Non-caloric sweeteners and methods for synthesizing

Patent metadata

Field	Value
Publication number	US-9567619-B2
Application number	US-201514873474-A
Country	US
Kind code	B2
Filing date	Oct 2, 2015
Priority date	Oct 3, 2014
Publication date	Feb 14, 2017
Grant date	Feb 14, 2017

Abstract

Disclosed are steviol glycosides referred to as rebaudioside V and rebaudioside W. Also disclosed are methods for producing rebaudioside M (Reb M), rebaudioside G (Reb G), rebaudioside KA (Reb KA), rebaudioside V (Reb V) and rebaudioside (Reb W).

First claim

What is claimed is:

1. A synthetic rebaudioside consisting of a chemical structure:

2. A method for synthesizing rebaudioside V from rebaudioside G, the method comprising:
preparing a reaction mixture comprising:
(a) rebaudioside G,
(b) substrates selected from the group consisting of sucrose, uridine diphosphate (UDP) and uridine diphosphate-glucose (UDP-glucose), and
(c) a HV1 UDP-glycosyltransferase having the amino acid sequence of SEQ ID NO: 5; and

incubating the reaction mixture for a sufficient time to produce rebaudioside V, wherein a glucose is covalently coupled to the rebaudioside G to produce rebaudioside V.

3. The method of claim 2 further comprising adding a sucrose synthase to the reaction mixture.

4. The method of claim 3 , wherein the sucrose synthase is an Arabidopsis thaliana sucrose synthase 1 having the amino acid sequence of SEQ ID NO: 7.

5. A method for synthesizing rebaudioside V from rebaudioside G, the method comprising:
preparing a reaction mixture comprising:
(a) rebaudioside G,
(b) substrates selected from the group consisting of sucrose, uridine diphosphate (UDP) and uridine diphosphate-glucose (UDP-glucose), and
(c) a uridine diphospho glycosyltransferase (UDP-glycosyltransferase) selected from the group consisting of an EUGT11 UDP-glycosyltransferase having the amino acid sequence of SEQ ID NO: 3 and a UDP-glycosyltransferase fusion enzyme having the amino acid sequence of SEQ ID NO: 11; and

incubating the reaction mixture for a sufficient time to produce rebaudioside V, wherein a glucose is covalently coupled to the rebaudioside G to produce rebaudioside V.

6. The method of claim 5 further comprising adding a sucrose synthase to the reaction mixture.

7. The method of claim 6 , wherein the sucrose synthase is an Arabidopsis thaliana sucrose synthase 1 having the amino acid sequence of SEQ ID NO: 7.

8. A method for synthesizing rebaudioside V from rebaudioside KA, the method comprising:
preparing a reaction mixture comprising:
(a) rebaudioside KA,
(b) substrates selected from the group consisting of sucrose, uridine diphosphate (UDP) and uridine diphosphate-glucose (UDP-glucose), and
(c) a uridine diphospho glycosyltransferase (UDP-glycosyltransferase) selected from the group consisting of a UGT76G1 UDP-glycosyltransferase having the amino acid sequence of SEQ ID NO: 1 and a UDP-glycosyltransferase fusion enzyme having the amino acid sequence of SEQ ID NO: 9; and

incubating the reaction mixture for a sufficient time to produce rebaudioside V, wherein a glucose is covalently coupled to the rebaudioside KA to produce rebaudioside V.

9. The method of claim 8 further comprising adding a sucrose synthase to the reaction mixture.

10. The method of claim 9 , wherein the sucrose synthase is an Arabidopsis thaliana sucrose synthase 1 having the amino acid sequence of SEQ ID NO: 7.

11. A method for synthesizing rebaudioside V from rubusoside, the method comprising:
preparing a reaction mixture comprising:
(a) rubusoside,
(b) substrates selected from the group consisting of sucrose, uridine diphosphate (UDP) and uridine diphosphate-glucose (UDP-glucose), and
(c) uridine diphospho glycosyltransferases (UDP-glycosyltransferases) selected from the group consisting of a UGT76G1 UDP-glycosyltransferase having the amino acid sequence of SEQ ID NO: 1, a HV1 UDP-glycosyltransferase having the amino acid sequence of SEQ ID NO: 5 and a UDP-glycosyltransferase fusion enzyme having the amino acid sequence of SEQ ID NO: 9; and incubating the reaction mixture for a sufficient time to produce rebaudioside V.

12. The method of claim 11 further comprising adding a sucrose synthase to the reaction mixture.

13. The method of claim 12 , wherein the sucrose synthase is an Arabidopsis thaliana sucrose synthase 1 having the amino acid sequence of SEQ ID NO: 7.

14. A method for synthesizing a rebaudioside V from rubusoside, the method comprising:
preparing a reaction mixture comprising:
(a) rubusoside,
(b) substrates selected from the group consisting of sucrose, uridine diphosphate (UDP) and uridine diphosphate-glucose (UDP-glucose), and
(c) uridine diphospho glycosyltransferases (UDP-glycosyltransferases) selected from the group consisting of a UGT76G1 UDP-glycosyltransferase having the amino acid sequence of SEQ ID NO: 1, EUGT11 UDP-glycosyltransferase having the amino acid sequence of SEQ ID NO: 3 and a UDP-glycosyltransferase fusion enzyme having the amino acid sequence of SEQ ID NO: 11; and incubating the reaction mixture for a sufficient time to produce rebaudioside V.

15. The method of claim 14 further comprising adding a sucrose synthase to the reaction mixture.

16. The method of claim 15 , wherein the sucrose synthase is an Arabidopsis thaliana sucrose synthase 1 having the amino acid sequence of SEQ ID NO: 7.

17. A sweetener comprising a synthetic rebaudioside, the synthetic rebaudioside consisting of a chemical structure:

18. The sweetener of claim 17 further comprising at least one of a filler, a bulking agent and an anticaking agent.

Assignees

• Conagen Inc

Inventors

• Mao Guohong
• Yu Xiaodan

Classifications

• C12N9/1062

  Sucrose synthase (2.4.1.13) · CPC title

• C12Y204/01

  Hexosyltransferases (2.4.1) · CPC title

• C07H1/00

  Processes for the preparation of sugar derivatives · CPC title

• A23L27/36

  Terpene glycosides · CPC title

• A23V2002/00

  Food compositions, function of food ingredients or processes for food or foodstuffs · CPC title