Highly Efficient Biosynthesis of Glycyrrhetinic Acid Glucosides by Coupling of Microbial Glycosyltransferase to Plant Sucrose Synthase

Glycyrrhetinic acid (GA) is a principal bioactive pentacyclic triterpenoid from Glycyrrhiza uralensis. Uridine diphosphate-dependent glycosyltransferases (UGTs) have been widely used to catalyze glycosylation of diverse nature products for the development of potential therapeutic compounds. In this study, we have characterized a UGT109A3 from Bacillus subtilis, which can glycosylate both the free C3 hydroxyl and C30 carboxyl groups of GA to yield a unique 3, 30-O-β-D-diglucoside-GA. By coupling the microbial UGT109A3 to plant sucrose synthase (SUS), GA-diglucoside could be biosynthesized in an efficient and economical way. With a fed-batch glycosylation, a large scale of GA-diglucoside (6.26 mM, 4.98 g/L in 8 h) could be enzymatically transformed from GA. The obtained GA-diglucoside showed a significant water solubility improvement of around 3.4 × 10³ fold compared with that of the parent GA (29 μM). Moreover, it also exhibited dose-dependent cytotoxicity toward human colon carcinoma Caco-2 cell line according to MTT assay, having an IC₅₀ at 160 μM. This study not only establishes efficient platform for producing GA-glucosides, but is also valuable for developing further the biosynthesis of other complex glycosylated natural products.