Recombinant production of steviol glycosides
AssigneeDanstar Ferment AG
InventorsGanesh M. Kishore, Michael Motion, Paula M. Hicks, Jorgen Hansen, Jens Houghton Larsen, Esben Halkjaer Hansen, Michael Dalgaard Mikkelsen, Sabina Tavares, Charlotte Blom
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Abstract
Recombinant microorganisms, plants, and plant cells are disclosed that have been engineered to express novel recombinant genes encoding steviol biosynthetic enzymes and UDP-glycosyltransferases (UGTs). Such microorganisms, plants, or plant cells can produce steviol or steviol glycosides, e.g., rubusoside or Rebaudioside A, which can be used as natural sweeteners in food products and dietary supplements.
Claims24
A method for transferring an additional sugar moiety to a C2′ position of a glucose in a steviol glycoside, comprising contacting the steviol glycoside with a recombinant polypeptide capable of beta 1,2 glycosylation of the C2′ of the 13-O-glucose, 19-O-glucose, or both 13-O-glucose and 19-O-glucose of a steviol glycoside and having at least 95% sequence identity to the amino acid sequence set forth in SEQ ID NO:
- 5 and a sugar moiety donor under suitable reaction conditions for the transfer of the additional sugar moiety to the steviol glycoside,
- wherein the steviol glycoside is steviol-13-O-glucoside, rubusoside, stevioside or rebaudioside A, and
- wherein a stevioside, rebaudioside E, rebaudioside D, steviol-1,2 bioside, steviol-1,2-xylobioside, steviol-1,2-rhamnobioside, an isomer thereof, and/or a steviol glycoside composition thereof is produced upon transfer of the additional sugar moiety.
A method of producing a steviol glycoside composition by transferring an additional sugar moiety from a sugar moiety donor to the C2′ of a glucose in a steviol glycoside with a recombinant polypeptide capable of beta 1,2 glycosylation of the C2′ of the 13-O-glucose, 19-O-glucose, or both 13-O-glucose and 19-O-glucose of a steviol glycoside and having at least 95% sequence identity to the amino acid sequence set forth in SEQ ID NO:5, in a whole cell of yeast S. cerevisiae;
- wherein the steviol glycoside is steviol-13-O-glucoside, rubusoside, stevioside or rebaudioside A;
- and
- wherein the steviol glycoside composition comprises stevioside, rebaudioside E, rebaudioside D, steviol-1,2 bioside, steviol-1,2-xylobioside, and/or steviol-1,2-rhamnobioside.
Dependencies are inferred from the claim text.