文献类型：期刊文献

英文题名：An Engineered Imine Reductase for Highly Diastereo- and Enantioselective Synthesis of β-Branched Amines with Contiguous Stereocenters

作者：Zhu, Zhen-Yu[1];Shi, Min[1];Li, Chen-Lin[1];Gao, Yun-Fei[1];Shen, Xin-Yuan[1];Ding, Xu-Wei[1];Chen, Fei-Fei[1];Xu, Jian-He[1];Chen, Qi[1];Zheng, Gao-Wei[1]

机构：[1]East China Univ Sci & Technol, Lab Biocatalysis & Synthet Biotechnol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2024

卷号：63

期号：47

外文期刊名：ANGEWANDTE CHEMIE-INTERNATIONAL EDITION

收录：;EI(收录号：20240206394);WOS:【SCI-EXPANDED(收录号:WOS:001338042200001)，CCR-EXPANDED(收录号:WOS:001338042200001)】；

基金：This work was financially supported by the National Natural Science Foundation of China (32371547 and 22008068) and the National Key Research and Development Program of China (2019YFA09005000 and 2021YFA0911400). We thank BL17B1/BL18 U1/BL19U1/beamline of National Facility for Protein Science in Shanghai (NFPS) at Shanghai Synchrotron Radiation Facility and Prof. R.-T. Guo from Hubei University for assistance during X-ray data collection. We also thank Dr. X.-X. Yuan, Prof. B.-B. Zeng, Dr. C. Ning, Dr. H.-G. Li, and Prof. Y.-T. Xie from East China University of Science and Technology for their kind assistance in chemical synthesis.

语种：英文

外文关键词：biocatalysis; dynamic kinetic resolution; imine reductase; tofacitinib; protein engineering

摘要：beta-Branched chiral amines with contiguous stereocenters are valuable building blocks for preparing various biologically active molecules. However, their asymmetric synthesis remains challenging. Herein, we report a highly diastereo- and enantioselective biocatalytic approach for preparing a broad range of beta-branched chiral amines starting from their corresponding racemic ketones. This involves a dynamic kinetic resolution-asymmetric reductive amination process catalyzed using only an imine reductase. Four rounds of protein engineering endowed wild-type PocIRED with higher reactivity, better stereoselectivity, and a broader substrate scope. Using the engineered enzyme, various chiral amine products were synthesized with up to >99.9 % ee, >99 : 1 dr, and >99 % conversion. The practicability of the developed biocatalytic method was confirmed by producing a key intermediate of tofacitinib in 74 % yield, >99.9 % ee, and 98 : 2 dr at a challenging substrate loading of 110 g L-1. Our study provides a highly capable imine reductase and a protocol for developing an efficient biocatalytic dynamic kinetic resolution-asymmetric reductive amination reaction system.