文献类型：期刊文献

英文题名：Reshaping the Active Pocket of Amine Dehydrogenases for Asymmetric Synthesis of Bulky Aliphatic Amines

作者：Chen, Fei-Fei[1];Zheng, Gao-Wei[1];Liu, Lei[1];Li, Hao[1];Chen, Qi[1];Li, Fu-Long[1];Li, Chun-Xiu[1];Xu, Jian-He[1]

机构：[1]East China Univ Sci & Technol, Shanghai Collaborat Innovat Ctr Biomfg, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2018

卷号：8

期号：3

起止页码：2622

外文期刊名：ACS CATALYSIS

收录：;EI(收录号：20181004882733);WOS:【SCI-EXPANDED(收录号:WOS:000426804100113)】；

基金：This work was financially supported by the National Natural Science Foundation of China (Nos. 21472045 and 21536004).

语种：英文

外文关键词：amine dehydrogenase; reductive amination; chiral amine; protein engineering; biocatalysis; substrate scope

摘要：The asymmetric reductive amination of ketones with ammonia using engineered amine dehydrogenases (AmDHs) is a particularly attractive and environmentally friendly method for the synthesis of chiral amines. However, one major challenge for these engineered AmDHs is their limited range of accepted substrates. Herein, several engineered AmDHs were developed through the evolution of naturally occurring leucine dehydrogenases, which displayed good amination activity toward aliphatic ketones but restricted catalytic scope for short-chain substrates. Computational analysis helped identify two residues, located at the distal end of the substrate-binding cavity, that generate steric hindrance and prevent the binding of bulky aliphatic ketones. By fine-tuning these two key hotspots, the resulting AmDH mutants are able to accept previously inaccessible bulky substrates. More importantly, the mutations were also proved applicable for expanding the substrate scope of other homologous AmDHs with sequence identities as low as 70%, indicating a broad effect on the development of AmDHs and the synthesis of structurally diverse chiral amines.