| Project/Area Number |
10556021
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
応用微生物学・応用生物化学
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| Research Institution | Tottori University |
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Principal Investigator |
IZUMI Yioshikazu Tottori University, Faculty of Engineearing, Professor, 工学部, 教授 (40026555)
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| Co-Investigator(Kenkyū-buntansha) |
YOSHIDA Toyokazu Gifu University, Faculty of Engineearing, Associate Professor, 工学部, 助教授 (90220657)
OHSHIRO Takashi Tottori University, Faculty of Engineearing, Research Associate, 工学部, 助手 (00233106)
下西 学 国立循環器病センター研究所, 薬理部, 研究員 (70300978)
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| Project Period (FY) |
1998 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥13,900,000 (Direct Cost: ¥13,900,000)
Fiscal Year 2000: ¥4,200,000 (Direct Cost: ¥4,200,000)
Fiscal Year 1999: ¥4,300,000 (Direct Cost: ¥4,300,000)
Fiscal Year 1998: ¥5,400,000 (Direct Cost: ¥5,400,000)
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| Keywords | C1 microorganism / serine pathway / isocitrate lyase / serine-glyoxylate aminotransferase / hydroxypyruvate reductase / 速度論的酵素の特性分析 / serin-glyoxylate aminotransfdeseDszA / 有用アミノ酸生産 / ヒドロキシピルビン酸レグクターゼ / 結晶化 / X線構造解析 / HPR遺伝子 / Hyphomicrobium methylovorum / メチロトローフ / メタノール資化性細菌 / L-セリン / メタノール脱水素酵素 / mxaF遺伝子 |
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| Research Abstract |
The research results of this study are summarized as follows.
(1) Using methanol as a carbon source and energy source, which is promissing and desirable for the microbial industries, we developd a resource-saving process for the production of useful amino acids by the C1 microorganisms whichh we have already isolated.
(2) Crystallization of novel enzymes specific for C1 microorganisms and X-ray crystallographic analyses : As for hydroxypyruvate reductase (HPR), a holo-enzyme which had the coenzyme NAD was crystallized (bipyramidal form) according to the crystallization procedures of the apo-enzyme which we have already established. As a result of X-ray crystallography of the holo-enzyme crystals. NAD was found to be bound to the site of the enzyme as we expected. Moreover, on the contrary to our expectation that there must be a change in angle of the cleft between the catalytic site and substrate binding site of the apo-enzyme, the structure of the holo-enzyme was unchaged as compared to that of the apo-enzyme. We succeeded in obtaining crystallization of the ternary complex of the enzyme (substrate-alalog- NAD and enzyme), so X-ray crystallographic analyses are now underway. As for the SGAT, we have obtained crystals of rhombus plate form by using the hanging drop method with Crystal Screen I.
(3) Kinetic studies of pa novel enzyme, SGAT, specific for C1 microorganisms : As for SGAT, kinetic studies were carried out by the stopped-flow method and by use of the isotope-labelled substrates, and revealed that the enzyme catalyzed the reaction through a different reaction mechanism form other known aminotransfereases.
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