Mogrol glycosyltransferase and gene encoding same

Patent metadata

Field	Value
Publication number	US-10689682-B2
Application number	US-201916538046-A
Country	US
Kind code	B2
Filing date	Aug 12, 2019
Priority date	Oct 17, 2014
Publication date	Jun 23, 2020
Grant date	Jun 23, 2020

Abstract

To provide a mogrol glycosyltransferase and a method for producing a mogrol glycoside using the enzyme. The present invention provides a mogrol glycosyltransferase and a method for producing a mogrol glycoside using the enzyme, and a transformant into which a mogrol glycosyltransferase gene is introduced and a method for preparing the transformant.

First claim

We claim:

1. A polynucleotide selected from:
(a) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 1;
(b) 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; and
(c) a polynucleotide encoding a protein consisting of an amino acid sequence in which 1 to 45 amino acids are deleted, substituted, inserted, and/or added in the amino acid sequence of SEQ ID NO: 2 and having an activity of adding a sugar molecule to an R 1 group at position 24 of a compound represented by the formula (I)

where R and R 1 each independently represent H, a monosaccharide, or a disaccharide.

2. A protein consisting of an amino acid sequence in which 1 to 45 amino acids are deleted, substituted, inserted, and/or added in the amino acid sequence of SEQ ID NO: 2 and having an activity of adding a sugar molecule to an R1 group at position 24 of a compound represented by formula (I)

where R and R 1 each independently represent H, a monosaccharide, or a disaccharide.

3. A vector comprising the polynucleotide of claim 1 .

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

5. The non-human transformant of claim 4 , which is a plant or a microorganism.

6. The non-human transformant of claim 4 , wherein the polynucleotide is inserted into an expression vector.

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

8. The method of claim 7 , further comprising:
producing a mogrol glycoside by reacting 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):

where R and R 1 each independently represent H, a monosaccharide, or a disaccharide.

9. A method of producing a mogrol glycoside, comprising:
reacting in vitro a protein consisting of the amino acid sequence of SEQ ID NO: 2 with a UDP-sugar and a compound represented by formula (I):

where R and R 1 each independently represent H, a monosaccharide, or a disaccharide.

10. A method comprising:
providing the non-human transformant of claim 4 or a culture thereof, and
obtaining an extract of the non-human transformant or of the culture of the non-human transformant.

11. A method comprising:
providing an extract of the non-human transformant of claim 4 or an extract of a culture of the non-human transformant of claim 4 ,
adding the extract to a raw material of a food, a pharmaceutical preparation or an industrial raw material, and
preparing a food, a pharmaceutical preparation or an industrial raw material.

12. The method of claim 8 , wherein the UDP-sugar is a UDP-hexose.

13. The method of claim 8 , wherein the sugar molecule is selected from the group consisting of UDP-glucose, UDP-mannose, and UDP-galactose.

14. The method of claim 8 , wherein R and R 1 is each independently H or a sugar residue of a glucose monomer or a glucose dimer.

15. The method of claim 8 , wherein the compound represented by formula (I) is mogrol or a mogrol glycoside.

16. The method of claim 8 , wherein the mogrol glycoside is mogroside IA, mogroside Ib, mogroside IE, mogroside IIA, mogroside IIA1, mogroside IIE, mogroside III, mogroside IIIA1, mogroside IIIA2, mogroside IIIE, mogroside IV, mogroside IVA, mogroside V, siamenoside I, 11-oxomogroside V, mogrol pentaglycoside, or a combination thereof.

17. The method of claim 9 , wherein the mogrol glycoside is mogroside IA, mogroside Ib, mogroside IE, mogroside IIA, mogroside IIA1, mogroside IIE, mogroside III, mogroside IIIA1, mogroside IIIA2, mogroside IIIE, mogroside IV, mogroside IVA, mogroside V, siamenoside I, 11-oxomogroside V, mogrol pentaglycoside, or a combination thereof.

18. The method of claim 11 , wherein the food is a beverage.

19. The method of claim 11 , wherein the food is selected from a juice drink, a soft drink, a sports beverage, an alcoholic beverage, a tea, a vinegar, and a soy sauce.

Assignees

• Suntory Holdings Ltd

Inventors

• Ono Eiichiro
• Ochiai Misa

Classifications

• C07J9/00 Primary

  Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane · CPC title

• C12N1/20

  Bacteria; Culture media therefor · CPC title

• C12Y204/01

  Hexosyltransferases (2.4.1) · CPC title

• C12P33/00

  Preparation of steroids · CPC title

• C12P19/18

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