Enzymatic method for preparing Rebaudioside C

Patent metadata

Field	Value
Publication number	US-11976312-B2
Application number	US-202217657307-A
Country	US
Kind code	B2
Filing date	Mar 30, 2022
Priority date	Oct 21, 2016
Publication date	May 7, 2024
Grant date	May 7, 2024

Abstract

Provided is a method for preparing Rebaudioside C using an enzymatic method, comprising using rubusoside or dulcoside A as a substrate, and making the substrate, in the presence of a glycosyl donor, react under the catalysis of UDP-glycosyltransferase-containing recombinant cell and/or UDP-glycosyltransferase prepared therefrom to generate Rebaudioside C.

First claim

What is claimed is:

1. A method for preparing Rebaudioside C, the method comprising: reacting dulcoside A with a glycosyl donor in a reaction system in the presence of recombinant cells comprising UDP-glycosyltransferase and/or UDP-glycosyltransferase prepared from the recombinant cells;
wherein the reaction system comprises toluene at a concentration by volume of 1-3%; and
wherein the UDP-glycosyltransferase has the amino acid sequence of SEQ ID NO: 2.

2. The method of claim 1 , wherein the glycosyl donor is UDP-glucose or a UDP-glucose regeneration system comprising sucrose, sucrose synthase, and UDP.

3. The method of claim 1 , wherein the UDP-glucosyltransferase comprises UGT-A from Stevia rebaudiana.

4. The method of claim 1 , wherein the reaction system comprises an aqueous system with a temperature of 35-45° C. and a pH of 7.5-8.5.

5. The method of claim 4 , wherein the reaction system comprises a phosphate buffer solution.

6. The method of claim 1 , wherein the recombinant cells are microorganism cells.

7. The method of claim 6 , wherein the microorganism is Escherichia coli, Saccharomyces cerevisiae or Pichia pastoris.

Assignees

• Pepsico Inc

Inventors

• Tao Alex
• Li Guoqing
• Wang Wenxia
• Zheng Leilei
• Zhu Chunlei
• Liang Xiaoliang
• Chan Kuikiu

Classifications

• C12P19/56 Primary

  having an oxygen atom of the saccharide radical directly bound to a condensed ring system having three or more carbocyclic rings, e.g. daunomycin, adriamycin · CPC title

• C12N9/10

  Transferases (2.) (ribonucleases C12N9/22) · CPC title

• C12Y204/01

  Hexosyltransferases (2.4.1) · CPC title

• A23L27/30 Primary

  Artificial sweetening agents · CPC title

• C12P19/18

  produced by the action of a glycosyl transferase, e.g. alpha-, beta- or gamma-cyclodextrins · CPC title