# US 8,569,035 B2 · Subtilase variants

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- Number: US 8,569,035 B2
- Document ID: US8569035B2
- Status: Granted
- Kind: B2
- Application number: 13/872,191
- Filed: 2013-04-29
- Published: 2013-08-29
- Granted: 2013-10-29
- Earliest claimed: 2001-07-12
- Assignees: Novozymes A/S
- Applicants: Novozymes A/S
- Inventors: Tina Sejersgard Fano; Claus Von Der Osten; Malene Kappen Kruger; Mads Norregaard-Madsen
- Web page: https://patentgrep.com/patent/US8569035B2

## CPC

- C12N 9/54: bacteria being Bacillus
- C11D 3/386: Preparations containing enzymes {, e.g. protease or amylase}
- C11D 3/38609: {Protease or amylase in solid compositions only}
- C11D 3/38618: {Protease or amylase in liquid compositions only}
- C11D 3/38627: {containing lipase}
- C11D 3/38636: {containing enzymes other than protease, amylase, lipase, cellulase, oxidase or reductase}
- C11D 3/38645: {containing cellulase}
- C11D 3/38654: {containing oxidase or reductase}
- C12N 9/52: derived from bacteria {or Archaea}
- C12Y 304/21062: Subtilisin (3.4.21.62)
- C11D 2111/12: Soft surfaces, e.g. textile
- C11D 2111/14: Hard surfaces
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C11D: DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C12Y: ENZYMES

## Abstract

The present invention relates to novel subtilase variants exhibiting alterations relative to the parent subtilase in one or more properties including: Wash performance, thermal stability, storage stability or catalytic activity. The variants of the invention are suitable for use in e.g. cleaning or detergent compositions, such as laundry detergent compositions and dishwash compositions, including automatic dishwash compositions.

## Claims

### Claim 1

A sub-group I-S2 subtilase variant having protease activity comprising at least one or more of the following alterations wherein each position corresponds to a position of the amino acid sequence set forth in SEQ ID NO:3: of subtilisin BPN′ and wherein the variant is:
V11I+I44V;
V11I+L96LA;
S9R+V11I+A15T+T22A;
V11I+L96LA+A108C+A138C;
V11I+N76D+A194P+A230V;
V11I+V84I;
V11I+V84I+K251R;
V11I+V30I;
V11I+V139L;
V30I;
V30I+V84I;
V30I+V139L;
S9R+A15T+T22A+V30I;
V30I+V244R;
V30I+K251R;
V30I+V139L+Y167A+R170S+A194P+N218S;
N76D+M175ML+A194P+A230V; and
N76D+M175MI+A194P+A230V.

### Claim 2 (depends on 1)

The variant subtilase according to claim 1, wherein the variant further comprises at least one modification at one or more of the positions corresponding to positions 27, 36, 56, 76, 87, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 120, 123, 159, 167, 170, 206, 218, 222, 224, 232, 235, 236, 245, 248, 252 or 274 of the amino acid sequence set forth in SEQ ID NO:3.

### Claim 3 (depends on 2)

The variant subtilase according to claim 2, wherein the variant further comprises the modification S101G+S103A+V1041+G159D+A232V+Q236H+Q245R+N248D+N252K.

### Claim 4 (depends on 1)

The variant subtilase according to claim 1, wherein the subtilase, before modification, is selected from the group consisting of BLS147, BLSAVI (Savinasee), BAPB92, TVTHER and BYSYAB.

### Claim 5 (depends on 4)

The variant subtilase according to claim 4, wherein the sub-group I-S2 subtilase, before modification has the amino acid sequence set forth in SEQ ID NO:4.

### Claim 6 (depends on 1)

A cleaning composition comprising the variant subtilase of claim 1.

### Claim 7 (depends on 6)

The composition according to claim 6 which is a laundry detergent composition or a dish wash detergent composition.

### Claim 8 (depends on 6)

A composition according to claim 6, which additionally comprises a cellulase, a lipase, a cutinase, an oxidoreductase, another protease, an amylase or a mixture thereof.