Method for producing steviol glycoside

Patent metadata

Field	Value
Publication number	US-10113154-B2
Application number	US-201314386934-A
Country	US
Kind code	B2
Filing date	Mar 14, 2013
Priority date	Mar 27, 2012
Publication date	Oct 30, 2018
Grant date	Oct 30, 2018

Abstract

The invention provides a method for producing steviol glycosides. The invention provides a transformant having introduced therein the steviol glucosyltransferase and a method for producing steviol glycosides using the transformant.

First claim

The invention claimed is:

1. A polynucleotide selected from:
a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 1; and
a polynucleotide comprising a heterologous regulatory element operably linked to a polynucleotide sequence encoding a protein consisting of the amino acid sequence of SEQ ID NO: 2.

2. A non-human transformant comprising the polynucleotide of claim 1 .

3. The non-human transformant according to claim 2 , which is a plant.

4. A method comprising culturing or cultivating the non-human transformant according to claim 2 to produce the protein consisting of the amino acid sequence of SEQ ID NO: 2.

5. The method of claim 4 , further comprising:
producing a steviol glycoside by contacting the protein consisting of the amino acid sequence of SEQ ID NO: 2 produced by the non-human transformant with a UDP-sugar and a compound represented by formula (I):

wherein R 1 represents H, a C 1 -C 20 alkyl, a C 2 -C 20 alkenyl, a C 2 -C 20 alkynyl, a C 4 -C 20 alkyldienyl, a C 6 -C 18 aryl, a C 6 -C 20 alkylaryl, a C 6 -C 20 arylalkyl, a C 4 -C 20 cycloalkyl, a C 4 -C 20 cycloalkenyl, a (C 3 -C 10 cycloalkyl) C 1 -C 10 alkyl, or a sugar residue.

6. A method for producing a steviol glycoside, comprising:
contacting, outside of a stevia plant, a protein consisting of the amino acid sequence of SEQ ID NO: 2 with a UDP-sugar and a compound represented by formula (I):

wherein R 1 represents H, a C 1 -C 20 alkyl, a C 2 -C 20 alkenyl, a C 2 -C 20 alkynyl, a C 4 -C 20 alkyldienyl, a C 6 -C 18 aryl, a C 6 -C 20 alkylaryl, a C 6 -C 20 arylalkyl, a C 4 -C 20 cycloalkyl, a C 4 -C 20 cycloalkenyl, a (C 3 -C 10 cycloalkyl) C 1 -C 10 alkyl, or a sugar residue.

7. The method according to claim 6 , wherein the sugar in the UDP-sugar is a hexose.

8. The method according to claim 6 , wherein the sugar in the UDP-sugar is selected from glucose, mannose, and galactose.

9. The method according to claim 6 , wherein R 1 is H or the sugar residue that is a glucose monomer, a glucose dimer, or a glucose trimer.

10. The method according to claim 6 , wherein the compound is steviol, steviolmonoside, or steviolbioside.

11. The method according to claim 6 , wherein the sugar in the UDP-sugar is glucose.

12. The method according to claim 6 , wherein the steviol glycoside is steviolbioside, stevioside, rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, or any combination thereof.

13. The non-human transformant according to claim 2 , wherein the polynucleotide is inserted into an expression vector.

14. A method for producing an extract of the non-human transformant according to claim 2 or an extract of a culture of the non-human transformant according to claim 2 , the method comprising:
providing the transformant according to claim 2 or the culture of the transformant, and
obtaining an extract of the transformant or of the culture of the transformant.

15. A method for producing a food, a pharmaceutical preparation, or an industrial raw material, the method comprising:
providing an extract of the transformant according to claim 2 or of a culture of the transformant,
adding the extract to a raw material of a food, a pharmaceutical preparation, or an industrial raw material, and
preparing the food, the pharmaceutical preparation, or the industrial raw material.

16. The method according to claim 15 , wherein the food is selected from fermented foods, fruit drinks, soft drinks, sports drinks, tea, bakery products, noodles, pastas, cooked rice, sweets, bean curd, ham, bacon, sausage, fish cake (kamaboko), deep-fried fish cake (ageten), and puffy fish cake (hanpen).

17. The method according to claim 16 , wherein the fermented foods are alcoholic beverages, tea, medicinal liquor, sweet cooking sherry (mirin), vinegar, soy sauce, miso (bean paste), and yogurt.

18. The method according to claim 15 , wherein the pharmaceutical preparation is selected from cream, gel, lipstick, facial pack, ointment, dentifrice, and cleansing foam.

Assignees

• Suntory Holdings Ltd

Inventors

• Ono Eiichiro

Classifications

• C12P19/44

  Preparation of O-glycosides, e.g. glucosides {(polysaccharides and not substituted disaccharides C12P19/04, C12P19/12)} · CPC title

• C07H15/256

  Polyterpene radicals · CPC title

• A61K2800/10

  General cosmetic use · CPC title

• 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

• C12Y204/01

  Hexosyltransferases (2.4.1) · CPC title