Stabilized humicola lanuginosa lipase variants in water-soluble films

Patent metadata

Field	Value
Publication number	US-10457920-B2
Application number	US-201414891199-A
Country	US
Kind code	B2
Filing date	May 14, 2014
Priority date	May 14, 2013
Publication date	Oct 29, 2019
Grant date	Oct 29, 2019

Abstract

The invention relates to a lipase containing water-soluble film, and a detergent pouch formed by the water-soluble film.

First claim

The invention claimed is:

1. A water-soluble film comprising a variant of a parent lipase, which variant has lipase activity, has at least 60% but less than 100% sequence identity with SEQ ID NO: 2,and comprises substitutions at positions corresponding to T231R+N233R and at least one or more of D96E, D111A, D254S, G163K, P256T, G91T and G38A of SEQ ID NO: 2.

2. The water-soluble film of claim 1 , wherein the variant further comprises substitutions at positions corresponding to D27R and/or N33Q of SEQ ID NO: 2.

3. The water-soluble film of claim 1 , wherein the variant comprises substitutions of SEQ ID NO: 2 selected from the group consisting of:
a) D96E+T231R+N233R;
b) N33Q+D96E+T231R+N233R;
c) N33Q+D111A+T231R+N233R;
d) N33Q+T231R+N233R+P256T;
e) N33Q+G38A+G91T+G163K+T231R+N233R+D254S;
f) N33Q+G38A+G91T+D96E+D111A+G163K+T231R+N233R+D254S+P256T;
g) D27R+N33Q+G38A+D96E+D111A+G163K+T231R+N233R+D254S+P256T;
h) D27R+N33Q+G38A+G91T+D96E+D111A+G163K+T231R+N233R+P256T;
i) D27R+N33Q+G38A+G91T+D96E+D111A+G163K+T231R+N233R+D254S;
j) D27R+G38A+G91T+D96E+D111A+G163K+T231R+N233R+D254S+P256T;
k) D96E+T231R+N233R+D254S;
I) T231R+N233R+D254S+P256T;
m) G163K+T231R+N233R+D254S;
n) D27R+N33Q+G38A+G91T+D96E+G163K+T231R+N233R+D254S+P256T;
o) D27R+G91T+D96E+D111A+G163K+T231R+N233R+D254S+P256T;
p) D96E+G163K+T231R+N233R+D254S;
q) D27R+G163K+T231R+N233R+D254S;
r) D27R+G38A+G91T+D96E+D111A+G163K+T231R+N233R+D254S;
s) D27R+G38A+G91T+D96E+G163K+T231R+N233R+D254S+P256T;
t) D27R+G38A+D96E+D111A+G163K+T231R+N233R+D254S+P256T:
u) D27R+D96E+G163K+T231R+N233R+D254S;
v) D27R+D96E+D111A+G163K+T231R+N233R+D254S+P256T;
w) D27R+G38A+D96E+G163K+T231R+N233R+D254S+P256T;
x) D111A+G163K+T231R+N233R+D254S+P256T;
y) D111A+T231R+N233R;
z) D111A+T231R+N233R+D254S+P256T;
aa) D27R+D96E+D111A+G163K+T231R+N233R;
bb) D27R+D96E+D111A+T231R+N233R;
cc) D27R+G38A+D96E+D111A+G163K+T231R+N233R+D254S+P256T;
dd) D27R+N33Q+G38A+D96E+D111A+T231R+N233R+D254S+P256T;
ee) D27R+G38A+D96E+D111A+G163K+E210Q+T231R+N233R+D254S+P256T;
ff) D27R+T231R+N233R+D254S+P256T;
gg) D96E+D111A+G163K+T231R+N233R;
hh) D96E+D111A+G163K+T231R+N233R+D254S+P256T;
ii) D96E+D111A+G163K+T231R+N233R+P256T;
jj) D96E+D111A+T231R+N233R;
kk) D96E+D111A+T231R+N233R+D254S;
II) D96E+D111A+T231R+N233R+D254S+P256T;
mm) D96E+D111A+T231R+N233R+P256T;
nn) D96E+G163K+T231R+N233R+D254S+P256T;
oo) D96E+T231R+N233R+D254S+P256T;
pp) D96E+T231R+N233R+P256T;
qq) G38A+D96E+D111A+T231R+N233R;
rr) G91T+D96E+D111A+G163K+T231R+N233R+D254S+P256T;
ss) G91T+D96E+D111A+T231R+N233R;
tt) G91T+D96E+T231R+N233R;
uu) G91T+T231R+N233R+D254S+P256T;
vv) N33Q+D96E+D111A+G163K+T231R+N233R+D254S+P256T;
ww) T231R+N233R+D254S+P256T; and
xx) T231R+N233R+P256T.

4. The water-soluble film of claim 1 , wherein the variant in comparison with the parent lipase has increased stability.

5. The water-soluble film of claim 4 , wherein the stability is stability under storage conditions, stability in the presence of surfactants; stability in the presence of protease, stability in the presence of protease and surfactants; stability in the presence of detergent components; chemical stability, oxidation stability, pH stability, and/or thermostability.

6. The water-soluble film of claim 1 , wherein the variant is selected from the group consisting of:
a) a polypeptide having at least 60%, but less than 100%, sequence identity to SEQ ID NO: 2;
b) a polypeptide encoded by a polynucleotide that hybridizes under low stringency conditions, medium stringency conditions, medium-high stringency conditions, high stringency conditions, or very high stringency conditions with (i) the polypeptide coding sequence of SEQ ID NO: 1 or (ii) the full-length complement of (i);
c) a polypeptide encoded by a polynucleotide having at least 60%, but less than 100% sequence identity to SEQ ID NO: 1; and
d) a fragment of the polypeptide of SEQ ID NO: 2.

7. The water-soluble film of claim 1 , wherein the variant is in a solid particulate form.

8. The water-soluble film of claim 1 , wherein the water-soluble film comprises from 35% to 90% of polyvinyl alcohol (PVOH) which has a degree of hydrolysis of from 75% to 99%.

9. The water-soluble film of claim 1 , wherein the water-soluble film comprises from 10% to 50% of polyols.

10. The water-soluble film of claim 1 , wherein the thickness of the water-soluble film is from 10 μm to 500 μm.

11. A detergent pouch, comprising a compartment formed by the water-soluble film of claim 1 , and a detergent composition containing a surfactant and/or a detergent builder.

12. The detergent pouch of claim 11 , wherein the detergent composition is a laundry or dish wash detergent composition.

13. The detergent pouch of claim 11 , wherein the detergent composition is a liquid detergent composition.

14. The detergent pouch of claim 11 , wherein the liquid detergent composition is substantially non-aqueous.

15. The detergent pouch of claim 11 , which comprises an additional enzyme selected from the group consisting of protease, lipase, cutinase, amylase, carbohydrase, cellulase, pectinase, mannanase, arabinase, galactanase, xylanase, DNAse, perhydrolase, and oxidase.

16. A method for preparing a detergent pouch, the method comprising encapsulating a detergent composition comprising a surfactant and/or a detergent builder with the water-soluble film of claim 1 .

Assignees

• Novozymes As
• Monosol Llc

Inventors

• Casella Victor
• Callisen Thomas Hoenger
• Rasmussen Tue
• Mikkelsen Lise Munch
• Simonsen Ole

Classifications

• C12N9/20

  Triglyceride splitting, e.g. by means of lipase · CPC title

• C11D3/38627

  containing lipase · CPC title

• C12N9/18 Primary

  Carboxylic ester hydrolases {(3.1.1)} · CPC title

• C11D17/043

  Liquid or thixotropic (gel) compositions · CPC title

• C12Y301/01003

  Triacylglycerol lipase (3.1.1.3) · CPC title