Enzymatic Synthesis of Novel Glycyrrhizic Acid Glucosides Using a Promiscuous Bacillus Glycosyltransferase

Abstract

Glycyrrhetinic acid (GA) and glycyrrhizin (GA-3-O-[β-d-glucuronopyranosyl-(1→2)-β-d-glucuronopyranoside], GL) are the major bioactive components of Glycyrrhiza uralensis and possess multifarious notable biological activities. UDP-glycosyltransferase (UGT)–catalyzed glycosylation remarkably extends the structural and functional diversification of GA-glycoside derivatives. In this study, six glucosides (1–6) of GA and GL were synthesized using a Bacillus subtilis 168–originated flexible UDP-glycosyltransferase Bs-YjiC. Bs-YjiC could transfer a glucosyl moiety from UDP-glucose to the free C3 hydroxyl and/or C30 carboxyl groups of GA and GL and further elongate the C30 glucosyl chain via a β-1-2-glycosidic bond. Glycosylation significantly increased the water solubility of these novel glucosides by 4–90 folds. In vitro assays showed that GA monoglucosides (1 and 2) showed stronger antiproliferative activity against human liver cancer cells HepG2 and breast cancer cells MCF-7 than that of GL and GL glucosides. These findings provide significant insights into the important role of promiscuous UGTs for the enzymatic synthesis of novel bioactive GA derivatives.