Polypeptides with lipase activity and polynucleotides encoding same

Patent metadata

Field	Value
Publication number	US-9631164-B2
Application number	US-201414770006-A
Country	US
Kind code	B2
Filing date	Mar 19, 2014
Priority date	Mar 21, 2013
Publication date	Apr 25, 2017
Grant date	Apr 25, 2017

Abstract

The present invention relates to isolated polypeptides with lipase activity, selected from the group consisting of: (a) a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 4; (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) SEQ ID NO: 3 or the full-length complement of (i); (c) a polypeptide encoded by a polynucleotide having at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 3; (d) a polypeptide which is a variant of SEQ ID NO: 4 comprising a substitution, deletion, and/or insertion at one or more (e.g. several) positions; and (e) a polypeptide which is a fragment of any of the polypeptides of (a), (b), (c) or (d). The invention also relates to polynucleotides encoding the polypeptides; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the polypeptides.

First claim

The invention is further defined in the following paragraphs:

1. An isolated polypeptide with lipase activity, selected from the group consisting of:
(a) a polypeptide having at least 96% sequence identity to SEQ ID NO: 4;
(b) a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions of 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide followed by washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 65° C. with (i) SEQ ID NO: 3 or the full-length complement of (i); and
(c) a fragment of (a) or (b) having lipase activity.

2. The polypeptide of claim 1 , comprising a substitution at one or more positions corresponding to positions 32, 40, 74, 82, 83, 86, 94, 138, 187, 289, 291, 364, 379, 381, and 384 of SEQ ID NO: 2.

3. The polypeptide of claim 2 , wherein the substitution at one or more positions corresponding to positions 32, 40, 74, 82, 83, 86, 94, 138, 187, 289, 291, 364, 379, 381, and 384 of SEQ ID NO: 3 is one or more substitutions selected from the group consisting of S32A, 540T, V74I, S82T, D83E, V86T, E94Q, T138A, I187V, D289I, R291T, N364S, S379T, T381 L, and L384F.

4. An isolated polypeptide with lipase activity, having at least 90% sequence identity to SEQ ID NO: 4, comprising the substitutions
S32A+S40T+V74I+S82T+D83E+V86T+E94Q+T138A+I187V+D289I+R291T+N364S+S379T+T 381L+L384F corresponding to positions of SEQ ID NO: 2.

5. A method for hydrolyzing a lipid, comprising contacting the lipid with the polypeptide of any of claim 1 .

6. A composition comprising the polypeptide of claim 1 and at least one surfactant, at least one surfactant system, at least one soap, or any mixtures thereof.

7. The composition of claim 6 , wherein the surfactant or surfactant systems are selected from anionic surfactants, cationic surfactants, non-ionic surfactants, ampholyttic surfactants, zwitterionic surfactants, semipolar nonionic surfactants or any mixtures thereof.

8. The composition of claim 6 , wherein the composition is a laundry cleaning composition, a dishwashing cleaning composition, a hard-surface cleaning composition and/or a personal care cleaning composition.

9. The composition of claim 6 , wherein the composition is formulated as a regular, compact or concentrated liquid; a gel; a paste; a soap bar; a regular or a compacted powder; a granulated solid; a homogenous or a multilayer tablet with two or more layers (same or different phases); a pouch having one or more compartments; a single or a multi-compartment unit dose form; or any combination thereof.

10. The composition of claim 6 , wherein the composition has cleaning activity from pH4 to pH11.5.

11. The composition of claim 6 , wherein the composition is effective in hydrolyzing a lipid at a temperature of from 10° C. to 90° C.

12. The composition of claim 6 , further comprising one or more enzymes selected from: hemicellulases, peroxidases, proteases, cellulases, xylanases, lipases, phospholipases, esterases, cutinases, pectinases, mannanases, pectate lyases, keratinases, reductases, oxidases, phenoloxidases, lipoxygenases, ligninases, pullulanases, tannases, pentosanases, malanases, β-glucanases, arabinosidases, hyaluronidase, chondroitinase, laccase, amylases, or mixtures thereof.

13. A method of producing a composition, comprising adding the polypeptide of claim 1 .

14. A method for cleaning a subject, comprising contacting a lipid stain present on the subject to be cleaned with the composition of claim 6 .

Assignees

• Novozymes As

Inventors

• Morant Marc Dominique
• Reiser Anna Verana
• Hansen Carsten Hoerslev
• Borch Kim
• Baunsgaard Lone
• Bhatia Vikram Kjoeller

Classifications

• C12P13/008

  containing a N-O bond, e.g. nitro (-NO2), nitroso (-NO) · CPC title

• C11D3/38627 Primary

  containing lipase · CPC title

• C11D3/38681

  Chemically modified or immobilised enzymes · CPC title

• C12N9/20

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