Biocatalysis for Rare Ginsenoside Rh2 Production in High Level with Co-Immobilized UDP-Glycosyltransferase Bs-YjiC Mutant and Sucrose Synthase AtSuSy

Rare ginsenoside Rh2 exhibits diverse pharmacological effects. UDP-glycosyltransferase (UGT) catalyzed glycosylation of protopanaxadiol (PPD) has been growing interest in recent years. Bs-YjiC coupling sucrose synthase one-pot reaction was successfully applied to biosynthesis with UDP-glucose regeneration from and UDP, which formed a green sustainable approach. In this study, the his-tagged mutant M315F AtSuSy were co-immobilized on heterofunctional supports. The affinity adsorption significantly improved capacity specific binding two recombinant enzymes, dual enzyme covalently cross-linked by acetaldehyde groups promoted stability immobilized bienzyme, allowing higher substrate concentration easing inhibition for coupled reaction. amount used is Bs-YjiC-M315F: = 18 mU/mL: 25.2 mU/mL, yield 79.2% achieved. coimmobilized M315F/AtSuSy had good operational repetitive usage 10 cycles, kept between 77.6% 81.3%. high titer cumulatively reached up 16.6 mM (10.3 g/L) using fed-batch technology, final 83.2%. This study established approach production level titer, provides promising candidates natural drug research development.