2023 Fiscal Year Final Research Report
Elucidating the inversion reaction mechanism of glycoside hydrolase by structure dynamics
| Project/Area Number |
21K06077
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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 43030:Functional biochemistry-related
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| Research Institution | Tohoku University |
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Principal Investigator |
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| Project Period (FY) |
2021-04-01 – 2024-03-31
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| Keywords | 糖加水分解酵素 / 反応中間体 / X線自由電子レーザー / シリアルフェムト秒結晶構造解析 / クライオトラップ法 / セルロース |
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| Outline of Final Research Achievements |
To elucidate the molecular mechanism of the enzymatic reaction of endoglucanase CeSZ, which is involved in the production of bacterial cellulose, we performed structure analysis of the reaction intermediate state. Under cryogenic conditions, the mutant enzyme in complex with the intermediate captured the state after hydrolysis, while the wild-type enzyme complexed with the other intermediate captured the structure before the transition state. This study unveiled the remaining two reaction intermediate states in the proposed reaction mechanism of CeSZ, which contributes to further understanding of the reaction mechanism. In addition, the structural analysis of the bifunctional alginate epimerase AlgE, which was not planned at the beginning of this research, was performed, and obtained insights into the recognition of substrates with different chain lengths.
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| Free Research Field |
構造生物学
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| Academic Significance and Societal Importance of the Research Achievements |
クライオトラップ法により，実験的にコンフォメーションが異なる糖の状態を可視化した本研究成果は，糖加水分解酵素の反応機構を詳細に解明するための有用な知見となる．そして，本研究で用いた手法は，異なるコンフォメーションの糖を経由する他の糖加水分解酵素に対して，中間状態を捉えるための一般的な実験指針となり得る．また，本研究で対象としたCeSZやAlgEはいずれも，オリゴ糖や多糖類の生産に関わる酵素であり，本研究により得られた構造情報をもとに酵素の高機能化が果たされれば，機能性糖質などの効率的生産につながることが期待される．
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