Non-caloric sweeteners and methods for synthesizing

Patent metadata

Field	Value
Publication number	US-2016153018-A1
Application number	US-201615000774-A
Country	US
Kind code	A1
Filing date	Jan 19, 2016
Priority date	Oct 3, 2014
Publication date	Jun 2, 2016
Grant date	—

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

1 .- 7 . (canceled)

8 . A method for synthesizing rebaudioside E from rubusoside, the method comprising:
preparing a reaction mixture comprising rubusoside, substrates selected from the group consisting of sucrose, uridine diphosphate (UDP) and uridine diphosphate-glucose (UDP-glucose), and a UDP-glycosyltransferase selected from the group consisting of HV1, EUGT11 and a UDP-glycosyltransferase fusion enzyme (EUS); and optionally, a sucrose synthase; and incubating the reaction mixture for a sufficient time to produce rebaudioside E, wherein a glucose is covalently coupled to rubusoside to produce rebaudioside KA or stcviosidc, and a glucose is covalently coupled to rebaudioside KA to produce rebaudioside E.

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 selected from the group consisting of an Arabidopsis sucrose synthase 1; an Arabidopsis sucrose synthase 3; and a Vigna radiate sucrose synthase.

11 . The method of claim 10 , wherein the sucrose synthase is an Arabidopsis thaliana sucrose synthase 1.

12 . The method of claim 8 , wherein the UDP-glycosyltransferase fusion enzyme is selected from the group consisting of an EUGT11 uridine diphospho glycosyltransferase domain coupled to a sucrose synthase domain.

13 . The method of claim 12 , wherein 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.

14 . The method of claim 13 , wherein the sucrose synthase domain is an Arabidopsis thaliana sucrose synthase 1.

15 .- 28 . (canceled)

Assignees

• Conagen Inc

Inventors

• Mao Guohong
• Luo Yang
• Lin Ying
• Yu Xiaodan

Classifications

• C12N9/1051

  Hexosyltransferases (2.4.1) · CPC title

• A23L27/36

  Terpene glycosides · CPC title

• C12Y204/01013

  Sucrose synthase (2.4.1.13) · CPC title

• C12P19/18

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

• A23L2/60

  Sweeteners · CPC title