• Title/Summary/Keyword: Batch culture

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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).

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.

Mass Production of Resting Eggs by the Freshwater Rotifer, Brachionus calyciflorus Pallas with Exchange of Culture Water and Supplemental Aeration (환수 및 공기공급에 따른 담수산 rotifer, Brachionus calyciflorus의 내구란 대량생산)

  • Kwon, O-Nam;Park, Heum-Gi
    • Journal of Aquaculture
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    • v.18 no.2
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    • pp.115-121
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    • 2005
  • This study was performed to investigate the formation and mass production of resting eggs by freshwater rotifer, Brachionus calyciflorus as influenced by different aeration supplements and exchange intervals of culture water in 15-L culture vessels and 500 L culture tanks. The maximum densities and mixis rates of the rotifers were not different between experimental group exposed to air or oxygen supplements. However, the fertilization rate and formation of resting eggs of the rotifers in the air-supplemented group were significantly higher than those in the oxygen supplemented group. In the experiment concerning exchange interval of culture water, the maximum density of the rotifers and formation of resting eggs in the batch culture were significantly higher than those in the semi-continuous culture with exchange of water every day. The formation of resting eggs per Chlorella dry weight was highest in the semi-continuous culture with exchange of water every day. The resting eggs of rotifers were produced at a density of $51.8{\sim}57.9{\times}10^6$ eggs in 500-L culture tanks. In this study, the batch culture with air is an effective method for mass production of resting eggs by the freshwater rotifer, B. calyciflorus, and the efficiency of mass production of resting eggs by this rotifer was similar to that of the seawater rotifers, B. plicatilis and B. rotundiformis.

Effect of Glycine on L-Ornithine Production by a Citrulline Auxotroph of Brevibacterium ketoglutamicum and Stoichiometric Analysis

  • Nam, Soo-Wan;Choi, Dae-Keon;Ryu, Wuk-Sang;Jang, Hyung-Wook;Chung, Bong-Hyun;Park, Young-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.4 no.2
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    • pp.95-101
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    • 1994
  • The effects of glycine on cell growth and L-omithine production were investigated in shake-flask and jar fermentor cultures of a citrulline auxotrophic mutant, Brevibacterium ketoglutamicum BK 1046. In the shake-flask culture, the optimal concentration of glycine for L-ornithine production was found to be 20 g/l. In the jar fermentor culture with the glycine at an initial concentration of 20 g/l, L-ornithine production increased by 28%, compared to that of the culture with no glycine added. 37 g/l of L-ornithine was produced when additional feeding of glycine (5 g/l) was made. This was a significant improvement in L-ornithine production compared to that (ca. 24 g/l) of the corresponding batch culture conducted without glycine. According to the stoichiometric analysis with the batch fermentation results, the experimental and theoretical L-ornithine yields based on the glucose consumption were 0.24 and 0.59, respectively. This indicates that the performance of L-ornithine fermentation can further be improved by the supplementation of glycine and the development of a mutant strain possessing a higher growth yield.

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Effect of Galactose and Dextrose on Human Lipocortin I Expression in Recombinant Saccharomyces cerevisiae Carrying Galactose-Regulated Expression System

  • Nam, Soo-Wan;Seo, Dong-Jin;Rhee, Sang-Ki;Park, Young-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.3 no.3
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    • pp.168-173
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    • 1993
  • The expression kinetics of human lipocortin I (LCI), a potential anti-inflammatory agent, was studied in the shake-flask and fermenter cultures of Saccharomyces cerevisiae carrying a galactose-inducible expression system. The cell growth, expression level of LCI, and the plasmid stability were investigted under various galactose induction conditions. The expression of LCI was repressed by the presence of a very small amount of dextrose in the culture medium, but it was induced by galactose after dextrose became completely depleted. The optimal ratio of dextrose to galactose for lipocortin I production was found to be 1.0 (10 g/l dextrose and 10 g/l galactose). With optimal D/G ratio of 1.0 and the addition of galactose prior to dextrose depletion, LCI of about 100~130 mg/l was produced. LCI at a concentration of 174 mg/l was porduced in the fed-batch culture, which was nearly a twice as much of that produced in the batch culture. The plasmid stability was very high in all culture cases, and thus was considered to be not an important parameter in the expression of LCI.

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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.

Automation of Glutamic Acid Fermentation (글루탐산 발효공정의 자동화)

  • Park, S.H.;Hong, K.T.;You, S.J.;Lee, J.H.;Bae, J.C.
    • Korean Journal of Food Science and Technology
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    • v.15 no.2
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    • pp.202-204
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    • 1983
  • A strategy for the automation of glutamic acid fermentation has been developed by the use of $CO_2$ analyzer together with a controller. It was found that a linear relationship existed between growth and $CO_2$ level in the exit gas. Therefore penicillin addition at an appropriate biomass concentration to excrete glutamate could be achieved automatically. In addition, an automatic batch feeding method (fed-batch culture) provided a means of overcoming substrate inhibition effects on growth and glutamic acid production in batch culture, thereby increasing productivity and product yield.

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In situ Separation of Lactic Acid by Electrodialysis in Batch Culture (회분식 발효에서의 전기투석에 의한 젖산의 동시분리)

  • Lee, Eun-Gyo;Chang, Yong-Keun;Chang, Ho-Nam;Kim, In-Ho
    • Microbiology and Biotechnology Letters
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    • v.23 no.5
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    • pp.609-616
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    • 1995
  • Effects of lactate concentration, temperature, counter ions, pH as well as voltage (current) in batch electrodialysis (ED) experiments with a 3-compartment unit were investigated. The applied voltage was found to be the most critical factor as expected. The electrodialysis rate increased with the lactate concentration of the source solution. The amount of lactate transferred was limited by the lactate concentration difference between cathode and permeate compartments. The electrodialysis rate did not heavily depend on temperature change. The electrodialysis rate of NH$_{4}$-lactate was faster than that of Na-lactate and both lactates showed the highest electrodialysis rate at a pH of 4.0. A little amount of non-ionic glucose diffused through the anionic membrane to the permeate compartment. To test the effectiveness of the in situ recovery of lactic acid from fermentation broth by ED, three cases of batch culture were carried out; pH control only, ED only, and pH control and ED. The case with both pH control and ED was more efficient than that with pH control only in the aspects of productivity and product yield.

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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.