SAMDI マススペクトロメトリーによる高通量スクリーニング
Adam J Pluchinsky, Daniel J Wackelin1, Xiongyi Huang1
1Division of Chemistry and Chemical Engineering MC 210-41, California Institute of Technology, Pasadena, California 91125, United States.
Journal of the American Chemical Society
|November 11, 2020
まとめ
研究者は,質量スペクトロメトリーを用いた酵素変異の高通量スクリーニング方法を開発しました. このアプローチにより,何千もの酵素触媒の迅速な評価が可能になり,化学的変換が改善され,酵素の進化が加速されます.
科学分野:
- 生物触媒
- 酵素工学
- 分析化学
背景:
- 誘導進化は酵素発見を進めるが,しばしば低通量スクリーニングを使用する.
- クロマトグラフィーベースの方法は,酵素変異の評価の規模と速度を制限します.
研究 の 目的:
- 酵素変異のスクリーニングのための高通量戦略を提示する.
- 何千もの酵素変種を直接評価して 活性化を図る
主な方法:
- 浄化されていない反応産物を自己組み立ての単層に固定する.
- 固定された製品の質量分析
- ランダムに変異したシトクロームP411の変異群への適用
主要な成果:
- 酵素変種をスクリーニングするための高通量戦略を示した.
- サイトクロームP411の変種を用いたC-Hアルキル化のための改善された触媒を成功裏に特定した.
- 数千の変異を 1時間以内に直接評価した
結論:
- 開発された方法は,酵素スクリーニングのスループットを大幅に増加させます.
- このテクニックは,様々な化学変換のための酵素活性を進化させるのに適応することができます.
- 新しく改良された生物触媒の発見を加速する
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