| Project/Area Number |
05660112
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
応用微生物学・応用生物化学
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| Research Institution | National Institute for Environmental Studies |
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Principal Investigator |
UCHIYAMA Hiroo Nat'l.Inst.Environ.Studies, Water & Soil Environ.Div., Senior researcher, 水土壌圈環境部, 主任研究員 (00185042)
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| Co-Investigator(Kenkyū-buntansha) |
IWASAKI Kazuhiro Nat'l.Inst.Environ.Studies, Regional Environ.Div., Senior researcher, 地域環境研究グループ, 主任研究員 (30193717)
TOMIOKA Noriko Nat'l.Inst.Environ.Studies, Water & Soil Environ.Div., Senior researcher, 水土壌圈環境部, 主任研究員 (40168399)
YAGI Osami Nat'l.Inst.Environ.Studies, Water & Soil Environ.Div., Chief, 水土壌圈環境部, 室長 (40132865)
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| Project Period (FY) |
1993 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1994: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1993: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | methane monooxygenase / methanotroph / trichloroethylene |
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| Research Abstract |
1. Soluble methane monooxygenase (sMMO) gene was cloned from a methane-utilizing bacterium, Methylocystis sp.M (strain M), and sequenced.
(1) sMMO structural gene (about 6kb) and its promotor region were sequenced, and those were confirmed to be sMMO genes by comparing them with complemental DNA sequences derived from N-terminal amino acid sequences of purified sMMO in strain M.
(2) Some characteristics were cleared as follows ;
・ sMMO from strain M was related closely with those of methylosinus trichosporium OB3b and Methylococcus capsulatus Bath in DNA sequences and putative amino acid sequences.
・ Metal-binding site in a active center of sMMO,high G/C content in the third letter of the used codons, and prothsetic groups binding site were confirmed in that DNA sequence.
2. Plasmid pWM3 coding sMMO genes was introduced into E.coli. Trichloroethylene degradation activity was not detected in E.coli harboring pWM3, and no difference in lysates was observed between E.coli harboring pWM3 and host by SDS-PAGE.Hydroxylase, one of the components of sMMO,consisted with alpha 2 beta 2 gamma 2 subunits and requires ferric ion in the active center, indicating that the precise assembly of those subunits and a metal would be required to express hydroxylase activity.
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