2020 Fiscal Year Final Research Report
Elucidation of the oxygen reduction mechanism of multicopper oxidase by structural analysis of reaction intermediates and its application.
| Project/Area Number |
18K05432
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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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| Review Section |
Basic Section 38030:Applied biochemistry-related
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| Research Institution | Kanazawa University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
山下 哲 金沢大学, 物質化学系, 准教授 (70361186)
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Keywords | マルチ銅オキシターゼ / 構造解析 / 反応機構 / 指向進化 |
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| Outline of Final Research Achievements |
Multicopper oxidases (MCOs) catalyze the oxidation of a wide range of substrates by reducing O2 to H2O without releasing activated oxygen species. In this study, we aimed to elucidate the structure of the 4-electron reduction intermediate of oxygen using the cuprous oxidase CueO from Escherichia coli. We succeeded in stabilizing the intermediate by introducing a triple mutation, and produced a high-quality crystal that can be used for neutron diffraction. In addition, directed evolution has yielded six very active mutants. Some also succeeded in structural analysis, and revealed that destabilization of the tower loop region of domain 3 leads to increased activity. These results provide a new methodology for controlling the activity of MCOs and contribute to their industrial application.
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| Free Research Field |
生化学
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| Academic Significance and Societal Importance of the Research Achievements |
マルチ銅オキシダーゼ（MCO）は様々な基質を電子供与体として利用できるのでその応用範囲が広い。例えばラッカーゼは製紙工業におけるパルプの脱リグニン・漂白工程，繊維産業などの脱色工程や排水処理に利用されている。他にもバイオエタノール生産のためのリファイニング工程や，医薬品の合成，バイオ燃料電池のカソード電極用触媒として利用や，環境中の金属イオンの濃縮・回収に利用できるなどMCOの応用範囲が拡大している。本研究では，MCOの反応機構を解析すると同時に，指向進化によりMCOの高機能化の新たな方法論を開発した。今後のMCOの産業応用に大いに貢献する成果と言える。
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