Method for preparing rebaudioside M by utilizing co-immobilized compound enzyme and application of rebaudioside M

Abstract

The invention relates to the technical field of biosynthesis and manufacturing, in particular to a method for preparing rebaudioside M by utilizing a co-immobilized compound enzyme and application of the rebaudioside M. A novel immobilized matrix material is developed, different reaction activity functional groups (such as maleimide, azide and tetrazine) are modified on the surface of a traditional chitosan matrix for respective immobilization of the three enzymes, and the rebaudioside M is prepared. The method is characterized in that a chitosan carrier material is used as a substrate, the immobilization reaction of each enzyme is ensured to be orthogonal to each other, so that the three enzymes are immobilized according to the optimal proportion, and specifically, in the implementation of the method, sucrose synthase, glycosyl transferase UGT1 and glycosyl transferase UGT2 required by the reaction are co-immobilized on the chitosan carrier material through a co-immobilization method; according to the present invention, the process from the substrate rebaudioside A to the substrate rebaudioside D to the substrate rebaudioside M is completed through the one-pot method, the sucrose is introduced, the cheap sucrose is adopted as the glucosyl group source, and the sucrose synthase is adopted to decompose the sucrose into glucose so as to be adopted as the active glucosyl group donor, such that the UDPG use problem is solved;

Images