定向进化
胺气处理
蛋白质工程
酶
生物催化
化学
组合化学
转氨酶
生物化学
有机化学
催化作用
基因
反应机理
突变体
作者
Shuke Wu,Chao Xiang,Yi Zhou,Mohammad Saiful Hasan Khan,Weidong Liu,C. Feiler,Ren Wei,Gert Weber,Matthias Höhne,Uwe T. Bornscheuer
标识
DOI:10.1038/s41467-022-35228-y
摘要
Fast screening of enzyme variants is crucial for tailoring biocatalysts for the asymmetric synthesis of non-natural chiral chemicals, such as amines. However, most existing screening methods either are limited by the throughput or require specialized equipment. Herein, we report a simple, high-throughput, low-equipment dependent, and generally applicable growth selection system for engineering amine-forming or converting enzymes and apply it to improve biocatalysts belonging to three different enzyme classes. This results in (i) an amine transaminase variant with 110-fold increased specific activity for the asymmetric synthesis of the chiral amine intermediate of Linagliptin; (ii) a 270-fold improved monoamine oxidase to prepare the chiral amine intermediate of Cinacalcet by deracemization; and (iii) an ammonia lyase variant with a 26-fold increased activity in the asymmetric synthesis of a non-natural amino acid. Our growth selection system is adaptable to different enzyme classes, varying levels of enzyme activities, and thus a flexible tool for various stages of an engineering campaign.
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