Bioresources and Bioprocessing ( IF 4.3 ) Pub Date : 2023-03-01 , DOI: 10.1186/s40643-023-00637-4 Man Zhang 1 , Zheng-Yu Huang 1 , Ying Su 1 , Fei-Fei Chen 1 , Qi Chen 1 , Jian-He Xu 1 , Gao-Wei Zheng 1
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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).
Graphical Abstract
中文翻译：
设计一种去甲乌云碱合成酶，用于一步合成 (S)-1-芳基-四氢异喹啉
四氢异喹啉生物碱 (THIQAs) 是一种普遍存在的化合物，具有重要的药物和生物活性。它们的关键N -杂环结构基序是由植物中的去甲乌云碱合酶 (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改进活性的来自黄杨的NCS L68T/M97V突变体。该突变体不仅对苯甲醛和多巴胺表现出比野生型酶更高的催化性能（> 96％转化率），而且还以高转化率（> 99％）催化大体积底物4-联苯醛和多巴胺的P-S反应用于有效合成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表现出高光学纯度（92-99%对映体过量）。
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