Carotenoid production method

Patent metadata

Field	Value
Publication number	US-11268121-B2
Application number	US-201716088058-A
Country	US
Kind code	B2
Filing date	Mar 29, 2017
Priority date	Mar 31, 2016
Publication date	Mar 8, 2022
Grant date	Mar 8, 2022

Abstract

A mutant carotenoidogenic bacterium, comprising any of genes (a)-(c) below: (a) a gene encoding a protein comprising a mutant amino acid sequence in which at least the 225th amino acid residue in the amino acid sequence of 1-deoxy-D-xylulose 5-phosphate synthase of a carotenoidogenic bacterium has been substituted with other amino acid residue; (b) a gene encoding a protein comprising a mutant amino acid sequence in which at least the 305th amino acid residue in the amino acid sequence of decaprenyl diphosphate synthase of a carotenoidogenic bacterium has been substituted with other amino acid residue; and (c) both of the genes (a) and (b) above.

First claim

The invention claimed is:

1. A mutant carotenoidogenic bacterium that produces a carotenoid, comprising a combination of
(i) a gene selected from (a)-(c), and
(ii) a gene selected from (d)-(f); which are:
(a) a gene encoding a protein comprising a mutant amino acid sequence in which at least glycine of the 225th amino acid residue in the amino acid sequence of SEQ ID NO:2 of 1-deoxy-D-xylulose 5-phosphate synthase (DXS) of a carotenoidogenic bacterium belonging to the genus Paracoccus has been substituted with aspartic acid;
(b) a gene encoding a protein comprising a mutant amino acid sequence in which glycine of the 225th amino acid residue in the amino acid sequence of SEQ ID NO:2 of DXS is substituted with aspartic acid and in which one to several amino acid residues other than the 225th amino acid residue have been deleted, substituted or added, and that has DXS activity;
(c) a gene consisting of DNA comprising the nucleotide sequence of SEQ ID NO:5;
(d) a gene encoding a protein comprising a mutant amino acid sequence in which at least alanine of the 305th amino acid residue in the amino acid sequence of SEQ ID NO:4 of decaprenyl diphosphate synthase (DPS) of a carotenoidogenic bacterium belonging to the genus Paracoccus has been substituted with valine;
(e) a gene encoding a protein comprising a mutant amino acid sequence in which alanine of the 305th amino acid residue in the amino acid sequence of SEQ ID NO:4 of DPS is substituted with valine and in which one to several amino acid residues other than the 305th amino acid residue have been deleted, substituted or added, and that has reduced DPS activity; and
(f) a gene consisting of DNA comprising the nucleotide sequence of SEQ ID NO:7.

2. The bacterium according to claim 1 , which has acquired carotenogenic capacity that is higher than the carotenogenic capacity of a carotenoidogenic bacterium without the gene encoding the protein comprising the mutant amino acid sequence.

3. The bacterium according to claim 2 , which has acquired carotenogenic capacity that is at least 5 times or more the carotenoid production amount of a carotenoidogenic bacterium without the gene encoding the protein comprising the mutant amino acid sequence.

4. The bacterium according to claim 1 , wherein the bacterium belonging to the genus Paracoccus is strain E-396.

5. The bacterium according to claim 1 , wherein the carotenoid is astaxanthin.

6. A method for producing a carotenoid, comprising culturing the bacterium according to claim 1 , and collecting the carotenoid from the resulting cultured product.

7. The method according to claim 6 , wherein the carotenoid production amount is at least 5 times or more the carotenoid production amount of a carotenoidogenic bacterium without the gene encoding the protein comprising the mutant amino acid sequence.

8. The method according to claim 1 , wherein the carotenoid is astaxanthin.

9. A recombinant vector comprising a combination of
(i) a gene selected from (a)-(c), and
(ii) a gene selected from (d)-(f); which are:
(a) a gene encoding a protein comprising a mutant amino acid sequence in which at least glycine of the 225th amino acid residue in the amino acid sequence of SEQ ID NO:2 of 1-deoxy-D-xylulose 5-phosphate synthase (DXS) of a carotenoidogenic bacterium belonging to the genus Paracoccus has been substituted with aspartic acid;
(b) a gene encoding a protein comprising a mutant amino acid sequence in which glycine of the 225th amino acid residue in the amino acid sequence of SEQ ID NO:2 of DXS is substituted with aspartic acid and in which one to several amino acid residues other than the 225th amino acid residue have been deleted, substituted or added, and that has DXS activity;
(c) a gene consisting of DNA comprising the nucleotide sequence of SEQ ID NO:5;
(d) a gene encoding a protein comprising a mutant amino acid sequence in which at least alanine of the 305th amino acid residue in the amino acid sequence of SEQ ID NO:4 of decaprenyl diphosphate synthase (DPS) of a carotenoidogenic bacterium belonging to the genus Paracoccus has been substituted with valine;
(e) a gene encoding a protein comprising a mutant amino acid sequence in which alanine of the 305th amino acid residue in the amino acid sequence of SEQ ID NO:4 of DPS is substituted with valine and in which one to several amino acid residues other than the 305th amino acid residue have been deleted, substituted or added, and that has reduced DPS activity; and
(f) a gene consisting of DNA comprising the nucleotide sequence of SEQ ID NO:7.

10. A transformant comprising the recombinant vector according to claim 9 .

11. A method for producing a carotenoid, comprising culturing the transformant according to claim 10 , and collecting a carotenoid from the resulting cultured product.

Assignees

• Jxtg Nippon Oil & Energy Corp
• Eneos Corp

Inventors

• Nagai Hidetada
• Sato Wataru
• Takahashi Toshiyuki
• Sato Harumi

Classifications

• C12P23/00 Primary

  Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes (containing heterorings C12P17/00) · CPC title

• C12Y205/01091

  All-trans-decaprenyl-diphosphate synthase (2.5.1.91) · CPC title

• C12N9/1022

  transferring aldehyde or ketonic groups (2.2) · CPC title

• C12N15/74 Primary

  Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora · CPC title

• C12Y205/01086

  Trans,polycis-decaprenyl diphosphate synthase (2.5.1.86) · CPC title