Highly Regioselective and Efficient Biosynthesis of Polydatin by an Engineered UGT BL 1- At SuSy Cascade Reaction

作者：Tianyi Chen, Ziyi Chen, Nan Wang, Jianlin Chu, Bo Fan, Cheng Cheng, Song Qin, Bingfang He · 发表于：Journal of Agricultural and Food Chemistry · 年份：2021 · DOI：10.1021/acs.jafc.1c02518 · 被引用次数：26 · 研究领域：Sirtuins and Resveratrol in Medicine、CRISPR and Genetic Engineering、Plant Gene Expression Analysis

Polydatin, resveratrol-3- O- β-glucoside, possesses various biological activities. However, the regioselective glucosylation of resveratrol by UDP-glycosyltransferases (UGTs) constitutes a persistent problem. In this study, semi-rational design and iterative combinatorial mutagenesis were carried out to screen the mutants of UGT BL 1 and the high specificity with the glycosylation of the 3-OH group of resveratrol was explored. The triple mutant I62G/M112D/K143G exhibited near-perfect control of polydatin synthesis (regioselectivity ∼ 99%), and the ratio of polydatin to resveratrol-4′- O- β-glucoside was finally enhanced by 786-fold. Molecular docking revealed that the mutant could form three H-bonds between 3-, 5-, and 4′-OH groups of resveratrol and the residues around the active center, resulting in the oriented-binding of resveratrol. Furthermore, UGT BL 1 mutant coupling sucrose synthase At SuSy can synthesize polydatin at an unprecedented high titer of 10.33 g/L, together with efficient UDPG regeneration (RC max = 54). This study provides an efficient approach for the regioselective biosynthesis of polydatin.