccharose and alcohols (methanol, ethanol, glycerol); and for growth in a medium with 33% glucose. The black yeast Aureobasidium pullulans showed the largest spectrum of activities of all the species tested. Almost 70% of the strains tested demonstrated some enzymatic activity, and more than 90% utilized one of the carbon compounds tested. Intraspecies variations were found for the species of the genera Cryptococcus and Pseudozyma. Interspecies differences of strains exhibiting some enzymatic activities and utilizing alcohols were also noted. The largest proportion of the yeasts exhibited β-glucosidase activity and assimilated alcohols independently of their origin. The highest number of strains positive for all activities tested was found among the yeasts associated with leaves. Yeasts isolated from blossoms assimilated saccharose and D-xylose the most frequently of all the yeasts tested. The majority of the fruit-inhabiting yeasts grew in the medium with higher osmotic pressure. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Extracellular polymer substance synthesized by a halophilic bacterium Chromohalobacter canadensis 28.

Science.gov (United States)

Radchenkova, Nadja; Boyadzhieva, Ivanka; Atanasova, Nikolina; Poli, Annarita; Finore, Ilaria; Di Donato, Paola; Nicolaus, Barbara; Panchev, Ivan; Kuncheva, Margarita; Kambourova, Margarita

2018-04-03

Halophilic microorganisms are producers of a lot of new compounds whose properties suggest promising perspectives for their biotechnological exploration. Moderate halophilic bacterium Chromohalobacter canadensis 28 was isolated from Pomorie salterns as an extracellular polymer substance (EP) producer. The best carbon source for extracellular polymer production was found to be lactose, a sugar received as a by-product from the dairy industry. After optimization of the culture medium and physicochemical conditions for cultivation, polymer biosynthesis increased more than 2-fold. The highest level of extracellular polymer synthesis by C. canadensis 28 was observed in an unusually high NaCl concentration (15% w/v). Chemical analysis of the purified polymer revealed the presence of an exopolysaccharide (EPS) fraction (14.3% w/w) and protein fraction (72% w/w). HPLC analysis of the protein fraction showed the main presence of polyglutamic acid (PGA) (75.7% w/w). EPS fraction analysis revealed the following sugar composition (% w/w): glucosamine 36.7, glucose 32.3, rhamnose 25.4, xylose 1.7, and not identified sugar 3.9. The hydrogel formed by PGA and EPS fractions showed high swelling behavior, very good emulsifying and stabilizing properties, and good foaming ability. This is the first report for halophilic bacterium able to synthesize a polymer containing PGA fraction. The synthesized biopolymer shows an extremely high hydrophilicity, due to the simultaneous presence of PGA and EPS. The analysis of its functional properties and the presence of glucosamine in the highest proportion in EPS fraction clearly determine the potential of EP synthesized by C. canadensis 28 for application in the cosmetics industry.

FED-R2: concept and magnet design of a low-cost FED

International Nuclear Information System (INIS)

Williams, J.E.C.; Becker, H.; Blackfield, D.; Bobrov, E.; Bromberg, L.; Cohn, D.R.; Diatchenko, N.; LeClaire, R.

1982-12-01

High performance resistive magnet technology was used to develop a design for a compact, low cost version of the fusion engineering device FED. We refer to this design as FED-R2, for FED-resistive magnet design 2 to distinguish it from the larger resistive magnet design for FED which uses demountable coils (FED-R1). The main objectives of FED-R2 are: (1) to demonstrate reliable, quasi-steady state (long pulse, high duty factor) operation with Q/sub p/ approx. 5; (2) to demonstrate Q/sub p/ > 5 operation for a limited number of pulses; (3) to provide high neutron flux for irradiation of nuclear test modules with a total area greater tha 20m 2 ; (4) to utilize steady-state RF current drive if this option appears promising. Based upon the costing codes at the Fusion Engineering Design Center and upon TFTR costs, the estimated direct costs of FED-R2 would be on the range 380 to 460M, a factor of about 2 below that of the baseline FED design

Microbe observation and cultivation array (MOCA) for cultivating and analyzing environmental microbiota.

Science.gov (United States)

Gao, Weimin; Navarroli, Dena; Naimark, Jared; Zhang, Weiwen; Chao, Shih-Hui; Meldrum, Deirdre R

2013-01-09

The use of culture-independent nucleic acid techniques, such as ribosomal RNA gene cloning library analysis, has unveiled the tremendous microbial diversity that exists in natural environments. In sharp contrast to this great achievement is the current difficulty in cultivating the majority of bacterial species or phylotypes revealed by molecular approaches. Although recent new technologies such as metagenomics and metatranscriptomics can provide more functionality information about the microbial communities, it is still important to develop the capacity to isolate and cultivate individual microbial species or strains in order to gain a better understanding of microbial physiology and to apply isolates for various biotechnological applications. We have developed a new system to cultivate bacteria in an array of droplets. The key component of the system is the microbe observation and cultivation array (MOCA), which consists of a Petri dish that contains an array of droplets as cultivation chambers. MOCA exploits the dominance of surface tension in small amounts of liquid to spontaneously trap cells in well-defined droplets on hydrophilic patterns. During cultivation, the growth of the bacterial cells across the droplet array can be monitored using an automated microscope, which can produce a real-time record of the growth. When bacterial cells grow to a visible microcolony level in the system, they can be transferred using a micropipette for further cultivation or analysis. MOCA is a flexible system that is easy to set up, and provides the sensitivity to monitor growth of single bacterial cells. It is a cost-efficient technical platform for bioassay screening and for cultivation and isolation of bacteria from natural environments.

FEDS

DEFF Research Database (Denmark)

Venable, John; Pries-Heje, Jan; Baskerville, Richard

2016-01-01

Evaluation of design artefacts and design theories is a key activity in Design Science Research (DSR), as it provides feedback for further development and (if done correctly) assures the rigour of the research. However, the extant DSR literature provides insufficient guidance on evaluation...... to enable Design Science Researchers to effectively design and incorporate evaluation activities into a DSR project that can achieve DSR goals and objectives. To address this research gap, this research paper develops, explicates, and provides evidence for the utility of a Framework for Evaluation in Design...... Science (FEDS) together with a process to guide design science researchers in developing a strategy for evaluating the artefacts they develop within a DSR project. A FEDS strategy considers why, when, how, and what to evaluate. FEDS includes a two-dimensional characterisation of DSR evaluation episodes...

Proteomic profiling of Bacillus licheniformis reveals a stress response mechanism in the synthesis of extracellular polymeric flocculants.

Science.gov (United States)

Yu, Wencheng; Chen, Zhen; Shen, Liang; Wang, Yuanpeng; Li, Qingbiao; Yan, Shan; Zhong, Chuan-Jian; He, Ning

2016-04-01

Some bioflocculants composed of extracellular polymeric substances are produced under peculiar conditions. Bacillus licheniformis CGMCC2876 is a microorganism that secretes both extracellular polysaccharides (EPS) and poly-gamma-glutamic acid (γ-PGA) under stress conditions. In this work, SWATH acquisition LC-MS/MS method was adopted for differential proteomic analysis of B. licheniformis, aiming at determining the bacterial stress mechanism. Compared with LB culture, 190 differentially expressed proteins were identified in B. licheniformis CGMCC2876 cultivated in EPS culture, including 117 up-regulated and 73 down-regulated proteins. In γ-PGA culture, 151 differentially expressed proteins, 89 up-regulated and 62 down-regulated, were found in the cells. Up-regulated proteins involved in amino acid biosynthesis were found to account for 43% and 41% of the proteomes in EPS and γ-PGA cultivated cells, respectively. Additionally, a series of proteins associated with amino acid degradation were found to be repressed under EPS and γ-PGA culture conditions. Transcriptional profiling via the qPCR detection of selected genes verified the proteomic analysis. Analysis of free amino acids in the bacterial cells further suggested the presence of amino acid starvation conditions. EPS or γ-PGA was synthesized to alleviate the effect of amino acid limitation in B. licheniformis. This study identified a stress response mechanism in the synthesis of macromolecules in B. licheniformis, providing potential culture strategies to improve the production of two promising bioflocculants. © 2015 Wiley Periodicals, Inc.

Queue Length and Server Content Distribution in an Infinite-Buffer Batch-Service Queue with Batch-Size-Dependent Service

Directory of Open Access Journals (Sweden)

U. C. Gupta

2015-01-01

Full Text Available We analyze an infinite-buffer batch-size-dependent batch-service queue with Poisson arrival and arbitrarily distributed service time. Using supplementary variable technique, we derive a bivariate probability generating function from which the joint distribution of queue and server content at departure epoch of a batch is extracted and presented in terms of roots of the characteristic equation. We also obtain the joint distribution of queue and server content at arbitrary epoch. Finally, the utility of analytical results is demonstrated by the inclusion of some numerical examples which also includes the investigation of multiple zeros.

Polynomial Batch Codes for Efficient IT-PIR

Directory of Open Access Journals (Sweden)

Henry Ryan

2016-10-01

Full Text Available Private information retrieval (PIR is a way for clients to query a remote database without the database holder learning the clients’ query terms or the responses they generate. Compelling applications for PIR are abound in the cryptographic and privacy research literature, yet existing PIR techniques are notoriously inefficient. Consequently, no such PIRbased application to date has seen real-world at-scale deployment. This paper proposes new “batch coding” techniques to help address PIR’s efficiency problem. The new techniques exploit the connection between ramp secret sharing schemes and efficient information-theoretically secure PIR (IT-PIR protocols. This connection was previously observed by Henry, Huang, and Goldberg (NDSS 2013, who used ramp schemes to construct efficient “batch queries” with which clients can fetch several database records for the same cost as fetching a single record using a standard, non-batch query. The new techniques in this paper generalize and extend those of Henry et al. to construct “batch codes” with which clients can fetch several records for only a fraction the cost of fetching a single record using a standard non-batch query over an unencoded database. The batch codes are highly tuneable, providing a means to trade off (i lower server-side computation cost, (ii lower server-side storage cost, and/or (iii lower uni- or bi-directional communication cost, in exchange for a comparatively modest decrease in resilience to Byzantine database servers.

Bioprocess development for extracellular production of recombinant human interleukin-3 (hIL-3) in Pichia pastoris.

Science.gov (United States)

Dagar, Vikas Kumar; Adivitiya; Devi, Nirmala; Khasa, Yogender Pal

2016-10-01

Human interleukin-3 (hIL-3) is a therapeutically important cytokine involved in the maturation and differentiation of various cells of the immune system. The codon-optimized hIL-3 gene was cloned in fusion with the N-terminus α-mating factor signal peptide of Saccharomyces cerevisiae under an inducible alcohol oxidase 1 (AOX1) and constitutive glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter. A Zeocin concentration up to 2000 mg/L was used to select hyper-producers. The shake flask cultivation studies in the Pichia pastoris GS115 host resulted a maximum recombinant hIL-3 expression level of 145 mg/L in the extracellular medium under the control of AOX1 promoter. The batch fermentation strategy allowed us to attain a fairly pure glycosylated hIL-3 protein in the culture supernatant at a final concentration of 475 mg/L with a high volumetric productivity of 4.39 mg/L/h. The volumetric product concentration achieved at bioreactor level was 3.28 folds greater than the shake flask results. The 6x His-tagged protein was purified using Ni-NTA affinity chromatography and confirmed further by western blot analysis using anti-6x His tag antibody. The glycosylation of recombinant hIL-3 protein was confirmed in a PNGase F deglycosylation reaction where it showed a molecular weight band pattern similar to E. coli produced non-glycosylated hIL-3 protein. The structural properties of recombinant hIL-3 protein were confirmed by CD and fluorescence spectroscopy where protein showed 40 % α-helix, 12 % β-sheets with an emission maxima at 343 nm. MALDI-TOF-TOF analysis was used to establish the protein identity. The biological activity of purified protein was confirmed by the human erythroleukemia TF-1 cell proliferation assay.

Impact of glycerol and nitrogen concentration on Enterobacter A47 growth and exopolysaccharide production.

Science.gov (United States)

Torres, Cristiana A V; Marques, Rodolfo; Ferreira, Ana R V; Antunes, Sílvia; Grandfils, Christian; Freitas, Filomena; Reis, Maria A M

2014-11-01

Enterobacter A47 produces a fucose-containing exopolysaccharide (EPS) by cultivation in mineral medium supplemented with glycerol. EPS synthesis by Enterobacter A47 was shown to be influenced by both the initial glycerol and nitrogen concentrations and by the nutrients' feeding rate during the fed-batch phase. Initial nitrogen concentrations above 1.05g/L were detrimental for EPS synthesis: the productivity was reduced to 0.35-0.62g/Ld (compared to 1.89-2.04g/Ld under lower nitrogen concentrations) and the polymer had lower fucose content (14-17%mol, compared to 36-38%mol under lower nitrogen concentrations). On the other hand, EPS productivity was improved to 5.66g/Ld by increasing the glycerol and nitrogen feeding rates during the fed-batch phase. However, the EPS thus obtained had lower fucose (26%mol) and higher galactose (34%mol) contents, as well as lower average molecular weight (7.2×10(5)). The ability of Enterobacter A47 to synthesize EPS with different physico-chemical characteristics may be useful for the generation of biopolymers with distinct functional properties suitable for different applications. Copyright © 2014 Elsevier B.V. All rights reserved.

Batch Computed Tomography Analysis of Projectiles

Science.gov (United States)

2016-05-01

ARL-TR-7681 ● MAY 2016 US Army Research Laboratory Batch Computed Tomography Analysis of Projectiles by Michael C Golt, Chris M...Laboratory Batch Computed Tomography Analysis of Projectiles by Michael C Golt and Matthew S Bratcher Weapons and Materials Research...values to account for projectile variability in the ballistic evaluation of armor. 15. SUBJECT TERMS computed tomography , CT, BS41, projectiles

Fungal cultivation on glass-beads

DEFF Research Database (Denmark)

Droce, Aida; Sørensen, Jens Laurids; Giese, Henriette

Transcription of various bioactive compounds and enzymes are dependent on fungal cultivation method. In this study we cultivate Fusarium graminearum and Fusarium solani on glass-beads with liquid media in petri dishes as an easy and inexpensive cultivation method, that resembles in secondary...... metabolite production to agar-cultivation but with an easier and more pure RNA-extraction of total fungal mycelia....

Increased kidney growth in formula-fed versus breast-fed healthy infants

DEFF Research Database (Denmark)

Schmidt, Ida M; Damgaard, Ida N; Boisen, Kirsten A

2004-01-01

versus breast feeding on kidney growth in a cohort of 631 healthy children examined at birth, and at 3 and 18 months of age. Kidney size was determined by ultrasonography and related to gender, age, body size, and feeding category (fully breast fed, partially breast fed, or fully formula fed at 3 months...

A canned food scheduling problem with batch due date

Science.gov (United States)

Chung, Tsui-Ping; Liao, Ching-Jong; Smith, Milton

2014-09-01

This article considers a canned food scheduling problem where jobs are grouped into several batches. Jobs can be sent to the next operation only when all the jobs in the same batch have finished their processing, i.e. jobs in a batch, have a common due date. This batch due date problem is quite common in canned food factories, but there is no efficient heuristic to solve the problem. The problem can be formulated as an identical parallel machine problem with batch due date to minimize the total tardiness. Since the problem is NP hard, two heuristics are proposed to find the near-optimal solution. Computational results comparing the effectiveness and efficiency of the two proposed heuristics with an existing heuristic are reported and discussed.

Prunus dulcis, Batch

African Journals Online (AJOL)

STORAGESEVER

2010-06-07

Jun 7, 2010 ... almond (Prunus dulcis, Batch) genotypes as revealed by PCR analysis. Yavar Sharafi1*, Jafar Hajilou1, Seyed AbolGhasem Mohammadi2, Mohammad Reza Dadpour1 and Sadollah Eskandari3. 1Department of Horticulture, Faculty of Agriculture, University of Tabriz, Tabriz, 5166614766, Iran.

40 CFR 63.1408 - Aggregate batch vent stream provisions.

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 11 2010-07-01 2010-07-01 true Aggregate batch vent stream provisions... § 63.1408 Aggregate batch vent stream provisions. (a) Emission standards. Owners or operators of aggregate batch vent streams at a new or existing affected source shall comply with either paragraph (a)(1...

Algae-facilitated chemical phosphorus removal during high-density Chlorella emersonii cultivation in a membrane bioreactor.

Science.gov (United States)

Xu, Meng; Bernards, Matthew; Hu, Zhiqiang

2014-02-01

An algae-based membrane bioreactor (A-MBR) was evaluated for high-density algae cultivation and phosphorus (P) removal. The A-MBR was seeded with Chlorella emersonii and operated at a hydraulic retention time of 1day with minimal biomass wastage for about 150days. The algae concentration increased from initially 385mg/L (or 315mg biomass COD/L) to a final of 4840mg/L (or 1664mg COD/L), yielding an average solids (algae biomass+minerals) production rate of 32.5gm(-3)d(-1) or 6.2gm(-2)d(-1). The A-MBR was able to remove 66±9% of the total P from the water while the algal biomass had an average of 7.5±0.2% extracellular P and 0.4% of intracellular P. The results suggest that algae-induced phosphate precipitation by algae is key to P removal and high-density algae cultivation produces P-rich algal biomass with excellent settling properties. Copyright © 2013 Elsevier Ltd. All rights reserved.

Transcriptomic and proteomic analyses of Desulfovibrio vulgaris biofilms: carbon and energy flow contribute to the distinct biofilm growth state.

Science.gov (United States)

Clark, Melinda E; He, Zhili; Redding, Alyssa M; Joachimiak, Marcin P; Keasling, Jay D; Zhou, Jizhong Z; Arkin, Adam P; Mukhopadhyay, Aindrila; Fields, Matthew W

2012-04-16

Desulfovibrio vulgaris Hildenborough is a sulfate-reducing bacterium (SRB) that is intensively studied in the context of metal corrosion and heavy-metal bioremediation, and SRB populations are commonly observed in pipe and subsurface environments as surface-associated populations. In order to elucidate physiological changes associated with biofilm growth at both the transcript and protein level, transcriptomic and proteomic analyses were done on mature biofilm cells and compared to both batch and reactor planktonic populations. The biofilms were cultivated with lactate and sulfate in a continuously fed biofilm reactor, and compared to both batch and reactor planktonic populations. The functional genomic analysis demonstrated that biofilm cells were different compared to planktonic cells, and the majority of altered abundances for genes and proteins were annotated as hypothetical (unknown function), energy conservation, amino acid metabolism, and signal transduction. Genes and proteins that showed similar trends in detected levels were particularly involved in energy conservation such as increases in an annotated ech hydrogenase, formate dehydrogenase, pyruvate:ferredoxin oxidoreductase, and rnf oxidoreductase, and the biofilm cells had elevated formate dehydrogenase activity. Several other hydrogenases and formate dehydrogenases also showed an increased protein level, while decreased transcript and protein levels were observed for putative coo hydrogenase as well as a lactate permease and hyp hydrogenases for biofilm cells. Genes annotated for amino acid synthesis and nitrogen utilization were also predominant changers within the biofilm state. Ribosomal transcripts and proteins were notably decreased within the biofilm cells compared to exponential-phase cells but were not as low as levels observed in planktonic, stationary-phase cells. Several putative, extracellular proteins (DVU1012, 1545) were also detected in the extracellular fraction from biofilm cells

Growth of aspergillus terreus and the production of itaconic acid in batch and continuous cultures. The influence of pH

Energy Technology Data Exchange (ETDEWEB)

Rychtera, M.; Wase, J.D.A.

1981-01-01

Aspergillus terreus (NRRL 1960) was cultivated in batch and in continuous single-stage culture. The influence of pH on the growth of the organism, on the formation of itaconic acid and on the kinetics of fermentation was studied under phosphate limitation, both at controlled ph values and also when the pH was allowed to decrease in a natural way. In the pH range 1.7-3.5, the ratio of undissociated:half-dissociated acid varied from 190:1 to 1.5:1. The amount of completely dissociated acid may be regarded as negligible. In batch systems operated without pH control, an initial pH of 3.1 proved to be the most effective. Product formation under such conditions started at a point where the exponential growth phase commenced and was described by a zero-order equation. The maximum itaconic acid production rate was shifted behind maximum growth rate. The continuous single-stage system was first order with respect to product formation. At pH greater than 3.1, a number of aberrant and pellet forms of the mould occurred, resulting in decreased acid production. (Refs. 41).

Effect of glass-batch makeup on the melting process

International Nuclear Information System (INIS)

Hrma, Pavel R.; Schweiger, Michael J.; Humrickhouse, Carissa J.; Moody, J. Adam; Tate, Rachel M.; Rainsdon, Timothy T.; Tegrotenhuis, Nathan E.; Arrigoni, Benjamin M.; Marcial, Jose; Rodriguez, Carmen P.; Tincher, Benjamin

2010-01-01

The response of a glass batch to heating is determined by the batch makeup and in turn determines the rate of melting. Batches formulated for a high-alumina nuclear waste to be vitrified in an all-electric melter were heated at a constant temperature-increase rate to determine changes in melting behavior in response to the selection of batch chemicals and silica grain-size as well as the addition of heat-generating reactants. The type of batch materials and the size of silica grains determine how much, if any, primary foam occurs during melting. Small quartz grains, 5 (micro)m in size, caused extensive foaming because their major portion dissolved at temperatures 800 C when batch gases no longer evolved. The exothermal reaction of nitrates with sucrose was ignited at a temperature as low as 160 C and caused a temporary jump in temperature of several hundred degrees. Secondary foam, the source of which is oxygen from redox reactions, occurred in all batches of a limited composition variation involving five oxides, B 2 O 3 , CaO, Li 2 O, MgO, and Na 2 O. The foam volume at the maximum volume-increase rate was a weak function of temperature and melt basicity. Neither the batch makeup nor the change in glass composition had a significant impact on the dissolution of silica grains. The impacts of primary foam generation on glass homogeneity and the rate of melting in large-scale continuous furnaces have yet to be established via mathematical modeling and melter experiments.

Effect Of Glass-Batch Makeup On The Melting Process

International Nuclear Information System (INIS)

Kruger, A.A.; Hrma, P.

2010-01-01

The response of a glass batch to heating is determined by the batch makeup and in turn determines the rate of melting. Batches formulated for a high-alumina nuclear waste to be vitrified in an all-electric melter were heated at a constant temperature-increase rate to determine changes in melting behavior in response to the selection of batch chemicals and silica grain-size as well as the addition of heat-generating reactants. The type of batch materials and the size of silica grains determine how much, if any, primary foam occurs during melting. Small quartz grains, 5 (micro)m in size, caused extensive foaming because their major portion dissolved at temperatures 800 C when batch gases no longer evolved. The exothermal reaction of nitrates with sucrose was ignited at a temperature as low as 160 C and caused a temporary jump in temperature of several hundred degrees. Secondary foam, the source of which is oxygen from redox reactions, occurred in all batches of a limited composition variation involving five oxides, B 2 O 3 , CaO, Li 2 O, MgO, and Na 2 O. The foam volume at the maximum volume-increase rate was a weak function of temperature and melt basicity. Neither the batch makeup nor the change in glass composition had a significant impact on the dissolution of silica grains. The impacts of primary foam generation on glass homogeneity and the rate of melting in large-scale continuous furnaces have yet to be established via mathematical modeling and melter experiments.

Simultaneous nitrogen, phosphorous, and hardness removal from reverse osmosis concentrate by microalgae cultivation.

Science.gov (United States)

Wang, Xiao-Xiong; Wu, Yin-Hu; Zhang, Tian-Yuan; Xu, Xue-Qiao; Dao, Guo-Hua; Hu, Hong-Ying

2016-05-01

While reverse osmosis (RO) is a promising technology for wastewater reclamation, RO concentrate (ROC) treatment and disposal are important issues to consider. Conventional chemical and physical treatment methods for ROC present certain limitations, such as relatively low nitrogen and phosphorus removal efficiencies as well as the requirement of an extra process for hardness removal. This study proposes a novel biological approach for simultaneous removal of nitrogen, phosphorus, and calcium (Ca(2+)) and magnesium (Mg(2+)) ions from the ROC of municipal wastewater treatment plants by microalgal cultivation and algal biomass production. Two microalgae strains, Chlorella sp. ZTY4 and Scenedesmus sp. LX1, were used for batch cultivation of 14-16 days. Both strains grew well in ROC with average biomass production of 318.7 mg/L and lipid contents up to 30.6%, and nitrogen and phosphorus could be effectively removed with efficiencies of up to 89.8% and 92.7%, respectively. Approximately 55.9%-83.7% Ca(2+) could be removed from the system using the cultured strains. Mg(2+) removal began when Ca(2+) precipitation ceased, and the removal efficiency of the ion could reach up to 56.0%. The most decisive factor influencing Ca(2+) and Mg(2+) removal was chemical precipitation with increases in pH caused by algal growth. The results of this study provide a new biological approach for removing nitrogen, phosphorous, and hardness from ROC. The results suggest that microalgal cultivation presents new opportunities for applying an algal process to ROC treatment. The proposed approach serves dual purposes of nutrient and hardness reduction and production of lipid rich micro-algal biomass. Copyright © 2016 Elsevier Ltd. All rights reserved.

Spatial and interannual variability in Baltic sprat batch fecundity

DEFF Research Database (Denmark)

Haslob, H.; Tomkiewicz, Jonna; Hinrichsen, H.H.

2011-01-01

in the central Baltic Sea, namely the Bornholm Basin, Gdansk Deep and Southern Gotland Basin. Environmental parameters such as hydrography, fish condition and stock density were tested in order to investigate the observed variability in sprat fecundity. Absolute batch fecundity was found to be positively related...... to fish length and weight. Significant differences in absolute and relative batch fecundity of Baltic sprat among areas and years were detected, and could partly be explained by hydrographic features of the investigated areas. A non-linear multiple regression model taking into account fish length...... and ambient temperature explained 70% of variability in absolute batch fecundity. Oxygen content and fish condition were not related to sprat batch fecundity. Additionally, a negative effect of stock size on sprat batch fecundity in the Bornholm Basin was revealed. The obtained data and results are important...

Fuzzy batch controller for granular materials

OpenAIRE

Zamyatin Nikolaj; Smirnov Gennadij; Fedorchuk Yuri; Rusina Olga

2018-01-01

The paper focuses on batch control of granular materials in production of building materials from fluorine anhydrite. Batching equipment is intended for smooth operation and timely feeding of supply hoppers at a required level. Level sensors and a controller of an asynchronous screw drive motor are used to control filling of the hopper with industrial anhydrite binders. The controller generates a required frequency and ensures required productivity of a feed conveyor. Mamdani-type fuzzy infer...

Research on AO/FO batch management technology in aircraft production

Directory of Open Access Journals (Sweden)

Yin Haijun

2018-01-01

Full Text Available Based on the analysis of the characteristics and significance of AO/FO in the process of aircraft production, this paper analyzes the format rules of AO/FO batch management from the perspective of technology realization, and details the AO/FO The change of the query and the change status tracking, introduces the AO/FO single-stand status display in the batch management, increases the structure definition of the attribute table in the batch management, and designs the relevant algorithm to store and calculate the batch information. Finally, based on the above theory support AO/FO batch management system successfully used in the production of a machine.

Multiplatform Mass Spectrometry-Based Approach Identifies Extracellular Glycolipids of the Yeast Rhodotorula babjevae UCDFST 04-877.

Science.gov (United States)

Cajka, Tomas; Garay, Luis A; Sitepu, Irnayuli R; Boundy-Mills, Kyria L; Fiehn, Oliver

2016-10-28

A multiplatform mass spectrometry-based approach was used for elucidating extracellular lipids with biosurfactant properties produced by the oleaginous yeast Rhodotorula babjevae UCDFST 04-877. This strain secreted 8.6 ± 0.1 g/L extracellular lipids when grown in a benchtop bioreactor fed with 100 g/L glucose in medium without addition of hydrophobic substrate, such as oleic acid. Untargeted reversed-phase liquid chromatography-quadrupole/time-of-flight mass spectrometry (QTOFMS) detected native glycolipid molecules with masses of 574-716 Da. After hydrolysis into the fatty acid and sugar components and hydrophilic interaction chromatography-QTOFMS analysis, the extracellular lipids were found to consist of hydroxy fatty acids and sugar alcohols. Derivatization and chiral separation gas chromatography-mass spectrometry (GC-MS) identified these components as d-arabitol, d-mannitol, (R)-3-hydroxymyristate, (R)-3-hydroxypalmitate, and (R)-3-hydroxystearate. In order to assemble these substructures back into intact glycolipids that were detected in the initial screen, potential structures were in-silico acetylated to match the observed molar masses and subsequently characterized by matching predicted and observed MS/MS fragmentation using the Mass Frontier software program. Eleven species of acetylated sugar alcohol esters of hydroxy fatty acids were characterized for this yeast strain.

Aerobic granulation strategy for bioaugmentation of a sequencing batch reactor (SBR) treating high strength pyridine wastewater

Energy Technology Data Exchange (ETDEWEB)

Liu, Xiaodong; Chen, Yan [Jiangsu Key Laboratory for Chemical Pollution Control and Resources Reuse, School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094, Jiangsu Province (China); Zhang, Xin [Jiangsu Key Laboratory for Chemical Pollution Control and Resources Reuse, School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094, Jiangsu Province (China); Suzhou Institute of Architectural Design Co., Ltd, Suzhou 215021, Jiangsu Province (China); Jiang, Xinbai; Wu, Shijing [Jiangsu Key Laboratory for Chemical Pollution Control and Resources Reuse, School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094, Jiangsu Province (China); Shen, Jinyou, E-mail: shenjinyou@mail.njust.edu.cn [Jiangsu Key Laboratory for Chemical Pollution Control and Resources Reuse, School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094, Jiangsu Province (China); Sun, Xiuyun; Li, Jiansheng; Lu, Lude [Jiangsu Key Laboratory for Chemical Pollution Control and Resources Reuse, School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094, Jiangsu Province (China); Wang, Lianjun, E-mail: wanglj@mail.njust.edu.cn [Jiangsu Key Laboratory for Chemical Pollution Control and Resources Reuse, School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094, Jiangsu Province (China)

2015-09-15

Abstract: Aerobic granules were successfully cultivated in a sequencing batch reactor (SBR), using a single bacterial strain Rhizobium sp. NJUST18 as the inoculum. NJUST18 presented as both a good pyridine degrader and an efficient autoaggregator. Stable granules with diameter of 0.5–1 mm, sludge volume index of 25.6 ± 3.6 mL g{sup −1} and settling velocity of 37.2 ± 2.7 m h{sup −1}, were formed in SBR following 120-day cultivation. These granules exhibited excellent pyridine degradation performance, with maximum volumetric degradation rate (V{sub max}) varied between 1164.5 mg L{sup −1} h{sup −1} and 1867.4 mg L{sup −1} h{sup −1}. High-throughput sequencing analysis exhibited a large shift in microbial community structure, since the SBR was operated under open condition. Paracoccus and Comamonas were found to be the most predominant species in the aerobic granule system after the system had stabilized. The initially inoculated Rhizobium sp. lost its dominance during aerobic granulation. However, the inoculation of Rhizobium sp. played a key role in the start-up process of this bioaugmentation system. This study demonstrated that, in addition to the hydraulic selection pressure during settling and effluent discharge, the selection of aggregating bacterial inocula is equally important for the formation of the aerobic granule.

Aerobic granulation strategy for bioaugmentation of a sequencing batch reactor (SBR) treating high strength pyridine wastewater

International Nuclear Information System (INIS)

Liu, Xiaodong; Chen, Yan; Zhang, Xin; Jiang, Xinbai; Wu, Shijing; Shen, Jinyou; Sun, Xiuyun; Li, Jiansheng; Lu, Lude; Wang, Lianjun

2015-01-01

Abstract: Aerobic granules were successfully cultivated in a sequencing batch reactor (SBR), using a single bacterial strain Rhizobium sp. NJUST18 as the inoculum. NJUST18 presented as both a good pyridine degrader and an efficient autoaggregator. Stable granules with diameter of 0.5–1 mm, sludge volume index of 25.6 ± 3.6 mL g −1 and settling velocity of 37.2 ± 2.7 m h −1 , were formed in SBR following 120-day cultivation. These granules exhibited excellent pyridine degradation performance, with maximum volumetric degradation rate (V max ) varied between 1164.5 mg L −1 h −1 and 1867.4 mg L −1 h −1 . High-throughput sequencing analysis exhibited a large shift in microbial community structure, since the SBR was operated under open condition. Paracoccus and Comamonas were found to be the most predominant species in the aerobic granule system after the system had stabilized. The initially inoculated Rhizobium sp. lost its dominance during aerobic granulation. However, the inoculation of Rhizobium sp. played a key role in the start-up process of this bioaugmentation system. This study demonstrated that, in addition to the hydraulic selection pressure during settling and effluent discharge, the selection of aggregating bacterial inocula is equally important for the formation of the aerobic granule

Variance bias analysis for the Gelbard's batch method

Energy Technology Data Exchange (ETDEWEB)

Seo, Jae Uk; Shim, Hyung Jin [Seoul National Univ., Seoul (Korea, Republic of)

2014-05-15

In this paper, variances and the bias will be derived analytically when the Gelbard's batch method is applied. And then, the real variance estimated from this bias will be compared with the real variance calculated from replicas. Variance and the bias were derived analytically when the batch method was applied. If the batch method was applied to calculate the sample variance, covariance terms between tallies which exist in the batch were eliminated from the bias. With the 2 by 2 fission matrix problem, we could calculate real variance regardless of whether or not the batch method was applied. However as batch size got larger, standard deviation of real variance was increased. When we perform a Monte Carlo estimation, we could get a sample variance as the statistical uncertainty of it. However, this value is smaller than the real variance of it because a sample variance is biased. To reduce this bias, Gelbard devised the method which is called the Gelbard's batch method. It has been certificated that a sample variance get closer to the real variance when the batch method is applied. In other words, the bias get reduced. This fact is well known to everyone in the MC field. However, so far, no one has given the analytical interpretation on it.

Biochemical studies of amylase, lipase and protease in Callosobruchus maculatus (Coleoptera: Chrysomelidae) populations fed with Vigna unguiculata grain cultivated with diazotrophic bacteria strains.

Science.gov (United States)

Silva, L B; Torres, É B; Nóbrega, R A S; Lopes, G N; Vogado, R F; Pavan, B E; Fernandes-Junior, P I

2017-12-01

The objective of this study was to evaluate the enzymatic activity of homogenates of insects fed on grain of cowpea, Vigna unguiculata (L.), cultivars grown with different nitrogen sources. For the experiment we used aliquots of the homogenate of 100 unsexed adult insects, emerged from 10 g of grain obtained from four cowpea cultivars: 'BRS Acauã', 'BRS Carijó', 'BRS Pujante', and 'BRS Tapaihum' grown under different regimes of nitrogen sources: mineral fertilizer, inoculation with strains of diazotrophs (BR 3267, BR 3262, BR 3299; INPA 03-11B, 03-84 UFLA, as well as the control (with soil nitrogen). The parameters evaluated were enzymatic activities of insect protease, amylase and lipase and the starch content of the grains. There were differences in the enzymatic activity of amylase, lipase and protease of insect homogenate according to the food source. A lower activity of the enzyme amylase from C. maculatus homogenate was observed when insects were fed grain of the cultivar BRS Carijó. A lower activity of lipase enzyme from C. maculatus homogenate was observed when the insects fed on grain from the interaction of the cultivar Tapaihum inoculated with BR 3262 diazotrophs. The lowest proteolytic activity was observed in homogenate of insects fed on interaction of 'BRS Carijó' inoculated with BR 3262 diazotrophs. Starch content correlated positively with the amylase activity of C. maculatus homogenate. The cultivar BRS Carijó had a different behavior from the other cultivars, according to the cluster analysis.

Enhanced biogas yield from energy crops with rumen anaerobic fungi

Energy Technology Data Exchange (ETDEWEB)

Prochazka, Jindrich; Zabranska, Jana; Dohanyos, Michal [Department of Water Technology and Environmental Engineering, Faculty of Environmental Technology, Institute of Chemical Technology in Prague, Prague (Czech Republic); Mrazek, Jakub; Strosova, Lenka; Fliegerova, Katerina [Laboratory of Anaerobic Microbiology, Institute of Animal Physiology and Genetics, CAS, v.v.i., Prague (Czech Republic)

2012-06-15

Anaerobic fungi (AF) are able to degrade crop substrates with higher efficiency than commonly used anaerobic bacteria. The aim of this study was to investigate ways of use of rumen AF to improve biogas production from energy crops under laboratory conditions. In this study, strains of AF isolated from feces or rumen fluid of cows and deer were tested for their ability to integrate into the anaerobic bacterial ecosystem used for biogas production, in order to improve degradation of substrate polysaccharides and consequently the biogas yield. Batch culture, fed batch culture, and semicontinuous experiments have been performed using anaerobic sludge from pig slurry fermentation and different kinds of substrates (celluloses, maize, and grass silage) inoculated by different genera of AF. All experiments showed a positive effect of AF on the biogas yield and quality. AF improved the biogas production by 4-22%, depending on the substrate and AF species used. However, all the cultivation experiments indicated that rumen fungi do not show long-term survival in fermenters with digestate from pig slurry. The best results were achieved during fed batch experiment with fungal culture Anaeromyces (KF8), in which biogas production was enhanced during the whole experimental period of 140 days. This result has not been achieved in semicontinuous experiment, where increment in biogas production in fungal enriched reactor was only 4% after 42 days. (copyright 2012 Wiley-VCH Verlag GmbH and Co. KGaA, Weinheim)

Bioaugmentation of aerobic sludge granules with a plasmid donor strain for enhanced degradation of 2,4-dichlorophenoxyacetic acid

International Nuclear Information System (INIS)

Quan Xiangchun; Tang Hua; Xiong Weicong; Yang Zhifeng

2010-01-01

Aerobic sludge granules pre-grown on glucose were bioaugmented with a plasmid pJP4 carrying strain Pseudomonas putida SM1443 in a fed-batch microcosm system and a lab-scale sequencing batch reactor (SBR) to enhance their degradation capacity to 2,4-dichlorophenoxyacetic acid (2,4-D). The fed-batch test results showed that the bioaugmented aerobic granule system gained 2,4-D degradation ability faster and maintained a more stable microbial community than the control in the presence of 2,4-D. 2,4-D at the initial concentration of about 160 mg/L was nearly completely removed by the bioaugmented granule system within 62 h, while the control system only removed 26% within 66 h. In the bioaugmented SBR which had been operated for 90 days, the seeded aerobic granules pre-grown on glucose successfully turned into 2,4-D degrading granules through bioaugmentation and stepwise increase of 2,4-D concentration from 8 to 385 mg/L. The granules showed a compact structure and good settling ability with the mean diameter of about 450 μm. The degradation kinetics of 2,4-D by the aerobic granules can be described with the Haldane kinetics model with V max = 31.1 mg 2,4-D/gVSS h, K i = 597.9 mg/L and K s = 257.3 mg/L, respectively. This study shows that plasmid mediated bioaugmentation is a feasible strategy to cultivate aerobic granules degrading recalcitrant pollutants.

40 CFR 63.1322 - Batch process vents-reference control technology.

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 11 2010-07-01 2010-07-01 true Batch process vents-reference control technology. 63.1322 Section 63.1322 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR... Batch process vents—reference control technology. (a) Batch process vents. The owner or operator of a...

Cashew apple juice as microbial cultivation medium for non-immunogenic hyaluronic acid production.

Science.gov (United States)

Oliveira, Adriano H; Ogrodowski, Cristiane C; de Macedo, André C; Santana, Maria Helena A; Gonçalves, Luciana R B

2013-12-01

In this work, natural cashew apple juice was used as cultivation medium as an alternative to substitute brain heart infusion medium. The effect of aeration and juice supplementation with yeast extract on the production of hyaluronic acid in batch fermentation was also investigated. Similar levels of cell mass were obtained in inoculum using cashew apple juice supplemented with yeast extract or the conventional brain heart infusion medium. Fermentation in Erlenmeyer flasks produced low biomass and hyaluronic acid concentrations. The hyaluronic acid concentration and viscosity increased from 0.15 g/L and 3.87 cP (no aeration or medium supplementation) to 1.76 g/L and 107 cP, when aeration (2 vvm) and 60 g/L of yeast extract were used. The results suggest the production of low-molecular weight hyaluronic acid oligomers instead of the high molecular weight polymer.

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...

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.

Abiotic factors in colony formation: effects of nutrition and light on extracellular polysaccharide production and cell aggregates of Microcystis aeruginosa

Science.gov (United States)

Yang, Zhen; Kong, Fanxiang

2013-07-01

Colony morphology is important for Microcystis to sustain a competitive advantage in eutrophic lakes. The mechanism of colony formation in Microcystis is currently unclear. Extracellular polysaccharide (EPS) has been reported to play an important role in cell aggregate formation of some phytoplankton. Microcystis aeruginosa was cultivated under varied abiotic conditions, including different nutrient, light, and temperature conditions, to investigate their effects on EPS production and morphological change. The results show that nutrient concentration and light intensity have great effects on EPS productionin M. aeruginosa. There was a considerable increase in EPS production after M. aeruginosa was cultivated in adjusted culture conditions similar to those present in the field (28.9 mg C/L, 1.98 mg N/L, 0.65 mg P/L, light intensity: 100 μmol/(m2 · s)). These results indicate that abiotic factors might be one of the triggers for colony formation in Microcystis.

Batch-to-Batch Quality Consistency Evaluation of Botanical Drug Products Using Multivariate Statistical Analysis of the Chromatographic Fingerprint

OpenAIRE

Xiong, Haoshu; Yu, Lawrence X.; Qu, Haibin

2013-01-01

Botanical drug products have batch-to-batch quality variability due to botanical raw materials and the current manufacturing process. The rational evaluation and control of product quality consistency are essential to ensure the efficacy and safety. Chromatographic fingerprinting is an important and widely used tool to characterize the chemical composition of botanical drug products. Multivariate statistical analysis has showed its efficacy and applicability in the quality evaluation of many ...

Fuzzy batch controller for granular materials

Directory of Open Access Journals (Sweden)

Zamyatin Nikolaj

2018-01-01

Full Text Available The paper focuses on batch control of granular materials in production of building materials from fluorine anhydrite. Batching equipment is intended for smooth operation and timely feeding of supply hoppers at a required level. Level sensors and a controller of an asynchronous screw drive motor are used to control filling of the hopper with industrial anhydrite binders. The controller generates a required frequency and ensures required productivity of a feed conveyor. Mamdani-type fuzzy inference is proposed for controlling the speed of the screw that feeds mixture components. As related to production of building materials based on fluoride anhydrite, this method is used for the first time. A fuzzy controller is proven to be effective in controlling the filling level of the supply hopper. In addition, the authors determined optimal parameters of the batching process to ensure smooth operation and production of fluorine anhydrite materials of specified properties that can compete with gypsum-based products.

Glass bead cultivation of fungi

DEFF Research Database (Denmark)

Droce, Aida; Sørensen, Jens Laurids; Giese, H.

2013-01-01

Production of bioactive compounds and enzymes from filamentous fungi is highly dependent on cultivation conditions. Here we present an easy way to cultivate filamentous fungi on glass beads that allow complete control of nutrient supply. Secondary metabolite production in Fusarium graminearum...... and Fusarium solani cultivated on agar plates, in shaking liquid culture or on glass beads was compared. Agar plate culture and glass bead cultivation yielded comparable results while liquid culture had lower production of secondary metabolites. RNA extraction from glass beads and liquid cultures was easier...... to specific nutrient factors. •Fungal growth on glass beads eases and improves fungal RNA extraction....

A disposable picolitre bioreactor for cultivation and investigation of industrially relevant bacteria on the single cell level.

Science.gov (United States)

Grünberger, Alexander; Paczia, Nicole; Probst, Christopher; Schendzielorz, Georg; Eggeling, Lothar; Noack, Stephan; Wiechert, Wolfgang; Kohlheyer, Dietrich

2012-05-08

In the continuously growing field of industrial biotechnology the scale-up from lab to industrial scale is still a major hurdle to develop competitive bioprocesses. During scale-up the productivity of single cells might be affected by bioreactor inhomogeneity and population heterogeneity. Currently, these complex interactions are difficult to investigate. In this report, design, fabrication and operation of a disposable picolitre cultivation system is described, in which environmental conditions can be well controlled on a short time scale and bacterial microcolony growth experiments can be observed by time-lapse microscopy. Three exemplary investigations will be discussed emphasizing the applicability and versatility of the device. Growth and analysis of industrially relevant bacteria with single cell resolution (in particular Escherichia coli and Corynebacterium glutamicum) starting from one single mother cell to densely packed cultures is demonstrated. Applying the picolitre bioreactor, 1.5-fold increased growth rates of C. glutamicum wild type cells were observed compared to typical 1 litre lab-scale batch cultivation. Moreover, the device was used to analyse and quantify the morphological changes of an industrially relevant l-lysine producer C. glutamicum after artificially inducing starvation conditions. Instead of a one week lab-scale experiment, only 1 h was sufficient to reveal the same information. Furthermore, time lapse microscopy during 24 h picolitre cultivation of an arginine producing strain containing a genetically encoded fluorescence sensor disclosed time dependent single cell productivity and growth, which was not possible with conventional methods.

Early-warning process/control for anaerobic digestion and biological nitrogen transformation processes: Batch, semi-continuous, and/or chemostat experiments. Final report

Energy Technology Data Exchange (ETDEWEB)

Hickey, R. [Science Applications International Corp., McLean, VA (United States)

1992-09-01

The objective of this project was to develop and test an early-warning/process control model for anaerobic sludge digestion (AD). The approach was to use batch and semi-continuously fed systems and to assemble system parameter data on a real-time basis. Specific goals were to produce a real-time early warning control model and computer code, tested for internal and external validity; to determine the minimum rate of data collection for maximum lag time to predict failure with a prescribed accuracy and confidence in the prediction; and to determine and characterize any trends in the real-time data collected in response to particular perturbations to feedstock quality. Trends in the response of trace gases carbon monoxide and hydrogen in batch experiments, were found to depend on toxicant type. For example, these trace gases respond differently for organic substances vs. heavy metals. In both batch and semi-continuously feed experiments, increased organic loading lead to proportionate increases in gas production rates as well as increases in CO and H{sub 2} concentration. An analysis of variance of gas parameters confirmed that CO was the most sensitive indicator variable by virtue of its relatively larger variance compared to the others. The other parameters evaluated including gas production, methane production, hydrogen, carbon monoxide, carbon dioxide and methane concentration. In addition, a relationship was hypothesized between gaseous CO concentration and acetate concentrations in the digester. The data from semicontinuous feed experiments were supportive.

Bioflocculation production from lower-molecular fatty acids as a novel strategy for utilization of sludge digestion liquor.

Science.gov (United States)

Fujita, M; Ike, M; Jang, J H; Kim, S M; Hirao, T

2001-01-01

We propose the bioproduction of a bioflocculant from lower-molecular fatty acids as an innovative strategy for utilizing waste sludge digestion liquor. Fundamental studies on the production, characterization and application of a novel bioflocculant were performed. Citrobactersp. TKF04 was screened out of 1,564 natural isolates as a bacterial strain capable of a bioflocculant from acetic and propionic acids. TKF04 produced the bioflocculant during the logarithmic growth in the batch cultivation, and it could be recovered from the culture supernatant by ethanol precipitation. The fed-batch cultivation with feeding of acetic acid: ammonium 10;1 (mole) to maintain pH 8.5 led to the hyper-production of the bioflocculant. The bioflocculant was found to be effective for flocculating a kaolin suspension, when added at a final concentration of 1-10 mg/l, over a wide range of pHs (2-8) and temperatures (3-95 degrees C), while the addition of cations was not required. It could flocculate a variety of inorganic and organic suspended particles including kaolin, diatomite, bentonite, activated carbon, soil and activated sludge. These indicated that the bioflocculant possesses flocculating activity comparable or superior to that of synthetic flocculants. The bioflocculation was identified as a chitosan-like biopolymer.

Extracellular Polymeric Substances Govern the Surface Charge of Biogenic Elemental Selenium Nanoparticles

KAUST Repository

Jain, Rohan

2015-02-03

© 2014 American Chemical Society. The origin of the organic layer covering colloidal biogenic elemental selenium nanoparticles (BioSeNPs) is not known, particularly in the case when they are synthesized by complex microbial communities. This study investigated the presence of extracellular polymeric substances (EPS) on BioSeNPs. The role of EPS in capping the extracellularly available BioSeNPs was also examined. Fourier transform infrared (FT-IR) spectroscopy and colorimetric measurements confirmed the presence of functional groups characteristic of proteins and carbohydrates on the BioSeNPs, suggesting the presence of EPS. Chemical synthesis of elemental selenium nanoparticles in the presence of EPS, extracted from selenite fed anaerobic granular sludge, yielded stable colloidal spherical selenium nanoparticles. Furthermore, extracted EPS, BioSeNPs, and chemically synthesized EPS-capped selenium nanoparticles had similar surface properties, as shown by ζ-potential versus pH profiles and isoelectric point measurements. This study shows that the EPS of anaerobic granular sludge form the organic layer present on the BioSeNPs synthesized by these granules. The EPS also govern the surface charge of these BioSeNPs, thereby contributing to their colloidal properties, hence affecting their fate in the environment and the efficiency of bioremediation technologies.

Treatment of opium alkaloid containing wastewater in sequencing batch reactor (SBR)-Effect of gamma irradiation

International Nuclear Information System (INIS)

Bural, Cavit B.; Demirer, Goksel N.; Kantoglu, Omer; Dilek, Filiz B.

2010-01-01

Aerobic biological treatment of opium alkaloid containing wastewater as well as the effect of gamma irradiation as pre-treatment was investigated. Biodegradability of raw wastewater was assessed in aerobic batch reactors and was found highly biodegradable (83-90% degradation). The effect of irradiation (40 and 140 kGy) on biodegradability was also evaluated in terms of BOD 5 /COD values and results revealed that irradiation imparted no further enhancement in the biodegradability. Despite the highly biodegradable nature of wastewater, further experiments in sequencing batch reactors (SBR) revealed that the treatment operation was not possible due to sludge settleability problem observed beyond an influent COD value of 2000 mg dm -3 . Possible reasons for this problem were investigated, and the high molecular weight, large size and aromatic structure of the organic pollutants present in wastewater was thought to contribute to poor settleability. Initial efforts to solve this problem by modifying the operational conditions, such as SRT reduction, failed. However, further operational modifications including addition of phosphate buffer cured the settleability problem and influent COD was increased up to 5000 mg dm -3 . Significant COD removal efficiencies (>70%) were obtained in both SBRs fed with original and irradiated wastewaters (by 40 kGy). However, pre-irradiated wastewater provided complete thebain removal and a better settling sludge, which was thought due to degradation of complex structure by radiation application. Degradation of the structure was observed by GC/MS analyses and enhancement in filterability tests.

Treatment of opium alkaloid containing wastewater in sequencing batch reactor (SBR)-Effect of gamma irradiation

Energy Technology Data Exchange (ETDEWEB)

Bural, Cavit B.; Demirer, Goksel N. [Middle East Technical University, Department of Environmental Engineering, 06531 Ankara (Turkey); Kantoglu, Omer [Turkish Atomic Energy Authority, Saraykoy Nuclear Research and Training Center, 06982, Kazan, Ankara (Turkey); Dilek, Filiz B., E-mail: fdilek@metu.edu.t [Middle East Technical University, Department of Environmental Engineering, 06531 Ankara (Turkey)

2010-04-15

Aerobic biological treatment of opium alkaloid containing wastewater as well as the effect of gamma irradiation as pre-treatment was investigated. Biodegradability of raw wastewater was assessed in aerobic batch reactors and was found highly biodegradable (83-90% degradation). The effect of irradiation (40 and 140 kGy) on biodegradability was also evaluated in terms of BOD{sub 5}/COD values and results revealed that irradiation imparted no further enhancement in the biodegradability. Despite the highly biodegradable nature of wastewater, further experiments in sequencing batch reactors (SBR) revealed that the treatment operation was not possible due to sludge settleability problem observed beyond an influent COD value of 2000 mg dm{sup -3}. Possible reasons for this problem were investigated, and the high molecular weight, large size and aromatic structure of the organic pollutants present in wastewater was thought to contribute to poor settleability. Initial efforts to solve this problem by modifying the operational conditions, such as SRT reduction, failed. However, further operational modifications including addition of phosphate buffer cured the settleability problem and influent COD was increased up to 5000 mg dm{sup -3}. Significant COD removal efficiencies (>70%) were obtained in both SBRs fed with original and irradiated wastewaters (by 40 kGy). However, pre-irradiated wastewater provided complete thebain removal and a better settling sludge, which was thought due to degradation of complex structure by radiation application. Degradation of the structure was observed by GC/MS analyses and enhancement in filterability tests.

History based batch method preserving tally means

International Nuclear Information System (INIS)

Shim, Hyung Jin; Choi, Sung Hoon

2012-01-01

In the Monte Carlo (MC) eigenvalue calculations, the sample variance of a tally mean calculated from its cycle-wise estimates is biased because of the inter-cycle correlations of the fission source distribution (FSD). Recently, we proposed a new real variance estimation method named the history-based batch method in which a MC run is treated as multiple runs with small number of histories per cycle to generate independent tally estimates. In this paper, the history-based batch method based on the weight correction is presented to preserve the tally mean from the original MC run. The effectiveness of the new method is examined for the weakly coupled fissile array problem as a function of the dominance ratio and the batch size, in comparison with other schemes available

Comparison between cultivated and total bacterial communities associated with Cucurbita pepo using cultivation-dependent techniques and 454 pyrosequencing.

Science.gov (United States)

Eevers, N; Beckers, B; Op de Beeck, M; White, J C; Vangronsveld, J; Weyens, N

2016-02-01

Endophytic bacteria often have beneficial effects on their host plants that can be exploited for bioremediation applications but, according to the literature, only 0.001-1% of all endophytic microbes should be cultivable. This study compared the cultivated endophytic communities of the roots and shoots of Cucurbita pepo with the total endophytic communities as determined by cultivation-dependent techniques and 454 pyrosequencing. The ten most abundant taxa of the total communities aligned well with the cultivated taxa; however, the abundance of these taxa in the two communities differed greatly. Enterobacter showed very low presence in the total communities, whereas they were dominantly present in the cultivated communities. Although Rhizobium dominated in total root and shoot communities, it was poorly cultivable and even then only in growth media containing plant extract. Since endophytes likely contribute to plant-growth promotion, cultivated bacterial strains were tested for their plant-growth promoting capabilities, and the results were correlated with their abundance in the total community. Bacillus and Pseudomonas showed promising results when considering cultivability, abundance in the total community and plant-growth promoting capability. This study demonstrated that, although a limited number of bacterial genera were cultivable, current cultivation-dependent techniques may be sufficient for further isolation and inoculation experiments that aim to improve phytoremediation efficiency. Copyright © 2015 Elsevier GmbH. All rights reserved.

Batch calculations in CalcHEP

International Nuclear Information System (INIS)

Pukhov, A.

2003-01-01

CalcHEP is a clone of the CompHEP project which is developed by the author outside of the CompHEP group. CompHEP/CalcHEP are packages for automatic calculations of elementary particle decay and collision properties in the lowest order of perturbation theory. The main idea prescribed into the packages is to make available passing on from the Lagrangian to the final distributions effectively with a high level of automation. According to this, the packages were created as a menu driven user friendly programs for calculations in the interactive mode. From the other side, long-time calculations should be done in the non-interactive regime. Thus, from the beginning CompHEP has a problem of batch calculations. In CompHEP 33.23 the batch session was realized by mean of interactive menu which allows to the user to formulate the task for batch. After that the not-interactive session was launched. This way is too restricted, not flexible, and leads to doubling in programming. In this article I discuss another approach how one can force an interactive program to work in non-interactive mode. This approach was realized in CalcHEP 2.1 disposed on http://theory.sinp.msu.ru/~pukhov/calchep.html

Saturated fatty acid palmitate induces extracellular release of histone H3: A possible mechanistic basis for high-fat diet-induced inflammation and thrombosis

Energy Technology Data Exchange (ETDEWEB)

Shrestha, Chandan [Department of Systems Biology in Thromboregulation, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima (Japan); Department of Laboratory and Vascular Medicine, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima (Japan); Ito, Takashi [Department of Systems Biology in Thromboregulation, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima (Japan); Kawahara, Ko-ichi [Department of Biomedical Engineering, Osaka Institute of Technology, Osaka (Japan); Shrestha, Binita; Yamakuchi, Munekazu; Hashiguchi, Teruto [Department of Laboratory and Vascular Medicine, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima (Japan); Maruyama, Ikuro, E-mail: rinken@m3.kufm.kagoshima-u.ac.jp [Department of Systems Biology in Thromboregulation, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima (Japan)

2013-08-09

Highlights: •High-fat diet feeding and palmitate induces the release of nuclear protein histone H3. •ROS production and JNK signaling mediates the release of histone H3. •Extracellular histones induces proinflammatory and procoagulant response. -- Abstract: Chronic low-grade inflammation is a key contributor to high-fat diet (HFD)-related diseases, such as type 2 diabetes, non-alcoholic steatohepatitis, and atherosclerosis. The inflammation is characterized by infiltration of inflammatory cells, particularly macrophages, into obese adipose tissue. However, the molecular mechanisms by which a HFD induces low-grade inflammation are poorly understood. Here, we show that histone H3, a major protein component of chromatin, is released into the extracellular space when mice are fed a HFD or macrophages are stimulated with the saturated fatty acid palmitate. In a murine macrophage cell line, RAW 264.7, palmitate activated reactive oxygen species (ROS) production and JNK signaling. Inhibitors of these pathways dampened palmitate-induced histone H3 release, suggesting that the extracellular release of histone H3 was mediated, in part, through ROS and JNK signaling. Extracellular histone activated endothelial cells toexpress the adhesion molecules ICAM-1 and VCAM-1 and the procoagulant molecule tissue factor, which are known to contribute to inflammatory cell recruitment and thrombosis. These results suggest the possible contribution of extracellular histone to the pathogenesis of HFD-induced inflammation and thrombosis.

Saturated fatty acid palmitate induces extracellular release of histone H3: A possible mechanistic basis for high-fat diet-induced inflammation and thrombosis

International Nuclear Information System (INIS)

Shrestha, Chandan; Ito, Takashi; Kawahara, Ko-ichi; Shrestha, Binita; Yamakuchi, Munekazu; Hashiguchi, Teruto; Maruyama, Ikuro

2013-01-01

Highlights: •High-fat diet feeding and palmitate induces the release of nuclear protein histone H3. •ROS production and JNK signaling mediates the release of histone H3. •Extracellular histones induces proinflammatory and procoagulant response. -- Abstract: Chronic low-grade inflammation is a key contributor to high-fat diet (HFD)-related diseases, such as type 2 diabetes, non-alcoholic steatohepatitis, and atherosclerosis. The inflammation is characterized by infiltration of inflammatory cells, particularly macrophages, into obese adipose tissue. However, the molecular mechanisms by which a HFD induces low-grade inflammation are poorly understood. Here, we show that histone H3, a major protein component of chromatin, is released into the extracellular space when mice are fed a HFD or macrophages are stimulated with the saturated fatty acid palmitate. In a murine macrophage cell line, RAW 264.7, palmitate activated reactive oxygen species (ROS) production and JNK signaling. Inhibitors of these pathways dampened palmitate-induced histone H3 release, suggesting that the extracellular release of histone H3 was mediated, in part, through ROS and JNK signaling. Extracellular histone activated endothelial cells toexpress the adhesion molecules ICAM-1 and VCAM-1 and the procoagulant molecule tissue factor, which are known to contribute to inflammatory cell recruitment and thrombosis. These results suggest the possible contribution of extracellular histone to the pathogenesis of HFD-induced inflammation and thrombosis

Influence of carbon source and the fermentation process on levan production by Zymomonas mobilis analyzed by the surface response method Influência da fonte de carbono e do processo fermentativo na produção de levana por Zymomonas mobilis analisada pela metodologia de superfície de resposta

Directory of Open Access Journals (Sweden)

Raquel Renan Jorge Borsari

2006-09-01

Full Text Available The aim of this study is to assess sugar cane juice and sucrose as substrates, the batch and fed batch processes and their interaction in the levan production using a complete factorial design. Zymomonas mobilis was cultivated in different sugar cane juice and sucrose concentrations in two fermentation processes at 25 °C for 20 h. A complete factorial design (2³ was used to analyze the effects of the type and concentration of the substrate, as well as the batch and fed batch processes. A complete second factorial design (2² was used to observe the importance of sugar cane juice. The results indicated that the batch process improved the levan production reaching 40.14 g/L. The addition of sugar cane juice was not statistically significant for levan formation, however sugar cane juice stimulated biomass, sorbitol and ethanol production. The best medium for levan production was 150 g/L sucrose in batch.O presente estudo avaliou caldo de cana de açúcar e sacarose como substratos e os processos batelada e batelada alimentada e suas interações na produção de levana. Zymomonas mobilis foi cultivada em diferentes concentrações de caldo de cana de açúcar e sacarose nos dois processos fermentativos a 25 °C por 20 h. Foi utilizado um delineamento fatorial completo (23 para analisar os efeitos do tipo e concentração de substratos e processos batelada e batelada alimentada. Um segundo delineamento fatorial completo (22 foi usado para confirmar a importância do caldo de cana de açúcar. Os resultados indicam que o processo batelada foi o melhor para a produção de levana, atingindo 40,14 g/L em 150 g/L de sacarose. A adição de caldo de cana de açúcar não foi estatisticamente significativa para formação de levana, porém o caldo estimulou a produção de biomassa, sorbitol e etanol.

Numerical modeling of batch formation in waste incineration plants

Directory of Open Access Journals (Sweden)

Obroučka Karel

2015-03-01

Full Text Available The aim of this paper is a mathematical description of algorithm for controlled assembly of incinerated batch of waste. The basis for formation of batch is selected parameters of incinerated waste as its calorific value or content of pollutants or the combination of both. The numerical model will allow, based on selected criteria, to compile batch of wastes which continuously follows the previous batch, which is a prerequisite for optimized operation of incinerator. The model was prepared as for waste storage in containers, as well as for waste storage in continuously refilled boxes. The mathematical model was developed into the computer program and its functionality was verified either by practical measurements or by numerical simulations. The proposed model can be used in incinerators for hazardous and municipal waste.

Monitoring a PVC batch process with multivariate statistical process control charts

NARCIS (Netherlands)

Tates, A. A.; Louwerse, D. J.; Smilde, A. K.; Koot, G. L. M.; Berndt, H.

1999-01-01

Multivariate statistical process control charts (MSPC charts) are developed for the industrial batch production process of poly(vinyl chloride) (PVC). With these MSPC charts different types of abnormal batch behavior were detected on-line. With batch contribution plots, the probable causes of these

Cashew apple juice as microbial cultivation medium for non-immunogenic hyaluronic acid production

Directory of Open Access Journals (Sweden)

Adriano H. Oliveira

2013-12-01

Full Text Available In this work, natural cashew apple juice was used as cultivation medium as an alternative to substitute brain heart infusion medium. The effect of aeration and juice supplementation with yeast extract on the production of hyaluronic acid in batch fermentation was also investigated. Similar levels of cell mass were obtained in inoculum using cashew apple juice supplemented with yeast extract or the conventional brain heart infusion medium. Fermentation in Erlenmeyer flasks produced low biomass and hyaluronic acid concentrations. The hyaluronic acid concentration and viscosity increased from 0.15 g/L and 3.87 cP (no aeration or medium supplementation to 1.76 g/L and 107 cP, when aeration (2 vvm and 60 g/L of yeast extract were used. The results suggest the production of low-molecular weight hyaluronic acid oligomers instead of the high molecular weight polymer.

The International Society for Extracellular Vesicles launches the first massive open online course on extracellular vesicles

OpenAIRE

L?sser, Cecilia; Th?ry, Clotilde; Buz?s, Edit I.; Mathivanan, Suresh; Zhao, Weian; Gho, Yong Song; L?tvall, Jan

2016-01-01

The International Society for Extracellular Vesicles (ISEV) has organised its first educational online course for students and beginners in the field of extracellular vesicles (EVs). This course, “Basics of Extracellular Vesicles,” uses recorded lectures from experts in the field and will be open for an unlimited number of participants. The course is divided into 5 modules and can be accessed at www.coursera.org/learn/extracellular-vesicles. The first module is an introduction to the field co...

Potential of biogas production with young bulls manure on batch biodigesters

Energy Technology Data Exchange (ETDEWEB)

Costa, Monica Sarolli S. de M.; Costa, Luiz A. de Mendonca [Universidade Estadual do Oeste do Parana (UNIOESTE), Cascavel, PR (Brazil)], E-mail: monicas@unioeste.br; Lucas Junior, Jorge de [Universidade Estadual Paulista (FCAV/UNESP), Jaboticabal, SP (Brazil). Faculdade de Ciencias Agrarias e Veterinarias

2008-07-01

The feedlot system concerning the young bull model allows that animals gain weight in a shorter time since there is higher daily intake of protein when compared to fiber. This change on animals' diet alters particularly manure characteristics and thus interferes on performance of biological systems of treatment. This study aimed at evaluating the potential of biogas production using manure of young bulls that received two different diets on batch biodigesters under three temperatures, with and without inoculum use. The results showed that manure from animals that received more protein on diet (80% concentrate + 20% roughage) had greater reductions on volatile solids when submitted to anaerobic biodigestion. Although the speed of biogas production was superior on treatments with inoculum, it was observed negative effect on inoculum use. There was no effect on temperature during biogas production. Regarding diet effect, manure of animals fed on diet with more protein produced larger amounts of biogas per kg of total added solids (0.2543) when compared to those who received less protein on diet (65% concentrate + 35% roughage), which meant 0.1001 m{sup 3} biogas/kg/total solids. (author)

The medicinal Agaricus mushroom cultivated in Brazil: biology, cultivation and non-medicinal valorisation.

Science.gov (United States)

Largeteau, Michèle L; Llarena-Hernández, Régulo Carlos; Regnault-Roger, Catherine; Savoie, Jean-Michel

2011-12-01

Sun mushroom is a cultivated mushroom extensively studied for its medicinal properties for several years and literature abounds on the topic. Besides, agronomical aspects were investigated in Brazil, the country the mushroom comes from, and some studies focus on the biology of the fungus. This review aimed to present an overview of the non-medicinal knowledge on the mushroom. Areas of commercial production and marketing trends are presented. Its specific fragrance, taste, nutritional value and potential use of extracts as food additives are compared to those of the most cultivated fungi and laboratory models. The interest of the mushroom for lignocellulosic enzyme production and source of biomolecules for the control of plant pathogens are shown. Investigation of genetic variability among cultivars is reported. Growing and storage of mycelium, as well as cultivation conditions (substrate and casing generally based on local products; indoor and outdoor cultivation; diseases and disorders) are described and compared to knowledge on Agaricus bisporus.

Competitive adsorption of heavy metal by extracellular polymeric substances (EPS) extracted from sulfate reducing bacteria.

Science.gov (United States)

Wang, Jin; Li, Qing; Li, Ming-Ming; Chen, Tian-Hu; Zhou, Yue-Fei; Yue, Zheng-Bo

2014-07-01

Competitive adsorption of heavy metals by extracellular polymeric substances (EPS) extracted from Desulfovibrio desulfuricans was investigated. Chemical analysis showed that different EPS compositions had different capacities for the adsorption of heavy metals which was investigated using Cu(2+) and Zn(2+). Batch adsorption tests indicated that EPS had a higher combined ability with Zn(2+) than Cu(2+). This was confirmed and explained by Fourier transform infrared (FTIR) and excitation-emission matrix (EEM) spectroscopy analysis. FTIR analysis showed that both polysaccharides and protein combined with Zn(2+) while only protein combined with Cu(2+). EEM spectra further revealed that tryptophan-like substances were the main compositions reacted with the heavy metals. Moreover, Zn(2+) had a higher fluorescence quenching ability than Cu(2+). Copyright © 2014 Elsevier Ltd. All rights reserved.

Phylogeography of the wild and cultivated stimulant plant qat (Catha edulis, Celastraceae) in areas of historical cultivation.

Science.gov (United States)

Tembrock, Luke R; Simmons, Mark P; Richards, Christopher M; Reeves, Patrick A; Reilley, Ann; Curto, Manuel A; Meimberg, Harald; Ngugi, Grace; Demissew, Sebsebe; Al Khulaidi, Abdul Wali; Al-Thobhani, Mansoor; Simpson, Sheron; Varisco, Daniel M

2017-04-01

Qat ( Catha edulis , Celastraceae) is a woody plant species cultivated for its stimulant alkaloids. Qat is important to the economy and culture in large regions of Ethiopia, Kenya, and Yemen. Despite the importance of this species, the wild origins and dispersal of cultivars have only been described in often contradictory historical documents. We examined the wild origins, human-mediated dispersal, and genetic divergence of cultivated qat compared to wild qat. We sampled 17 SSR markers and 1561 wild and cultivated individuals across the historical areas of qat cultivation. On the basis of genetic structure inferred using Bayesian and nonparametric methods, two centers of origin in Kenya and one in Ethiopia were found for cultivated qat. The centers of origin in Ethiopia and northeast of Mt. Kenya are the primary sources of cultivated qat genotypes. Qat cultivated in Yemen is derived from Ethiopian genotypes rather than Yemeni wild populations. Cultivated qat with a wild Kenyan origin has not spread to Ethiopia or Yemen, whereas a small minority of qat cultivated in Kenya originated in Ethiopia. Hybrid genotypes with both Ethiopian and Kenyan parentage are present in northern Kenya. Ethiopian cultivars have diverged from their wild relatives, whereas Kenyan qat has diverged less. This pattern of divergence could be caused by the extinction of the wild-source qat populations in Ethiopia due to deforestation, undersampling, and/or artificial selection for agronomically important traits. © 2017 Tembrock et al. Published by the Botanical Society of America. This work is licensed under a Creative Commons public domain license (CC0 1.0).

Failure of the cultivated mushroom (Agaricus bisporus) to induce tumors in the A/J mouse lung tumor model

DEFF Research Database (Denmark)

Pilegaard, Kirsten; Kristiansen, E.; Meyer, Otto A.

1997-01-01

We studied whether the cultivated mushroom (Agaricus bisporus) or 4-(carboxy)phenylhydrazine (CP) induce lung adenomas in the A/J mouse lung tumor model. For 26 weeks female mice were fed a semisynthetic diet where 11 or 22% of the diet was replaced by freeze-dried mushrooms. The intake...... of the mushroom diets was equivalent to an intake of agaritine, the major phenylhydrazine derivative occurring in the mushroom, of 92 or 166 mg/kg body weight per day. The intake of CP was 106 mg/kg body weight per day. Neither the;freeze-dried mushroom nor CP induced statistically significant increased numbers...

Look-ahead strategies for controlling batch operations in industry - An overview

NARCIS (Netherlands)

Zee, Durk-Jouke van der; Chick, SE; Sanchez, PJ; Ferrin, D; Morrice, DJ

2003-01-01

Batching jobs in a manufacturing system is a very common policy in most industries. Main reasons for batching are avoidance of set ups and/or facilitation of material handling. Examples of batch-wise production systems are ovens found in aircraft industry and in semiconductor manufacturing. Starting

Effects of salinity, commercial salts, and water type on cultivation of the cryptophyte microalgae Rhodomonas salina and the calanoid Copepod Acartia tonsa

DEFF Research Database (Denmark)

Jepsen, Per Meyer; Thoisen, Christina V.; Carron-Cabaret, Thibaut

2018-01-01

Marine aquaculture facilities positioned far from the sea need access to seawater (SW); hence, commercial salts are often the chosen solution. In marine hatcheries, most fish larvae require live feed (zooplankton) that are in turn fed with microalgae. The objective of this research was to investi......Marine aquaculture facilities positioned far from the sea need access to seawater (SW); hence, commercial salts are often the chosen solution. In marine hatcheries, most fish larvae require live feed (zooplankton) that are in turn fed with microalgae. The objective of this research...... was to investigate the applicability of commercial salts and clarify the potential effects on the cultivation of the microalga Rhodomonas salina and the copepod Acartia tonsa. Three commercial salts were tested, Red Sea Salt (RS), Red Sea – Coral Pro Salt (CP), and Blue Treasure Salt. R. salina was cultured...

Solving a chemical batch scheduling problem by local search

NARCIS (Netherlands)

Brucker, P.; Hurink, Johann L.

1999-01-01

A chemical batch scheduling problem is modelled in two different ways as a discrete optimization problem. Both models are used to solve the batch scheduling problem in a two-phase tabu search procedure. The method is tested on real-world data.

Study on Serum Lipoprotein Profile of Exclusive Breast Fed, Mixed Fed and Formula Fed Preterm Infants

Directory of Open Access Journals (Sweden)

Vineet Jaiswal

2017-10-01

Full Text Available Introduction: Breast feeding is protective for atherosclerotic cardiovascular disease, obesity, Diabetes Mellitus (DM and hypertension. Serum lipoprotein is principal risk factor for atherosclerosis. There is growing evidence that risk of Coronary Heart Disease (CHD begins to emerge from infancy. Lipoprotein level is affected by different feeding pattern during infancy. Aim: To compare serum lipoprotein profile of exclusively breast fed, mixed fed and formula fed preterm infant. Materials and Methods: A total of two fifty preterm newborn were recruited at birth and divided into three groups. Group A were Exclusively Breast Fed (EBF, Group B were Mixed Fed (MF and Group C were Formula/bovine milk Fed (FF infants. Preterm newborns with severe sepsis, hypoglycemia, Hypoxic Ischemic Encephalopathy (HIE stage II and III, meconium stained amniotic fluid, pathological jaundice, Hyaline Membrane Disease (HMD, less than 28 weeks gestation, with major congenital anomaly and infants born to mothers with DM, gestational diabetes, hypertension, pre-eclampsia, eclampsia or on long term medications were excluded from the study. Lipoprotein profile estimation was done at four weeks and again at 16 weeks of age. Results: At four weeks of age, Total Cholesterol (TC, Triglyceride (TG, Low Density Lipoprotein (LDL and Very Low Density Lipoprotein (VLDL were higher in EBF infants as compared to MF and FF infants. For TC, difference was significant between EBF vs. MF (p<0.001, EBF vs. FF (p<0.001 and MF vs. FF (p=0.005 infants. At 16 weeks also, TC and HDL were higher in EBF infants as compared to MF and FF infants. For TC, this difference was significant between EBF vs. MF (p<0.001 and EBF vs. FF (p<0.001 infants. When infants were followed up to 16 weeks of age, TC and LDL level fell significantly (p<0.001 in EBF and MF group, a significant (p<0.05 rise for TC was seen in FF group. At 16 weeks of age, there was no significant rise in HDL in EBF infants, but

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

Science.gov (United States)

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

2009-04-01

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

Treatment of opium alkaloid containing wastewater in sequencing batch reactor (SBR)—Effect of gamma irradiation

Science.gov (United States)

Bural, Cavit B.; Demirer, Goksel N.; Kantoglu, Omer; Dilek, Filiz B.

2010-04-01

Aerobic biological treatment of opium alkaloid containing wastewater as well as the effect of gamma irradiation as pre-treatment was investigated. Biodegradability of raw wastewater was assessed in aerobic batch reactors and was found highly biodegradable (83-90% degradation). The effect of irradiation (40 and 140 kGy) on biodegradability was also evaluated in terms of BOD 5/COD values and results revealed that irradiation imparted no further enhancement in the biodegradability. Despite the highly biodegradable nature of wastewater, further experiments in sequencing batch reactors (SBR) revealed that the treatment operation was not possible due to sludge settleability problem observed beyond an influent COD value of 2000 mg dm -3. Possible reasons for this problem were investigated, and the high molecular weight, large size and aromatic structure of the organic pollutants present in wastewater was thought to contribute to poor settleability. Initial efforts to solve this problem by modifying the operational conditions, such as SRT reduction, failed. However, further operational modifications including addition of phosphate buffer cured the settleability problem and influent COD was increased up to 5000 mg dm -3. Significant COD removal efficiencies (>70%) were obtained in both SBRs fed with original and irradiated wastewaters (by 40 kGy). However, pre-irradiated wastewater provided complete thebain removal and a better settling sludge, which was thought due to degradation of complex structure by radiation application. Degradation of the structure was observed by GC/MS analyses and enhancement in filterability tests.

Acceptance Test Data for Candidate AGR-5/6/7 TRISO Particle Batches BWXT Coater Batches 93165 93172 Defective IPyC Fraction and Pyrocarbon Anisotropy

Energy Technology Data Exchange (ETDEWEB)

Helmreich, Grant W. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Hunn, John D. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Skitt, Darren J. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Dyer, John A. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Schumacher, Austin T. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

2017-03-01

Coated particle fuel batches J52O-16-93165, 93166, 93168, 93169, 93170, and 93172 were produced by Babcock and Wilcox Technologies (BWXT) for possible selection as fuel for the Advanced Gas Reactor Fuel Development and Qualification (AGR) Program’s AGR-5/6/7 irradiation test in the Idaho National Laboratory (INL) Advanced Test Reactor (ATR). Some of these batches may alternately be used as demonstration coated particle fuel for other experiments. Each batch was coated in a 150-mm-diameter production-scale fluidized-bed chemical vapor deposition (CVD) furnace. Tristructural isotropic (TRISO) coatings were deposited on 425-μm-nominal-diameter spherical kernels from BWXT lot J52R-16-69317 containing a mixture of 15.5%-enriched uranium carbide and uranium oxide (UCO). The TRISO coatings consisted of four consecutive CVD layers: a ~50% dense carbon buffer layer with 100-μm-nominal thickness, a dense inner pyrolytic carbon (IPyC) layer with 40-μm-nominal thickness, a silicon carbide (SiC) layer with 35-μm-nominal thickness, and a dense outer pyrolytic carbon (OPyC) layer with 40-μmnominal thickness. The TRISO-coated particle batches were sieved to upgrade the particles by removing over-sized and under-sized material, and the upgraded batches were designated by appending the letter A to the end of the batch number (e.g., 93165A).

PROOF on a Batch System

International Nuclear Information System (INIS)

Behrenhoff, W; Ehrenfeld, W; Samson, J; Stadie, H

2011-01-01

The 'parallel ROOT facility' (PROOF) from the ROOT framework provides a mechanism to distribute the load of interactive and non-interactive ROOT sessions on a set of worker nodes optimising the overall execution time. While PROOF is designed to work on a dedicated PROOF cluster, the benefits of PROOF can also be used on top of another batch scheduling system with the help of temporary per user PROOF clusters. We will present a lightweight tool which starts a temporary PROOF cluster on a SGE based batch cluster or, via a plugin mechanism, e.g. on a set of bare desktops via ssh. Further, we will present the result of benchmarks which compare the data throughput for different data storage back ends available at the German National Analysis Facility (NAF) at DESY.

From 32 ounces to zero: a medical geographic study of dispensing a cultivated batch of "plum" cannabis flowers to medical marijuana patients in Washington State.

Science.gov (United States)

Aggarwal, Sunil K; Carter, Gregory T; Zumbrunnen, Craig; Morrill, Richard; Sullivan, Mark; Mayer, Jonathan D

2013-01-01

The medicinal use of cannabis is a growing phenomenon in the U.S. predicated on the success of overcoming specific spatial challenges and establishing particular human-environment relationships. This article takes a medical geographic "snapshot" of an urban site in Washington State where qualifying chronically ill and debilitated patients are delivered locally produced botanical cannabis for medical use. Using interview, survey, and observation, this medical geographic research project collected information on the social space of the particular delivery site and tracked the production cost, reach, and health value of a 32-ounce batch of strain-specific medical cannabis named "Plum" dispensed over a four-day period. A convenience sample of 37 qualifying patients delivered this batch of cannabis botanical medicine was recruited and prospectively studied with survey instruments. Results provide insight into patients' self-rated health, human-plant relationships, and travel-to-clinic distances. An overall systematic geographic understanding of the medical cannabis delivery system gives a grounded understanding of the lengths that patients and care providers go, despite multiple hurdles, to receive and deliver treatment with botanical cannabis that relieves diverse symptoms and improves health-related quality-of-life.

Comparative study of trapping parameters of LiF(TLD-100) from different production batches

Energy Technology Data Exchange (ETDEWEB)

Bos, A.J.J.; Piters, T.M.; Vries, W. de; Hoogenboom, J.E. (Delft Univ. of Technology (Netherlands). Interfaculty Reactor Institute)

1990-01-01

Computerised glow curve analysis has been used to determine the trapping parameters of the main peaks of the thermoluminescent (TL) material LiF(TLD-100). The TL material (solid state chips) originated from six different production batches with at least 19 chips per batch. The maxima of glow peaks 2 to 5 are found at the same temperature within very small limits. The activation energy and frequency factor of the main glow peak (peak 5) of TLD-100 originating from two batches differ significantly from those of the other four investigated batches. Nevertheless, the sensitivity of glow peak 5 is more or less the same for all batches. The trapping parameters of glow peaks 2 to 4 of TLD-100 vary little from batch to batch. The measured half-life of peak 2 differed strongly from batch to batch. For all investigated peaks no correlation has been found between glow peak sensitivity and trapping parameters. The results of this study suggest that both defect concentration and nature of the trapping centres vary from batch to batch. It would appear that as a consequence of selection by the manufacturer, the differences between the batches in terms of total light output are small. (author).

Decolorization of complex dyes and textile effluent by extracellular enzymes of Cyathus bulleri cultivated on agro-residues/domestic wastes and proposed pathway of degradation of Kiton blue A and reactive orange 16.

Science.gov (United States)

Vats, Arpita; Mishra, Saroj

2017-04-01

In this study, the white-rot fungus Cyathus bulleri was cultivated on low-cost agro-residues, namely wheat bran (WB), wheat straw (WS), and domestic waste orange peel (OP) for production of ligninolytic enzymes. Of the three substrates, WB and OP served as good materials for the production of laccase with no requirement of additional carbon or nitrogen source. Specific laccase activity of 94.4 U mg -1 extracellular protein and 21.01 U mg -1 protein was obtained on WB and OP, respectively. Maximum decolorization rate of 13.6 μmol h -1  U -1 laccase for reactive black 5 and 22.68 μmol h -1  U -1 laccase for reactive orange 16 (RO) was obtained with the WB culture filtrate, and 11.7 μmol h -1  U -1 laccase for reactive violet 5 was observed with OP culture filtrate. Importantly, Kiton blue A (KB), reported not to be amenable to enzymatic degradation, was degraded by culture filtrate borne activities. Products of degradation of KB and RO were identified by mass spectrometry, and a pathway of degradation proposed. WB-grown culture filtrate decolorized and detoxified real and simulated textile effluents by about 40%. The study highlights the use of inexpensive materials for the production of enzymes effective on dyes and effluents.

A novel biosorbent for dye removal: Extracellular polymeric substance (EPS) of Proteus mirabilis TJ-1

Energy Technology Data Exchange (ETDEWEB)

Zhang Zhiqiang [State Key Laboratory of Pollution Control and Resource Reuse, Key Laboratory of Yangtze River Water Environment of Ministry of Education, College of Environmental Science and Engineering, Tongji University, Shanghai 200092 (China); Laboratoire de Sciences Analytiques (UMR CNRS 5180), Universite Claude Bernard Lyon 1, Universite de Lyon, 69622 Villeurbanne Cedex (France); Xia Siqing [State Key Laboratory of Pollution Control and Resource Reuse, Key Laboratory of Yangtze River Water Environment of Ministry of Education, College of Environmental Science and Engineering, Tongji University, Shanghai 200092 (China)], E-mail: siqingxia@mail.tongji.edu.cn; Wang Xuejiang; Yang Aming; Xu Bin; Chen Ling; Zhu Zhiliang; Zhao Jianfu [State Key Laboratory of Pollution Control and Resource Reuse, Key Laboratory of Yangtze River Water Environment of Ministry of Education, College of Environmental Science and Engineering, Tongji University, Shanghai 200092 (China); Jaffrezic-Renault, Nicole; Leonard, Didier [Laboratoire de Sciences Analytiques (UMR CNRS 5180), Universite Claude Bernard Lyon 1, Universite de Lyon, 69622 Villeurbanne Cedex (France)

2009-04-15

This paper deals with the extracellular polymeric substance (EPS) of Proteus mirabilis TJ-1 used as a novel biosorbent to remove dye from aqueous solution in batch systems. As a widely used and hazardous dye, basic blue 54 (BB54) was chosen as the model dye to examine the adsorption performance of the EPS. The effects of pH, initial dye concentration, contact time and temperature on the sorption of BB54 to the EPS were examined. At various initial dye concentrations (50-400 mg/L), the batch sorption equilibrium can be obtained in only 5 min. Kinetic studies suggested that the sorption followed the internal transport mechanism. According to the Langmuir model, the maximum BB54 uptake of 2.005 g/g was obtained. Chemical analysis of the EPS indicated the presence of protein (30.9%, w/w) and acid polysaccharide (63.1%, w/w). Scanning electron microscopy (SEM) images showed that the EPS with a crystal-linear structure was whole enwrapped by adsorbed dye molecules. FTIR spectrum result revealed the presence of adsorbing groups such as carboxyl, hydroxyl and amino groups in the EPS. High-molecular weight of the EPS with more binding-sites and stronger van der Waals forces together with its specific construct leads to the excellent performance of dye adsorption. The EPS shows potential board application as a biosorbent for both environmental protection and dye recovery.

A novel biosorbent for dye removal: Extracellular polymeric substance (EPS) of Proteus mirabilis TJ-1

International Nuclear Information System (INIS)

Zhang Zhiqiang; Xia Siqing; Wang Xuejiang; Yang Aming; Xu Bin; Chen Ling; Zhu Zhiliang; Zhao Jianfu; Jaffrezic-Renault, Nicole; Leonard, Didier

2009-01-01

This paper deals with the extracellular polymeric substance (EPS) of Proteus mirabilis TJ-1 used as a novel biosorbent to remove dye from aqueous solution in batch systems. As a widely used and hazardous dye, basic blue 54 (BB54) was chosen as the model dye to examine the adsorption performance of the EPS. The effects of pH, initial dye concentration, contact time and temperature on the sorption of BB54 to the EPS were examined. At various initial dye concentrations (50-400 mg/L), the batch sorption equilibrium can be obtained in only 5 min. Kinetic studies suggested that the sorption followed the internal transport mechanism. According to the Langmuir model, the maximum BB54 uptake of 2.005 g/g was obtained. Chemical analysis of the EPS indicated the presence of protein (30.9%, w/w) and acid polysaccharide (63.1%, w/w). Scanning electron microscopy (SEM) images showed that the EPS with a crystal-linear structure was whole enwrapped by adsorbed dye molecules. FTIR spectrum result revealed the presence of adsorbing groups such as carboxyl, hydroxyl and amino groups in the EPS. High-molecular weight of the EPS with more binding-sites and stronger van der Waals forces together with its specific construct leads to the excellent performance of dye adsorption. The EPS shows potential board application as a biosorbent for both environmental protection and dye recovery

Nitrate removal from high strength nitrate-bearing wastes in granular sludge sequencing batch reactors.

Science.gov (United States)

Krishna Mohan, Tulasi Venkata; Renu, Kadali; Nancharaiah, Yarlagadda Venkata; Satya Sai, Pedapati Murali; Venugopalan, Vayalam Purath

2016-02-01

A 6-L sequencing batch reactor (SBR) was operated for development of granular sludge capable of denitrification of high strength nitrates. Complete and stable denitrification of up to 5420 mg L(-1) nitrate-N (2710 mg L(-1) nitrate-N in reactor) was achieved by feeding simulated nitrate waste at a C/N ratio of 3. Compact and dense denitrifying granular sludge with relatively stable microbial community was developed during reactor operation. Accumulation of large amounts of nitrite due to incomplete denitrification occurred when the SBR was fed with 5420 mg L(-1) NO3-N at a C/N ratio of 2. Complete denitrification could not be achieved at this C/N ratio, even after one week of reactor operation as the nitrite levels continued to accumulate. In order to improve denitrification performance, the reactor was fed with nitrate concentrations of 1354 mg L(-1), while keeping C/N ratio at 2. Subsequently, nitrate concentration in the feed was increased in a step-wise manner to establish complete denitrification of 5420 mg L(-1) NO3-N at a C/N ratio of 2. The results show that substrate concentration plays an important role in denitrification of high strength nitrate by influencing nitrite accumulation. Complete denitrification of high strength nitrates can be achieved at lower substrate concentrations, by an appropriate acclimatization strategy. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

A study on the comparison of antioxidant effects among wild ginseng, cultivated wild ginseng, and cultivated ginseng extracts

Directory of Open Access Journals (Sweden)

Hae Young, Jang

2008-09-01

Full Text Available Objective : The objective of this study was to compare the antioxidant effects among wild ginseng, cultivated wild ginseng, and ginseng extracts. Methods : In vitro antioxidant activities were examined by total antioxidant capacity (TAC, oxygen radical scavenging capacity(ORAC, total phenolic content, 1, 1-Diphenyl-2-picrylhydrazyl (DPPH radical scavenging activity, inhibition of induced lipid peroxidation using liver mitochondria, reactive oxygen species(ROS scavenging effect using 2’, 7’-dichlorofluorescein(DCF fluorescence. Results : 1. TAC of 1.5 and 3.75 mg extracts was highest in cultivated wild ginseng, followed by wild ginseng and lowest in ginseng. 2. ORAC of 2, 10, and 20 μg extracts was highest in cultivated wild ginseng, followed by wild ginseng and lowest in ginseng. 3. Total phenolic content of 0.375, 0.938, and 1.875 mg extracts was highest in cultivated wild ginseng, followed by wild ginseng and lowest in ginseng. 4. DPPH(1, 1 -Diphenyl-2-picrylhydrazyl scavenging activity between wild ginseng and cultivated wild ginseng did not differ significantly (p>0.05. 5. Induced lipid peroxidation, measured by TBARS concentration in solution containing rat liver mitochondria incubated in the presence of FeSO4/ascorbic acid was inhibited as amounts of wild ginseng, cultivated wild ginseng, and ginseng extracts increased. TBARS concentration of ginseng extracts were significantly (p<0.05 higher than wild ginseng or cultivated wild ginseng extracts. 6. DCF fluorescence intensity was decreased as concentrations of wild ginseng, cultivated wild ginseng, and ginseng extracts increased, demonstrating that ROS generation was inhibited in a concentrationdependent manner. Conclusions : In summary, the results of this study demonstrate that cultivated wild ginseng extracts had similar antioxidant activities to wild ginseng extracts and greater that of cultivated ginseng extracts.

Inorganic fouling mitigation by salinity cycling in batch reverse osmosis

OpenAIRE

Maswadeh, Laith A.; Warsinger, David Elan Martin; Tow, Emily W.; Connors, Grace B.; Swaminathan, Jaichander; Lienhard, John H

2018-01-01

Enhanced fouling resistance has been observed in recent variants of reverse osmosis (RO) desalination which use time-varying batch or semi-batch processes, such as closed-circuit RO (CCRO) and pulse flow RO (PFRO). However, the mechanisms of batch processes' fouling resistance are not well-understood, and models have not been developed for prediction of their fouling performance. Here, a framework for predicting reverse osmosis fouling is developed by comparing the fluid residence time in bat...

Dynamic Scheduling Of Batch Operations With Non-Identical Machines

NARCIS (Netherlands)

van der Zee, D.J.; van Harten, A.; Schuur, P.C.

1997-01-01

Batch-wise production is found in many industries. A good example of production systems which process products batch-wise are the ovens found in aircraft industry and in semiconductor manufacturing. These systems mostly consist of multiple machines of different types, given the range and volumes of

Dynamic scheduling of batch operations with non-identical machines

NARCIS (Netherlands)

van der Zee, D.J.; van Harten, Aart; Schuur, Peter

1997-01-01

Batch-wise production is found in many industries. A good example of production systems which process products batch-wise are the ovens found in aircraft industry and in semiconductor manufacturing. These systems mostly consist of multiple machines of different types, given the range and volumes of

Some performance measures for vacation models with a batch Markovian arrival process

Directory of Open Access Journals (Sweden)

Sadrac K. Matendo

1994-01-01

Full Text Available We consider a single server infinite capacity queueing system, where the arrival process is a batch Markovian arrival process (BMAP. Particular BMAPs are the batch Poisson arrival process, the Markovian arrival process (MAP, many batch arrival processes with correlated interarrival times and batch sizes, and superpositions of these processes. We note that the MAP includes phase-type (PH renewal processes and non-renewal processes such as the Markov modulated Poisson process (MMPP.

Isolation and Cultivation of Anaerobes

DEFF Research Database (Denmark)

Aragao Börner, Rosa

2016-01-01

Anaerobic microorganisms play important roles in different biotechnological processes. Their complex metabolism and special cultivation requirements have led to less isolated representatives in comparison to their aerobic counterparts.In view of that, the isolation and cultivation of anaerobic...

Flux Balance Analysis Inspired Bioprocess Upgrading for Lycopene Production by a Metabolically Engineered Strain of Yarrowia lipolytica

Directory of Open Access Journals (Sweden)

Komi Nambou

2015-12-01

Full Text Available Genome-scale metabolic models embody a significant advantage of systems biology since their applications as metabolic flux simulation models enable predictions for the production of industrially-interesting metabolites. The biotechnological production of lycopene from Yarrowia lipolytica is an emerging scope that has not been fully scrutinized, especially for what concerns cultivation conditions of newly generated engineered strains. In this study, by combining flux balance analysis (FBA and Plackett-Burman design, we screened chemicals for lycopene production from a metabolically engineered strain of Y. lipolytica. Lycopene concentrations of 126 and 242 mg/L were achieved correspondingly from the FBA-independent and the FBA-assisted designed media in fed-batch cultivation mode. Transcriptional studies revealed upregulations of heterologous genes in media designed according to FBA, thus implying the efficiency of model predictions. Our study will potentially support upgraded lycopene and other terpenoids production from existing or prospect bioengineered strains of Y. lipolytica and/or closely related yeast species.

21 CFR 80.37 - Treatment of batch pending certification.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 1 2010-04-01 2010-04-01 false Treatment of batch pending certification. 80.37 Section 80.37 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL COLOR ADDITIVE CERTIFICATION Certification Procedures § 80.37 Treatment of batch pending certification...

Lingonberries alter the gut microbiota and prevent low-grade inflammation in high-fat diet fed mice

Directory of Open Access Journals (Sweden)

Lovisa Heyman-Lindén

2016-04-01

Full Text Available Background: The gut microbiota plays an important role in the development of obesity and obesity-associated impairments such as low-grade inflammation. Lingonberries have been shown to prevent diet-induced obesity and low-grade inflammation. However, it is not known whether the effect of lingonberry supplementation is related to modifications of the gut microbiota. The aim of the present study was to describe whether consumption of different batches of lingonberries alters the composition of the gut microbiota, which could be relevant for the protective effect against high fat (HF-induced metabolic alterations. Methods: Three groups of C57BL/6J mice were fed HF diet with or without a supplement of 20% lingonberries from two different batches (Lingon1 and Lingon2 during 11 weeks. The composition and functionality of the cecal microbiota were assessed by 16S rRNA sequencing and PICRUSt. In addition, parameters related to obesity, insulin sensitivity, hepatic steatosis, inflammation and gut barrier function were examined. Results: HF-induced obesity was only prevented by the Lingon1 diet, whereas both batches of lingonberries reduced plasma levels of markers of inflammation and endotoxemia (SAA and LBP as well as modified the composition and functionality of the gut microbiota, compared to the HF control group. The relative abundance of Akkermansia and Faecalibacterium, genera associated with healthy gut mucosa and anti-inflammation, was found to increase in response to lingonberry intake. Conclusions: Our results show that supplementation with lingonberries to an HF diet prevents low-grade inflammation and is associated with significant changes of the microbiota composition. Notably, the anti-inflammatory properties of lingonberries seem to be independent of effects on body weight gain.

Influence of coal batch preparation on the quality of metallurgical соkе

Directory of Open Access Journals (Sweden)

Катерина Олегівна Шмельцер

2015-10-01

Full Text Available To study the influence of coal batch properties on coke strength we have considered the quality of the coke produced at the plant in Krivoy Rog from 2008 till 2012. Such factors as the large number of coal suppliers, imprecise selection of the optimal degree of batch crushing result in the decline in coke quality, the batch density and contents of the lean class (<0,5 mm are not optimum; poor blending of the batch after crushing; increased moisture and ash content of the coking batch; and extreme fluctuation in the coal and batch characteristics. It was found that high humidity of coal batch and its large fluctuations has most profound effect on the mechanical properties of coke. Under deteriorating resource base the quality of the coking batch preparation is important, To have batch of proper quality the following key aspects must be taken into account: the batch must be crushed to an optimum degree that will result in leaning components decrease and increased contents of vitrivite in it which improves the sinterability and coking, and hence the quality of coke; the degree of mixing of the coking batch in all indices must be up to 98-99%, for uneven distribution in the coal chamber worsens the quality of coke

Beyond Batch Processing: Towards Real-Time and Streaming Big Data

OpenAIRE

Shahrivari, Saeed; Jalili, Saeed

2014-01-01

Today, big data is generated from many sources and there is a huge demand for storing, managing, processing, and querying on big data. The MapReduce model and its counterpart open source implementation Hadoop, has proven itself as the de facto solution to big data processing. Hadoop is inherently designed for batch and high throughput processing jobs. Although Hadoop is very suitable for batch jobs but there is an increasing demand for non-batch processes on big data like: interactive jobs, r...

Efficacy and immunogenicity of live-attenuated human rotavirus vaccine in breast-fed and formula-fed European infants.

Science.gov (United States)

Vesikari, Timo; Prymula, Roman; Schuster, Volker; Tejedor, Juan-C; Cohen, Robert; Bouckenooghe, Alain; Damaso, Silvia; Han, Htay Htay

2012-05-01

Rotavirus is the main cause of severe gastroenteritis and diarrhea in infants and young children less than 5 years of age. Potential impact of breast-feeding on the efficacy and immunogenicity of human rotavirus G1P[8] vaccine was examined in this exploratory analysis. Healthy infants (N = 3994) aged 6-14 weeks who received 2 doses of human rotavirus vaccine/placebo according to a 0-1 or 0-2 month schedule were followed for rotavirus gastroenteritis during 2 epidemic seasons. Rotavirus IgA seroconversion rate (anti-IgA antibody concentration ≥ 20 mIU/mL) and geometric mean concentrations were measured prevaccination and 1-2 months post-dose 2. Vaccine efficacy against any and severe rotavirus gastroenteritis was analyzed according to the infants being breast-fed or exclusively formula-fed at the time of vaccination. Antirotavirus IgA seroconversion rate was 85.5% (95% confidence interval [CI]: 82.4-88.3) in breast-fed and 89.2% (95% CI: 84.2-93) in exclusively formula-fed infants; geometric mean concentrations in the respective groups were 185.8 U/mL (95% CI: 161.4-213.9) and 231.5 U/mL (95% CI: 185.9-288.2). Vaccine efficacy was equally high in breast-fed and exclusively formula-fed children in the first season but fell in breast-fed infants in the second rotavirus season. During the combined 2-year efficacy follow-up period, vaccine efficacy against any rotavirus gastroenteritis was 76.2% (95% CI: 68.7-82.1) and 89.8% (95% CI: 77.6-95.9) and against severe rotavirus gastroenteritis 88.4% (95% CI: 81.6-93) and 98.1% (95% CI: 88.2-100) in the breast-fed and exclusively formula-fed infants, respectively. The difference in immunogenicity of human rotavirus vaccine in breast-fed and exclusively formula-fed infants was small. Vaccine efficacy was equally high in breast-fed and exclusively formula-fed children in the first season. Breast-feeding seemed to reduce slightly the efficacy in the second season.

Micrometeorological principles of protected cultivation

Science.gov (United States)

Protected cultivation is a broad term commonly used among producers of specialty crops. Techniques can range from complex fixed structures to field site selection, to straightforward cultural practices in the field. This introduction to the ASHS workshop "Protected cultivation for fruit crops" consi...

Application of solid waste from anaerobic digestion of poultry litter in Agrocybe aegerita cultivation: mushroom production, lignocellulolytic enzymes activity and substrate utilization.

Science.gov (United States)

Isikhuemhen, Omoanghe S; Mikiashvili, Nona A; Kelkar, Vinaya

2009-06-01

The degradation and utilization of solid waste (SW) from anaerobic digestion of poultry litter by Agrocybe aegerita was evaluated through mushroom production, loss of organic matter (LOM), lignocellulolytic enzymes activity, lignocellulose degradation and mushroom nutrients content. Among the substrate combinations (SCs) tested, substrates composed of 10-20% SW, 70-80% wheat straw and 10% millet was found to produce the highest mushroom yield (770.5 and 642.9 g per 1.5 kg of substrate). LOM in all SCs tested varied between 8.8 and 48.2%. A. aegerita appears to degrade macromolecule components (0.6-21.8% lignin, 33.1-55.2% cellulose and 14-53.9% hemicellulose) during cultivation on the different SCs. Among the seven extracellular enzymes monitored, laccase, peroxidase and CMCase activities were higher before fruiting; while xylanase showed higher activities after fruiting. A source of carbohydrates (e.g., millet) in the substrate is needed in order to obtain yield and biological efficiency comparable to other commercially cultivated exotic mushrooms.