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US 9,783,566 B2Granted
Non-caloric sweeteners and methods for synthesizing
AssigneeConagen Inc.
InventorsGuohong Mao, Xiaodan Yu
Life of this patent
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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).
Claims7
A method for synthesizing rebaudioside E from rebaudioside KA, the method comprising:
- adding to an in vitro reaction mixture:
- (a) rebaudioside KA,
- (b) substrates selected from the group consisting of sucrose, uridine diphosphate (UDP) and uridine diphosphate-glucose (UDP-glucose), and
- (c) a UDP-glycosyltransferase selected from the group consisting of an HV1 having the amino acid sequence of SEQ ID NO:5, an EUGT11 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 E, wherein a glucose is covalently coupled to the CT 13-O-glucose of rebaudioside KA to produce rebaudioside E.
5 more elements
A method for synthesizing rebaudioside D3 from rubusoside, the method comprising:
- (i) adding to an in vitro reaction mixture:
- (a) rubusoside,
- (b) substrates selected from the group consisting of sucrose, uridine diphosphate (UDP) and uridine diphosphate-glucose (UDP-glucose), and
- (c) a UDP-glycosyltransferase fusion enzyme having the amino acid sequence of SEQ ID NO:
- 11;
- (ii) incubating the reaction mixture for a sufficient time to produce a mixture of stevioside and rebaudioside KA, wherein a first glucose is covalently coupled to the rubusoside to produce a mixture of stevioside and rebaudioside KA;
- (iii) further incubating the mixture of stevioside and rebaudioside KA with the substrates and an EUGT11 having the amino acid sequence of SEQ ID NO:
- 3 or the UDP-glycosyltransferase fusion enzyme to produce rebaudioside E, wherein a second glucose is covalently coupled to the stevioside and the rebaudioside KA to produce rebaudioside E;
- and
- (iv) further incubating the rebaudioside E with the substrates and the EUGT11 or the UDP-glycosyltransferase fusion enzyme to produce rebaudioside D3, wherein a third glucose is covalently coupled to the rebaudioside E to produce rebaudioside D3.
7 more elements
A method for synthesizing rebaudioside D3 from rebaudioside KA, the method comprising:
- adding to an in vitro reaction mixture:
- (a) rebaudioside KA,
- (b) substrates selected from the group consisting of sucrose, uridine diphosphate (UDP) and uridine diphosphate-glucose (UDP-glucose), and
- (c) a UDP-glycosyltransferase selected from the group consisting of an EUGT11 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 D3, wherein a first glucose is covalently coupled to the rebaudioside KA to produce rebaudioside E and a second glucose is covalently coupled to the rebaudioside E to produce rebaudioside D3.
5 more elements
Dependencies are inferred from the claim text.