Lipase variants and polynucleotides encoding same

Patent metadata

Field	Value
Publication number	US-11725197-B2
Application number	US-201816765247-A
Country	US
Kind code	B2
Filing date	Dec 3, 2018
Priority date	Dec 4, 2017
Publication date	Aug 15, 2023
Grant date	Aug 15, 2023

Abstract

The present invention relates to a variant of a parent lipase, wherein said variant has at least 60% but less than 100% sequence identity with SEQ ID NO: 2; has lipase activity; has substitutions corresponding to G91A and N92D in the lid region located from position G82 to position 1100 of SEQ ID NO: 2; has one or more additional substitutions in the lid region; and has at least mutations in positions corresponding to position T231 and position N233 of SEQ ID NO: 2. The present invention also relates to compositions, methods for cleaning a surface using a variant or a composition of the invention; polynucleotides encoding the variants; nucleic acid constructs, vectors, host cells comprising the polynucleotides; methods of using the variants; and methods for obtaining the lipase variant.

First claim

The invention claimed is:

1. A variant of a parent lipase, wherein said variant has:
a) at least 90% but less than 100% sequence identity with SEQ ID NO: 2;
b) lipase activity;
c) substitutions corresponding to G91A, N92D, and K98I in the region located from position G82 to position 1100 of SEQ ID NO: 2; and
d) substitutions corresponding to T231R and N233R of SEQ ID NO: 2.

2. The variant of claim 1 , further comprising one or more additional substitutions in the lid region corresponding to 583T; R84N; I86L,P; 190V; L93F; N94E; D96I,V,H,L; or I100Y of SEQ ID NO: 2.

3. The variant of claim 1 , wherein the variant further comprises one or more additional substitutions corresponding to Q4E, R; D27R; G38A; K46Q; T50K; F51I, V; S54T, E56R, Q, R; T72R; K74E; D111A; R118F; G163K; G212R; Y220F; R232Q; K237V; T244E; D254A,G,L,S,T; 1255G,L,T,V; P256G,L,N,R,S,T,Y; L264A,P; and/or L269V in SEQ ID NO: 2.

4. The variant of claim 1 , wherein the variant is selected from the group consisting of:
(a) a polypeptide having at least 95% but less than 100% sequence identity to SEQ ID NO: 2;
(b) a polypeptide encoded by a polynucleotide having at least 85% but less than 100% sequence identity to the polypeptide coding sequence of SEQ ID NO: 1.

5. The variant of claim 1 , wherein the parent lipase comprises or consists of SEQ ID NO: 2 or (a) a polypeptide having at least 90% sequence identity to the polypeptide of SEQ ID NO: 2; or (b) a polypeptide encoded by the polynucleotide having at least 85% sequence identity to the polypeptide coding sequence of SEQ ID NO: 1.

6. The variant of claim 1 , comprising or consisting of one of the following set of mutations (using SEQ ID NO: 2 for numbering) selected from the group consisting of:
F51I E56R G91A N92D D96I K98I R118F Y220F T231R N233R T244E D254S L264A;
E56R G91A N92D D96I K98I R118F Y220F T231R N233R T244E D254S L264A;
F51I E56R S83T G91A N92D D96I K98I R118F Y220F T231R N233R T244E D254S L264A;
E56R G91A N92D D96I K98I R118F T231R N233R D254G I255V L264A;
E56R G91A N92D D96I K98I R118F T231R N233R D254S P256Q L264A
E56R G91A N92D D96I K98I R118F T231R N233R D254S I255V L264A;
E56R G91A N92D D96I K98I R118F T231R N233R D254S P256S L264A;
E56R G91A N92D D96I K98I R118F T231R N233R D254S P256T L264A;
E56R G91A N92D D96I K98I R118F T231R N233R D254S P256L L264A;
E56R G91A N92D D96I K98I R118F T231R N233R D254S P256R L264A;
E56R G91A N92D N94E D96I K98I R118F T231R N233R D254S L264A;
E56R G91A N92D D96I K98I R118F T231R N233R D254S I255G P256S L264A;
E56R G91A N92D D96I K98I R118F T231R N233R D254S I255G P256A L264A;
E56R G91A N92D D96I K98I R118F T231R N233R D254S I255M P256S L264A;
E56R G91A N92D D96I K98I R118F T231R N233R D254S I255S P256V L264A;
E56R G91A N92D D96I K98I R118F T231R N233R D254S L264P;
E56R G91A N92D D96I K98I R118F T231R N233R D254S P256G L264A;
E56R G91A N92D D96I K98I R118F T231R N233R D254L P256S L264A;
E56R G91A N92D D96I K98I R118F T231R N233R D254A P256Y L264A;
E56R G91A N92D D96I K98I R118F T231R N233R D254G P256R L264A
E56R G91A N92D D96I K98I R118F T231R N233R D254T P256N L264A;
E56R G91A N92D D96I K98I R118F T231R N233R D254G I255L L264A;
Q4E F51I E56R S83T G91A N92D D96I K98I R118F Y220F T231R N233R T244E D254S L264A;
Q4R F51I E56R S83T G91A N92D D96I K98I R118F Y220F T231R N233R T244E D254S L264A;
K46Q F51I E56R S83T G91A N92D D96I K98I R118F Y220F T231R N233R T244E D254S L264A;
F51I E56R K74E S83T G91A N92D D96I K98I R118F Y220F T231R N233R T244E D254S L264A;
F51I E56R T72R S83T G91A N92D D96I K98I R118F Y220F T231R N233R T244E D254S L264A;
F51I E56R N71R S83T G91A N92D D96I K98I R118F Y220F T231R N233R T244E D254S L264A;
Q4E F51I E56R S83T G91A N92D N94D D96I K98I R118F Y220F T231R N233R T244E D254S L264A;
Q4R F51I E56R G91A N92D D96I K98I R118F Y220F T231R N233R T244E D254S L264A;
E56R G91A N92D D96I K98I R118F Y220F T231R N233R T244E D254S L264A;
F51I E56R G91A N92D D96I K98I R118F Y220F T231R N233R T244E D254S L264A.

7. The variant of claim 1 , which has a Relative Wash Performance (RP(Wash)) improvement factor of above 1.00 compared to a lipase variant of SEQ ID NO: 2 with substitutions T231R N233R, when tested in Automatic Mechanical Stress Assay (AMSA) assay using Model B detergent.

8. A composition comprising the variant of claim 1 .

9. The composition of claim 8 , further comprising a surfactant, wherein the composition comprises an anionic surfactant and/or wherein the composition comprises a nonionic surfactant.

10. The composition of claim 8 , wherein the composition comprises one or more anionic and/or one or more nonionic surfactants.

11. A method for cleaning a surface comprising the step of contacting said surface with the variant of claim 1 .

Assignees

• Novozymes As

Inventors

• Svendsen Allan
• Christensen Sune Fang
• Christensen Lars Lehman Hylling
• Bhatia Vikram Kjoller
• Vind Jesper
• Nielsen Vibeke Skovsgaard

Classifications

• C11D2111/12

  Soft surfaces, e.g. textile · CPC title

• C12N9/20 Primary

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

• C11D1/83

  Mixtures of non-ionic with anionic compounds · CPC title

• C11D3/38627

  containing lipase · CPC title

• C11D11/0017

  Chemistry & Metallurgy · mapped topic