Acyl-ACP thioesterase

Patent metadata

Field	Value
Publication number	US-9828613-B2
Application number	US-201414897049-A
Country	US
Kind code	B2
Filing date	Jun 27, 2014
Priority date	Jul 12, 2013
Publication date	Nov 28, 2017
Grant date	Nov 28, 2017

Abstract

An acyl-ACP thioesterase consisting of an amino acid sequence of the 72 nd to 233 rd amino acids set forth in SEQ ID NO: 1, an acyl-ACP thioesterase gene encoding the protein, a transformant having the gene, and a method of producing a lipid using the transformant.

First claim

What is claimed is:

1. A method of producing a lipid, comprising the steps of:
culturing a transformant in a culture medium under conditions suitable for production of lipid to produce a cultured product, and
collecting the lipid from the resulting cultured product,
wherein the transformant is a host organism transformed with a heterologous gene encoding a protein selected from the group consisting of the following (a) to (c), wherein the host organism is a microorganism or a plant cell:
(a) A protein consisting of the amino acid sequence of:
the 72nd to 233rd amino acids set forth in SEQ ID NO: 1;
the amino acid sequence set forth in SEQ ID NO: 33: or
the 2nd to 167th amino acids set forth in SEQ ID NO: 33,

(b) A protein having acyl-ACP thioesterase activity and consisting of an amino acid sequence with 90% or more sequence identity to the amino acid sequence of the protein of (a); and
(c) A protein comprising the amino acid sequence of the protein of (a) or (b), and having acyl-ACP thioesterase activity.

2. The method of producing a lipid according to claim 1 wherein the protein is protein (c) and protein (c) is
a protein consisting of the amino acid sequence set forth in SEQ ID NO: 1,
a protein consisting of the amino acid sequence of the 32nd to 233rd amino acids set forth in SEQ ID NO: 1,
a protein consisting of the amino acid sequence of the 52nd to 233rd amino acids set forth in SEQ ID NO: 1,
a protein consisting of the amino acid sequence set forth in SEQ ID NO: 33, or
a protein consisting of the amino acid sequence of the 2nd to 167th amino acids set forth in SEQ ID NO: 33.

3. The method of producing a lipid according to claim 1 , wherein the lipid comprises fatty acids having 12 or more and 14 or less carbon atoms.

4. The method of producing a lipid according to claim 1 wherein the protein is encoded by a DNA selected from the group consisting of the following (d) to (j):
(d) A DNA consisting of the nucleotide sequence of the 214th to 699th nucleotides set forth in SEQ ID NO: 2;
(e) A DNA consisting of the nucleotide sequence set forth in SEQ ID NO: 2;
(f) A DNA consisting of the nucleotide sequence of the 94th to 699th nucleotides set forth in SEQ ID NO: 2;
(g) A DNA consisting of the nucleotide sequence of the 154th to 699th nucleotides set forth in SEQ ID NO: 2;
(h) A DNA consisting of the nucleotide sequence set forth in SEQ ID NO: 34;
(i) A DNA encoding a protein having acyl-ACP thioesterase activity and consisting of a nucleotide sequence with 90% or more sequence identity to the nucleotide sequence of a DNA selected from the group consisting of the DNAs of (d) to (g); and
(j) A DNA comprising the nucleotide sequence of a DNA selected from the group consisting of the DNAs of (d) to (i), and encoding a protein having acyl-ACP thioesterase activity.

5. The method of producing a lipid according to claim 1 wherein the host organism is a microorganism.

6. A method of modifying the fatty acid composition of a lipid, comprising
(i) transforming a host organism with a gene encoding a protein selected from the group consisting of the following (a) to (c), wherein the host organism is a microorganism or a plant cell:
(a) A protein consisting of the amino acid sequence of:
the 72nd to 233rd amino acids set forth in SEQ ID NO: 1;
the amino acid sequence set forth in SEQ ID NO: 33; or
the 2nd to 167th amino acids set forth in SEQ ID NO: 33;

(b) A protein having acyl-ACP thioesterase activity and consisting of an amino acid sequence with 90% or more sequence identity to the amino acid sequence of the protein of (a); and
(c) A protein comprising the amino acid sequence of the protein of (a) or (b), and having acyl-ACP thioesterase activity,

(ii) culturing the transformed host organism of part (i) in a culture medium;
(iii) expressing the protein encoded by the gene in the transformed host organism; and
(iv) collecting lipid produced by the transformed host organism during the culturing from the transformed host organism or from the culture medium;
wherein the fatty acid composition of the lipid that is collected in part (iv) is modified, as compared to that of a control host organism that is not transformed with the gene.

7. The method of modifying the fatty acid composition of a lipid according to claim 6 wherein the protein is protein (c) and protein (c) is
a protein consisting of the amino acid sequence set forth in SEQ ID NO: 1,
a protein consisting of the amino acid sequence of the 32nd to 233rd amino acids set forth in SEQ ID NO: 1,
a protein consisting of the amino acid sequence of the 52nd to 233rd amino acids set forth in SEQ ID NO: 1,
a protein consisting of the amino acid sequence set forth in SEQ ID NO: 33, or
a protein consisting of the amino acid sequence of the 2nd to 167th amino acids set forth in SEQ ID NO: 33.

8. The method of modifying the fatty acid composition of a lipid according to claim 6 wherein the protein is encoded by a DNA selected from the group consisting of the following (d) to (j):
(d) A DNA consisting of the nucleotide sequence of the 214th to 699th nucleotides set forth in SEQ ID NO: 2;
(e) A DNA consisting of the nucleotide sequence set forth in SEQ ID NO: 2;
(f) A DNA consisting of the nucleotide sequence of the 94th to 699th nucleotides set forth in SEQ ID NO: 2;
(g) A DNA consisting of the nucleotide sequence of the 154th to 699th nucleotides set forth in SEQ ID NO: 2;
(h) A DNA consisting of the nucleotide sequence set forth in SEQ ID NO: 34;
(i) A DNA encoding a protein having acyl-ACP thioesterase activity and consisting of a nucleotide sequence with 90% or more sequence identity to the nucleotide sequence of a DNA selected from the group consisting of the DNAs of (d) to (g); and
(j) A DNA comprising the nucleotide sequence of a DNA selected from the group consisting of the DNAs of (d) to (i), and encoding a protein having acyl-ACP thioesterase activity.

9. The method of modifying the fatty acid composition of a lipid according to claim 6 wherein the host organism is a microorganism.

10. A method of enhancing productivity of a lipid, comprising
(i) transforming a host organism with a gene encoding a protein selected from the group consisting of the following (a) to (c), wherein the host organism is a microorganism or a plant cell:
(a) A protein consisting of the amino acid sequence of:
the 72nd to 233rd amino acids set forth in SEQ ID NO: 1;
the amino acid sequence set forth in SEQ ID NO: 33; or
the 2nd to 167th amino acids set forth in SEQ ID NO: 33;

(b) A protein having acyl-ACP thioesterase activity and consisting of an amino acid sequence with 90% or more sequence identity to the amino acid sequence of the protein of (a); and
(c) A protein comprising the amino acid sequence of the protein of (a) or (b), and having acyl-ACP thioesterase activity,

(ii) culturing the transformed host organism of part (i) in a culture medium;
(iii) expressing the protein encoded by the gene in the transformed host organism; and
(iv) collecting lipid produced by the transformed host organism during the culturing from the transformed host organism or from the culture medium;
wherein production of the lipid that is collected in part (iv) is increased, as compared to that of a control host organism that is not transformed with the gene.

11. The method of enhancing productivity of a lipid according to claim 10 wherein the protein is protein (c) and protein (c) is
a protein consisting of the amino acid sequence set forth in SEQ ID NO: 1,
a protein consisting of the amino acid sequence of the 32nd to 233rd amino acids set forth in SEQ ID NO: 1,
a protein consisting of the amino a

Assignees

• Kao Corp

Inventors

• Ozaki Tatsuro

Classifications

• C12Y301/02014

  Oleoyl-[acyl-carrier-protein] hydrolase (3.1.2.14), i.e. ACP-thioesterase · CPC title

• C12N9/16 Primary

  acting on ester bonds (3.1) · CPC title

• C12P7/6409 Primary

  Fatty acids · CPC title

• C12P7/6463

  obtained from glyceride producing microorganisms, e.g. single cell oil · CPC title