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非正規の有機触媒機構を持つ酵素の設計と進化
Ashleigh J Burke1, Sarah L Lovelock1, Amina Frese1
1Manchester Institute of Biotechnology, School of Chemistry, University of Manchester, Manchester, UK.
Nature
|May 28, 2019
まとめ
研究者はコンピュータ設計と 実験室での進化を用いて 新しい酵素を設計しました この酵素は,改変されたアミノ酸であるNδ-メチルヒスティジンを利用して,エステル水解を効率的に触媒化し,標準的な遺伝子コードの限界を克服します.
科学分野:
- 生物触媒と酵素工学
- 合成生物学
- コンピュータ化学
背景:
- 遺伝子コードの限界は 新しい触媒機能のための酵素設計を制限します
- 酵素活性部位にあるカノニカルヌクレオフィルは非反応性中間物質を形成し,触媒効率を阻害する.
- 小分子有機触媒は酵素設計のためのメカニズム的戦略を提供します.
研究 の 目的:
- 非正規のアミノ酸を触媒核愛者として用いて水解酵素を開発する.
- Nδ-メチルヒスティジンを用いることで,酵素設計における正規のヌクレオフィルの限界を克服する.
- 遺伝子コードの拡張を通じて 酵素触媒反応のレパートリーを拡大する
主な方法:
- コンピュータによる酵素設計と 実験室での進化
- Nδ-メチルヒスティジンを触媒性ヌクレオフィルとして組み込む.
- 進化のプロトコルは 拡張された遺伝子コードに適応した
- 触媒メカニズムの解明のための結晶分析.
主要な成果:
- Nδ-メチルヒスティディンを利用した水分解酵素の生成.
- 自由なNδ-メチルヒスティジンと比較して,エステル水解の触媒効率の9000倍以上の増加を達成した.
- 誘導進化の過程で結晶学的な研究によって重要な触媒的適応を特定した.
- Nδ-メチルヒスティジンは,ジメチラミノピリジンの遺伝的にコード可能な代用物として実証されています.
結論:
- Nδ-メチルヒスティジンは,アチル酵素の中間生成を防ぐことで効率的な触媒を可能にします.
- このアプローチは 価値ある化学変換のための酵素設計の範囲を拡大します
- 遺伝子でコードされたNδ-メチルヒスティジンは,新しい生物触媒を設計するための多用途のプラットフォームを提供します.
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