Comprehensive engineering of novel glycosyltransferase for efficient, donor-promiscuous, and regioselective glycosylation of flavonoids.

ABSTRACT: Flavonoid O-glycosylation, catalyzed by uridine diphosphate (UDP)-glycosyltransferases, is crucial for their therapeutic efficacy. However, most UDP-glycosyltransferases encounter three major limitations: low activity, poor regioselectivity, and restricted substrate availability, hindering their pharmaceutical applications. To address these challenges, we conducted protein engineering on a previously unidentified glycosyltransferase, UGT75AJ2, which had 3',7-O-glycosylation capabilities. Our approach involved three strategies: (i) development of a tailored focused rational iterative site-specific mutagenesis strategy, augmented by virtual screening and iterative mutagenesis, to design mutant Mut4-1 (S367A/V274A/F82V/I132T) with a 128-fold enhancement in relative catalytic act

SUBMITTER: Lu Y 

PROVIDER: S-EPMC12494026 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature