in vivo メタテシスのための人工メタロ酵素の指向的進化
Markus Jeschek1, Raphael Reuter2, Tillmann Heinisch2
1Department of Biosystems Science and Engineering, ETH Zurich, Basel CH-4058, Switzerland.
Nature
|August 30, 2016
まとめ
研究者は,E. coliの周回体内でオレフィン転移のための人工メタロ酵素を設計し,非自然な反応のための生物触媒の進化と拡張を可能にしました.
科学分野:
- 生物触媒と合成生物学
- タンパク質工学と導かれた進化
- 有機金属化学
背景:
- 生物触媒は伝統的に天然の酵素を用いるが,指向進化と人工の金属酵素は新しい反応の能力を拡張している.
- 人工メタロ酵素は 移行金属と酵素の構造を組み合わせて 汎用的で工学的な触媒を 作り出します
- 人工メタロ酵素の既存の方法は,しばしばタンパク質の浄化を必要とするため,in vivoの応用が制限されています.
研究 の 目的:
- アビオティック反応のための人工メタロ酵素の in vivo 進化のための方法を開発する.
- オレフィン転移のための人工メタロ酵素を設計する 自然界にはない反応だ
- 人工メタロ酵素の開発と適用のためのタンパク質浄化の限界を克服する.
主な方法:
- エシェリキア・コライの周回体内の人工メタロ酵素の分割.
- 人工メタロ酵素の指向的な進化により,活性と安定性が強化される.
- 細胞成分であるグルタチオンの阻害を緩和するために 周回血質を反応区として利用する.
主要な成果:
- オレフィンメタテシスのための機能的な人工メタ酵素 ("メタテース") の in vivo アセンブリと進化の成功.
- 精製されたタンパク質を用いた従来の方法と比較して,指向的な進化のために著しく高いスループットを達成した.
- 進化したメタタゼは,商業的触媒と好ましい比較と,様々な基板との活性を示した.
結論:
- 周辺プラズマ区分化戦略は,無生物反応のための人工金属酵素の効率的なin vivo進化を可能にします.
- このアプローチは,非自然な代謝と生物触媒の応用のための人工金属酵素の可能性を拡大します.
- 開発されたワークフローは,多様な触媒機能のための人工金属酵素のさらなる設計のための堅固なプラットフォームを提供します.
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