Directed Evolution of an Enantioselective Lipase with Broad Substrate Scope for Hydrolysis of α-Substituted Esters
Directed Evolution of an Enantioselective Lipase with Broad Substrate Scope for Hydrolysis of α-Substituted Esters
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DOI：
10.1021/ja100593j
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发表时间：
2010-05-26
影响因子：
15
通讯作者：
Backvall, Jan-E.
中科院分区：
文献类型：
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作者：
Engstrom, Karin;Nyhlen, Jonas;Backvall, Jan-E.
利用定向进化方法，开发了一种南极念珠菌脂肪酶A (CalA)，用于水解A -取代对硝基苯酯。该变异对7种不同酯的E值为45-276，与野生型的2-20相比有很大的提高。该酶变体的底物范围广泛，可用于合成，并且在所测底物的水解过程中对映体过量在95-99%之间。大多数底物的活性也增加了30倍。开发的酶变体显示(R)选择性，与对大多数底物具有(S)选择性的野生型相反。
A variant of Candida antarctica lipase A (CalA) was developed for the hydrolysis of a-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.