• Title/Summary/Keyword: overproduction.

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Overproduction of Bacillus macerans Cyclodextrin Glucanotransferase in E. coli by Coexpression of GroEL/ES Chaperone

  • Kwon, Mi-Jung;So-Lim Park;Sung-Koo Kim;Soo-Wan Nam
    • Journal of Microbiology and Biotechnology
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    • v.12 no.6
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    • pp.1002-1005
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    • 2002
  • The effects of GroEL/ES chaperone on the production of soluble form of B. macerans cyclodextrin glucanotransferase (CGTase) in recombinant E. coli were investigated. The cgt gene and groEL/ES genes are under the control of T7 promoter and Pzt-1 promoter, respectively. The optimal concentrations of inducers, IPTG and tetracycline, were found to be 1.0 mM and 10 ng/ml, respectively. When tetracycline and IPTG were added at the early exponential phase (2h) and exponential phase (3h) of growth, respectively, about 1.5-fold increase of soluble CGTase activity and 1.6-fold increase of soluble CGTase protein were obtained. An SDS-PAGE analysis revealed that about $37.2\%$ of total CGTase protein was in the soluble fraction when GroEL/ES chaperone was overexpressed.

Overproduction of Cellulose in Acetobacter xylinum KCCM 10100 Defective in GDP-Mannosyltransferase

  • Park Sang-Tae;Kim Eung-Bin;Kim Young-Min
    • Journal of Microbiology and Biotechnology
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    • v.16 no.6
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    • pp.961-964
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    • 2006
  • GDP-mannosyltransferase (GMT) is an enzyme responsible for the addition of a mannose to glucose ($\alpha$[1$\rightarrow$3]) during biosynthesis of the water-soluble branched polysaccharide acetan in Acefobacter species. In an effort to obtain a cellulose-overproducing bacterium, a mutant defective in GMT of Acetobacter xylinum KCCM 10100 was constructed by single crossover homologous recombination using part of the aceA gene encoding GMT amplified by polymerase chain reaction. The GMT-disrupted mutant produced 23% more cellulose, but 16% less water-soluble polysaccharide than those of the wild-type strain. Analysis of the sugar composition by gel permeation chromatography revealed that water-soluble polysaccharides produced by the GMT-defective mutant contained no mannose molecule.

Overexpression of Escherichia coli Thiol Peroxidase in the Periplasmic Space

  • Kim, Sung-Jin;Cha, Mee-Kyung;Kim, Il-Han;Kim, Ha-Kun
    • Journal of Microbiology and Biotechnology
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    • v.8 no.1
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    • pp.92-95
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    • 1998
  • Overproduction of Escherichia coli thiol peroxidase in the periplasmic space was achieved by locating the appropriate gene on a downstream region of the strong T7 promoter. E. coli strain BL21 carrying the recombinant plasmid pSK-TPX was induced by IPTG, lysed, and analyzed by SDS-polyacrylamide gel electrophoresis. A large amount of the overexpressed thiol peroxidase was located in the periplasmic space. A homogeneous thiol peroxidase was obtained from E. coli osmotic shock fluid by simple one-step gel permeation chromatography.

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High Production of L-Crnithine by L-Citrulline Auxotroph of Brevibacterium ketoglutamicum : PART I : Selection of Stable Mutant Strains with Less Revertant Formation (Brevibacterium ketoglutamicum을 이용한 L-Ornithine 생산 연구 PART I : L-Ornithine 생산 고역가 안정화 균주 선발)

  • 류욱상;장형욱;이홍원;정준기;장순재;유연우;박영훈
    • Microbiology and Biotechnology Letters
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    • v.27 no.4
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    • pp.292-297
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    • 1999
  • Overproduction of L-ornithine by mutant strains isolated from Brevibacterium detoglutamicum BK1046 was investigated. The strain was a L-ornithine auxotroph and exhibited culture instability during fermentation. Through a sequential screening effort, a highly stable strain with lless revertant formation was finally selected and designated B. ketoglutamicum BK52 (KCTC0141BP). It prouduced L-ornithine at a high concentration (above 9 g/L) independent of subculture or cultivation time, and also had a very low tendency of revertant formation. In a long-term storage, this strain maintained its cell stability and productivity of L-ornithine to a reasonable range.

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The Research on the Fish Aquiculture Structure of Fishing Community (어촌계 가두리 양식어장의 행사실태분석을 통한 정책방향 고찰)

  • Ock, Young-Soo
    • The Journal of Fisheries Business Administration
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    • v.36 no.2 s.68
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    • pp.25-52
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    • 2005
  • Since late 1990's, the Fish Aquiculture Households belong Fishing Community have been faced very serious pressure of management. The surface factors of this situation have been understood to overproduction and too much imports from China, but basic factor was caused structural problem, that is considered small scale production system. Then, We should enlarge scale of the Fish Aquiculture Households belong Fishing Community to overcoming profitability aggravation. To the Enlargement of Scale, We should consider to change the regime of Fishing Community that means two practical programs. First, it need to enlarge the scale of Fishing Community than now, and Second, it need to incorporate the Fishing Community for exclusive Management of Fish Aquiculture Rights.

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재조합 Escherichia coli 시스템을 이용한 폐흡충 cystein proteinase의 생산 연구

  • Hong, Seong-Hui;Lee, Gil-Hwan;Jeon, Hui-Jin;Hwang, Hyeon-A;Park, Seong-Ryeol;Hwang, Yeong-Bo;Park, Hyeon
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.651-654
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    • 2000
  • A fermentation study of the recombinant Escherichia coli system has been tired to overproduce a recombinant Paragonimus westermani cysteine proteinase, rPwCP1. Using the modified LB media, main cultures and chemical induction with IPTG were carried out to examine the possibility of secretory protein overproduction at high cell density culture. As a result, the target protein of rPwCP1, purified by metal affinity chromatography and gel filtration, has been shown at 50.8 kDa on SDS-PAGE, and its final concentration turns out to be 350mg/L.

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Bioconversion of Aniline to Acetaminophen and Overproduction of Acetaminophen by Streptomyces spp.

  • Jin, Hyung-Jong;Park, Ae-Kyung;Lee, Sang-Sup
    • Archives of Pharmacal Research
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    • v.15 no.1
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    • pp.41-47
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    • 1992
  • In order to obtain acetaminophen, a popular analgesic-antipyretic, though microbial p-hydroxylation and N-acetylation of aniline, various Streptomyces strains were screened. Aniline N-acetylation activity was rather ubiquitous but-hydroxylation activity was selective. Microbial conversion pathway of aniline to acetaminophen was considered to be through N-acetylation and p-hydroxylation or vice versa. However, depending on species used, o-hydroxylation and its degradation activity (S. fradiae) and acetaminophen degradation activity (S. coelicolar) were also detected. Among the screened Streptomyces strains, S fradiae NRRL 2702 showed the highest acetanilide p-hydroxylation activity (203% conversion rate). Furthermore, in S. fradiae carbon source and its concentration, phosphate ion concentration and pH of growth medium were found to play the crucial roles in p-hydroxylation activity. Through the proper combination of factors mentioned above, the ten times more activity (26-30% conversion rate) was attained.

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Inhibitors of Inducible Nitric Oxide Synthase Expression from Artemisia iwayomogi

  • Ahn, Hanna;Kim, Ji-Yeon;Lee, Hwa-Jin;Kim, Yong-Kyun;Ryu, Jae-Ha
    • Archives of Pharmacal Research
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    • v.26 no.4
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    • pp.301-305
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    • 2003
  • Nitric oxide (NO) is an important bioactive agent that mediates a wide variety of physiological and pathophysiological events. NO overproduction by inducible nitric oxide synthase (iNOS) results in severe hypotension and inflammation. This investigation is part of a study to discover new iNOS inhibitors from medicinal plants using a macrophage cell culture system. Two sesquiterpenes (1 and 2) were isolated from Artemisia iwayomogi (Compositae) and were found to inhibit NO synthesis ($IC_{50} 3.64 \mu g/mL and 2.81 \mu$g/mL, respectively) in lipopolysaccharide (LPS)-activated RAW 264.7 cells. Their structures were identified as 3-Ο-methyl-iso-secotanapartholide (1) and iso-secotanapartholide (2). Compounds 1 and 2 inhibited the LPS-induced expression of the iNOS enzyme in the RAW 264.7 cells. The inhibition of NO production via the down regulation of iNOS expression may substantially modulate the inflammatory responses.

Inhibitors of Nitric Oxide Synthesis from Ginseng in Activated Macrophages (활성화한 RAW 264.7 세 포주에서 인삼 Polyacetylene류의 Nitric Oxide 생성저해)

  • 류재하;장세란
    • Journal of Ginseng Research
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    • v.22 no.3
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    • pp.181-187
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    • 1998
  • Nitric Oxide (NO), derived from L-arginine, is produced by two types (constitutive and inducible) of nitric oxide synthase (NOS). The NO produced in large amounts by the inducible NOS is known to be responsible for the vasodilation and hypotension observed in septic shock. We have found three polyacetylene compounds which inhibited the production of NO in LPS-activated RAW 264.7 cells. Their structures were identified as panauynol, ginsenoyne A and PQ-6 by the spec- troscopic analysis (IC50 values were 32.3 $\mu$M, 2.3 $\mu$M, 1.5 $\mu$M, respectively). These polyacetylenes may be useful candidates for the development of new drug to treat endotoxemia and inflammation accompanied by the overproduction of NO.

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Monascus Red Pigment Overproduction by Coculture with Recombinant Saccharomyces cerevisiae Secreting Glucoamylase

  • Lim, Ho-Soo;Yoo, Seung-Ku;Shin, Chul-Soo;Hyun, Young-Min
    • Journal of Microbiology
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    • v.38 no.1
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    • pp.48-51
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    • 2000
  • In liquid cultures using sucrose media, the coculture of Monascus with recombinant Saccharomyces cerevisiae expressing the glucoamylase gene from Aspergillus niger enhanced red pigment production by approx. 19%, compared with the coculture of wild type S. cerevisiae. Coculture with recombinant S. cerevisiae was more effective than with wild type S. cerevisiae for Monascus red pigment production. Cocultures of Monascus with commercial amylases of Aspergillus also induced high production of pigment and morphological changes in a solid culture using sucrose media.

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