Biocatalytic stereoselective synthesis of methyl mandelates by engineering a cytochrome P450 hydroxylase
作者机构：Key Laboratory of Biocatalysis&Chiral Drug Synthesis of Guizhou ProvinceGeneric Drug Research Center of Guizhou ProvinceGreen Pharmaceuticals Engineering Research Center of Guizhou ProvinceSchool of PharmacyZunyi Medical UniversityZunyi 563000China Key Laboratory of Basic Pharmacology of Ministry of Education and Joint International Research Laboratory of Ethnomedicine of Ministry of EducationZunyi Medical UniversityZunyi 563000China Department of Chemical and Biomolecular Engineering National University of Singapore4 Engineering Drive 4Singapore 117585Singapore
出 版 物：《Green Synthesis and Catalysis》 (绿色合成与催化（英文）)
年 卷 期：2024年第5卷第2期
页 面：88-93页
学科分类：081704[工学-应用化学] 07[理学] 08[工学] 0817[工学-化学工程与技术] 070303[理学-有机化学] 0703[理学-化学]
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)
主 题：Bio-hydroxylation Directed evolution Cytochrome P450 High-throughput screening Methyl mandelates
摘 要：Chiral methyl mandelates are useful synthons in organic transformation and pharmaceutical *** synthesis of these valuable compounds by direct C–H activating oxidative hydroxylation has attracted keen *** 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 *** 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 *** beneficial variants with enhanced bio-hydroxylation activity were created by combining random mutagenesis and site-saturated/directed mutagenesis ***-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 and99%ee.
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