Highly Regioselective and Efficient Biosynthesis of Polydatin by an Engineered UGTBL1-AtSuSy Cascade Reaction.
J Agric Food Chem
; 69(31): 8695-8702, 2021 Aug 11.
Article
em En
| MEDLINE
| ID: mdl-34319737
ABSTRACT
Polydatin, resveratrol-3-O-ß-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 UGTBL1 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 â¼ 99%), and the ratio of polydatin to resveratrol-4'-O-ß-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, UGTBL1 mutant coupling sucrose synthase AtSuSy can synthesize polydatin at an unprecedented high titer of 10.33 g/L, together with efficient UDPG regeneration (RCmax = 54). This study provides an efficient approach for the regioselective biosynthesis of polydatin.
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Base de dados:
MEDLINE
Assunto principal:
Estilbenos
/
Glucosídeos
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article