7XG6

Crystal structure of an (R)-selective omega-transaminase mutant from Aspergillus terreus with covalently bound PLP

7XG6 の概要

• エントリーDOI: 10.2210/pdb7xg6/pdb
• 分子名称: omega-transaminase (2 entities in total)
• 機能のキーワード: transferase, llp
• 由来する生物種: Aspergillus terreus (strain NIH 2624 / FGSC A1156)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 72040.42

構造登録者

Xiang, C.,Wu, S.K.,Weber, G.,Liu, W.D.,Wei, R.,Bornscheuer, U.T. (登録日: 2022-04-03, 公開日: 2022-12-14, 最終更新日: 2023-11-29)

主引用文献

Wu, S.,Xiang, C.,Zhou, Y.,Khan, M.S.H.,Liu, W.,Feiler, C.G.,Wei, R.,Weber, G.,Hohne, M.,Bornscheuer, U.T.
A growth selection system for the directed evolution of amine-forming or converting enzymes.
Nat Commun, 13:7458-7458, 2022

PubMed Abstract: 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.

PubMed: 36460668
DOI: 10.1038/s41467-022-35228-y

実験手法

X-RAY DIFFRACTION (1.32 Å)