METHOD FOR OBTAINING REBAUDIOSIDE M USING THE ENZYME METHOD Russian patent published in 2018 - IPC C12P19/18 C12P19/56 C12N1/15 C12N1/21 A23L27/30 

FIELD: biotechnology.

SUBSTANCE: group of inventions relates to biotechnology. Method for the preparation of rebaudioside M is proposed. Method comprises reacting rebaudioside D with a glucosyl donor in the presence of UDP-glucosyltransferase or recombinant cells, containing UDP-glucosyltransferase, to obtain rebaudioside M. Glucosyl donor is UDP-glucose or a UDP-glucose regeneration system comprising sucrose, sucrose synthetase and UDP, UDP-glucosyltransferase has an amino acid sequence, having at least 90% identity to SEQ ID NO:2. Recombinant cells are microbial cells selected from the group consisting of Escherichia coli cells, Saccharomyces cerevisiae or Pichia pastoris. Method for the preparation of rebaudioside M, comprising interaction of rebaudioside A with a glucosyl donor in the presence of the first UDP-glucosyltransferase or recombinant cells containing the first UDP-glucosyltransferase with the formation of rebaudioside D, and the interaction of rebaudioside D with a glucosyl donor in the presence of the second UDP-glucosyltransferase or recombinant cells, containing the second UDP-glucosyltransferase, to obtain rebaudioside M. First UDP-glucosyltransferase has an amino acid sequence, having at least 90% identity with SEQ ID NO: 4, and the second UDP-glucosyltransferase has an amino acid sequence having at least 90% identity to SEQ ID NO: 2.

EFFECT: group of inventions ensures the conversion of at least 40% of rebaudioside D to rebaudioside M after interaction for two hours.

11 cl, 1 dwg, 10 ex