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A growth selection system for the directed evolution of amine-forming or converting enzymes
- 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.
Author: | Shuke Wu, Chao Xiang, Yi Zhou, Mohammad Saiful Hasan Khan, Weidong Liu, Christian G. Feiler, Ren Wei, Gert Weber, Matthias Höhne, Uwe T. BornscheuerORCiD |
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URN: | urn:nbn:de:gbv:9-opus-105230 |
DOI: | https://doi.org/10.1038/s41467-022-35228-y |
ISSN: | 2041-1723 |
Parent Title (English): | Nature Communications |
Publisher: | Nature Publishing Group |
Place of publication: | London |
Document Type: | Article |
Language: | English |
Date of first Publication: | 2022/12/03 |
Release Date: | 2024/01/22 |
Volume: | 13 |
Article Number: | 7458 |
Page Number: | 10 |
Faculties: | Mathematisch-Naturwissenschaftliche Fakultät / Institut für Biochemie |
Collections: | Artikel aus DFG-gefördertem Publikationsfonds |
Licence (German): | Creative Commons - Namensnennung |