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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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Production of GABA (gamma amino butyric acid) by Lactic Acid Bacteria

  • Kook, Moo-Chang;Cho, Seok-Cheol
    • Food Science of Animal Resources
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    • v.33 no.3
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    • pp.377-389
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    • 2013
  • Gamma-amino butyric acid (GABA) is a kind of pharmacological and biological component and its application is wide and useful in Korea specially, becoming aging society in the near feature. GABA is request special dose for the purposed biological effect but the production of concentrated GABA is very difficult due to low concentration of glutamic acid existed in the fermentation broth. To increase GABA concentrate using fermentation technology, high content of glutamic acid is required. For this reason, various strains which have the glutamic acid decarboxylase (GAD) and can convert glutamic acid to GABA, were isolated from various fermented foods. Most of GABA producing strains are lactic acid bacteria isolated from kimchi, especially added monosodium glutamate (MSG) as a taste enhancer. Optimizing the formulation of culture media and the culture condition, GABA conversion yield and amounts were increased. Finally GABA concentration of fermentation broth in batch or fed batch fermentation reached 660 mM or 1000 mM, respectively. Furthermore formulation of culture media for GABA production developed commercially. Many studies about GABA-rich product have been continued, so GABA-rich kimchi, cheese, yogurt, black raspberry juice and tomato juices has been also developed. In Korea many biological effects of GABA are evaluated recently and GABA will be expected to be used in multipurpose.

Efficient Use of Lactose for Production of the Soluble Recombinant Human Epidermal Growth Factor in Escherichia coli. (대장균에서 lactose를 이용한 수용성 재조합 인간 상피 세포 성장 인자의 생산)

  • 박세철;권태종;고인영;유광현
    • Microbiology and Biotechnology Letters
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    • v.26 no.1
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    • pp.61-67
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    • 1998
  • Recombinant human epidermal growth factor (rhEGF) was produced by E. coli BL2l (DE3) harboring a plasmid pYHB101. The production of rhEGF was 44.5 mg/L when the E. coli BL2l (pYHB101) was cultured at 27$^{\circ}C$ for 48 hr in the modified MBL medium containing 10 $\mu\textrm{g}$/L glucose with 10 $\mu\textrm{m}$ IPTG/lactose induction at 2 hr after inoculation. It was shown that lactose is able to induce the rhEGF expression of E. coli BL2l (pYHB101) with the same efficiency as IPTG. In the batch culture system, when induced with 10 $\mu\textrm{m}$ lactose, E. coli BL2l (pYHB101) produced maximum 45 mg/L of the rhEGF at 28 hr culture in the modified MBL medium containing 10 g/L glucose. In the semi-fed batch culture system, the volumetric yield was 160 mg/L when the culture was added with 0.5% (w/v) lactose and 0.25% (w/v) yeast extract in the late logarithmic phase and 94.3% of rhEGF was secreted as soluble form. However, when the culture was added with them in the early logarithmic phase, the volumetric yield was 120 mg/L and 20.9% of rhEGF was found in cytoplasmic insoluble aggregates. It was found that the addition time of lactose was important for production of soluble rhEGF from E. coli BL21 (pYHB101).

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Ethanol Production from Raw Starch by Co-Immobilized Mixed Rhizopus japonicus and zymomonas mobilis (Rhizopus japonicus와 Zymomonas mobilis의 혼합고정화 배양계에 의한 생전분으로부터 에탄올 생산)

  • 최수철;이상원;박석규;성찬기;손봉수;성낙계
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.25 no.4
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    • pp.708-714
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    • 1996
  • Ethanol production from raw starch was performed using the co-immobilized culture system of Rhizopu japonicus and zymomonas mobilis(R-Z). Glucose Production in immobilized R. japonicus culture was 2-fold higher than that in free cell culture. Ethanol production was 1.67g/L(Yp/s, 0.094) and 6.549/L(Yp/s, 0.38) in R-Z and R-Z 24 culture system, respectively. R-Z system was modified and designated as R-Z 24 system by replacing cotton plug with silicon check valve after 24h fermentation with R-Z system. Optimal substrate concentration for ethanol production in batch culture was 5%(w/v) and ethanol concentration produced was 15.02g/L(Yp/s, 0.36). Ethanol yield(Yp/s, 0.38) in fed-batch culture of 5 times with 2%(w/v) substrate was equal to that in batch culture of 2%(w/v) substrate.

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Kinetics of Converting Single Chain Urokinase Type Plasminogen Activator into Two Chain Plasminogen Activator in Cultivating HEK Cells with Low Serum Containing Medium (저혈청 배지에서 생산된 scu-PA의 tc-PA로의 전환에 관한 연구)

  • 김영남;김현구
    • KSBB Journal
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    • v.9 no.1
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    • pp.48-54
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    • 1994
  • A modified amidolytic assay and a fibrin plate method were used to accurately measure the concentration of single chain urokinase type plasminogen activator (scu-PA) and two-chain urokinase type plasminogen activator (tc-PA) in the spent media. $1.65{\times}10^6$(viable cells/ml) of maximum cell density and 1670(IU/ml) of scu-PA concentration were obtained in 1% serum containing medium. The overall conversion ratio from scu-PA to tc-PA was less than 10%. In the results of batch cultivation in a spinner vessel, $4.43{\times}10^6(total cells/ml)$ of maximum cell density and 1560(IU/ml) of scu-PA concentration was observed. The maximum scu-PA concentration and specific scu-PA Productivity were obtained in 1760(IU/ml) and $3.13{\times}10^{-4}(IU/cell)$, respectively, from perfusion cultivation. The conveysion ratios from batch, fed-batch and perfusion cultivations were less than 12%, which means that about 90% of scu-PA secreted from the cells can be maintained during the cultivations.

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High xylitol production rate of osmophilic yeast Candida tropicalis by long-term cell-recycle fermentation in a submerged membrane bioreactor

  • Kwon, Seun-Gyu;Park, Seung-Won;Oh, Deok-Kun
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.272-276
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    • 2005
  • Candida tropicalis, an osmophilic strain isolated from honeycomb, produced xylitol at a maximal volumetric production rate of 3.5 g $l^{-1}$ $h^{-1}$ from an initial xylose concentration of 200 g $l^{-1}$. Even with a very high xylose concentration, e.g., 350 g $l^{-1}$, this strain produced xylitol at a moderate rate of 2.07 g $l^{-1}$ $h^{-1}$. In a fed-batch fermentation of xylose and glucose, 260 g $l^{-1}$ of xylose was added, and xylitol production was 234 g $l^{-1}$ for 48 h, corresponding to a rate of 4.88 g $l^{-1}$ $h^{-1}$. To increase the xylitol production rate, cells were recycled in a submerged membrane bioreactor with suction pressure and air sparging. In cell-recycle fermentation, the average concentration of xylitol produced per recycle round, total fermentation time, volumetric production rate, and product yield for ten rounds were 180 g $l^{-1}$, 195 h, 8.5 g $l^{-1}$ $h^{-1}$, and 85%, respectively. When cell-recycle fermentation was started with the cell mass contratrated two-fold after batch fermentation and was performed for ten recycle rounds, we achieved a very high production rate of 12 g $l^{-1}$ $h^{-1}$. The production rate and total amount of xylitol produced in cell-recycle fermentation were 3.4 and 11 times higher than in batch fermentation, respectively.

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Process Kinetics of Nisin Production in Batch and Continuous Culture (회분식 및 연속식 배양시 Nisin의 생산특성)

  • Yoo, Jin-Young;Park, Shin-Yang;Jin, Young-Ok;Koo, Young-Jo;Chung, Kun-Sub
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.504-509
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    • 1989
  • Fermentation condition of Streptococcus lactis IFO 12007 for nisin production was examined. The optimal glucose concentration was 60g/ι. The pH and temperature optimum were 6.5 and 31$^{\circ}C$, respectively. The maximum nisin activity in batch culture was 2000IU/$m\ell$. The fermentation quotients after 7 hours of fermentation in batch culture were; specific glucose uptake rate:0.59g/g/h , specific nisin productivity: 34924IU/g/h, product yield: 5944IU/g, growth yield:0.24, biomass:4.81g/ι. The specific growth rate was affected by pH and temperature and the activation energy for growth was 1.35kcal/mole. pH control was essential for nisin production. Fed-batch culture using 20g/$\ell$ glucose medium produced 1420IU/$m\ell$ after 14 hours. The continuous culture could be operated at below 0.38h$^{-1}$ for nisin production. The steady state nisin concentration and specific nisin productivity were 740IU/$m\ell$ and 45000IU/g/h. The growth yield and maintenance energy were 0.144 and 207mg glucose/g-cell/h.

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$\small{D}$-Lactic Acid Production by Sporolactobacillus inulinus Y2-8 Immobilized in Fibrous Bed Bioreactor Using Corn Flour Hydrolyzate

  • Zhao, Ting;Liu, Dong;Ren, Hengfei;Shi, Xinchi;Zhao, Nan;Chen, Yong;Ying, Hanjie
    • Journal of Microbiology and Biotechnology
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    • v.24 no.12
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    • pp.1664-1672
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    • 2014
  • In this study, a fibrous bed bioreactor (FBB) was used for $\small{D}$-lactic acid ($\small{D}$-LA) production by Sporolactobacillus inulinus Y2-8. Corn flour hydrolyzed with ${\alpha}$-amylase and saccharifying enzyme was used as a cost-efficient and nutrient-rich substrate for $\small{D}$-LA production. A maximal starch conversion rate of 93.78% was obtained. The optimum pH for $\small{D}$-LA production was determined to be 6.5. Ammonia water was determined to be an ideal neutralizing agent, which improved the $\small{D}$-LA production and purification processes. Batch fermentation and fed-batch fermentation, with both free cells and immobilized cells, were compared to highlight the advantages of FBB fermentation. In batch mode, the $\small{D}$-LA production rate of FBB fermentation was 1.62 g/l/h, which was 37.29% higher than that of free-cell fermentation, and the $\small{D}$-LA optical purities of the two fermentation methods were above 99.00%. In fe$\small{D}$-batch mode, the maximum $\small{D}$-LA concentration attained by FBB fermentation was 218.8 g/l, which was 37.67% higher than that of free-cell fermentation. Repeate$\small{D}$-batch fermentation was performed to determine the long-term performance of the FBB system, and the data indicated that the average $\small{D}$-LA production rate was 1.62 g/l/h and the average yield was 0.98 g/g. Thus, hydrolyzed corn flour fermented by S. inulinus Y2-8 in a FBB may be used for improving $\small{D}$-LA fermentation by using ammonia water as the neutralizing agent.

재조합 효모를 이용한 항혈전 단백질 히루딘 발효 생산공정의 최적화

  • Kim, Myeong-Dong;Gang, Hyeon-A;Lee, Sang-Gi;Seo, Jin-Ho
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.99-102
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    • 2001
  • Recombinant Saccharomyces cerevisiae strains harboring various copy numbers of hirudin gene were developed to study dependency of hirudin expression level on its gene copy number. A linear relationship between the copy number of hirudin expression cassette and hirudin expression level was observed up to 10 copies. A double <5-integration vector truncated wi 디 1 the unnecessary bacterial genes before yeast transformation showed a four-fold increase in transformation efficiency and a 1.3-fold enhancement in hirudin expression level compared with a single <5 system. Gratuitous hirudin expression strain was developed by disrupting the GALl gene of S. cerevisiae. Glucose that was fed in a limited manner effectively supported cell growth and hi겨din expression by the gratuitous strain. Effects of methanol concentrations on hirudin production in recombinant Hansenula polymorpha were investigated in continuous and fed-batch cultures. At a steady-state of continuous culture, an optimum methanol concentration of 1.7 g/L was determined at a dilution rate of 0.18 $h^{-1}$ with 1.8 mg/L ${\cdot}$ h hirudin productivity.

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A Study on Denitrification Efficiency with External Carbon Sources in Pilot-scale Modified SBR (Pilot-scale modified SBR 공정에서 외부탄소원의 종류에 따른 탈질효율 비교 연구)

  • 서인석;김병군;홍성택;정위득
    • Journal of environmental and Sanitary engineering
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    • v.17 no.3
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    • pp.52-59
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    • 2002
  • Pilot-scale PSSBR(Phase Separated Sequencing Batch Reactor) was operated to evaluate requirement of external carbon sources(${\Delta}gCOD/{\Delta}gNO_3^{-}-N$) in denitrification. Methanol and fermented food waste were used as external carbon sources. Methanol and fermented food waste were fed to the anoxic state of first reactor and concentration were 50 and 40 mgCOD/L on the basis of concentration in reactor, respectively. In case that external carbon source was not used, average $NO_3^{-}-N$ concentration in effluent was 22.49 mg/L. When methanol and fermented food waste were fed, average $NO_3^{-}-N$ concentration in effluent were 10.13 mg/L and 6.3 mg/L, respectively and requirement of external carbon sources were 4.04 and 2.5 ${\Delta}gCOD/{\Delta}gNO_3^{-}-N$, respectively. Fermented food waste was better than methanol in denitrification efficiency. Therefore fermented food waste could be one of the excellent external carbon sources for nitrogen removal in biological nutrient removal process.