高通量筛选策略在细胞色素P450单加氧酶定向进化中的应用

细胞色素P450单加氧酶具有催化活性混杂性的特点,可以催化多种氧化反应,因而在生物催化领域受到了极大的关注。然而P450单加氧酶往往存在催化活性低、稳定性差、区域和立体选择性不理想等问题,从而限制了其在生物催化领域的广泛运用。蛋白质定向进化的发展与运用为改善P450单加氧酶的催化性能提供了有效的途径,而一种高效的高通量筛选策略是保证酶蛋白定向进化成功实施的关键。本文综述了P450单加氧酶定向进化过程中高通量筛选策略的最新进展。

Application of High-throughput Screening Strategy Indirected Evolution of Cytochrome P450 Monooxygenase

Cytochrome P450 monooxygenases have promiscuous catalytic activity and catalyze many oxidation reactions,which have received great attention in biocatalysis.However,P450 monooxygenases usually suffer from low catalytic activity,poor stability,unreasonable regio-and stereoselectivity and thus,its wide application in biocatalysis is limited.The development and application of protein directed evolution provides a powerful tool to improve the P450 s’catalytic performances,while an efficient high-throughput screening assay is very important for the successful directed evolution of enzymes.This review focus on the recent advances of high-throughput screening strategies in the directed evolution of P450 monooxygenases.