文献类型：期刊文献

英文题名：Asymmetric Reductive Amination of Structurally Diverse Ketones with Ammonia Using a Spectrum-Extended Amine Dehydrogenase

作者：Wang, Dong-Hao[1];Chen, Qi[1];Yin, Sai-Nan[1];Ding, Xu-Wei[1];Zheng, Yu-Cong[1];Zhang, Zhi[1];Zhang, Yu-Hui[1];Chen, Fei-Fei[1];Xu, Jian-He[1,2];Zheng, Gao-Wei[1,2]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai Collaborat Innovat Ctr Biomfg, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Frontiers Sci Ctr Mat & Dynam Chem, Shanghai 200237, Peoples R China

年份：2021

卷号：11

期号：22

起止页码：14274

外文期刊名：ACS CATALYSIS

收录：;EI(收录号：20214811219536);WOS:【SCI-EXPANDED(收录号:WOS:000758014300007)】；

基金：The work was financially supported by the National Natural Science Foundation of China (21878085, 31971380, 21472045 and 21871085), the National Key Research and Development Program of China (2019YFA09005000), and the Fundamental Research Funds for the Central Universities (22221818014).

语种：英文

外文关键词：< em > reductive amination <; em >; < em > amine dehydrogenase <; em >; < em > protein engineering <; em >; < em > substrate scope <; em >; < em > chiral amine <; em >

摘要：Amine dehydrogenase-catalyzed reductive amination of prochiral ketones with ammonia is a promising method for the synthesis of optically pure amines in the pharmaceutical and fine chemical industries. However, previously reported amine dehydrogenases show restricted catalytic capacity toward bulky ketones, which limits their widespread applications toward the production of chiral amines. Herein, we expanded the substrate scope of an engineered amine dehydrogenase GkAmDH from Geobacillus kaustophilus via laboratory evolution for the reductive amination of an extensive set of ketones. Several beneficial mutants were identified with a up to 2.2 U mg-1 activity toward bulky benzylacetone, 110-fold higher than that of M0. Using the engineered M3 and M8, structurally diverse bulky chiral amines could be synthesized with up to >99% conversion, >99% ee, and up to 18,900 TON. Among them, two key chiral intermediates used in the synthesis of the drugs medroxalol and dilevalol were produced on a gram scale in up to 85% yield and >99% ee. Additionally, the engineered enzymes M3 and M8 displayed considerable thermostability with a half-life of more than three days at 50 degrees C. These results demonstrate that these engineered amine dehydrogenases are promising biocatalysts for the synthesis of chiral amines. Molecular dynamics simulations provide insights into how mutations improve the amination activity toward bulky ketones and the thermostability. Superscript/Subscript Available