Abstract
For a set of flavonoid O- and C-β-glycosyltransferases, we show that β-glucosyl fluoride can function as substrate for an enzymatic reaction wherein uridine 5′-diphosphate (UDP) α-glucose is synthesized in the presence of UDP. In pH and mutagenesis studies of the C-glycosyltransferase from rice, we show that reaction with the β-glucosyl fluoride can serve to identify the acid-base catalytic residue of the enzyme (His24). We also show that β-glucosyl fluoride can rescue activity in an enzyme variant (Ile121Asp) strongly impaired in the canonical reaction wherein flavonoid acceptor is glucosylated from UDP-glucose. Coupling of this variant with the wildtype C-glycosyltransferase in a one-pot reaction enabled efficient 3′-β-C-glucosylation of phloretin from β-glucosyl fluoride in the presence of substochiometric amounts of UDP.
Original language | English |
---|---|
Pages (from-to) | 9148-9153 |
Number of pages | 6 |
Journal | ACS Catalysis |
Volume | 8 |
Issue number | 10 |
DOIs | |
Publication status | Published - 5 Oct 2018 |
Keywords
- general acid-base catalysis
- glycosyl fluoride
- glycosylation
- glycosyltransferase
- inverting
- retaining
ASJC Scopus subject areas
- Catalysis
- Chemistry(all)