Method for utilizing monoterpene glycosyltransferase

Patent metadata

Field	Value
Publication number	US-9518282-B2
Application number	US-201113997683-A
Country	US
Kind code	B2
Filing date	Dec 27, 2011
Priority date	Dec 28, 2010
Publication date	Dec 13, 2016
Grant date	Dec 13, 2016

Abstract

The object of the present invention is to provide a novel method for producing a terpene 8-glycoside.
The present invention provides a method for producing a terpene 8-glycoside by means of glycosyltransferase acting on the 8-position of terpenes. The present invention provides a transformant transformed with a gene for the glycosyltransferase acting on the 8-position of terpenes and a method for producing such a transformant. The present invention provides a plant modified to suppress the expression of a protein having glycosylation activity on the 8-position of a monoterpene compound.

First claim

The invention claimed is:

1. A method for producing a monoterpene 8-glycoside, which comprises:
(i) reacting a protein of any one selected from the group consisting of (a) to (c) shown below, a UDP-sugar, and a monoterpene compound to cause glycosylation at the 8-position of the monoterpene compound; wherein the monoterpene compound is 8-hydroxygeraniol or 8-hydroxylinalool:
(a) a protein which consists of the amino acid sequence shown in SEQ ID NO: 2 or 9;
(b) a protein which consists of an amino acid sequence with deletion, substitution, insertion and/or addition of 1 to 10 amino acids in the amino acid sequence shown in SEQ ID NO: 2 or 9 and which has glycosylation activity on the 8-position of a monoterpene compound; and
(c) a protein which has an amino acid sequence sharing a sequence identity of 90% or more with the amino acid sequence shown in SEQ ID NO: 2 or 9 and which has glycosylation activity on the 8-position of a monoterpene compound; and
(ii) purifying the 8-glycoside of the monoterpene compound generated in step (i).

2. The method according to claim 1 , wherein the UDP-sugar is UDP-glucose.

3. A non-human transformant transformed with a polynucleotide of any one selected from the group consisting of (a) to (e):
(a) a polynucleotide containing the nucleotide sequence shown in SEQ ID NO: 1, 3, 8 or 10;
(b) a polynucleotide encoding a protein which consists of the amino acid sequence shown in SEQ ID NO: 2 or 9;
(c) a polynucleotide encoding a protein which consists of an amino acid sequence with a deletion, substitution, insertion and/or addition of 1 to 10 amino acids in the amino acid sequence shown in SEQ ID NO: 2 or 9 and which has glycosylation activity on the 8-position of a monoterpene compound; and
(d) a polynucleotide encoding a protein which has an amino acid sequence sharing a sequence identity of 90% or more with the amino acid sequence shown in SEQ ID NO: 2 or 9 and which has glycosylation activity on the 8-position of a monoterpene compound.

4. The transformant according to claim 3 , which contains the nucleotide sequence shown in SEQ ID NO: 1, 3, 8 or 10.

5. The transformant according to claim 3 , wherein the polynucleotide is inserted into an expression vector.

6. The transformant according to claim 3 , which is a plant.

7. An extract of the transformant according to claim 3 , wherein the extract is prepared by homogenizing the transformant to obtain a homogenate, centrifuging the homogenate to obtain a supernatant, and collecting the supernatant as the extract.

8. A food, an aromatic, a pharmaceutical preparation or an industrial raw material, which comprises the extract according to claim 7 .

9. A method for producing a protein having glycosylation activity on the 8-position of a monoterpene compound, which comprises culturing the non-human transformant according to claim 3 , wherein the monoterpene compound is 8-hydroxygeraniol or 8-hydroxylinalool.

Assignees

• Ono Eiichiro
• Tsuruoka Nobuo
• Suntory Holdings Ltd

Inventors

• Ono Eiichiro
• Tsuruoka Nobuo

Classifications

• C12P19/44 Primary

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

• C12N15/8257

  for the production of primary gene products, e.g. pharmaceutical products, interferon · CPC title

• A61Q19/00

  Preparations for care of the skin · CPC title

• C12N9/1051

  Hexosyltransferases (2.4.1) · CPC title

• A61K8/602

  Glycosides, e.g. rutin · CPC title