Lipase variants, polynucleotides encoding same and the use thereof

Patent metadata

Field	Value
Publication number	US-11326152-B2
Application number	US-201716316458-A
Country	US
Kind code	B2
Filing date	Jul 14, 2017
Priority date	Jul 18, 2016
Publication date	May 10, 2022
Grant date	May 10, 2022

Abstract

The present invention relates to lipase variants. 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.

First claim

The invention claimed is:

1. A lipase variant of a parent lipase, which comprises
(a) a modification at a position corresponding to position 225 of the polypeptide of SEQ ID NO: 1, wherein the modification is a deletion, an insertion of Arg, or a substitution with Arg or Lys, and
(b) a modification at a position corresponding to position 231 of the polypeptide of SEQ ID NO: 1, wherein the modification is a substitution with Arg,

wherein the lipase variant has at least 90% sequence identity and less than 100% sequence identity to the polypeptide of SEQ ID NO: 1 and the lipase variant has lipase activity.

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

3. The lipase variant of claim 1 , which has at least 97% sequence identity to the polypeptide of SEQ ID NO: 1.

4. The lipase variant of claim 1 , wherein said lipase variant further comprises one or more substitutions that correspond to substitutions in the polypeptide of SEQ ID NO: 1, selected from the group consisting of R81Q; S83T; R84H; S85T; I186L, I86P, I86V, I86W; E87A, E87I, E87K, E87T, E87V; N88Q; I90L, I90M; G91A, G91L, G91N, G91Q, G91T; N92D, N92K; L93F; N94D, N94K, N94R; F95A, F95L, F95Y; D96E, D961, D96L, D96T: L97F, L97M, L97P; K98D, K98E, K981, K98Q; and E99K.

5. The lipase variant of claim 1 , which further comprises one or more substitutions at positions corresponding to positions selected from: 27, 33, 38, 51, 56, 57, 58, 60, 69, 101, 106, 111, 131, 149, 150, 163, 198, 210, 211, 216, 220, 254, 255, 256, 263, 264, 265, 266, 267, and 269 of the polypeptide of SEQ ID NO: 1.

6. The lipase variant of claim 5 , which further comprises one or more substitutions corresponding to substitutions in the polypeptide of SEQ ID NO: 1 selected from the group consisting of: 27N, 27R; 33K, 33Q; 38A; 51I, 51L, 51V; 56K, 56Q, 56R, 56S; 57G, 57N; 58A; 60S; 69R; 101R; 106K, 106R; 111A; 131T; 149D; 150G; 163K, 163S; 198S; 210K, 210Q; 211L; 216P; 220F; 254S; 255A, 255T; 256K, 256T, 256V; 263Q; 264A, 264F, 264W; 265T; 266D; 267A; and 269N.

7. The lipase variant of claim 1 , wherein said variant comprises one-set of modifications that correspond to modifications in the polypeptide of SEQ ID NO: 1 selected from the group consisting of:

N200R + G225R,

S224K + G225K,

S224K + G225K + L227K,

S224R + G225R,

S224R + G225R + L227R + V228*,

S224* + G225* + T226R + L227R + V228*,

S224* + G225* + T226* + L227R + V228R + V230*,

S224* + G225* + T226* + L227R + P229R,

G225GR,

G225R + T226R,

G225R + L227R,

G225R + V228R,

G225R + P229R,

G225R + T231R,

G225R + N233R, and

G225* + T226* + L227R + V228R.

8. The lipase variant of claim 1 , wherein said variant has improved wash performance, a higher Benefit Risk Factor (BRF), higher relative wash performance (RP(Wash)) or lower relative odor (RP(Odor)) compared to the lipase of SEQ ID NO: 1.

9. A composition comprising the lipase variant of claim 1 and a surfactant.

10. The lipase variant of claim 1 , wherein said variant comprises a deletion or an insertion of Arg at the position corresponding to position 225 of the polypeptide of SEQ ID NO: 1.

11. The lipase variant of claim 1 , wherein said variant comprises modifications that correspond to modifications G225R+T231R in the polypeptide of SEQ ID NO: 1.

12. A method for cleaning a surface, comprising contacting the surface with the lipase variant of claim 1 .

13. A method of hydrolyzing a lipase substrate, comprising treating the lipase substrate with the lipase variant of claim 1 .

Assignees

• Novozymes As

Inventors

• Toscano Miguel Duarte Guilherme
• Svendsen Allan
• Vind Jesper
• Fulton Alexander David
• Baunsgaard Lone

Classifications

• C11D3/38627

  containing lipase · CPC title

• C12Y301/01003

  Triacylglycerol lipase (3.1.1.3) · CPC title

• C12N9/20 Primary

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

• C12N9/18

  Carboxylic ester hydrolases {(3.1.1)} · CPC title