Glycosyltransferase derived from Dolwoe and use thereof

Patent metadata

Field	Value
Publication number	US-9896710-B2
Application number	US-201514857703-A
Country	US
Kind code	B2
Filing date	Sep 17, 2015
Priority date	Sep 19, 2014
Publication date	Feb 20, 2018
Grant date	Feb 20, 2018

Abstract

Provided are a novel UDP-glycosyltransferase (uridine diphosphate glycosyltransferase) protein having glycosyltransfer activity for glucose linked by a glycosidic bond at the C-20 position of PPD (protopanaxadiol)-type or PPT (protopanaxatriol)-type ginsenoside, and use thereof.

First claim

What is claimed is:

1. A method of preparing a glycosylated protopanaxadiol (PPD)-type ginsenoside or a glycosylated protopanaxatriol (PPT)-type ginsenoside from a PPD-type ginsenoside or a PPT-type ginsenoside, wherein the glycosylated PPT-type ginsenoside and the PPD-type ginsenoside have a glucose that is linked by a glyosidic bond at the C-20 position, the method comprising:
contacting the PPD-type ginsenoside or the PPT-type ginsenoside and UDP-glucose with:
(i) an isolated Uridine diphosphate (UDP)-glycosyltransferase protein comprising an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 1, and which has activity for transferring a glucose moiety of UDP-glucose to the C-20 position of the PPD-type ginsenoside or the PPT-type ginsenoside, or
(ii) an isolated host cell transformed with an expression vector, wherein the expression vector comprises a polynucleotide encoding a UDP-glycosyltransferase protein comprising an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 1, and which has glycosyltransfer activity for transferring a glucose moiety of UDP-glucose to the C-20 position of the PPD-type ginsenoside or the PPT-type ginsenoside or
(iii) a culture comprising the host cell of (ii),

to thereby prepare the glycosylated PPD-type ginsenoside or the glycosylated PPT-type ginsenoside.

2. The method of claim 1 , wherein the PPD-type ginsenoside or the PPT-type ginsenoside is selected from the group consisting of compound K (CK), F2, Rd, F1, Rg 1 and Re.

3. The method of claim 2 , wherein the UDP-glycosyltransferase protein catalyzes the conversion of CK into gypenoside LXXV, the conversion of F2 into gypenoside XVII, the conversion of Rd into Rb 1 , the conversion of F1 into notoginsenoside U, the conversion of Rg 1 into notoginsenoside R3, and/or the conversion of Re into gluco-Re.

4. The method of claim 1 , wherein the PPD-type ginsenoside and the PPT-type ginsenoside are chemically synthesized, or contained in an extract of ginseng or red ginseng.

5. The method of claim 1 , wherein the UDP-glycosyltransferase protein comprises the amino acid sequence of SEQ ID NO: 1.

6. The method of claim 5 , wherein the PPD-type ginsenoside or the PPT-type ginsenoside is selected from the group consisting of CK, F2, Rd, F1, Rg 1 and Re.

7. The method of claim 5 , wherein the UDP-glycosyltransferase protein catalyzes the glycosylation of CK into gypenoside LXXV, the glycosylation of F2 into gypenoside XVII, the glycosylation of Rd into Rb 1 , the glycosylation of F1 into notoginsenoside U, the glycosylation of Rg 1 into notoginsenoside R3, and/or the glycosylation of Re into gluco-Re.

8. The method of claim 5 , wherein the PPD-type ginsenoside and the PPT-type ginsenoside are chemically synthesized, or contained in an extract of ginseng or red ginseng.

9. An expression vector comprising a polynucleotide encoding a UDP-glycosyltransferase protein and a heterologous promoter operably linked to the polynucleotide, wherein the UDP-glycosyltransferase protein comprises an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 1, and which has glycosyltransfer activity for transferring a glucose moiety of UDP-glucose to the C-20 position of a PPD-type ginsenoside or a PPT-type ginsenoside.

10. An isolated host cell transformed with the expression vector of claim 9 .

11. The method of claim 1 , wherein the isolated UDP-glycosyltransferase protein has been purified by affinity chromatography.

12. The method of claim 5 , wherein the isolated UDP-glycosyltransferase protein has been purified by affinity chromatography.

Assignees

• Korea Advanced Inst Sci & Tech
• Intelligent Synthetic Biology Ct

Inventors

• Kim Sun Chang
• Choi Giltsu
• Jung Suk Chae
• Kim Woohyun
• Lim Soohwan
• Im Wan Taek

Classifications

• C12Y204/01

  Hexosyltransferases (2.4.1) · CPC title

• C12N9/1051

  Hexosyltransferases (2.4.1) · CPC title

• C12P19/56 Primary

  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

• C12P33/20 Primary

  containing heterocyclic rings · CPC title

• C12P33/00

  Preparation of steroids · CPC title