• Title/Summary/Keyword: 대사공학

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Regulation of Sulfur Metabolism in Cephalosporium acremonium (Cephalosporium acremonium에서 황화합물 대사의 조절)

  • Lee, Kyoung;Park, Sang-Ho;Lee, Jung-Joon;Mheen, Tae-Ick
    • Microbiology and Biotechnology Letters
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    • v.15 no.5
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    • pp.361-367
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    • 1987
  • A DL-seleno-methionine resistant mutant, Cephalosporium acremonium MS-92 showed increased activities of sulfate and L-methionine uptake than the parent strain, and accumulated excess methionine and S-adenosylmethionine (SAM) intracellularly. And the sulfate uptake system was severely inhibited by L-cysteine. In crude enzyme extracts, the mutant MS-92 showed lower L-serine sulfhydrylase (identical with cystathionine $\beta$-synthase) activity than the parent. Also, cysteine desulfhydrylase activity, an index of intracellular L-cysteine concentration, of the mutant MS-92 was decreased by about 50% as com-pared with that of the parent. Thus, it was supposed that the mutant MS-92 should have n lower level of L-cysteine than the parent. In C. acremonium like A. nidulans, the enzymes related to the biosynthesis of methionine might be regulated by L-cysteine, but not by methionine or SAM.

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Inhibition of $\alpha$-Glucosidase Activity by Quercetin (Quercetin에 의한 $\alpha$-glucosidase 활성 저해)

  • Lee, Dong-Sun;Kim, Jong-Guk;Lee, Sang-Han
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.368-372
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    • 2006
  • Quercetin is a flavonoid molecule that is known to tie in various sources of natural products such as vegetables and fruits. It has been proven that quercetin plays a crucial role in the prevention of colon cancer as well as homeostasis as radical scavenger in human body. It is also well-known that glycosidases, including $\alpha$-glucosidase, are involved in a variety of degenerative metabolic disorders. In the course of screening useful $\alpha$-glucosidase inhibitors, we screened out quercetin as a $\alpha$-glucosidase inhibitor from chemical libraries. Quercetin was shown to be a reversible, slow-binding, and noncompetitive inhibitor of yeast a-glucosidase with a K$_i$ value of $6.3\times10^{-8}$ M when it was included with an enzyme mixture. Together, these results show that quercetin has potential in treating disorders including diabetes, although the further mechanistic study is needed.

Effect of NaCl on Halomonas subglaciescola DH-1 Incapable of Growing at Non-Salinity (Halomonas subglaciescola DH-1의 생장에 미치는 염화나트륨의 영향)

  • Na, Byung-Kwan;Yoo, Young-Sun;Park, Doo-Hyun
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.298-303
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    • 2007
  • A halophilic bacterium, H. subglaciescola DH-1, grew at 2.0 M salinity, but did not grow at 0.8 M salinity when cultivated at higher temperature ($40^{\circ}C$) than optimum ($30^{\circ}C$). When the cell extract of strain DH-1 was heated at $50^{\circ}C$ for 60 min in the absence of NaCl, isocitrate dehydrogenase and malate dehydrogenase lost their activities, but when it was heated in the presence of 2.0 M NaCl, the activity was maintained. Meanwhile, the cell extract of E. coli did not catalyze the reduction of $NAD^+$ to NADH coupled with the oxidation of isocitrate and malate at higher salinities than 1.0 M. The pH range for DH-1 was 7 to 10, and that for E. coli was 5 to 9. DH-1 was not grown in conditions with sodium salts other than NaCl.

Development of High-yielding Mutants of Streptomyces avermitilis for Avermectin B_{1a} Production through Protoplast Fusion. (원형질체 융합에 의한 Avermectina B_{1a} 고생산성 Streptomyces avermitilis 균주 개발)

  • 김경희;송성기;정연호;정용섭;전계택
    • Microbiology and Biotechnology Letters
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    • v.32 no.2
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    • pp.101-109
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    • 2004
  • In order to enhance the productivity of AVM $B_{la}$ produced by Streptomyces avermitilis as a secondary metabolite, we established a basic protocol necessary for protoplast fusion with high-producing strains as a fusion partner, and then obtained various kinds offusants by adopting a massive strain-development procedure (a miniaturized strain screening system). An alternative fusion method using UV and/or NTG mutation of protoplasts was developed to screen genetic recombinants without specific selectable markers. In this method, the mutants obtained by protoplast fusion after UV and/or NTG treatment (95% death rate) of the respective fusion partner (protoplasts of the respective mutants resistant against L-isoleucine antimetabolites such as O-methylthreonine and/or azaleucine) were regarded as DNA-recombined protoplast fusants. Notably it was demonstrated that most of the protoplast recombinants obtained by the UV mutation method were able to biosynthesize higher amount of AVM $B_{la}$ , reaching almost three times higher level (almost equal to the industrial productivity), compared to the average AVM Bla amount of the parallel mother strains.

Metabolic Control of Maintenance for the Production of pro-Urokinase from Human Thyroid cells (인체 갑상세포의 대사조절에 의한 프로 유로카이나제의 생산)

  • 강재구;최석규;이현용
    • Microbiology and Biotechnology Letters
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    • v.18 no.4
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    • pp.401-405
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    • 1990
  • Maximum specific pro-UK production rate was achieved at 15 mllmin of perfusion rate as $5.7 \times 10^{-8}$ $\mu g$/h/cell by cytostatic cultivation of human cell line 579 with DMEM and 590 FBS. As perfusion rates were increased, glutamine uptake rates were also increased but ammonium production rates remained relatively constant, which resulted in low ratio of ammonium to glutamine at high perfusion rate. Partially it quantitatively explains why the productivity is increased in perfusion cultivations. At maintenance period of 15 days by controlling metabolic process, such as 5 mM of glucose, 2 mM of gtutamine, 10% of air saturation and pH 6.2, high speecific product production rate and product yield on substrate were obtained as $12\times 10^{-8}$$\mu g$/h/cell abd 0.226 mglg of glucose, respetively. This product yield corresponds to 0.223 mg/day of productivity at 10 mllmin of perfusion rate.

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Effects nit Mineral Salts on the Improvements of Sisomicin field (무기질 염이 Sisomicin 발효 수율의 증가에 미치는 영향)

  • Shin, Chul-S;Sang H. Han;Lee, Sang H.
    • Microbiology and Biotechnology Letters
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    • v.17 no.3
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    • pp.247-251
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    • 1989
  • Effects of mineral salts on sisomicin fermentation were investigated. The optimal concentration of CoCl$_2$for accomplishing a high antibiotic yield was found to be 16.8 $\mu$M at which it could function as a cofactor. At this level the other mineral salts tested had no effect. On the other hand, at much higher concentration levels (above 1 mM), four mineral salts such as ZnSO$_4$, KH$_2$PO$_4$, FeSO$_4$and MgSO$_4$were used in order to liberate the intracellular sisomicin out-side the cells, because the sisomicin accumulated mostly in cells and it was supposed to limit the improvement of antibiotic yield. ZnSO$_4$and KH$_2$PO$_4$had no effect at all, and FeSO$_4$brought about some improvement. However, by keeping the concentration of MgSO$_4$to be 25 mM or higher in culture broths, the antibiotic yield could be improved by more than 100%, partially due to the enhanced liberation of the intracellular antibiotic.

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The Mechanisms for Xylose Transport into Yeasts (효모내로의 Xylose 운반 기작)

  • Han, Ji-Hye;Choi, Gi-Wook;Chung, Bong-Woo;Min, Ji-Ho
    • Microbiology and Biotechnology Letters
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    • v.38 no.1
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    • pp.7-12
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    • 2010
  • The biochemical study of sugar uptake in yeasts started five decades ago and led to the early production of abundant kinetic and mechanistic data. However, the first accurate overview of the underlying sugar transporter genes was obtained relatively late, due mainly to the genetic complexity of hexose uptake in the model yeast, Saccharomyces cerevisiae. The genomic era generated in turn a massive amount of information, allowing the identification of a multitude of putative sugar transporter and sensor-encoding genes in yeast genomes, many of which are phylogenetically related. This review aims to briefly summarize our current knowledges on the biochemical and molecular features of the transporters of pentoses in yeasts, when possible establishing links between previous kinetic studies and genomic data currently available. Emphasis is given to recent developments concerning the identification of D-xylose transporter genes, which are thought to be key players in the optimization of S. cerevisiae for bioethanol production from lignocellulose hydrolysates.

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

  • Park, Young-Jin
    • Microbiology and Biotechnology Letters
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    • v.13 no.2
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    • pp.119-127
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    • 1985
  • A series of Escherichia coli mutants were exmined for ability to convert glucose and ammonium salts into phenylalanine. This enabled the biochemical changes having major. effects on phenylaianine yield, and interactions between mutations, to be identified. Changes to the common pathway of aromatic biosynthesis having a major effects include desensitization of the first enzyme (3-deoxy-D-arabinoheptulosonate synthase) to end-product inhibition, and removal of repression of enzyme synthesis. It is suggested that the 3-deoxy-D-arabino-heptulosonate synthase Phe isoenzyme has a more important effect on yield. Similarly, removal of repression and end-product inhibition on the phenylalanine terminal pathway increased yield, and changes to both common and branch pathways were synergistic. Blockage of the typrosine and tryptophan pathways had minor effects on phenylalanine yield, and a mutation affecting aramatic amino acid transport (aroP) decreased yield. With multiple-mutation strains hish specific rates of product formation (ie 0.1-0.17g phenylalanine/g cells/h) were obtained.

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Enhanced Prodiginines Production in Streptomyces coelicolor M511 by Stress of Acidic pH Shock (산성 pH 충격 스트레스에 의한 스트랩토마이세스 시에리컬러 M511의 프로디지닌 생산 증대)

  • Mo, Sang-Joon
    • Microbiology and Biotechnology Letters
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    • v.38 no.3
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    • pp.273-277
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    • 2010
  • Undecylprodiginine and streptorubin B are red-pigmented antibiotics produced by Streptomyces coelicolor A3(2). In this study, we investigated the correlation between productivity of these red-pigmented antibiotics and stress of pH shock. Biosynthesis of these red-pigmented antibiotics is enhanced at acidic pH shock on solid R2YE medium. The optimal pH shock is pH 4 which led to 1.6 fold and two-fold increase in the production of undecylprodiginine and streptorubin B as compared with control, respectively. In addition, the extract of pH 4 shocked cells exhibited a remarkable activity against Trichophyton mentagrophytes. However, neutral and basic pH shock did not give raise to promote a production of these red-pigmented antibiotics as well as antifungal activity. Thus, although the acidic pH shock is simple and easy method, it should be extremely effective approach to enhance a productivity of these red-pigmented antibiotics and other secondary metabolites.

Hydrogen Evolution from Biomass-Derived Carbohydrates by Clostridia (Clostridia에 의한 Biomass 구성당으로부터의 수소생성)

  • Bae, Moo;Yi, Hye-Joo
    • Microbiology and Biotechnology Letters
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    • v.18 no.1
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    • pp.6-11
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    • 1990
  • Hydrogen evolution from biomass-derived carbohydrates by some Clostridia and optimal culture conditions for hydrogen evolution were investigated. Among the organisms tested, Clostridium butyricum was efficient hydrogen producer with starch, xylan, pectin, cellobiose and xylose. In batch fermentation of Cl. butyricum, optimal conditions for hydrogen evolution were achieved at pH 7.0-8.5, 10-50 mM phosphate, and 2% (w/v) glucose. Total amount of molecular hydrogen evolved by the organism slightly increased at the presence of acetate (<150 mM) or butyrate (<20 mM) in the initial fermentation medium. Especially, in case of more than the above concentration of butyrate, growth and hydrogen evolution were dramatically inhibited. In the conditions were described here, 70 mmole of molecular hydrogen per mg of DCW was produced with 1%(w/v) glucose by the organism.

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