Protease variants and polynucleotides encoding same

Patent metadata

Field	Value
Publication number	US-9719077-B2
Application number	US-201414899242-A
Country	US
Kind code	B2
Filing date	Jul 28, 2014
Priority date	Jul 29, 2013
Publication date	Aug 1, 2017
Grant date	Aug 1, 2017

Abstract

The present invention relates to protease variants and methods for obtaining protease variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

First claim

The invention claimed is:

1. A protease variant, comprising a substitution at the position corresponding to position 297 of SEQ ID NO: 3 with Pro or Glu, wherein
the variant has a sequence identity to SEQ ID NO: 3 of at least 90% and less than 100%, and
the variant has protease activity.

2. The variant of claim 1 , wherein the substitution at the position corresponding to position 297 of SEQ ID NO: 3 is with Glu.

3. The variant of claim 1 , wherein the substitution at the position corresponding to position 297 of SEQ ID NO: 3 is with Pro.

4. The variant of claim 1 , has a sequence identity to SEQ ID NO: 3 of at least 95% and less than 100%.

5. The variant of claim 1 , which comprises one or more of the following substitutions: Y39D; T40{D,P}; Q70N; T74M; L81{F,H,V}; A102T; I121{S,C,V,T}; V124A; G132{l,E}; I137{E,C,S,A,M,T,Q,G}; S144{Q,R}; D155N; G159S; V162{W,R}; S171{W,K,E,N}; S173{P,Y}; G174{S,T,K}; S175{A,V,P}; N176G; T177S; G179{C,V,Q,S,T,E,H,K,M,N,Y,A}; F180Y; T241P; I247M; H256F; S274I or V286Q.

6. The variant of claim 1 , which is:
I137E S171N S173P G174K S175P N176G T177S F180Y T297P;
I137E S173P G174K S175P N176G T177S F180Y T297P;
I137E S173P G174T V175P T177S F180Y T297P;
I137E S173P G174T S175V T177S F180Y T297P;
I137E S173Y G174S S175A F180Y T297P; or
S173P G174K S175P N176G T177S F180Y T297P.

7. The variant of claim 1 , which has an improved detergent stability compared to the protease of SEQ ID NO: 3.

8. The variant of claim 1 , wherein the variant is encoded by a polynucleotide that hybridizes under high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) a sequence encoding the mature polypeptide of SEQ ID NO: 2, or (iii) the full-length complement of (i) or (ii).

9. The variant of claim 1 , wherein the variant is encoded by a polynucleotide having at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or a sequence encoding the mature polypeptide of SEQ ID NO: 2.

10. The variant of claim 1 , wherein the variant is a fragment of the mature polypeptide of SEQ ID NO: 2, which has protease activity.

11. The variant of claim 1 , wherein the total number of alterations compared to SEQ ID NO: 3 is 1-20.

12. A cleaning composition comprising a surfactant and a variant of claim 1 .

13. The composition of claim 12 , further comprising one or more additional enzymes selected from the group consisting of amylase, arabinase, carbohydrase, cellulase, cutinase, galactanase, lipase, mannanase, oxidase, pectinase, peroxidase, and/or xylanase.

14. The composition of claim 12 , which is the form of a bar, a homogenous tablet, a tablet having two or more layers, a pouch having one or more compartments, a regular or compact powder, a granule, a paste, a gel, or a regular, compact or concentrated liquid.

15. A method for removing a stain from a surface which comprises contacting the surface with a composition according to claim 12 .

Assignees

• Novozymes As

Inventors

• Rasmussen Frank Winther
• Lenhard Rolf Thomas
• Toscano Miguel Duarte Guilherme Perreira
• Friis Esben Peter
• Larsen Signe Eskildsen
• Knotzel Jurgen Carsten Franz
• Bauer Michael

Classifications

• C12Y304/21062

  Subtilisin (3.4.21.62) · CPC title

• C12N9/54 Primary

  bacteria being Bacillus · CPC title

• C11D3/38681

  Chemically modified or immobilised enzymes · CPC title

• C11D3/386

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