Modified lipase and use thereof

Patent metadata

Field	Value
Publication number	US-10870840-B2
Application number	US-201916522803-A
Country	US
Kind code	B2
Filing date	Jul 26, 2019
Priority date	Dec 10, 2013
Publication date	Dec 22, 2020
Grant date	Dec 22, 2020

Abstract

The present invention addresses a problem of providing a lipase derived from a microorganism that is specific for short-chain to medium-chain fatty acids. A modified lipase is obtained by making a substitution in the amino acid sequence of a Candida cylindracea derived lipase, wherein the substitution is (1) a substitution of asparagine for an amino acid corresponding to the amino acid at position 428 in the amino acid sequence set forth in SEQ ID NO: 1; or (2) a substitution of phenylalanine, methionine, or isoleucine for an amino acid corresponding to the amino acid at position 429 in the amino acid sequence set forth in SEQ ID NO: 1.

First claim

The invention claimed is:

1. A modified lipase consisting of an amino acid sequence with a substitution made in the amino acid sequence of a lipase, wherein the substitution is:
(1) a substitution of asparagine for an amino acid corresponding to the amino acid at position 428 in the amino acid sequence set forth in SEQ ID NO: 1; or
(2) a substitution of phenylalanine, methionine, or isoleucine for an amino acid corresponding to the amino acid at position 429 in the amino acid sequence set forth in SEQ ID NO:1,
wherein the amino acid sequence of the lipase is an amino acid sequence that is 90% or more identical to the amino acid sequence of SEQ ID NO. 3 and wherein the substitution is the substitution represented in (1), or
wherein the amino acid sequence of the lipase is an amino acid sequence that is 90% or more identical to the amino acid sequence of SEQ ID NO. 3 and wherein the substitution is the substitution represented in (2).

2. The modified lipase according to claim 1 , wherein the amino acid sequence of the lipase is an amino acid sequence of SEQ ID NO. 3.

3. A gene encoding the modified lipase according to claim 1 .

4. The gene according to claim 3 , comprising the polynucleotide sequence set forth in SEQ ID No. 14.

5. A recombinant DNA comprising the gene according to claim 3 .

6. A microorganism carrying the recombinant DNA according to claim 5 .

7. The microorganism according to claim 6 , wherein the host is Escherichia coli, Candida cylindracea, Aspergillus oryzae, Bacillus subtilis , or Pichia pastoris.

8. An enzyme preparation comprising the modified lipase according to claim 1 .

9. A method for improving the flavor of a food product or food raw material, wherein the enzyme according to claim 1 is allowed to act on the food product or food raw material.

10. A method for producing a food product, wherein the enzyme according to claim 1 is allowed to act on a food raw material or intermediate product.

11. The method according to claim 9 , wherein the food product is a dairy product.

12. A flavor-improving agent that is allowed to act on a food product or food raw material, comprising the modified lipase according to claim 1 .

13. A food product or food raw material obtained by treatment with the enzyme according to claim 1 .

14. The method according to claim 10 , wherein the food product is a dairy product.

15. A flavor-improving agent that is allowed to act on a food product or food raw material, comprising the enzyme preparation according to claim 8 .

16. A modified lipase consisting of an amino acid sequence with a substitution made in the amino acid sequence of a lipase, wherein the substitution is:
(1) a substitution of asparagine for an amino acid corresponding to the amino acid at position 428 in the amino acid sequence set forth in SEQ ID NO: 1; or
(2) a substitution of phenylalanine, methionine, or isoleucine for an amino acid corresponding to the amino acid at position 429 in the amino acid sequence set forth in SEQ ID NO:1,
wherein the amino acid sequence of the lipase is an amino acid sequence that is 95% or more identical to the amino acid sequence of SEQ ID NO: 2 and wherein the substitution is the substitution represented in (1), or
wherein the amino acid sequence of the lipase is an amino acid sequence that is 95% or more identical to the amino acid sequence of SEQ ID NO: 2 and wherein the substitution is the substitution represented in (2).

Assignees

• Amano Enzyme Inc

Inventors

• Ishigaki Yuki
• Ishihara Satoru
• Tanaka Shun-Ichi

Classifications

• C12Y301/01003

  Triacylglycerol lipase (3.1.1.3) · CPC title

• A23V2002/00

  Food compositions, function of food ingredients or processes for food or foodstuffs · CPC title

• C12N9/20 Primary

  Triglyceride splitting, e.g. by means of lipase · CPC title

• A23C9/1216

  Other enzymes · CPC title

• A23L27/25

  Dairy flavours · CPC title