• Title/Summary/Keyword: Batch-culture system

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Factors Indicating Culture Status During Cultivation of Spirulina (Arthrospira) platensis

  • Kim, Choong-Jae;Jung, Yun-Ho;Oh, Hee-Mock
    • Journal of Microbiology
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    • v.45 no.2
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    • pp.122-127
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    • 2007
  • Factors indicating culture status of two Spirulina platensis strains were monitored in a batch mode cultivation for 36 days. Changing mode in all factors showed a common turning point, indicating shift of cell or culture status. Mean biomass productivity was highly sustained until day 22, chlorophyll a concentration peaked on day 22, pH value was > 12 on day 22, coil number was abruptly shortened on day 22, and floating activity was sustained at greater than 79% after day 22, indicating that day 22 is a criterion reflecting phase-transfer in cell physiology in a batch culture system. Many of these changes may have been caused by increased pH, suggesting that pH control is essential for mass production of S. platensis. Fluctuations in floating activity were likely induced by the number of cellular gas vacuoles. Consequently, coil number per trichome and floating activity of S. platensis could readily act as simple indicators for determination of culture status or harvesting time of cells.

Batch and Fed-batch Production of Hyperthermostable $\alpha$-L-Arabinofuranosidase of Thermotoga maritima in Recombinant Escherichia coli by Using Constitutive and Inducible Promoters

  • Song, Jae-Yong;Keum, In-Kyung;Jin, Qing;Park, Jung-Mi;Kim, Beom-Soo;Jung, Bong-Hwan;Kim, Tae-Jip;Han, Nam-Soo
    • Food Science and Biotechnology
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    • v.17 no.5
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    • pp.990-995
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    • 2008
  • A thermostable $\alpha$-L-arabinofuranosidases ($\alpha$-L-AFase) is an industrially important enzyme for recovery of L-arabinose from hemicellulose. The recombinant $\alpha$-L-AFase from Thermotoga maritima was expressed in Escherichia coli by using a constitutive pHCE or an inducible pRSET vectors. In batch fermentation, the constitutive expression system resulted in slightly faster growth rate (0.78 vs. 0.74/hr) but lower enzyme activity (2,553 vs. 3,723 units/L) than those of the induction system. When fed-batch fermentation was performed, biomass and enzyme activity reached the highest levels of 36 g/L and 9,152 units/L, respectively. The fed batch cultures performed superior results than batch culture in terms of biomass yield (4.62-5.42 folds) and enzyme synthesis (3.39-4.00 folds). In addition, the fed-batch induction strategy at high cell density resulted in the best productivity in cell growth as well as enzyme activity rather than the induction method at low cell density or the constitutive expression.

Monitoring Anaerobic Reductive Dechlorination of TCE by Biofilm-Type Culture in Continuous-Flow System (연속흐름반응조에서 바이오필름형태의 탈염소화 미생물에 의한 TCE분해 모니터링)

  • Park, Sunhwa;Han, Kyungjin;Hong, Uijeon;Ahn, Hongil;Kim, Namhee;Kim, Hyunkoo;Kim, Taeseung;Kim, Young
    • Journal of Soil and Groundwater Environment
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    • v.17 no.5
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    • pp.49-55
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    • 2012
  • A 1.28 L-batch reactor and continuous-flow stirred tank reactor (CFSTR) fed with formate and trichloroethene (TCE) were operated for 120 days and 56 days, respectively, to study the effect of formate as electron donor on anaerobic reductive dechlorination (ARD) of TCE to cis-1,2-dichloroethylene (c-DCE), vinyl chloride (VC), and ethylene (ETH). In batch reactor, injected 60 ${\mu}mol$ TCE was completely degraded in the presence of 20% hydrogen gas ($H_2$) in less than 8 days by anaerobic dechlorination mixed-culture (300 mg-soluble protein), Evanite Culture with ability to completely degrade tetrachloroethene (PCE) and -TCE to ETH under anaerobic conditions. Once the formate was used as electron donor instead of hydrogen gas in batch or chemostat system, the TCE-dechlorination rate decreased and acetate production rate increased. It indicates that the concentration of hydrogen produced in both systems is possibly more close to threshold for homoacetogenesis process. Soluble protein concentration of Evanite culture during the batch test increased from 300 mg to 688 mg for 120 days. Through the protein monitoring, we confirmed an increase of microbial population during the reactor operation. In CFSTR test, TCE was fed continuously at 9.9 ppm (75.38 ${\mu}mol/L$) and the influent formate feed concentration increased stepwise from 1.3 mmol/L to 14.3 mmol/L. Injected TCE was accumulated at 18 days of HRT, but TCE was completely degraded at 36 days of HRT without accumulation of the injected-TCE during the left of experiment period, getting $H_2$ from fermentative hydrogen production of injected formate. Although c-DCE was also accumulated for 23 days after beginning of CFSTR operation, it reached steady-state in the presence of excessive formate. We also evaluated microbial dynamic of the culture at different chemical state in the reactor by DGGE (denaturing gradient gel electrophoresis).

Mass Production of Poly(3-Hydroxybutyrate) by Fed-Batch Cultures of Ralstonia eutropha with Nitrogen and Phosphate Limitation

  • Ryu, Hee-Wook;Cho, Kyung-Suk;Kim, Beom-Soo;Chang, Yong-Keun;Chang, Ho-Nam;Shim, Hyun-Joo
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.751-756
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    • 1999
  • For mass production of poly(3-hydroxybutyrate) (PHB), high cell density cultures of Ralstonia eutropha were carried out in 2.5-1 and 60-1 fermentors by two fed-batch culture techniques of nitrogen and phosphate limitation. When the nitrogen limitation technique was employed using both an on-line glucose monitoring and control system, a high concentration level of PHB (121g/l) was obtained in the small-scale fermentor of 2.5 1. However, the PHB concentration obtained in a large-scale fermentor of 60 1 only turned out to be 60g/l. In contrast, when another fed-batch culture technique of the phosphate-limitation employing dissolved oxygen (DO) stat glucose feeding was used, a large amount of PHB was successfully produced in both 60-1 and 2.5-1 fermentors. In a 2.5-1 fermentor, concentrations of PHB and cells obtained in 58 h were 175 and 210 g/l, respectively, which corresponded to the PHB productivity level of 3.02 g/l/h. In a 60-1 fermentor, a final cell concentration of 221 g/l and a PHB concentration of 180 g/l with PHB productivity level of 3.75 g/l/h were obtained in 48h. PHB content and yield from glucose were 81% and 0.38g PHB/g glucose, respectively. These data suggest that the phosphate limitation technique is more effective compared to nitrogen limitation in the mass production of PHB by R. eutropha of a large scale.

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Kinetics of nitrification and acrylamide biodegradation by Enterobacter aerogenes and mixed culture bacteria in sequencing batch reactor wastewater treatment systems

  • Madmanang, Romsan;Jangkorn, Siriprapha;Charoenpanich, Jittima;Sriwiriyarat, Tongchai
    • Environmental Engineering Research
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    • v.24 no.2
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    • pp.309-317
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    • 2019
  • This study evaluated the kinetics of acrylamide (AM) biodegradation by mixed culture bacteria and Enterobacter aerogenes (E. aerogenes) in sequencing batch reactor (SBR) systems with AQUASIM and linear regression. The zero-order, first-order, and Monod kinetic models were used to evaluate the kinetic parameters of both autotrophic and heterotrophic nitrifications and both AM and chemical oxygen demand (COD) removals at different AM concentrations of 100, 200, 300, and 400 mg AM/L. The results revealed that both autotrophic and heterotrophic nitrifications and both AM and COD removals followed the Monod kinetics. High AM loadings resulted in the transformation of Monod kinetics to the first-order reaction for AM and COD removals as the results of the compositions of mixed substrates and the inhibition of the free ammonia nitrogen (FAN). The kinetic parameters indicated that E. aerogenes degraded AM and COD at higher rates than mixed culture bacteria. The FAN from the AM biodegradation increased both heterotrophic and autotrophic nitrification rates at the AM concentrations of 100-300 mg AM/L. At higher AM concentrations, the FAN accumulated in the SBR system inhibited the autotrophic nitrification of mixed culture bacteria. The accumulation of intracellular polyphosphate caused the heterotrophic nitrification of E. aerogenes to follow the first-order approximation.

Optimization of Culture Conditions for D-Ribose Production by Transketolase-Deficient Bacillus subtilis JY1

  • Park, Yong-Cheol;Seo, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.665-672
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    • 2004
  • D-Ribose is a five-carbon sugar used for the commercial synthesis of riboflavin, antiviral agents, and flavor enhancers. Batch fermentations with transketolase-deficient B. subtilis JY1 were carried out to optimize the production of D-ribose from xylose. The best results for the fermentation were obtained with a temperature of $37^{\circ}C$ and an initial pH of 7.0. Among various sugars and sugar alcohols tested, glucose and sucrose were found to be the most effective for both cell growth and D-ribose production. The addition of 15 g/l xylose and 15 g/l glucose improved the fermentation performance, presumably due to the adequate supply of ATP in the xylose metabolism from D-xylulose to D-xylulose-5-phosphate. A batch culture in a 3.7-1 jar fermentor with 14.9 g/l xylose and 13.1 g/l glucose resulted in 10.1 g/l D-ribose concentration with a yield of 0.62 g D-ribose/g sugar consumed, and 0.25 g/l-h of productivity. Furthermore, the sugar utilization profile, indicating the simultaneous consumption of xylose and glucose, and respiratory parameters for the glucose and sucrose media suggested that the transketolase-deficient B. subtilis JY1 lost the glucose-specific enzyme II of the phosphoenolpyruvate transferase system.

Shikimate Metabolic Pathway Engineering in Corynebacterium glutamicum

  • Park, Eunhwi;Kim, Hye-Jin;Seo, Seung-Yeul;Lee, Han-Na;Choi, Si-Sun;Lee, Sang Joung;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.31 no.9
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    • pp.1305-1310
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    • 2021
  • Shikimate is a key high-demand metabolite for synthesizing valuable antiviral drugs, such as the anti-influenza drug, oseltamivir (Tamiflu). Microbial-based strategies for shikimate production have been developed to overcome the unstable and expensive supply of shikimate derived from traditional plant extraction processes. In this study, a microbial cell factory using Corynebacterium glutamicum was designed to overproduce shikimate in a fed-batch culture system. First, the shikimate kinase gene (aroK) responsible for converting shikimate to the next step was disrupted to facilitate the accumulation of shikimate. Several genes encoding the shikimate bypass route, such as dehydroshikimate dehydratase (QsuB), pyruvate kinase (Pyk1), and quinate/shikimate dehydrogenase (QsuD), were disrupted sequentially. An artificial operon containing several shikimate pathway genes, including aroE, aroB, aroF, and aroG were overexpressed to maximize the glucose uptake and intermediate flux. The rationally designed shikimate-overproducing C. glutamicum strain grown in an optimized medium produced approximately 37.3 g/l of shikimate in 7-L fed-batch fermentation. Overall, rational cell factory design and culture process optimization for the microbial-based production of shikimate will play a key role in complementing traditional plant-derived shikimate production processes.

The preliminary batch study for evaluating biobarrier application on sequential degradation of TCE products

  • 이재선;이시진;장순웅
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2003.09a
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    • pp.454-457
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    • 2003
  • A new approach for groundwater treatment combines a permeable Fe(0) barrier to breakdown higher chlorinated solvents like PCE and TCE with a downgradient aerobic biological treatment system to biotransform less chlorinated solvents, such as DCE and vinyl chloride (VC). The expected bacterial performance downgradient of an Fe(0) barrier was evaluated through laboratory batch experiments with a toluene-degrading mixed culture that cometabolically transforms cis-1,2-DCE and VC. The amount of cis-1,2-DCE (initially at 2,000 ppb) and VC (initially at 2,000 ppb) transformed was controlled by the initial toluene (20,000 ppb) concentration. VC was removed much more effectively than Cis-1,2-DCE, and a higher toluene concentration in comparison to the co-substrate concentrations was needed for complete co-substrate removal. Overall, the coupling of an Fe(0) barrier and subsequent biodegradation appears feasible for remediation of complex mixtures of chlorinated solvents and petroleum hydrocarbons in groundwater

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Ethanol Production from Tapioca Hydrolysate by Batch and Continuous Cell Retention Cultures (회분 및 연속세포유지 배양에 의한 타피오카당화액으로부터의 에탄올생산)

  • 이용석;이우기
    • KSBB Journal
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    • v.10 no.5
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    • pp.598-603
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    • 1995
  • Batch and continuous cell retention cultures were carried out using tapioca hydrolysate. In batch culture, reducing sugar of about 180g/$\ell$ was almost consumed in about 36 hours, and the concentration of ethanol produced was about 84g/$\ell$ making the ethanol yield 0.48 g-ethanol/g-(reducing sugar). The final yeast concentration was 8.5${\times}$107 cells/ml(about 2.1g/$\ell$). In a total cell retention culture operated with a dilution rate of 0.18h-1, the yeast concentration, the residual reducing sugar concentration, the ethanol concentration, and the volumetric ethanol productivity were about 40g/$\ell$, about 15g/$\ell$, 81.4g/$\ell$, and 14.7g/$\ell$-h, respectively. In another cell retention culture operated with a dilution rate and a bleed ratio of 0.2h-1 and 0.14, respectively, the yeast concentration increased to 22g/$\ell$ and the ethanol concentration oscillated around 68g/$\ell$. The volumetric ethanol productivity was about 13.6g/$\ell$-h and the residual reducing sugar concentration about 12g/$\ell$ containing glucose of about 4.5g/$\ell$. According to the results of batch fermentation using the solid residue from hydrolysate filtration as the substrate, it seemed to have a certain value. Thus, development of an effective reactor system to produce ethanol from this solid residue is in need.

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