# US 11,078,445 B2 · Compositions comprising lipase and sulfite

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- Number: US 11,078,445 B2
- Document ID: US11078445B2
- Status: Granted
- Kind: B2
- Application number: 16/608,694
- Filed: 2018-05-04
- Published: 2020-02-20
- Granted: 2021-08-03
- Earliest claimed: 2017-05-05
- Assignees: Novozymes A/S
- Applicants: Novozymes A/S
- Inventors: Jesper Vind; Thomas A. Poulsen; Miguel D G P Toscano
- Web page: https://patentgrep.com/patent/US11078445B2

## CPC

- C11D 3/38663: {Stabilised liquid enzyme compositions}
- C11D 3/046: {Salts}
- C11D 3/38627: {containing lipase}
- C12N 9/20: Triglyceride splitting, e.g. by means of lipase
- C12Y 301/01003: Triacylglycerol lipase (3.1.1.3)
- C11D 2111/12: Soft surfaces, e.g. textile
- C11D: DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES

## Abstract

The present invention relates to a composition comprising: (a) a source of sulfite; and (b) a lipase variant of a parent lipase, wherein one or more amino acids in the parent lipase within 20 angstrom (Å) of a cysteine bridge are replaced with a more negatively charged amino acid.

## Claims

### Claim 1

A composition comprising:
(a) a source of sulfite; and
(b) a lipase variant which
(i) comprises a substitution at one or more positions corresponding to positions 35, 39, 42, and 105, of SEQ ID NO: 2 with a more negatively charged amino acid;
(ii) has at least 80% but less than 100% sequence identity to SEQ ID NO: 2; and
(iii) has lipase activity.

### Claim 2 (depends on 1)

The composition of claim 1, wherein the lipase variant has at least 85% sequence identity to SEQ ID NO: 2.

### Claim 3 (depends on 1)

The composition of claim 1, wherein the lipase variant has at least 90% sequence identity to SEQ ID NO: 2.

### Claim 4 (depends on 1)

The composition of claim 1, wherein the lipase variant has at least 95% sequence identity to SEQ ID NO: 2.

### Claim 5 (depends on 1)

The composition of claim 1, wherein the lipase variant comprises one or more of the following substitutions: T35D, E; N39D, E; P42D, E; or S105D, E of SEQ ID NO: 2.

### Claim 6 (depends on 1)

The composition of claim 1, wherein the lipase variant comprises T35D.

### Claim 7 (depends on 1)

The composition of claim 1, wherein the lipase variant comprises N39E.

### Claim 8 (depends on 1)

The composition of claim 1, wherein the lipase variant comprises P42D.

### Claim 9 (depends on 1)

The composition of claim 1, wherein the lipase variant comprises S105E.

### Claim 10 (depends on 1)

The composition of claim 1, wherein the lipase variant further comprises a substitution at one or more positions corresponding to positions 4, 27, 38, 57, 58, 60, 83, 86, 91, 94, 97, 99, 111, 150, 163, 210, 216, 225, 227, 231, 233, 249, 254, 255, 256, 263, 264, 265, 266, 267, and 269 of SEQ ID NO: 2.

### Claim 11 (depends on 10)

The composition of claim 10, wherein the substitution is selected from the group consisting of 4V, 27R, 38A, 57G, S58A, 60S, 83T, 86V, 91A/N/Q, 94K/R, 97M, 99K, 111A, 150G, 163K, 210K/Q, 216P, 225R, L227G, 231R, 233R, 249R, 254S, 255A, 256K/T/V, 263Q, 264A, 265T, 266D, 267A, and 269N.

### Claim 12 (depends on 1)

The composition of claim 1, wherein the lipase variant has one or more cysteine bridges at positions corresponding to positions C22-C268, C36-C41, and C104-C107 of SEQ ID NO: 2.

### Claim 13 (depends on 1)

The composition of claim 1, wherein the lipase variant has improved stability in the presence of the source of sulfite as compared to the lipase of SEQ ID NO: 2.

### Claim 14 (depends on 1)

The composition of claim 1, wherein the source of sulfite is sodium sulfite, potassium sulfite; calcium sulfite; potassium bisulfite sodium bisulfite, calcium bisulfite; potassium metabisulfite, sodium metabisulfite, calcium metabisulfite; or any combination thereof.

### Claim 15 (depends on 14)

The composition of claim 14, wherein the amount of sulfite (as SO32−) or metabisulfite (as S2O52−) is from 0.1 wt. % to 3 wt. %.

### Claim 16 (depends on 1)

The composition of claim 1, further comprising a surfactant.

### Claim 17 (depends on 1)

A method of cleaning a surface, comprising treating the surface with a composition of claim 1.

### Claim 18 (depends on 17)

The method of claim 17, wherein the surface is a textile or hard surface.