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

• Published in: Bioresources and bioprocessing Vol. 10; no. 1; pp. 15 - 10
• Main Authors: Zhang, Man, Huang, Zheng-Yu, Su, Ying, Chen, Fei-Fei, Chen, Qi, Xu, Jian-He, Zheng, Gao-Wei
• Format: Journal Article
• Language: English
• Published: Singapore Springer Nature Singapore 01.03.2023; Springer Nature B.V; SpringerOpen
• Subjects: Alkaloids; Anticancer properties; Aromatic compounds; Benzaldehyde; Biocatalysis; Biochemical Engineering; Biological activity; Catalytic converters; Chemistry; Chemistry and Materials Science; Dopamine; Environmental Engineering/Biotechnology; Enzymes; Industrial and Production Engineering; Mutagenesis; Mutants; Norcoclaurine synthase; Pictet–Spengler reaction; Protein engineering; Protein expression; Proteins; Stereoselectivity; Substrates; Synthesis; Tetrahydroisoquinoline; Tetrahydroisoquinoline alkaloids; Protein engineering; Norcoclaurine synthase; Biocatalysis; Pictet–Spengler reaction; Tetrahydroisoquinoline alkaloids
• ISSN: 2197-4365; 2197-4365
• DOI: 10.1186/s40643-023-00637-4

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