亚胺还原酶的改造及其催化合成1-苯基-1,2,3,4-四氢异喹啉的性能  

Modification and Characterization of Imine Reductases for Synthesis of 1-Phenyl-1,2,3,4-Tetrahydro-Isoquinoline

作　　者：曹文斌 李昊 陈飞飞 潘江[1] 郑高伟 许建和[1] CAO Wenbin;LI Hao;CHEN Feifei;PAN Jiang;ZHENG Gaowei;XU Jianhe(Shanghai Collaborative Innovation Center for Biomanufacturing,School of Biotechnology,East China University of Science and Technology,Shanghai 200237,China)

机构地区：[1]华东理工大学生物工程学院,省部共建上海生物制造产业协同创新中心,上海200237

出　　处：《华东理工大学学报（自然科学版）》2022年第4期511-518,共8页Journal of East China University of Science and Technology

基　　金：国家自然科学基金(21878085,21871085)。

摘　　要：对亚胺还原酶酶库的筛选获得了性能优良的候选酶AdIR1,基于蛋白结构分析对AdIR1进行分子改造,获得了催化效率更高的突变体酶F172Y,并对其进行动力学与热稳定性表征;最后利用所开发的突变酶实现了(S)-1-苯基-1,2,3,4-四氢异喹啉((S)-1-Ph-THIQ)克级制备,证明了该酶法合成路线的实用性。1,2,3,4-Tetrahydro-isoquinolines are a significant class of building blocks used in the pharmaceutical and agrochemical industries,and exist widely in a variety of chiral amine drugs.Among them,(S)-1-Phenyl-1,2,3,4-tetrahydro-isoquinoline((S)-1-Ph-THIQ)is the key precursor for the synthesis of solifenacin,a drug for the treatment of overactive bladder.Imine reductase(IRED)-catalyzed asymmetric reduction of 1-phenyl-3,4-dihydroisoquinoline(1-Ph-DHIQ)is a green and promising route towards chiral 1-Ph-THIQ.However,currently there is only a limited number of reported IREDs that can catalyze the synthesis of chiral 1-Ph-THIQ from 1-Ph-DHIQ,and they may suffer from issues including low activity,poor stereoselectivity,and substrate inhibition.In this study,we first discovered an IRED AdIR1 with considerable properties by screening a panel of IREDs and identified key residues which might affect the activity via homo-modelling and structure comparison.Protein engineering was performed to generate mutant F172Y with elevated catalytic efficiency,which was then characterized in terms of kinetic parameters and thermostability.Finally,preparative synthesis of(S)-1-Ph-THIQ on gram-scale was achieved employing mutant F172Y,demonstrating the considerable applicability of this biocatalytic route in the synthesis of(S)-1-Ph-THIQ.

关 键 词：四氢异喹啉 手性胺 亚胺还原酶 不对称合成 生物催化 

分 类 号：Q81[生物学—生物工程]