| Project/Area Number |
17580069
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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 |
Applied microbiology
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| Research Institution | Faculty of Engineering, Toyama Prefectural University |
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Principal Investigator |
KATO Yasuo Toyama Prefectural University, Faculty of Engineering, Associate Professor, 工学部, 助教授 (20254237)
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| Co-Investigator(Kenkyū-buntansha) |
ASANO Yasuhisa Toyama Prefectural University, Faculty of Engineering, Professor, 工学部, 教授 (00222589)
KOMEDA Hidenobu Toyama Prefectural University, Faculty of Engineering, Assistant Professor, 工学部, 講師 (50285160)
AONO Shigetoshi Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences, Professor, 岡崎統合バイオサイエンスセンター, 教授 (60183729)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2006: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2005: ¥2,800,000 (Direct Cost: ¥2,800,000)
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| Keywords | aldoxime / nitrile / aldoxime-nitrile pathway / enzyme / gene / screening / microorganisms / reaction mechanisms |
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| Research Abstract |
We examined the following experiments in order to clarify 1) reaction mechanism of aldoxime dehydratase (Oxd), 2) diversity of aldoxime dehydratase, and 3) diversity and physiological function of "aldoxime-nitrile pathway".
1. We constructed an overexpression system of nitrilase (Nit)-linked oxd gene from Bacillus sp. OxB-1 in a recombinant E. coli and obtained large quantities of Oxd protein (OxdB) in highly pure and homogenous form.
2. We analyzed OxdB protein spectrophotometrically to identify its heme environment and functional residue during enzyme catalysis.
3. We screened oxd genes genetically from stock cultures and cloned whole lengths oxd genes.
4. We overexpressed new oxds in E.coli and examined their characters. Based on the findings, physiological functions of Oxds are studied.
5. We genetically screened nitrile-degrading enzymes in strains having "aldoxime-nitrile pathway" and cloned their genes.
6. We compared the characters and primary structures of the nitrile-degrading enzymes and studied the relationship between Oxds enzymologically and genetically. Physiology and evolutions of the pathway are also discussed.
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