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Publication Abstract from PubMed

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.

A growth selection system for the directed evolution of amine-forming or converting enzymes.,Wu S, Xiang C, Zhou Y, Khan MSH, Liu W, Feiler CG, Wei R, Weber G, Hohne M, Bornscheuer UT Nat Commun. 2022 Dec 3;13(1):7458. doi: 10.1038/s41467-022-35228-y. PMID:36460668^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.