Detergent composition comprising protease variants

Patent metadata

Field	Value
Publication number	US-10072236-B2
Application number	US-201414907593-A
Country	US
Kind code	B2
Filing date	Jul 28, 2014
Priority date	Jul 29, 2013
Publication date	Sep 11, 2018
Grant date	Sep 11, 2018

Abstract

The present invention relates to detergent compositions comprising protease variants and methods for obtaining such detergent compositions. The present invention also relates to the use of such detergent compositions, especially in laundry or in hard surface cleaning applications.

First claim

The invention claimed is:

1. A detergent composition comprising a protease variant, the protease variant comprising a substitution of an amino acid in the loop corresponding to position 173 of SEQ ID NO: 3, wherein
(a) the variant has a sequence identity to SEQ ID NO: 3 of at least 70% and less than 100%, and
(b) the variant has protease activity, and
(c) the amino acid at the position corresponding to position 173 of SEQ ID NO: 3 is Pro.

2. The detergent composition of claim 1 , wherein the protease comprises substitutions at two positions corresponding to position 173 and any one of position 171, 175, 179, and 180.

3. The detergent composition of claim 2 , wherein in the protease variant
a) the amino acid at the position corresponding to position 171 of SEQ ID NO: 3 is selected from the group consisting of Trp, Lys, Glu, or
b) the amino acid at the position corresponding to position 175 of SEQ ID NO: 3 is Ala, Val, Pro, or
c) the amino acid at the position corresponding to position 179 of SEQ ID NO: 3 is selected from the group consisting of Cys, Val, Gln, Ser, Thr, Glu, His, Lys, Met, Asn.

4. The detergent composition of claim 2 , wherein in the protease variant the amino acid at the position corresponding to position 180 of SEQ ID NO: 3 is Tyr.

5. The detergent composition of claim 1 , wherein the protease variant comprises one or more substitutions selected from the group consisting of Y39D; T40{D,P}; Q7ON; T74M; L81{F,H,V}; A102T; I121{V,T}; G132 {I,E}; I137{M; E}; S144{Q,R}; D155N; G159S; V162R; G174{S,T}; N176G; T177S; T241P; I247M; H256F; S274I; V286Q; T297P.

6. The detergent composition of claim 1 , wherein the protease variant is selected from the variants that comprise any of the following substitutions compared to SEQ ID NO. 3:
S173P S274I
I137M S173P
S171N S173P
L81F S173P
L81H S173P
A102T S173P
S144Q S173P
I137E S173P
S173P S175P F180Y
S173P G174T S175V T177S F180Y
S173P G174K S175P N176G T177S F180Y
S173P T241P
V162R S173P G174T S175V T177S F180Y
I121V S173P G174T S175V T177S F180Y
L81V S173P G174T S175V T177S F180Y
A102T S173P G174T S175V T177S F180Y
S173P G174T S175V T177S F180Y T241P
I137E S173P G174T S175V T177S F180Y
S144Q S173P G174T S175V T177S F180Y
Q70N S173P G174T S175V T177S F180Y
D155N G159S S173P G174T S175V T177S F180Y
S173P G174T S175V T177S F180Y S274I
S173P G174T S175V T177S F180Y V286Q
T40D S173P G174T S175V T177S F180Y
S171N S173P G174T S175V T177S F180Y
V162R S173P G174K S175P N176G T177S F180Y
I121V S173P G174K S175P N176G T177S F180Y
L81V S173P G174K S175P N176G T177S F180Y
A102T S173P G174K S175P N176G T177S F180Y
S173P G174K S175P N176G T177S F180Y T241P
I137E S173P G174K S175P N176G T177S F180Y
S144Q S173P G174K S175P N176G T177S F180Y
Q70N S173P G174K S175P N176G T177S F180Y
D155N G159S S173P G174K S175P N176G T177S F180Y
S173P G174K S175P N176G T177S F180Y S274I
S173P G174K S175P N176G T177S F180Y V286Q
T40D S173P G174K S175P N176G T177S F180Y
S171N S173P G174K S175P N176G T177S F180Y
I121V I137E S173P G174T S175V T177S F180Y
L81V I137E S173P G174T S175V T177S F180Y
I137E S173P G174T S175V T177S F180Y T241P
Q70N I137E S173P G174T S175V T177S F180Y
I137E S173P G174T S175V T177S F180Y V286Q
I137E S173P G174T S175V T177S F180Y T297P
I137E S171N S173P G174T S175V T177S F180Y
I137E S173P G174T S175A T177S F180Y
I121V I137E S173P G174K S175P N176G T177S F180Y
Q70N I137E S173P G174K S175P N176G T177S F180Y
I137E S173P G174K S175P N176G T177S F180Y S274I
I137E S173P G174K S175P N176G T177S F180Y V286Q
I137E S173P G174K S175P N176G T177S F180Y T297P
I137E S171N S173P G174K S175P N176G T177S F180Y
I137E S173P G174K S175A N176G T177S F180Y
G1321 S173P G174T S175V T177S F180Y
S173P G174T S175V T177S F180Y T297P
S173P G174T S175A T177S F180Y
S173P S175A
I137E S173P S175V T177S F180Y
I137E S173P S175P N176G T177S F180Y
S173P G174K S175A N176G T177S F180Y
I137E S173P G174T S175V T177S F180Y S274I
T74M I137E S173P
T79I I137E S173P
L81G S173P
L34I I137E S173P
Y39D I137E S173P
T40P I137E S173P
I137E S173P I247M
I137E S173P H256F
I137E S144Q S173P G174T S175V T177S F180Y
I137E S144Q S173P G174K S175P N176G T177S F180Y
S173P G174T V175P T177S F180Y
I137E S173P G174T V175P T177S F180Y
I137E S173P G174T V175P T177S F180Y T297P
V162R S171N S173P G174K S175P N176G T177S F180Y
I137E S171N S173P G174K S175P N176G T177S F180Y T241P
I137E S171N S173P G174K S175P N176G T177S F180Y T297P
I137E S173P S175P F180Y
V162R S173P S175P F180Y
S173P S175P F180Y S274I
S173P S175P F180Y T241P
S173P S175P F180Y T297P
S173P S175P F180Y V286Q.

7. The detergent composition of claim 6 , wherein the protease variant has an improved detergent stability compared to the protease with SEQ ID NO: 3.

8. The detergent composition of claim 7 , the composition further comprising a borate and/or 4-formylphenyl boronic acid.

9. The detergent composition of claim 8 , wherein the protease variant is selected from the group consisting of:
a) a polypeptide having at least 70% sequence identity to the mature polypeptide of SEQ ID NO: 2;
b) a polypeptide encoded by a polynucleotide that hybridizes under medium, or 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);
c) a polypeptide encoded by a polynucleotide having at least 60% identity to the mature polypeptide coding sequence of SEQ ID NO: 1, or a sequence encoding the mature polypeptide of SEQ ID NO: 2; and
d) a fragment of the mature polypeptide of SEQ ID NO: 2, which has protease activity.

10. The detergent composition of claim 9 , wherein
(a) the protease variant has at least 75%, but less than 100%, sequence identity to the mature polypeptide of SEQ ID NO: 3; and
(b) in the protease the total number of alterations is from 1 to 20.

11. The detergent composition of claim 9 , wherein
(a) the protease variant has at least 85%, but less than 100%, sequence identity to the mature polypeptide of SEQ ID NO: 3; and
(b) in the protease the total number of alterations is from 1 to 10.

12. The detergent composition of claim 9 , wherein
(a) the protease variant has at least 90%, but less than 100%, sequence identity to the mature polypeptide of SEQ ID NO: 3; and
(b) in the protease the total number of alterations is from 1 to 5.

13. The detergent composition of claim 9 , wherein
(a) the protease variant has at least 95%, but less than 100%, sequence identity to the mature polypeptide of SEQ ID NO: 3; and
(b) in the protease the total number of alterations is from 1 to 5.

14. The detergent composition of claim 9 , wherein
(a) the protease variant has at least 99%, but less than 100%, sequence identity to the mature polypeptide of SEQ ID NO: 3; and
(b) in the protease the total number of alterations is from 1 to 5.

15. The detergent composition of claim 9 , wherein
(a) the composition is in 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; and/or
(b) the composition further comprises one or more additional enzymes selected from protease, lipase, cutinase, amylase, carbohydrase, cellulase, pectinase, mannanase, arabinase, galactanase, xylanase, oxidase, laccase, and peroxidase.

16. The detergent composition of claim 9 , wherein the composition is a laundry detergent compos

Assignees

• Henkel Ag & Co Kgaa

Inventors

• O'Connell Timothy
• Tondera Susanne
• Hellmuth Hendrik
• Weber Thomas
• Rasmussen Frank Winther
• Lenhard Rolf Thomas
• Toscano Miguel Duarte Guilherme Pereira
• Friis Esben
• Larsen Signe Eskildsen
• Knoetzel Juergen Carsten Franz
• Bauer Mikael

Classifications

• C12Q1/37

  involving peptidase or proteinase · CPC title

• C12Y304/21062

  Subtilisin (3.4.21.62) · CPC title

• C12N9/54

  bacteria being Bacillus · CPC title

• C12Y304/21

  Serine endopeptidases (3.4.21) · CPC title

• C11D3/386

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