工程改造去甲乌药碱合酶用于一步合成(S)-1-芳基-四氢异喹啉。

Engineering a norcoclaurine synthase for one-step synthesis of (S)-1-aryl-tetrahydroisoquinolines.

作者信息

Zhang Man, Huang Zheng-Yu, Su Ying, Chen Fei-Fei, Chen Qi, Xu Jian-He, Zheng Gao-Wei

机构信息

State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Centre for Biomanufacturing, College of Biotechnology, East China University of Science and Technology, Shanghai, 200237, People's Republic of China.

出版信息

Bioresour Bioprocess. 2023 Mar 1;10(1):15. doi: 10.1186/s40643-023-00637-4.

DOI:10.1186/s40643-023-00637-4

PMID:38647611

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10992437/

Abstract

Tetrahydroisoquinoline alkaloids (THIQAs) are ubiquitous compounds with important pharmaceutical and biological activity. Their key N-heterocyclic structural motifs are synthesised via Pictet-Spengler (P-S) reaction by norcoclaurine synthases (NCS) in plants. The synthesis of 1-aryl-tetrahydroisoquinoline alkaloids has attracted increasing attention due to their antitumor and antivirus activities. Herein, the L68T/M97V mutant of NCS from Thalictrum flavum with improved activity was developed by semi-rational design. This mutant not only showed higher catalytic performance (> 96% conversion) toward benzaldehyde and dopamine over the wild-type enzyme, but also catalysed the P-S reaction of the bulky substrate 4-biphenylaldehyde and dopamine with high conversion (> 99%) for the effective synthesis of 1-aryl-THIQA. In terms of stereoselectivity, all products synthesised by the L68T/M97V mutant showed high optical purity (92-99% enantiomeric excess).

摘要

四氢异喹啉生物碱（THIQAs）是普遍存在的化合物，具有重要的药学和生物活性。它们关键的N-杂环结构基序是植物中的去甲乌药碱合酶（NCS）通过Pictet-Spengler（P-S）反应合成的。1-芳基-四氢异喹啉生物碱的合成因其抗肿瘤和抗病毒活性而受到越来越多的关注。在此，通过半理性设计开发了具有更高活性的黄连NCS的L68T/M97V突变体。该突变体不仅对苯甲醛和多巴胺的催化性能比野生型酶更高（转化率>96%），而且还能以高转化率（>99%）催化大体积底物4-联苯甲醛和多巴胺的P-S反应，从而有效合成1-芳基-THIQA。在立体选择性方面，L68T/M97V突变体合成的所有产物均显示出高光学纯度（对映体过量92-99%）。