2018 Fiscal Year Final Research Report
Isolation and characterization of highly stable, highly catalytic and broad substrate specificity D-amino acid oxidase of thermophilic fungi
| Project/Area Number |
16K07660
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Applied microbiology
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| Research Institution | Nagaoka University of Technology |
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Principal Investigator |
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Keywords | D-アミノ酸オキシダーゼ / Rasamsonia emersonii / 好熱性真菌 / 耐熱性 / クローニング / 酵素学的緒特性 |
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| Outline of Final Research Achievements |
D-Amino acid oxidase (DAO), which is a D-amino acid-degrading enzyme, is a useful enzyme for detecting and quantifying D-amino acids involved in neuropsychiatric disorders and producing pharmaceutical raw materials. This study, therefore, aimed to obtain a unique DAO with high stability, high activity, and broad substrate specificity. We isolated a thermophilic fungus that can utilize D-amino acids for growth from compost and identified the fungus as a strain (named strain YA) of Rasamsonia emersonii. We then identified and isolated DAO gene (ReDAO) of the fungus. ReDAO produced in E. coli utilized not only various D-amino acids but also cephalosporin C, a pharmaceutical material, as a substrate. ReDAO also exhibited high thermal stability and activity. These results showed that we successfully obtained the desired DAO that has high stability and high activity with broad substrate specificity.
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| Free Research Field |
応用微生物学
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| Academic Significance and Societal Importance of the Research Achievements |
D-アミノ酸酸化酵素（DAO）は，D-アミノ酸の検出・定量，医薬品合成，病気の診断や治療など広範な産業分野での利用が期待されている．しかし，工業的には安定性の低いDAOが利用されており，高コストであることからDAOの応用的利用は制限されている．本研究の成果により，DAOを利用した物質生産プロセスや分析システムのコストを大幅に低減させることでDAOの多様な分野における応用的利用を可能にし，広範な産業分野の発展が期待される．さらに，今後の解析により，酵素の基質特異性や安定化メカニズムに新たな学術的な知見をもたらし，他の酵素の基質特異性改変技術や安定化技術の開発につながると期待される．
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