| Project/Area Number |
08660123
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
応用微生物学・応用生物化学
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| Research Institution | KUMAMOTO INSTITUTE OF TECHNOLOGY |
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Principal Investigator |
SHIN Takashi KUMAMOTO INSTITUTE OF TECHNOLOGY,DEPARTMENT OF APPLIED MICROBIOLOGY,PROFESSOR, 工学部, 教授 (50179066)
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| Co-Investigator(Kenkyū-buntansha) |
OYAMA Hiroshi KUMAMOTO INSTITUTE OF TECHNOLOGY,DEPARTMENT OF APPLIED MICROBIOLOGY,ASSOCIATED P, 工学部, 助教授 (50221700)
MURAO Sawao KUMAMOTO INSTITUTE OF TECHNOLOGY,DEPARTMENT OF APPLIED MICROBIOLOGY,PROFESSOR, 工学部, 教授 (00081472)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1997: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1996: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | 5-oxoprolinase / Alkaligens faecalis / ピログルタミン酸 |
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| Research Abstract |
A screening test was undertaken to isolate a microorganism that produced 5-oxoprolinase (without ATP-hydrolyzing). The 5-oxoprolinase (without ATP-hydrolyzing) activity (decyclization activity toward L-pyroglutamate) was found in a cell-free extract of Alkaligenes faecalis N-38A,newly isolated from a soil sample. The enzyme was purified as a homogeneous preparation. The molecular weight of the enzyme was estimated to be 47,000. The decyclization activity was specific for L-pyroglutamate, and independent of ATP and metal ions. The reaction was a reversible one, i.e., cyclization reaction of L-glutamate to yield pyroglutamate was identified. Cloning of enzyme gene was undertaken, plasmid B8 bearing 1.5kb enzyme gene was idetified. The base sequence of the plasmid was evaluated, no homologus protein was found. Expression of enzyme gene in E.Coli JM-109 was achieved, expressed activity enzyme protein was purified to crystalline state. But large amout expression was not established due to the formation of inclusion body in host cell.
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