Method for efficient biosynthesis of Reb D by glycosyltransferase

Abstract

The disclosure discloses a method for efficient biosynthesis of Reb D by glycosyltransferase, belonging to the field of biocatalytic synthesis. According to the disclosure, a glycosyltransferase having an activity to catalyze synthesis of Reb D from Reb A is obtained, and a mutant YojK-1241T/G327N with high catalytic activity is obtained through directed evolution. The glycosyltransferase mutant YojK-1241T/G327N and a sucrose synthase AtSuSy derived from Arabidopsis thaliana are used for constructing a coupling reaction to realize efficient catalytic synthesis of Reb D with Reb A as a substrate. The reaction is carried out by using 19.32 g/L (20 mmol/L) of Reb A as the substrate for 15 h to efficiently synthesize 20.59 g/L of Reb D, and the yield of Reb D reaches 91.29%, which provides an efficient and green new pathway for production of Reb D.

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