2011 Fiscal Year Final Research Report
Molecular bleeding of the enzymes catalyzing the asymmetric reaction : Construction of a chemical library of unnatural amino acids
| Project/Area Number |
20780076
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Applied biochemistry
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| Research Institution | Kochi University |
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Principal Investigator |
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| Project Period (FY) |
2008 – 2011
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| Keywords | 不斉合成 / タンパク質の耐熱化 / 非タンパク質性アミノ酸 / N-メチル-L-アミノ酸脱水素酵素 / Pseudomonas putida |
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| Research Abstract |
The dpkA gene from Pseudomonas putida was introduced random mutation by error-prone PCR and the thermostabilized mutant enzymes, V117M and Q302R were obtained. The wildtype enzyme, V117M, Q302R and V117M/ Q302R were purified and characterized. The mutant enzymes showed higher thermal and pH stability than wildtype enzyme, but substrate specificities of mutant enzymes did not change. Therefore, the thermostabilized mutant enzymes were useful for synthesis of various N-methyl-L-amino acids.
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