文献类型：期刊文献

英文题名：Direct reductive amination of ketones with amines by reductive aminases

作者：Li, Bo-Bo[1];Zhang, Jing[1];Chen, Fei-Fei[1];Chen, Qi[1];Xu, Jian-He;Zheng, Gao-Wei[1,2]

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

年份：2021

卷号：2

期号：4

起止页码：345

外文期刊名：GREEN SYNTHESIS AND CATALYSIS

收录：WOS:【ESCI(收录号:WOS:001022690000001)】；

基金：The work was fi nancially supported by the National Key Research and Development Program of China (No. 2019YFA09005000) , the National Natural Science Foundation of China (Nos. 21878085, 31971380, 21472045, 21871085) , and the Fundamental Research Funds for the Central Universities (No. 22221818014) .

语种：英文

外文关键词：Biocatalysis; Reductive amination; Reductive aminases; Protein engineering; Chiral amines

摘要：Direct reductive amination of ketones by reductive aminases (RedAms) is a promising method for the synthesis of primary, secondary and tertiary amines. In this work, five naturally occurring RedAms possessing reductive amination activity were identified using a structure-guided genome mining (SGGM) approach, which was based on highly conserved substrate binding and catalysis motif alignment. This RedAm toolbox facilitated the reductive amination of cyclohexanone and 4-fluoropropiophenone with various amine nucleophiles. In a preparative biotransformation (100 mg) with low stoichiometric ammonia donor and low catalyst loading, and three types of N-alkylcyclohexylamine were efficiently synthesized with a space-time yield up to 64.2 g L-1 d-1, demonstrating the potential of the RedAm enzymes as a biocatalyst toolbox for the synthesis of primary and secondary amines. Additionally, through protein engineering, the W205F variant of KfRedAm from Kribbella flavida was obtained with a 2-fold increased catalytic efficiency (kcat/Km), and the general applicability of the variant was verified, providing useful guidance for further evolution of RedAms within this family.