Biocatalytic stereoselective synthesis of methyl mandelates by engineering a cytochrome P450 hydroxylase  

作　　者：Lingzhi Xie Yun Zhang Ruyue Zhang Haibo Cui Baodong Cui Wenyong Han Nanwei Wan Zhi Li Yongzheng Chen 

机构地区：[1]Key Laboratory of Biocatalysis&Chiral Drug Synthesis of Guizhou Province,Generic Drug Research Center of Guizhou Province,Green Pharmaceuticals Engineering Research Center of Guizhou Province,School of Pharmacy,Zunyi Medical University,Zunyi 563000,China [2]Key Laboratory of Basic Pharmacology of Ministry of Education and Joint International Research Laboratory of Ethnomedicine of Ministry of Education,Zunyi Medical University,Zunyi 563000,China [3]Department of Chemical and Biomolecular Engineering National University of Singapore,4 Engineering Drive 4,Singapore 117585,Singapore

出　　处：《Green Synthesis and Catalysis》2024年第2期88-93,共6页绿色合成与催化（英文）

基　　金：supported by the National Natural Science Foundation of China(Nos.21961048 and 32271537);Science and Technology Department of Zunyi(Nos.ZSKRPT-2020-5,ZSKH-2018-3 and ZSKRPT-2021-5).

摘　　要：Chiral methyl mandelates are useful synthons in organic transformation and pharmaceutical synthesis.Green synthesis of these valuable compounds by direct C–H activating oxidative hydroxylation has attracted keen interest.Described herein is achieving the stereoselective and efficient bio-hydroxylation of methyl 2-phenylacetates to the chiral methyl mandelates by directed evolution of the cytochrome P450DA hydroxylase.In the present study,a new colorimetric high-throughput screening assay was successfully developed based on a dualenzyme cascade for the engineering of the P450DA's hydroxylation activity.Several beneficial variants with enhanced bio-hydroxylation activity were created by combining random mutagenesis and site-saturated/directed mutagenesis strategies.Whole-cell bio-hydroxylation of various methyl 2-phenylacetates using the best septupletmutant P450DA-11 yielded the corresponding chiral methyl mandelates in up to 92%isolated yields and>99%ee.

关 键 词：Bio-hydroxylation Directed evolution Cytochrome P450 High-throughput screening Methyl mandelates 

分 类 号：O62[理学—有机化学]