A biocatalytic synthesis of diosgenyl-β-d-glucopyranoside by the use of four recombinant enzymes in one pot

A system for the one-pot synthesis of diosgenyl-β-d-glucopyranoside (trillin) using multiple recombinant enzymes is developed. The enzymes maltodextrin phosphorylase (E1), glucose-1-phosphate thymidylyltransferase (E2), inorganic pyrophosphatase (E3), and solanidine glucosyltransferase (E4) involved in the work have been cloned and expressed in Escherichia coli. Under the optimized reaction conditions, the yield of trillin reached 28% (ca. 15.8 mg/l). The recovery yield of trillin after purification was 89%.