Non-caloric sweeteners and methods for synthesizing

Patent metadata

Field	Value
Publication number	US-9908913-B2
Application number	US-201615000774-A
Country	US
Kind code	B2
Filing date	Jan 19, 2016
Priority date	Oct 3, 2014
Publication date	Mar 6, 2018
Grant date	Mar 6, 2018

Abstract

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

First claim

What is claimed is:

1. A method for synthesizing rebaudioside E from rubusoside, the method comprising:
(a) preparing a reaction mixture comprising;
(i) rubusoside,
(ii) substrates selected from the group consisting of sucrose, uridine diphosphate (UDP) and uridine diphosphate-glucose (UDP-glucose), and
(iii) a HV1 UDP-glycosyltransferase having the amino acid sequence of SEQ ID NO: 5 or a UDP-glycosyltransferase fusion enzyme comprising a EUGT11 uridine diphospho glycosyltransferase domain coupled to a sucrose synthase domain having the amino acid sequence of SEQ ID NO: 11; and

(b) incubating the reaction mixture for a sufficient time to produce rebaudioside E, wherein a glucose is covalently coupled to rubusoside to produce rebaudioside KA, and a glucose is covalently coupled to rebaudioside KA to produce rebaudioside E.

2. The method of claim 1 , wherein the reaction mixture comprises the HV1 UDP-glycosyltransferase and a sucrose synthase.

3. The method of claim 2 , wherein the sucrose synthase is selected from the group consisting of an Arabidopsis sucrose synthase 1; an Arabidopsis sucrose synthase 3; and a Vigna radiate sucrose synthase.

4. The method of claim 3 , wherein the sucrose synthase is an Arabidopsis thaliana sucrose synthase 1.

5. The method of claim 1 , wherein the reaction mixture comprises the UDP-glycosyltransferase fusion enzyme and the sucrose synthase domain is selected from the group consisting of an Arabidopsis sucrose synthase 1; an Arabidopsis sucrose synthase 3; and a Vigna radiate sucrose synthase.

6. The method of claim 5 , wherein the sucrose synthase domain is an Arabidopsis thaliana sucrose synthase 1.

7. The method of claim 1 , wherein the reaction mixture comprises the HV1 UDP-glycosyltransferase.

Assignees

• Conagen Inc

Inventors

• Mao Guohong
• Yu Xiaodan

Classifications

• C12P19/56

  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

• A23L2/60

  Sweeteners · CPC title

• C07H15/256

  Polyterpene radicals · CPC title

• A23L27/36

  Terpene glycosides · CPC title

• C07H15/24 Primary

  Condensed ring systems having three or more rings · CPC title