• 제목/요약/키워드: Continuous cultures

검색결과 143건 처리시간 0.027초

Lovastatin 고생산성 변이주의 신속 선별을 위해 통계적 방법을 적용한 Sporulation 배지 개발 및 Miniature 배양 방법 개발 (Mass Screening of Lovastatin High-yielding Mutants through Statistical Optimization of Sporulation Medium and Application of Miniaturized Fungal Cell Cultures)

  • 안현정;정용섭;김평현;전계택
    • KSBB Journal
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    • 제22권5호
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    • pp.297-304
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    • 2007
  • Lovastatin은 근사형성 균류인 Aspergillus terreus가 생합성하는 이차대사산물로 강력한 고지혈증 치료제로 널리 이용되는 물질이다. 본 연구에서는 lovastatin 고생산변이주를 이용하여 포자배지 최적화를 통한 miniature 배양 방법을 확립하고자 하였다. 우선 miniature 배양에 필수적인 효과적인 포자 형성 방법을 개발하고자 포자 형성 배지의 통계학적 배지 최적화를 수행하였다. Miniature 배양의 inoculum으로 이용되는 대량의 포자를 획득하기 위해 Plackett-Burman 실험법을 이용하여 포자 형성을 향상시키는 성분을 조사한 결과, glucose, sucrose, yeast extract 그리고 $KH_2PO_4$가 주목할 만한 효과를 보였다. 상기 성분의 최적 농도를 확인하기 위해 반응표면분석법 (RSM)을 이용한 결과, PDA 포자 형성 배지와 비교하여 볼 때, 최적 성분 농도에서 포자 형성이 약 190배 증가하였다. 최적화된 포자형성 배지를 이용하여 lovastatin 고생산성 변이주의 대량 선별을 위한 miniature 배양 방법을 확립하기 위해 기존의 실험 과정에 'PaB (adaptation)'라는 한 번의 계대배양을 더 추가한 결과 생산균주의 안정성과 재현성이 큰 폭으로 증가하는 주목할 만한 결과를 얻을 수 있었다. 단기간에 가능한 한 다량의 균주를 스크리닝하기 위해 성장배양과 생산배양 모두 조업부피가 7 ml인 tube를 이용해 miniature 배양을 반복 수행하여, lovastatin 생산성과 배양형태가 훌륭한 변이주를 선별할 수 있었는데, 이 균주는 7 ml tube배양과 250 ml flask배양 (조업부피 50 ml) 모두에서 생산성이 높은 것으로 보아 산소 의존도가 비교적 낮고 생산 안정성이 높은 균주인 것으로 판단되었다. 한편 miniature 배양을 이용해서 lovastatin 고생산성을 보이는 균주를 신속 선별하기 위해서는 균주의 적절한 배양형태 유도가 매우 중요한 것으로 관찰되었다. 즉 생산배양으로의 고활성 균주의 접종을 위해서, 또한 생산배양에서 pellet의 배양형태 유도를 위해서 성장배양 시에는 반드시 고농도의 균사모양을, 생산배양 시에는 직경 1 mm 이하의 pellet모양의 배양 형태를 유지해야만, 생산균주가 lovastatin을 안정적으로 고생산할 수 있는 것으로 관찰되었다. 초기에 선별된 균주를 이용하여 miniature 배양에 의해 고속 균주선별 실험을 반복함으로써 고생산성 균주들을 다량 선별할 수 있었는데, 이들의 lovastatin 생산성을 조사한 결과, 기존의 flask 배양대비 오차범위가 $\pm$20% 이내의 생산성을 보이는 균주가 초기 선별시의 32%에 비해 81%로 크게 증가함을 확인할 수 있었다. 이와 같은 결과는 lovastatin 고생산성, 고안정성 균주의 고속 스크리닝을 위해서 본 연구에서 개발한 tube를 이용한 miniature 배양이 기존의 flask 배양을 대체할 수 있는 훌륭한 배양방법임을 제시해 준다.

막분리를 이용한 미생물 재순환 연속 알콜발효에 관한 연구 (A Study on Continuous Alcohol Fermentation with Cell Recycle by Means of Membrane Separation)

  • 이준형;목영일허병기
    • KSBB Journal
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    • 제7권2호
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    • pp.139-143
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    • 1992
  • 본 연구에서는 균주 S. cerevisiae ATCC 24858을 막분리를 이용한 미생물 재순환 연속에탄올 발효시스템에 적용하여 희석비 (0.3, 0.5, $0.7hr^{-1}$)와 주입기질의 당농도 (50, 100, 150, 200g/l)에 따른 균체농도 및 알콜생산성의 변화관계를 규명하여 보았다. 주입기질 당농도50g/l, 희석비$0.3hr^{-1}$ 내지 당농도200g/l, 희석비 $0.7hr^{-1}$ 사이의 실험범위에서 균체농도는 12g/l내지 107g/l 사이에, 알콜생산성은 7.0g/l 내지 55.6g/l 사이에 분포되었다. 더욱이 알콜생산성은 회분식 발효의 생산성에 비하여 최소 307배 내지 최대 29.3배 까지 증가하였다. 또한 본 연구의 실험범위에 대한 에탄올 평균수율은 0.42로써 이론수율의 82%였으며 회분식 수율보다 약간 상회하였다.

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Evaluating Carriers for Immobilizing Saccharomyces cerevisiae for Ethanol Production in a Continuous Column Reactor

  • Cha, Hye-Geun;Kim, Yi-Ok;Choi, Woon Yong;Kang, Do-Hyung;Lee, Hyeon-Yong;Jung, Kyung-Hwan
    • Mycobiology
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    • 제42권3호
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    • pp.249-255
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    • 2014
  • We evaluated a more practical and cost-effective immobilization carriers for ethanol production using the yeast Saccharomyces cerevisiae. Three candidate materials-rice hull, rice straw, and sawdust-were tested for their cell-adsorption capacity and operational durability. Derivatizations of rice hull, rice straw, and sawdust with the optimal concentration of 0.5 M of 2-(diethylamino)ethyl chloride hydrochloride (DEAE HCl) resulted in > 95% adsorption of the initial yeast cells at 2 hr for DEAE-rice hull and DEAE-sawdust and in only approximately 80% adsorption for DEAE-rice straw. In addition, DEAE-sawdust was found to be a more practical carrier for immobilizing yeast cells in terms of operational durability in shaking flask cultures with two different speeds of 60 and 150 rpm. Furthermore, the biosorption isotherms of DEAE-rice hull, -rice straw, and -sawdust for yeast cells revealed that the $Q_{max}$ of DEAE-sawdust (82.6 mg/g) was greater than that of DEAE-rice hull and DEAE-rice straw. During the 404-hr of continuous column reactor operation using yeast cells immobilized on DEAE-sawdust, no serious detachment of the yeast cells from the DEAE-sawdust was recorded. Ethanol yield of approximately 3.04 g/L was produced steadily, and glucose was completely converted to ethanol at a yield of 0.375 g-ethanol/g-glucose (73.4% of the theoretical value). Thus, sawdust is a promising practical immobilization carrier for ethanol production, with significance in the production of bioethanol as a biofuel.

소프트웨어 개발 프로젝트에서 수행의 지속행위에 관한 연구 : 문화적 차이의 관점에서 (A Study on Escalation of Commitment Behavior in Software Projects : The Perspective of Cultural Differences)

  • 김인재;심형섭;김종은
    • Journal of Information Technology Applications and Management
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    • 제11권2호
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    • pp.1-14
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    • 2004
  • The level of sunk cost and risk-taking theory have been offered as one explanation for the escalation of commitment behavior. This Study attempted to replicate Keil's study in Korea. Keil examined the level of sunk cost associated with the risk propensity and risk perception of decision-makers, and these factors are assessed for cross-cultural robustness using matching laboratory experiments carried out in three countries. The level of sunk cost and the risk perception of decision-makers contributed significantly to their continuous willingness to their project. Moreover, the risk propensity of decision-makers was inversely related to risk perception, and this inverse relationship was significantly more weak in Korea than in Singapore. These results show that the sunk-cost effect exists across cultures, and that the risk-taking behaviors are partially mediated by cultural factors.

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The Effect of Light on the Production of Reserpine in Cultured Rauwolfia serpentina Cells

  • Yamamoto, Osamu
    • Natural Product Sciences
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    • 제2권2호
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    • pp.90-95
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    • 1996
  • When reserpine-producing cell strains of Rauwolfia serpentina were transferred from the dark to the light irradiation, the production of reserpine was extremely enhanced whereas the cell growth was suppressed. In an incubation period of 20 days, the most effective culture condition for reserpine production was the combination of 8 days of dark culture and following 12 days of light culture. The time courses of both cell growth and reserpine production were measured in vitro in order to clarify the effect of wave length range of light on the biosynthesis of reserpine. Although the growth of cultured cells which had been incubated under continuous red, yellow, and green lights, respectively, was similar to that of the cultured cells subcultured in the dark. The cells cultured under red light irradiation produced less reserpine than dark-grown cultures. Both blue and near-ultraviolet light inhibited the growth of cultured cells. The production of reserpine was strikingly enhanced by blue light, but was strongly inhibited by near-ultraviolet light.

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Functional Cardiomyocytes Formation Derived from Parthenogenetic Mouse Embryonic Stem Cells

  • Sin Hyeon-A;Kim Eun-Yeong;Lee Geum-Sil;Park Eun-Mi;Park Se-Pil;Im Jin-Ho
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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    • pp.20-20
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    • 2002
  • This study was to establish the use of parthenogenetic mouse ES (P-mES02) cells as a reproducible differentiation system for mouse cardiomyocytes. To induce differentiation, P-mES02 cells were dispersed by dissociation and the formation of ES cell aggregates in differentiation medium. After 7 days in differentiation culture, the embryoid bodies (EBs) were plated onto gelatin-coated dish. Cultures were observed daily using an inverted light microscope to determine the day of contraction onset and total duration of continuous contractile activity for each contracting focus. (omitted)

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Influence of Growth Rate on Biosorption of Heavy Metals by Nocardia amarae

  • Kim, Dong Wook;Daniel K. Cha;Hyung-Joon Seo;Jong Bok Bak
    • Journal of Microbiology and Biotechnology
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    • 제12권6호
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    • pp.878-881
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    • 2002
  • The goal of the current research was to assess the influence of the growth rate of Nocardia amarae on its overall metal binding capacity. Batch sorption isotherms for cadmium (Cd), copper (Cu), and nickel (Ni) showed that Nocardia cells harvested from chemostat cultures at a dilution rate of $0.33d^-1$ had a significantly higher metal sorption capacity than cells grown at 0.5 and $1d^-1$. The cell surface area estimated using a dye technique indicated that pure N. amarae cells grown at a lower growth rate had a significantly more specific surface area than cells harvested from a higher growth rate operation. Accordingly, this difference in the specific surface area seemed to indicate that the higher metal sorption capacity of the slowly growing Nocardia cells was due to their higher specific surface area.

Zymomonas mobilis Plasmid Vector의 숙주세포 내에서의 안정성에 관한 연구 (Stabilities of Plasmid Vectors in Zymomonas mobilis)

  • 이상기;박은숙;황덕주;박무영
    • 한국미생물·생명공학회지
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    • 제15권5호
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    • pp.328-333
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    • 1987
  • 고효율 에타놀 생성 균주인 Zymomonas mobilis의 plasmid vector가 숙주세포 내에서 안정하게 유지되는지의 여부를 회분 및 연속배양 방법을 통하여 조사하였다. Z. mobilis 숙주세포는 전이된 plasmid의 크기와 배지의 조성에 따라 증식속도에 영향을 받음을 알 수 있었고 작은 크기의 plasmid를 비 선택성 (non-selective) 배지에서 배양한 경우 더 빠른 성장을 보였다. 또한 사용한 4종류의 plasmid vector 모두 숙주세포 내에서 30세대 이상에 걸쳐 93% 이상의 높은 안정성을 나타내면서 독립적으로 증식함이 확인되었다.

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연속배양에서 고핵산 효모 Saccharomyces cerevisiae MTY62 균주의 배양 ${\cdot}$ 생리학적 특성

  • 김재범;남수완
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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    • pp.346-349
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    • 2000
  • In order to investigate the characteristics for RNA accumulation, continuous cultures of S. cerevisiae MTY62 under the carbon-limitation or carbon-phosphate-limitation were performed. In the carbon-limited chemostat, the highest RNA content of 144 mg-RNA/g-dry cell was observed at the dilution rate of $0.30\;hr^{-1}$. In the carbon and phosphate-limited chemostat, the culture condition with the dilution rate of $0.25\;hr^{-1}$ gave the maximal RNA content of 209 mg-RNA/g-dry cell. The accumulation rate of RNA $(mg-RNA/L\;{\cdot}\;hr)$ was decreased at higher dilution rate in the carbon and phosphate-limited chemostat.

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발효소시지의 생산과 미생물적 특성 (Production and Microbiological Characteristics of Fermented Sausages)

  • ;이주연
    • 한국축산식품학회지
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    • 제23권4호
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    • pp.361-375
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    • 2003
  • In this study, significant factors influencing on the quality and stability of fermented sausage, such as materials, processing conditions, and microbiological characteristics as well as topography during ripening, were documented. Since most fermented sausages are not heated during manufacture or before consumption, a strict control of the growth of pathogens and the selection of favourable conditions that encourage the specific growth and development of desirable microflora are particularly important. With respect to microbiological safety, hurdles, i.e., preservations(nitrite), redox potential, competitive flora, acidity(pH), and water activity($a_{w}$) are matters of importance to prevent proliferation of bacterial pathogens. Today, for ensuring the safety and quality of the final product, the application of starter cultures in combination with the proper processing is subsequently used in practice. For improving the efficiency of microbiological utility in the production of fermented sausages, the understanding of their topography is essential. The documented different points must be taken into account when HACCP systems set up for the manufacture of fermented sausages. There are continuous researches concerning desirable improvements to sausage fermentation with health enhancing properties.