US 2020/0270650 A1 · Method of Producing Lipid

Abstract

A method of producing lipids, containing the steps of: culturing a transformant into which a gene encoding at least one protein selected from the group consisting of the following proteins (A) to (C) is introduced; and producing fatty acids or lipids containing the same as components: (A) a protein consisting of an amino acid sequence having at least one amino acid substitution selected from the group consisting of the following (A-1) to (A-11) in the amino acid sequence set forth in SEQ ID NO: 1, and having acyl-ACP thioesterase activity; (B) a protein consisting of an amino acid sequence having at least one amino acid substitution selected from the group consisting of the following (B-1) to (B-11) in an amino acid sequence having 85% or more identity with the amino acid sequence 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: (A-1) substitution of isoleucine for leucine at position 257 of the amino acid sequence set forth in SEQ ID NO: 1; (A-2) substitution of arginine for threonine at position 251 of the amino acid sequence set forth in SEQ ID NO: 1; (A-3) substitution of lysine for threonine at position 251 of the amino acid sequence set forth in SEQ ID NO: 1; (A-4) substitution of histidine for threonine at position 251 of the amino acid sequence set forth in SEQ ID NO: 1; (A-5) substitution of isoleucine for tryptophan at position 254 of the amino acid sequence set forth in SEQ ID NO: 1; (A-6) substitution of tyrosine for tryptophan at position 254 of the amino acid sequence set forth in SEQ ID NO: 1; (A-7) substitution of methionine for leucine at position 257 of the amino acid sequence set forth in SEQ ID NO: 1; (A-8) substitution of valine for leucine at position 257 of the amino acid sequence set forth in SEQ ID NO: 1; (A-9) substitution of phenylalanine for leucine at position 257 of the amino acid sequence set forth in SEQ ID NO: 1; (A-10) substitution of cysteine for valine at position 266 of the amino acid sequence set forth in SEQ ID NO: 1; (A-11) substitution of tyrosine for tryptophan at position 271 of the amino acid sequence set forth in SEQ ID NO: 1; (B-1) substitution of isoleucine for an amino acid at a position corresponding to position 257 of the amino acid sequence set forth in SEQ ID NO: 1; (B-2) substitution of arginine for an amino acid at a position corresponding to position 251 of the amino acid sequence set forth in SEQ ID NO: 1; (B-3) substitution of lysine for an amino acid at a position corresponding to position 251 of the amino acid sequence set forth in SEQ ID NO: 1; (B-4) substitution of histidine for an amino acid at a position corresponding to position 251 of the amino acid sequence set forth in SEQ ID NO: 1; (B-5) substitution of isoleucine for an amino acid at a position corresponding to position 254 of the amino acid sequence set forth in SEQ ID NO: 1; (B-6) substitution of tyrosine for an amino acid at a position corresponding to position 254 of the amino acid sequence set forth in SEQ ID NO: 1; (B-7) substitution of methionine for an amino acid at a position corresponding to position 257 of the amino acid sequence set forth in SEQ ID NO: 1; (B-8) substitution of valine for an amino acid at a position corresponding to position 257 of the amino acid sequence set forth in SEQ ID NO: 1; (B-9) substitution of phenylalanine for an amino acid at a position corresponding to position 257 of the amino acid sequence set forth in SEQ ID NO: 1; (B-10) substitution of cysteine for an amino acid at a position corresponding to position 266 of the amino acid sequence set forth in SEQ ID NO: 1; and (B-11) substitution of tyrosine for an amino acid at a position corresponding to position 271 of the amino acid sequence set forth in SEQ ID NO: 1.

Claims18

Dependent claims

A method of producing lipids, comprising the steps of:

culturing a transformant into which a gene encoding at least one protein selected from the group consisting of the following proteins (A) to (C) is introduced;
and
producing fatty acids or lipids containing the same as components:
(A) a protein consisting of an amino acid sequence having at least one amino acid substitution selected from the group consisting of the following (A-1) to (A-11) in the amino acid sequence set forth in SEQ ID NO:
1, and having acyl-ACP thioesterase activity;
(B) a protein consisting of an amino acid sequence having at least one amino acid substitution selected from the group consisting of the following (B-1) to (B-11) in an amino acid sequence having 85% or more identity with the amino acid sequence 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:
(A-1) substitution of isoleucine for leucine at position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-2) substitution of arginine for threonine at position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-3) substitution of lysine for threonine at position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-4) substitution of histidine for threonine at position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-5) substitution of isoleucine for tryptophan at position 254 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-6) substitution of tyrosine for tryptophan at position 254 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-7) substitution of methionine for leucine at position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-8) substitution of valine for leucine at position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-9) substitution of phenylalanine for leucine at position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-10) substitution of cysteine for valine at position 266 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-11) substitution of tyrosine for tryptophan at position 271 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-1) substitution of isoleucine for an amino acid at a position corresponding to position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-2) substitution of arginine for an amino acid at a position corresponding to position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-3) substitution of lysine for an amino acid at a position corresponding to position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-4) substitution of histidine for an amino acid at a position corresponding to position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-5) substitution of isoleucine for an amino acid at a position corresponding to position 254 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-6) substitution of tyrosine for an amino acid at a position corresponding to position 254 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-7) substitution of methionine for an amino acid at a position corresponding to position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-8) substitution of valine for an amino acid at a position corresponding to position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-9) substitution of phenylalanine for an amino acid at a position corresponding to position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-10) substitution of cysteine for an amino acid at a position corresponding to position 266 of the amino acid sequence set forth in SEQ ID NO:
1;
and
(B-11) substitution of tyrosine for an amino acid at a position corresponding to position 271 of the amino acid sequence set forth in SEQ ID NO:
1.

51 more elements

A protein, selected from the following proteins (A) to (C):

(A) a protein consisting of an amino acid sequence having at least one amino acid substitution selected from the group consisting of the following (A-1) to (A-11) in the amino acid sequence set forth in SEQ ID NO:
1, and having acyl-ACP thioesterase activity;
(B) a protein consisting of an amino acid sequence having at least one amino acid substitution selected from the group consisting of the following (B-1) to (B-11) in an amino acid sequence having 85% or more identity with the amino acid sequence 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:
(A-1) substitution of isoleucine for leucine at position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-2) substitution of arginine for threonine at position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-3) substitution of lysine for threonine at position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-4) substitution of histidine for threonine at position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-5) substitution of isoleucine for tryptophan at position 254 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-6) substitution of tyrosine for tryptophan at position 254 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-7) substitution of methionine for leucine at position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-8) substitution of valine for leucine at position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-9) substitution of phenylalanine for leucine at position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-10) substitution of cysteine for valine at position 266 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-11) substitution of tyrosine for tryptophan at position 271 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-1) substitution of isoleucine for an amino acid at a position corresponding to position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-2) substitution of arginine for an amino acid at a position corresponding to position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-3) substitution of lysine for an amino acid at a position corresponding to position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-4) substitution of histidine for an amino acid at a position corresponding to position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-5) substitution of isoleucine for an amino acid at a position corresponding to position 254 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-6) substitution of tyrosine for an amino acid at a position corresponding to position 254 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-7) substitution of methionine for an amino acid at a position corresponding to position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-8) substitution of valine for an amino acid at a position corresponding to position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-9) substitution of phenylalanine for an amino acid at a position corresponding to position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-10) substitution of cysteine for an amino acid at a position corresponding to position 266 of the amino acid sequence set forth in SEQ ID NO:
1;
and
(B-11) substitution of tyrosine for an amino acid at a position corresponding to position 271 of the amino acid sequence set forth in SEQ ID NO:
1.

48 more elements

A gene encoding a protein selected from the following proteins (A) to (C):

(A) a protein consisting of an amino acid sequence having at least one amino acid substitution selected from the group consisting of the following (A-1) to (A-11) in the amino acid sequence set forth in SEQ ID NO:
1, and having acyl-ACP thioesterase activity;
(B) a protein consisting of an amino acid sequence having at least one amino acid substitution selected from the group consisting of the following (B-1) to (B-11) in an amino acid sequence having 85% or more identity with the amino acid sequence 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:
(A-1) substitution of isoleucine for leucine at position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-2) substitution of arginine for threonine at position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-3) substitution of lysine for threonine at position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-4) substitution of histidine for threonine at position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-5) substitution of isoleucine for tryptophan at position 254 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-6) substitution of tyrosine for tryptophan at position 254 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-7) substitution of methionine for leucine at position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-8) substitution of valine for leucine at position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-9) substitution of phenylalanine for leucine at position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-10) substitution of cysteine for valine at position 266 of the amino acid sequence set forth in SEQ ID NO:
1;
(A-11) substitution of tyrosine for tryptophan at position 271 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-1) substitution of isoleucine for an amino acid at a position corresponding to position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-2) substitution of arginine for an amino acid at a position corresponding to position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-3) substitution of lysine for an amino acid at a position corresponding to position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-4) substitution of histidine for an amino acid at a position corresponding to position 251 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-5) substitution of isoleucine for an amino acid at a position corresponding to position 254 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-6) substitution of tyrosine for an amino acid at a position corresponding to position 254 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-7) substitution of methionine for an amino acid at a position corresponding to position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-8) substitution of valine for an amino acid at a position corresponding to position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-9) substitution of phenylalanine for an amino acid at a position corresponding to position 257 of the amino acid sequence set forth in SEQ ID NO:
1;
(B-10) substitution of cysteine for an amino acid at a position corresponding to position 266 of the amino acid sequence set forth in SEQ ID NO:
1;
and
(B-11) substitution of tyrosine for an amino acid at a position corresponding to position 271 of the amino acid sequence set forth in SEQ ID NO:
1.

48 more elements

Dependencies are inferred from the claim text.

Method of Producing Lipid

Life of this patent

Method of Producing Lipid

Life of this patent

Abstract

Claims18