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

被引：3

作者：

Xie, Lingzhi ^{[1
,2
,3
]}

Zhang, Yun ^{[1
,2
,3
]}

Zhang, Ruyue ^{[1
,2
,3
]}

Cui, Haibo ^{[1
,2
,3
]}

Cui, Baodong ^{[1
,2
,3
]}

Han, Wenyong ^{[1
,2
,3
]}

Wan, Nanwei ^{[1
,2
,3
]}

Li, Zhi ^{[4
]}

Chen, Yongzheng ^{[1
,2
,3
]}

机构：

[1] Zunyi Med Univ, Sch Pharm, Green Pharmaceut Engn Res Ctr Guizhou Prov, Key Lab Biocatalysis & Chiral Drug Synth Guizhou P, Zunyi 563000, Peoples R China

[2] Zunyi Med Univ, Key Lab Basic Pharmacol, Minist Educ, Zunyi 563000, Peoples R China

[3] Zunyi Med Univ, Joint Int Res Lab Ethnomed, Minist Educ, Zunyi 563000, Peoples R China

[4] Natl Univ Singapore, Dept Chem & Biomol Engn, 4 Engn Dr 4, Singapore 117585, Singapore

来源：

GREEN SYNTHESIS AND CATALYSIS | 2024年 / 5卷 / 02期

基金：

中国国家自然科学基金;

关键词：

Bio-hydroxylation; Directed evolution; Cytochrome P450; High-throughput screening; Methyl mandelates; DYNAMIC KINETIC RESOLUTION; H BOND FUNCTIONALIZATION; DIRECTED EVOLUTION; ACID-DERIVATIVES; ENANTIOSELECTIVE SYNTHESIS; ASYMMETRIC HYDROGENATION; AEROBIC OXIDATION; CATALYSTS; LIPASE; ESTERS;

D O I：

10.1016/j.gresc.2023.01.005

中图分类号：

O6 [化学];

学科分类号：

0703 ;

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.

引用

页码：88 / 93

页数：6

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