• Title/Summary/Keyword: Batch-culture system

검색결과 131건 처리시간 0.034초

SUC2 Gene을 갖는 재조합 Saccharomyces cerebisiae의 Invertase 발현특성 (Expression of Invertase in Recombinant Saccharomyces cerebisiae Containing SUC2 Gene)

  • 정상철;장재권;김인규;변유량
    • 한국미생물·생명공학회지
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    • 제17권3호
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    • pp.263-268
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    • 1989
  • 유전자의 재조합 균주의 생산성을 향상시키기 위한 발효시스템 개발을 목적으로 regulated promoter 인 SUC2 gene 갖는 유전자 재조합 S. cerebisiae를 모델로하여 유전자 산물인 Invertase 발현에 미치는 글루코오스 농도의 영향, 발효 중 플라스미드의 불안정성, 생육특성 및 continuous fed batch system을 연구하였다. 유전자 재조합 균주는 biphasic growth 현상을 보였으며 글루코오스 농도가 0.9g/$\ell$에서 2.2g/L로 증가함에 따라 비증식 속도는 0.224 h$^{-1}$에서 0.226 h$^{-1}$로 증가했으며 숙주효모보다 낮은 값을 나타내었다. 유전자 재조합 균주의 invertase 생산은 발효조내의 글루코오스 농도에 크게 영향을 받아 글루코오스 농도가 0.25~0.4g/L로 감소될 때 invertase 생산이 시작되었으며 회분발효중 플라스미드의 분리는 글루코오스 자화기간 동안에 빈번히 일어났으나 에탄을 자화기간에는 완만해지는 경향을 보였다. 또한 통기를 해주지 않고 배지의 용존산소만으로 배양시킨 결과 통기를 한 경우에 비하여 invertase의 비활성과 총활성이 각각 1.5 및 1.3 배 증가되었다. 균체의 증식단계와 유전자의 발현단계로 구분하여 발효시키기 위하여 영양분을 함유한 글루코오스 용액을 48m1/h(0.096g 글루코오스/48L의 유량으로 12시간 연속적으로 공급하여 fed batch 배양한 결과 invertase의 비활성과 총활성이 비통기적 회분배양 보다 각각 1.74, 2.74배 증가되었다.

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Ethylene Glycol 분해균주의 순수분리.동전 및 배양특성 (Pure Isolation, Identification and Culture Characteristics of Ethylene Glycol Degrading Bacteria)

  • 류원률;최장승;주무환
    • KSBB Journal
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    • 제13권5호
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    • pp.518-523
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    • 1998
  • A large quantity of ethylene glycol(EG) is remained in the effluent after pretreating polyester weight-loss wastewater physicochemically in the fist stage and must be treated biologically in the second stage. Therefore, an excellent EG-utilizing bacteria strain was isolated from the natural system and the optimal culture conditions of the strain were investigated. The optimal culture conditions of temperature, pH, and nitrogen source were found to be 35$^{\circ}C$, 7.5 and ammonium chloride, respectively, when CODCr removal efficiency was more than 90%. The growth of stains and EG removal efficiency was slightly improved by adding elements such as niacin and biotin. With increasing inoculation size in a batch culture, the removal efficiency of EG was conspicuously increased. Growth rate was inhibited when the initial concentration of EG was more then 30g/L. The strain was identified as Pseudomonas sp. based on morphological and biological characteristics and named as Pseudomonas sp. EG1.

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Effects of the $Na^+/K^+$ ratios on hybridoma cell growth

  • 박준영;박홍우
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2000년도 춘계학술발표대회
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    • pp.161-164
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    • 2000
  • In hybridoma cell culture, $NH_4{^+}$ is the most important toxic byproduct so far identified. It has been postulated that $NH_4{^+}$, which is similar to $K^+$ in size, is taken up non-specifically by the cells through a potassium transport system, and that the addition of $K^+$ to the culture medium may have a detoxifying effect of $NH_4{^+}$. Thus, in this article the effects of high $K^+$ concentrations in the range of 10 mM to 60mM on hybridoma cell growth and metabolism were investigated. No significant differences in growth were found for $K^+$ concentrations up to 40 mM, but cell death in the death phase was slightly delayed in the cultures with $K^+$ addition. At 60mM, growth was initially poor but the cells could be adapted after approximately 13 passages. With similar growth levels for high $K^+$ concentrations having been Identified in batch cultivations using basal medium, we are currently investigating how such high levels of $K^+$ will affect cell growth in fortified batch cultures where the accumulation of $NH_4{^+}$ is more problematical.

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Effect of Galactose Feeding Strategy on Heterologous Human Lipocortin-I Production in the Fed-Batch Culture of Saccharomyces cerevisiae Controlled by the GAL10 Promoter

  • Chung, Bong-Hyun;Kim, Byung-Moon;Rhee, Sang-Ki;Park, Young-Hoon;Nam, Soo-Wan
    • Journal of Microbiology and Biotechnology
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    • 제5권4호
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    • pp.224-228
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    • 1995
  • Fed-batch fermentations were conducted to produce human lipocortin-I (LC1), a potential anti-inflammatory agent, from recombinant Sacchromyces cerevisiae carrying a galactose-inducible expression system. The cell growth, expression level of LC1, and the plasmid stability were investigated under various LC1 induction modes performed by three different galactose feeding strategies. Galactoe was fed to induce the expression of LCl from the beginning (initial induction) of culture or when the cell concentration reached 120 OD (mid-phase induction) or 300 OD (late induction). Among the three galactose-induction modes tested, the initial induction mode yielded the best result with respect to a final expression level of LC1. Fedbatch fermentation with initial induction mode produced LC1 at a conentration of 220 mg/l, which corresponded to 1.38- and 1.53-fold increases over those produced by mid-phase and late induction modes.

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Effect of 3,3',4',5-Tetrachlorosalicylanilide on Reduction of Excess Sludge and Nitrogen Removal in Biological Wastewater Treatment Process

  • Rho, Sang-Chul;Nam, Gil-Nam;Shin, Jee-Young;Jahng, Deok-Jin
    • Journal of Microbiology and Biotechnology
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    • 제17권7호
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    • pp.1183-1190
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    • 2007
  • A metabolic uncoupler, 3,3',4',5-tetrachlorosalicylanilide (TCS), was used to reduce excess sludge production in biological wastewater treatment processes. Batch experiments confirmed that 0.4 mg/l of TCS reduced the aerobic growth yield of activated sludge by over 60%. However, the growth yield remained virtually constant even at the increased concentrations of TCS when cultivations were carried out under the anoxic condition. Reduction of sludge production yield was confirmed in a laboratory-scale anoxic-oxic process operated for 6 months. However, it was found that ammonia oxidation efficiency was reduced by as much as 77% in the presence of 0.8 mg/l of TCS in the batch culture. Similar results were also obtained through batch inhibition tests with activated sludges and by bioluminescence assays using a recombinant Nitrosomonas europaea (pMJ217). Because of this inhibitory effect of TCS on nitrification, the TCS-fed continuous system failed to remove ammonia in the influent. When TCS feeding was stopped, the nitrification yield of the process was resumed. Therefore, it seems to be necessary to assess the nitrogen content of wastewater if TCS is used for reducing sludge generation.

Continuous Production of Succinic Acid Using an External Membrane Cell Recycle System

  • Kim, Moon-Il;Kim, Nag-Jong;Shang, Longan;Chang, Yong-Keun;Lee, Sang-Yup;Chang, Ho-Nam
    • Journal of Microbiology and Biotechnology
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    • 제19권11호
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    • pp.1369-1373
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    • 2009
  • Succinic acid was produced by continuous fermentation of Actinobacillus succinogenes sp. 130Z in an external membrane cell recycle reactor to improve viable cell concentration and productivity. Using this system, cell concentration increased to 16.4 g/l at the dilution rate $0.2\;h^{-1}$, up to 3 times higher than that of batch culture, and the volumetric productivity of succinic acid increased up to 6.63 g/l/h at the dilution rate $0.5\;h^{-1}$, 5 times higher than that of batch fermentation. However, in the continuous culture using a high dilution rate, operational problems including severe membrane fouling and contamination by lactic acid producer were observed. Another succinic acid producer, Mannheimia succiniciproducens MBEL55E, was also utilized in this system, and the cell concentration and productivity of succinic acid at the dilution rate of $0.3\;h^{-1}$ were found to be above 3 and 2.3 times higher, respectively, compared with those obtained at the dilution rate of $0.1\;h^{-1}$. These observations give a deep insight into the process design for a continuous succinic acid production by microorganisms.

평판형 광생물반응기의 미세조류 연속배양을 위한 On-off 제어 (On-off Control for Continuous Culture of Microalgae in Flat Panel Photobioreactor)

  • 김재혁;윤충만;정상화
    • 한국생산제조학회지
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    • 제25권3호
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    • pp.237-243
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    • 2016
  • Recently, technologies that produce biofuels from microalgae are being studied worldwide. It is necessary to significantly reduce the production costs of biofuels from microalgae for economic reasons. In this study, the growth curve of the microalgae was obtained using the batch-culture method, and the specific growth rate was predicted using the regression method. Based on the culture conditions of the estimated specific growth rate, the turbidity of the microalgae in the flat panel photobioreactor (PBR) was measured. Furthermore, an on-off control scheme was applied to the flat panel PBR in order to culture the microalgae continuously on the basis of turbidity. The parameters of the on-off control system were displayed by LabView. The on-off scheme of peristaltic pump was controlled based on the turbidity in the PBR. In addition, the turbidity values of growth curves were compared and analyzed in the continuous culture process using the on-off controller.

Production of Lipocortin-1$_{1-185}$ Using A Recombinant of Escherichia coli.

  • Lee, Kyung-Il;Oh, Kyung-Hee;Lee, Jung-Hyun;Na, Do-Sun;Lee, Kye-Joon
    • Journal of Microbiology
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    • 제35권2호
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    • pp.123-126
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    • 1997
  • The aim of the present study was to optimize culture condition for the expression of lipocortin-1$_{1-185}$ in a recombinant of Escherichia coli using batch system. Plasmid (pHT22) carrying lipocortin-1$_{1-185}$ gene was well maintained in the recombinant with the addition of amplicillin as a selection pressures. Optimum temperature was 28$^{\circ}C$ for seed culture and 4$0^{\circ}C$ for main culture and the optimum pH was 7.0. The production of Lipocortin-1$_{1-185}$ was closely associated with cell growth and related to plasmid amplification.

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마이크로컴퓨터를 이용한 고농도 유가배양시스템 (Microcomputer-aided Fermentation System for High Density Fed-Batch Cultivation)

  • 이형준;이계호허윤행
    • KSBB Journal
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    • 제5권3호
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    • pp.307-313
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    • 1990
  • 배양기와 16비트 마이크로컴퓨터를 접속하여 호기성 미생물의 고농도 유가배양시스템을 구성하였다. 기질공급을 위한 제어변수로는 용존산소(DO)를 이용하였다. 용존산소측정기의 출력신호를 컴퓨터에 입력시켜 측정한 DO값에 근거하여 교반모터의 교반속도와 산소유량을 제어함으로써 배양액의 DO를 일정하게 유지하였으며, 연동펌프의 제어에 의해 기질공급을 안정하게 수행하였다. 기질공급의 제어와 DO의 제어를 한가지의 하아드웨어 및 소프트 웨어로 수행할 수 있었다. 제어시스템을 이용하여 메탄올이용균인 Methylobacillus sp. SKI을 유가배양한 결과 배양액의 DO제어와 메탄올 공급의 제어가 무리없이 수행되었으며, 배양 10시간만에 16.53g/l의 균체농도에 도달하였다.

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Speculation on the Identity of Bacteria Named TFOs Occurring in the Inefficient P-Removal Phase of a Biological Phosphorus Removal System

  • Lee, Young-Ok;Ahn, Chang-Hoon;Park, Jae-Kwang
    • Environmental Engineering Research
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    • 제15권1호
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    • pp.3-7
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    • 2010
  • To better understand the ecology of tetrade forming organisms (TFOs) floating in a large amount of dairy wastewater treatment plant (WWTP) effluent (sequencing batch reactor [SBR]) during the inefficient phosphorus (P) removal process of an enhanced biological P removal system, the TFOs from the effluent of a full scale WWTP were separated and attempts made to culture the TFOs in presence/absence of oxygen. The intact TFOs only grew aerobically in the form of unicellular short-rods. Furthermore, to identify the intact TFOs and unicellular short-rods the DNAs of both were extracted, analyzed using their denaturing gradient gel electrophoresis (DGGE)-profiles and then sequenced. The TFOs and unicellular short-rods exhibited the same banding pattern in their DGGE-profiles, and those sequencing data resulted in their identification as Acinetobacter sp. The intact TFOs appeared in clumps and packages of tetrade cells, and were identified as Acinetobacter sp., which are known as strict aerobes and efficient P-removers. The thick layer of extracellular polymeric substance surrounding Acinetobacter sp. may inhibit phosphate uptake, and the cell morphology of TFOs might subsequently be connected with their survival strategy under the anaerobic regime of the SBR system.