| 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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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2017: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2016: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
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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 / 構造機能相関 / 大腸菌発現 / 精製 / 高安定性 / 広基質特異性 / 高活性 / D-アミノ酸資化 / Thermomyces dupontii / DAOホモログ遺伝子 / 大腸菌発現ベクター / 微生物 / 酵素 / 遺伝子 / バイオテクノロジー / 応用微生物 |
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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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| 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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