# US 10,119,128 B2 · Lipase variants and polynucleotides encoding same

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- Number: US 10,119,128 B2
- Document ID: US10119128B2
- Status: Granted
- Kind: B2
- Application number: 15/177,879
- Filed: 2016-06-09
- Published: 2016-09-29
- Granted: 2018-11-06
- Earliest claimed: 2012-02-03
- Assignees: Novozymes A/S
- Applicants: Novozymes A/S
- Inventors: Luise Erlandsen; Carsten Hoerslev Hansen; Jesper Vind; Allan Svendsen; Carsten Peter Sonksen
- Web page: https://patentgrep.com/patent/US10119128B2

## CPC

- C12N 9/20: Triglyceride splitting, e.g. by means of lipase
- C11D 3/38627: {containing lipase}
- C11D 3/392: {Heterocyclic compounds, e.g. cyclic imides or lactames}
- C12N 9/18: Carboxylic ester hydrolases {(3.1.1)}
- C12Y 301/01: Carboxylic ester hydrolases (3.1.1)
- 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 lipase variants and methods of obtaining them. 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.

## Claims

### Claim 1

A lipase variant, comprising a substitution at a position corresponding to position G91A of the mature polypeptide of SEQ ID NO: 2, wherein the variant has at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 2, has lipase activity and, in comparison with the parent lipase, has improved performance in the presence of an organic catalyst selected from the group consisting of organic catalysts having the formulae (a)-(c):
and
c) mixtures thereof;
wherein each R1 is independently a branched alkyl group containing from 3 to 24 carbons or a linear alkyl group containing from 1 to 24 carbons; and
wherein said lipase variant is selected from the group consisting of the substitutions (d)-(g):
d) T37R+N39R+G91A+D96G+T231R+N233R;
e) G91A+D96G+T231R+N233R;
f) G91A+D96G+G225R+T231R+N233R; and
g) G91A+D96G+A150G+T231R+N233R.

### Claim 2 (depends on 1)

The variant of claim 1, which has at least 95% sequence identity to the amino acid sequence of the parent lipase.

### Claim 3 (depends on 1)

The variant of claim 1, which has at least 97% sequence identity to the amino acid sequence of the parent lipase.

### Claim 4 (depends on 1)

The variant of claim 1, which has at least 98% sequence identity to the amino acid sequence of the parent lipase.

### Claim 5 (depends on 1)

The variant of claim 1, which further comprises a substitution at one or more positions corresponding to positions D96, T143, A150, E210, G225, T231, N233 and P250.

### Claim 6 (depends on 1)

The variant of claim 1, wherein the substitution is selected from D96G, T143A, A150G, E210Q, G225R, T231R, N233R and P250R.

### Claim 7 (depends on 1)

The variant of claim 1, wherein R1 is independently selected from the group consisting of 2-propylheptyl, 2-butyloctyl, 2-pentylnonyl, 2-hexyldecyl, n-dodecyl, n-tetradecyl, n-hexadecyl, n-octadecyl, iso-nonyl, iso-decyl, iso-tridecyl, and iso-pantadecyl.