定向进化具有广泛底物范围的对映选择性脂肪酶，用于水解α-取代酯。

Directed evolution of an enantioselective lipase with broad substrate scope for hydrolysis of alpha-substituted esters.

机构信息

Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, SE-106 91 Stockholm, Sweden.

出版信息

J Am Chem Soc. 2010 May 26;132(20):7038-42. doi: 10.1021/ja100593j.

PMID:20450151

Abstract

A variant of Candida antarctica lipase A (CalA) was developed for the hydrolysis of alpha-substituted p-nitrophenyl esters by directed evolution. The E values of this variant for 7 different esters was 45-276, which is a large improvement compared to 2-20 for the wild type. The broad substrate scope of this enzyme variant is of synthetic use, and hydrolysis of the tested substrates proceeded with an enantiomeric excess between 95-99%. A 30-fold increase in activity was also observed for most substrates. The developed enzyme variant shows (R)-selectivity, which is reversed compared to the wild type that is (S)-selective for most substrates.

摘要

通过定向进化，开发了一种新型南极假丝酵母脂肪酶 A（CalA）变体，用于水解α-取代的对硝基苯酯。与野生型相比，该变体对 7 种不同酯的 E 值提高了 45-276 倍。该酶变体具有广泛的底物范围，可用于合成，并且测试的底物的水解产物的对映体过量值在 95-99%之间。大多数底物的活性也提高了 30 倍。与野生型相比，开发的酶变体具有（R）选择性，而野生型对大多数底物具有（S）选择性。