Monoterpene glycosyltransferase originating from hop and method for using same

Patent metadata

Field	Value
Publication number	US-9574182-B2
Application number	US-201314376463-A
Country	US
Kind code	B2
Filing date	Feb 5, 2013
Priority date	Feb 6, 2012
Publication date	Feb 21, 2017
Grant date	Feb 21, 2017

Abstract

The object of the present invention is to provide Humulus lupulus -derived monoterpene glycosyltransferase and a method for producing a monoterpene glycoside by means of this enzyme.
The present invention provides Humulus lupulus -derived monoterpene glycosyltransferase and a method for producing a monoterpene glycoside by means of this enzyme. The present invention provides a transformant transformed with a gene for Humulus lupulus -derived monoterpene glycosyltransferase and a method for producing such a transformant.

First claim

The invention claimed is:

1. A recombinant protein
which consists of an amino acid sequence with substitution, insertion and/or addition of 1 to 70 amino acids in the amino acid sequence shown in SEQ ID NO: 2, 4 or 6 and which has glycosylation activity on a monoterpene compound.

2. The recombinant protein according to claim 1 , wherein the monoterpene compound is geraniol or linalool.

3. A polynucleotide of any one selected from the group consisting of (a) to (e) shown below:
(a) a polynucleotide containing the nucleotide sequence shown in SEQ ID NO: 1, 3 or 5;
(b) a polynucleotide encoding a recombinant protein which consists of the amino acid sequence shown in SEQ ID NO: 2, 4 or 6;
(c) a polynucleotide encoding a recombinant protein which consists of an amino acid sequence with deletion, substitution, insertion and/or addition of 1 to 70 amino acids in the amino acid sequence shown in SEQ ID NO: 2, 4 or 6 and which has glycosylation activity on a monoterpene compound;
(d) a polynucleotide encoding a recombinant protein which has an amino acid sequence sharing a sequence identity of 85% or more with the amino acid sequence shown in SEQ ID NO: 2, 4 or 6 and which has glycosylation activity on a monoterpene compound; and
(e) a polynucleotide which is hybridizable under high stringent conditions with a polynucleotide consisting of a nucleotide sequence complementary to the nucleotide sequence shown in SEQ ID NO: 1, 3 or 5 and which encodes a recombinant protein having glycosylation activity on a monoterpene compound,
wherein the polynucleotide is inserted into an expression vector.

4. A non-human transformant transformed with the polynucleotide according to claim 3 .

5. The transformant according to claim 4 , which is a whole plant.

6. An extract of the transformant according to claim 4 , wherein the extract contains monoterpene glycosides at higher concentrations than a wild-type counterpart.

7. A food, an aromatic, a pharmaceutical preparation or an industrial raw material, which comprises the extract according to claim 6 .

8. A method for producing a protein having glycosylation activity on a monoterpene compound, which comprises culturing the non-human transformant according to claim 4 .

9. A method for producing a monoterpene glycoside, which comprises reacting
(1) a recombinant protein of any one selected from the group consisting of (a) to (c) shown below:
(a) a recombinant protein which consists of the amino acid sequence shown in SEQ ID NO: 2, 4 or 6;
(b) a recombinant protein which consists of an amino acid sequence with deletion, substitution, insertion and/or addition of 1 to 70 amino acids in the amino acid sequence shown in SEQ ID NO: 2, 4 or 6 and which has glycosylation activity on a monoterpene compound; and
(c) a recombinant protein which has an amino acid sequence sharing a sequence identity of 85% or more with the amino acid sequence shown in SEQ ID NO: 2, 4 or 6 and which has glycosylation activity on a monoterpene compound,

(2) a UDP-sugar, and
(3) a monoterpene compound

to cause glycosylation of the monoterpene compound.

10. The method according to claim 9 , wherein the UDP-sugar is UDP-glucose.

11. The method according to claim 9 , wherein the monoterpene compound is geraniol or linalool.

Assignees

• Suntory Holdings Ltd

Inventors

• Ono Eiichiro
• Tsuruoka Nobuo

Classifications

• A61K8/64

  Proteins; Peptides; Derivatives or degradation products thereof · CPC title

• A61K31/7028

  Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages · CPC title

• A23L33/17

  Amino acids, peptides or proteins · CPC title

• C12N15/8243

  involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine · CPC title

• C12P19/18

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