Method of Producing Lipid Using Acyl-ACP Thioesterase

Patent metadata

Field	Value
Publication number	US-2017335353-A1
Application number	US-201515520146-A
Country	US
Kind code	A1
Filing date	Nov 6, 2015
Priority date	Dec 5, 2014
Publication date	Nov 23, 2017
Grant date	—

Abstract

[Problems] To provide a method of producing a lipid, containing enhancing productivity of medium chain fatty acids or the lipid containing these medium chain fatty acids as components.
[Means to solve] A method of producing a lipid, containing the steps of:
culturing a transformant in which a gene encoding any one of the following proteins (A) to (C) is introduced into a host, and collecting a lipid from the cultured product: (A) a protein consisting of the amino acid sequence of the 611th to 772nd positions set forth in SEQ ID NO: 1; (B) a protein consisting of an amino acid sequence having 80% or more identity with the amino acid sequence of the 611th to 772nd positions set forth in SEQ ID NO: 1, and having acyl-ACP thioesterase activity; and (C) a protein containing the amino acid sequence of the protein (A) or (B), and having acyl-ACP thioesterase activity.

[Selected Drawing] None

First claim

What is claimed is:

1 . A method of producing a lipid, comprising the steps of:
culturing a transformant in which a gene encoding any one of the following proteins (A) to (C) is introduced into a host, and collecting a lipid from the cultured product:
(A) a protein consisting of the amino acid sequence of the 611th to 772nd positions set forth in SEQ ID NO: 1;
(B) a protein consisting of an amino acid sequence having 80% or more identity with the amino acid sequence of the 611th to 772nd positions set forth in SEQ ID NO: 1, and having acyl-ACP thioesterase activity; and
(C) a protein containing the amino acid sequence of the protein (A) or (B), and having acyl-ACP thioesterase activity.

2 . (canceled)

3 . A method of modifying the composition of a lipid, comprising the steps of:
introducing a gene encoding any one of the following proteins (A) to (C) into a host, and thereby obtaining a transformant, and enhancing productivity of medium chain fatty acids or a lipid containing the fatty acids as components produced in a cell of the transformant, to modify the composition of fatty acids or a lipid in all fatty acids or all lipids to be produced:
(A) a protein consisting of the amino acid sequence of the 611th to 772nd positions set forth in SEQ ID NO: 1;
(B) a protein consisting of an amino acid sequence having 80% or more identity with the amino acid sequence of the 611th to 772nd positions set forth in SEQ ID NO: 1, and having acyl-ACP thioesterase activity; and
(C) a protein containing the amino acid sequence of the protein (A) or (B), and having acyl-ACP thioesterase activity.

4 . The method according to claim 1 , wherein the protein (C) is any one of the following proteins (C1) to (C20):
(C1) a protein consisting of the amino acid sequence of the 1st to 772nd positions set forth in SEQ ID NO: 1; (C2) a protein consisting of the amino acid sequence of the 487th to 772nd positions set forth in SEQ ID NO: 1; (C3) a protein consisting of the amino acid sequence of the 488th to 772nd positions set forth in SEQ ID NO: 1; (C4) a protein consisting of the amino acid sequence of the 497th to 772nd positions set forth in SEQ ID NO: 1; (C5) a protein consisting of the amino acid sequence of the 507th to 772nd positions set forth in SEQ ID NO: 1; (C6) a protein consisting of the amino acid sequence of the 517th to 772nd positions set forth in SEQ ID NO: 1; (C7) a protein consisting of the amino acid sequence of the 527th to 772nd positions set forth in SEQ ID NO: 1; (C8) a protein consisting of the amino acid sequence of the 537th to 772nd positions set forth in SEQ ID NO: 1; (C9) a protein consisting of the amino acid sequence of the 547th to 772nd positions set forth in SEQ ID NO: 1; (C10) a protein consisting of the amino acid sequence of the 557th to 772nd positions set forth in SEQ ID NO: 1; (C11) a protein consisting of the amino acid sequence of the 567th to 772nd positions set forth in SEQ ID NO: 1; (C12) a protein consisting of the amino acid sequence of the 577th to 772nd positions set forth in SEQ ID NO: 1; (C13) a protein consisting of the amino acid sequence of the 587th to 772nd positions set forth in SEQ ID NO: 1; (C14) a protein consisting of the amino acid sequence of the 597th to 772nd positions set forth in SEQ ID NO: 1; (C15) a protein consisting of the amino acid sequence of the 607th to 772nd positions set forth in SEQ ID NO: 1; (C16) a protein consisting of the amino acid sequence of the 608th to 772nd positions set forth in SEQ ID NO: 1; (C17) a protein consisting of the amino acid sequence of the 609th to 772nd positions set forth in SEQ ID NO: 1; (C18) a protein consisting of the amino acid sequence of the 610th to 772nd positions set forth in SEQ ID NO: 1; (C19) a protein consisting of an amino acid sequence having 80% or more identity with the amino acid sequence of any one of the proteins (C1) to (C18), and having acyl-ACP thioesterase activity; and (C20) a protein consisting of an amino acid sequence in which 1 or several amino acids are deleted, substituted, inserted or added to the amino acid sequence of any one of the proteins (C1) to (C18), and having acyl-ACP thioesterase activity.

5 . The method according to claim 3 , wherein the protein (C) is any one of the following proteins (C1) to C20):
(C1) a protein consisting of the amino acid sequence of the 1st to 772nd positions set forth in SEQ ID NO: 1; (C2) a protein consisting of the amino acid sequence of the 487th to 772nd positions set forth in SEQ ID NO: 1; (C3) a protein consisting of the amino acid sequence of the 488th to 772nd positions set forth in SEQ ID NO: 1;
(C4) a protein consisting of the amino acid sequence of the 497th to 772nd positions set forth in SEQ ID NO: 1;
(C5) a protein consisting of the amino acid sequence of the 507th to 772nd positions set forth in SEQ ID NO: 1;
(C6) a protein consisting of the amino acid sequence of the 517th to 772nd positions set forth in SEQ ID NO: 1;
(C7) a protein consisting of the amino acid sequence of the 527th to 772nd positions set forth in SEQ ID NO: 1;
(C8) a protein consisting of the amino acid sequence of the 537th to 772nd positions set forth in SEQ ID NO: 1;
(C9) a protein consisting of the amino acid sequence of the 547th to 772nd positions set forth in SEQ ID NO: 1;
(C10) a protein consisting of the amino acid sequence of the 557th to 772nd positions set forth in SEQ ID NO: 1;
(C11) a protein consisting of the amino acid sequence of the 567th to 772nd positions set forth in SEQ ID NO: 1;
(C12) a protein consisting of the amino acid sequence of the 577th to 772nd positions set forth in SEQ ID NO: 1;
(C13) a protein consisting of the amino acid sequence of the 587th to 772nd positions set forth in SEQ ID NP: 1;
(C14) a protein consisting of the amino acid sequence of the 597th to 772nd positions set forth in SEQ ID NO: 1;
(C15) a protein consisting of the amino acid sequence of the 607th to 772nd positions set forth in SEQ ID NO: 1;
(C16) a protein consisting of the amino acid sequence of the 608th to 772nd positions set forth in SEQ ID NO: 1;
(C17) a protein consisting of the amino acid sequence of the 609th to 772nd positions set forth in SEQ ID NO: 1;
(C18) a protein consisting of the amino acid sequence of the 610th to 772nd positions set forth in SEQ ID NO: 1;
(C19) a protein consisting of an amino acid sequence having 80% or more identity with the amino acid sequence of any one of the proteins (C1) to (C18) and having acyl-ACP thioesterase activity; and
(C20) a protein consisting of an amino acid sequence in which 1 or several amino acids are deleted, substituted, inserted or added to the amino acid sequence of any one of the proteins (C1) to (C18) and having acyl-ACP thioesterase activity.

6 . The method according to claim 1 , wherein the host is Escherichia coli.

7 . The method according to claim 3 , wherein the host is Escherichia coli.

8 . The method according to claim 1 , wherein the host is a microalga.

9 . The method according to claim 8 , wherein the microalga is an alga belonging to the genus Nannochloropsis.

10 . The method according to claim 1 , wherein the lipid contains a fatty acid having 12 carbon atoms or a fatty acid ester compound thereof.

11 . The method according to claim 3 , wherein the host is a microalga.

12 . The method protein according to claim 11 , wherein the microalga is an alga belonging to the genus Nannochloropsis.

13 . The method according to claim 3 , wherein the lipid contains a fatty acid having 12 carbon atoms or a fatty acid ester compound thereof.

Assignees

• Kao Corp

Inventors

• Ozaki Tatsuro

Classifications

• C12N9/16 Primary

  acting on ester bonds (3.1) · CPC title

• C12Y301/02

  Thioester hydrolases (3.1.2) · CPC title

• C12P7/64 Primary

  Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats · CPC title

• C12P7/6409

  Fatty acids · CPC title

• C12Y301/02014

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