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검색결과 14건 처리시간 0.021초

Fermentation and Metabolic Pathway Optimization to De Novo Synthesize (2S)-Naringenin in Escherichia coli

  • Zhou, Shenghu;Hao, Tingting;Zhou, Jingwen
    • Journal of Microbiology and Biotechnology
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    • 제30권10호
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    • pp.1574-1582
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    • 2020
  • Flavonoids have diverse biological functions in human health. All flavonoids contain a common 2-phenyl chromone structure (C6-C3-C6) as a scaffold. Hence, in using such a scaffold, plenty of high-value-added flavonoids can be synthesized by chemical or biological catalyzation approaches. (2S)-Naringenin is one of the most commonly used flavonoid scaffolds. However, biosynthesizing (2S)-naringenin has been restricted not only by low production but also by the expensive precursors and inducers that are used. Herein, we established an induction-free system to de novo biosynthesize (2S)-naringenin in Escherichia coli. The tyrosine synthesis pathway was enhanced by overexpressing feedback inhibition-resistant genes (aroGfbr and tyrAfbr) and knocking out a repressor gene (tyrR). After optimizing the fermentation medium and conditions, we found that glycerol, glucose, fatty acids, potassium acetate, temperature, and initial pH are important for producing (2S)-naringenin. Using the optimum fermentation medium and conditions, our best strain, Nar-17LM1, could produce 588 mg/l (2S)-naringenin from glucose in a 5-L bioreactor, the highest titer reported to date in E. coli.

도시오픈스페이스로서의 도심지 소공원 디자인에 관한 연구 (A Study on the Pocket Park as the urban Open Space)

  • 이강일
    • 디자인학연구
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    • 제5권1호
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    • pp.7-20
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    • 1992
  • Since 80th, in the monetary markets due to the escalation of creditable promptness dealing of financial goods, Korea monetary business has been confronted with structual contradictions. The first like a men power reduction, inorder to meet and satisfying with cost reduction policy of the holding firms and secondly, escalation and expention of business networks or facilities inorder to meet and satis~'ing with customers continuously seeking for prompt, acurate, and helpfulness service nearly like one similar to a door to door service. Accordingly, the firms eventually poured mere sweaty money into procuring such devices for upgrading. On the contrary, however, regardless the escalation monetary expenditure may be, inorder to promote quality of the same devices are keen necessary because present days customer groups were tended to receives ample service almost like door to door service without moving too$$\mu$h apart from the immediate spot where they belongs tl or veryspot of where actually they spend instead of deep consideration of natures, quality or sttractiveness of service medias what they can received from. Under the present circumstance, the financial business firms, however, would highly suffering and recei\ulcornerves pressures from imblalance natures ofincoming and out go of the business budget due to excessive inc1ease of managerial expences. For these reasons, therefor, in order to solves, compensates, improves, and developes most effective fina\ulcornerncial service for customers can use under most pleasant environment, the establishment of adequate standardization of such service devices based on the foregoing environmental circumstances. This analytical study, therefor, mainly aimed and emphasized based on pre- estimatimation of proper de\ulcornervlopment and use of the unattended servicing devices such as Cash Dispenser and Auto Teller Machine bygeneralization, automating. and innovation of design to meet compactness and door to door like convenient service for customers which can be achieved by performing constant comparative analysis and continual study toward development of such devices those of which produced by Five (5) major domestic producers aro~nd the country.

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Expression, Purification and Properties of Shikimate Dehydrogenase from Mycobacterium Tuberculosis

  • Zhang, Xuelian;Zhang, Shunbao;Hao, Fang;Lai, Xuhui;Yu, Haidong;Huang, Yishu;Wang, Honghai
    • BMB Reports
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    • 제38권5호
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    • pp.624-631
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    • 2005
  • Tuberculosis, caused by Mycobacterium tuberculosis, continues to be one of the main diseases to mankind. It is urgent to discover novel drug targets for appropriate antimicrobial agents against this human pathogen. The shikimate pathway is onsidered as an attractive target for the discovery of novel antibiotics for its essentiality in bacteria and absence in mammalian cells. The Mycobacterium tuberculosis aroE-encoded shikimate dehydrogenase was cloned, expressed and purified. Sequence alignment analysis shows that shikimate dehydrogenase of Mycobacterium tuberculosis exhibit the pattern of G-X-(N/S)-V-(T/S)-X-PX-K, which is highly conserved within the shikimate dehydrogenase family. The recombinant shikimate dehydrogenase spectrum determined by CD spectroscopy showed that the percentages for $\alpha$-helix, $\beta$-sheet, $\beta$-turn, and random coil were 29.2%, 9.3%, 32.7%, and 28.8%, respectively. The enzymatic characterization demonstrates that it appears to be fully active at pH from 9.0 to 12, and temperature $63^{\circ}C$. The apparent Michaelis constant for shikimic acid and $NADP^+$ were calculated to be about $29.5\;{\mu}M$ and $63\;{\mu}M$. The recombinant shikimate dehydrogenase catalyzes the substrate in the presence of $NADP^+$ with an enzyme turnover number of $399\;s^{-1}$. Zymological studies suggest that the cloned shikimate dehydrogenase from M. tuberculosis has a pretty activity, and the work should help in the discovery of enzyme inhibitors and further of possible antimicrobial agents against Mycobacterium tuberculosis.

Escherichia coli에 의한 방향족 아미노산 생산에 관한 연구 (A Study on the Production of Aromatic Amino Acids by Escherichia coli.)

  • Park, Young-Jin
    • 한국미생물·생명공학회지
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    • 제13권2호
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    • pp.119-127
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    • 1985
  • 여러 가지 Escherchia coli 변이주의, glucose 와 ammonium염과 같은 간단한 기질로부터 방향족 아미노산 특히 phenylanine을 생합성하는 능력을 비교 검토한 결과 방향족 아미노산 생합성과정중 common pathway의 첫 번째 반응이 phenylanine 생합성에 가장 큰 영향을 준다는 것을 확인하였다. 따라서 관계효소인 DAHP synthase의 효소활성과 생합성에 관련된 각종 대사 제어작용을 효과적으로 제거시킴으로서 phenylalanine 생산량을 크게 높일 수 있었으며 더욱이 phenylalanine terminal pathway의 첫 단계 반응을 촉매하는 prephenate de-hydratase의 효소활성과 효소생합성에 관련된 제어 작용도 동시에 제거하면 phenylalanine생산이 상승적으로 증가됨을 보였다. 한편 방향족아미노산의 transport system에 관계하는 arop유전자의 변이는 phenylalanine생산을 크게 저하시키는 효과를 나타내었다.

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