Amylase variants

Patent metadata

Field	Value
Publication number	US-9856437-B2
Application number	US-201514619909-A
Country	US
Kind code	B2
Filing date	Feb 11, 2015
Priority date	Mar 8, 2000
Publication date	Jan 2, 2018
Grant date	Jan 2, 2018

Abstract

The present invention relates to variants (mutants) of polypeptides, in particular Termamyl-like alpha-amylases, which variant has alpha-amylase activity and exhibits an alteration in at least one of the following properties relative to said parent alpha-amylase: substrate specificity, substrate binding, substrate cleavage pattern, thermal stability, pH/activity profile, pH/stability profile, stability towards oxidation, Ca 2+ dependency, specific activity, and solubility, in particular under production conditions.

First claim

The invention claimed is:

1. An isolated variant alpha-amylase, comprising an alteration at one or more positions selected from the group consisting of:
R28, R118, N174; R181, G182, W189, N195, Y298, N299, K302, S303, N306, R310, N314; R320, H324, E345, Y396, R400, W439, R444, N445, K446, Q449, R458, N471, and N484,
wherein
(a) the alteration(s) are independently
(i) an insertion of an amino acid downstream of the amino acid which occupies the position;
(ii) a deletion of the amino acid which occupies the position; or
(iii) a substitution of the amino acid which occupies the position with a different amino acid;

(b) the variant has alpha-amylase activity and at least 90% sequence identity to SEQ ID NO: 12; and
(c) each position is numbered according to the amino acid sequence of SEQ ID NO: 12.

2. The isolated variant of claim 1 , which further comprises one or more mutations selected from the group consisting of:
Delta G184; Delta (R181-G182); Delta (D183-G184); R28N,K; S94K; R118K; N125A,R,K; N174D; R181Q,E,K; G186R; W189R,K; N195F; M202L; Y298H,F; N299A; K302R, 5303Q, N306G,D,R,K; R310A, K,Q, E, H, D, N; N314D; R320K; H324K; E345R, D, K, N; Y396F; R400T,K; W439R; R444K; N445K,Q; K446N; Q449E; R458K; N471E and N484Q.

3. The isolated variant of claim 1 , which further comprises a mutation at M202.

4. The isolated variant of claim 3 , wherein the mutation is M202L or M202T.

5. The isolated variant of claim 1 , wherein the mutation is at N195.

6. The isolated variant of claim 5 , wherein the mutation is N195F.

7. The isolated variant of claim 1 , wherein the mutation is at G186.

8. The isolated variant of claim 7 , wherein the mutation is G186R.

9. The isolated variant of claim 1 , wherein the mutation is at R181.

10. The isolated variant of claim 9 , wherein the mutation is R181Q.

11. The isolated variant of claim 1 , wherein the parent Termamyl-like alpha-amylase has an amino acid sequence of SEQ ID NOS: 2, 4, 6, 8, 10, 12, or 13.

12. The isolated variant of claim 1 , which variant has altered solubility under washing conditions.

13. A composition comprising the isolated variant of claim 1 .

14. A variant of a parent alpha-amylase, wherein: (a) the variant has at least 90% sequence identity to SEQ ID NO: 12, (b) the variant comprises a substitution at position 439 relative to the parent alpha-amylase, using the amino acid sequence of SEQ ID NO: 12 for determining position numbering, and (c) the variant has alpha-amylase activity.

15. The variant of claim 14 , wherein the variant has at least 95% sequence identity to SEQ ID NO: 12.

16. The variant of claim 14 , wherein the variant has at least 99% sequence identity to SEQ ID NO: 12.

17. The composition of claim 13 , which is a detergent composition.

Assignees

• Novozymes As

Inventors

• Andersen Carsten
• Borchert Torben Vedel
• Nielsen Bjarne Ronfeldt

Classifications

• C11D3/38681

  Chemically modified or immobilised enzymes · CPC title

• C12N9/2417

  from microbiological source · CPC title

• C11D3/386 Primary

  Preparations containing enzymes {, e.g. protease or amylase} · CPC title

• C11D3/38672

  Granulated or coated enzymes · CPC title