Polypeptides having lysozyme activity and polynucleotides encoding same

Patent metadata

Field	Value
Publication number	US-9701952-B2
Application number	US-201214360529-A
Country	US
Kind code	B2
Filing date	Nov 23, 2012
Priority date	Nov 25, 2011
Publication date	Jul 11, 2017
Grant date	Jul 11, 2017

Abstract

The present invention relates to isolated polypeptides having lysozyme activity and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

First claim

The invention claimed is:

1. A method of isolating DNA from bacteria, comprising:
(a) treating cell walls of the bacteria with a polypeptide having lysozyme activity, wherein the polypeptide has at least 90% sequence identity to the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8; and
(b) recovering the DNA.

2. The method of claim 1 , wherein the polypeptide has at least 92% sequence identity to the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8.

3. The method of claim 1 , wherein the polypeptide has at least 95% sequence identity to the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8.

4. The method of claim 1 , wherein the polypeptide has at least 97% sequence identity to the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8.

5. The method of claim 1 , wherein the polypeptide is encoded by a polynucleotide that hybridizes under high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 3 or SEQ ID NO: 7, or the full-length complement of thereof, and has at least 90% sequence identity to the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8.

6. The method of claim 1 , wherein the polypeptide is a variant of the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8, comprising a substitution, deletion, and/or insertion at one or more positions, and has at least 90% sequence identity to the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8.

7. The method of claim 1 , wherein the polypeptide is a fragment of the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8, which has lysozyme activity and has at least 90% sequence identity to the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8.

8. The method of claim 1 , wherein the polypeptide comprises or consists of amino acids 1 to 208 of SEQ ID NO: 8.

9. A method of producing a polypeptide having lysozyme activity, comprising:
(a) cultivating a recombinant host cell under conditions conducive for production of the polypeptide, wherein
(i) the polypeptide has at least 90% sequence identity to the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8; and
(ii) the recombinant host cell comprises a polynucleotide which encodes the polypeptide and which is operably linked to one or more control sequences that direct the production of the polypeptide in the recombinant host cell; and

(b) recovering the polypeptide.

10. The method of claim 9 , wherein the polypeptide has at least 92% sequence identity to the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8.

11. The method of claim 9 , wherein the polypeptide has at least 95% sequence identity to the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8.

12. The method of claim 9 , wherein the polypeptide has at least 97% sequence identity to the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8.

13. The method of claim 9 , wherein the polypeptide is encoded by a polynucleotide that hybridizes under high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 3 or SEQ ID NO: 7, or the full-length complement of thereof, and has at least 90% sequence identity to the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8.

14. The method of claim 9 , wherein the polypeptide is a variant of the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8, comprising a substitution, deletion, and/or insertion at one or more positions, and has at least 90% sequence identity to the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8.

15. The method of claim 9 , wherein the polypeptide is a fragment of the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8, which has lysozyme activity and has at least 90% sequence identity to the polypeptide of amino acids 1 to 208 of SEQ ID NO: 8.

16. The method of claim 9 , wherein the polypeptide comprises or consists of amino acids 1 to 208 of SEQ ID NO: 8.

17. The method of claim 9 , wherein the recombinant host cell is a recombinant filamentous fungal host cell.

18. The method of claim 17 , wherein the recombinant filamentous fungal host cell is selected from the group consisting of Aspergillus awamori, Aspergillus foetidus, Aspergillus fumigatus, Aspergillus japonicus, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Bjerkandera adusta, Ceriporiopsis aneirina, Ceriporiopsis caregiea, Ceriporiopsis gilvescens, Ceriporiopsis pannocinta, Ceriporiopsis rivulosa, Ceriporiopsis subrufa, Ceriporiopsis subvermispora, Chrysosporium inops, Chrysosporium keratinophilum, Chrysosporium lucknowense, Chrysosporium merdarium, Chrysosporium pannicola, Chrysosporium queenslandicum, Chrysosporium tropicum, Chrysosporium zonatum, Coprinus cinereus, Coriolus hirsutus, Fusarium bactridioides, Fusarium cerealis, Fusarium crookwellense, Fusarium culmorum, Fusarium graminearum, Fusarium graminum, Fusarium heterosporum, Fusarium negundi, Fusarium oxysporum, Fusarium reticulatum, Fusarium roseum, Fusarium sambucinum, Fusarium sarcochroum, Fusarium sporotrichioides, Fusarium suiphureum, Fusarium torulosum, Fusarium trichothecioides, Fusarium venenatum, Humicola insolens, Humicola lanuginosa, Mucor miehei, Myceliophthora thermophila, Neurospora crassa, Penicillium purpurogenum, Phanerochaete chrysosporium, Phiebia radiata, Pleurotus eryngii, Thielavia terrestris, Trametes villosa, Trametes versicolor, Trichoderma harzianum, Trichoderma Trichoderma longibrachiatum, Trichoderma reesei , and Trichoderma viride recombinant host cells.

19. The method of claim 10 , wherein the recombinant host cell is a recombinant filamentous fungal host cell.

20. The method of claim 19 , wherein the recombinant filamentous fungal host cell is selected from the group consisting of Aspergillus awamori, Aspergillus foetidus, Aspergillus fumigatus, Aspergillus japonicus, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Bjerkandera adusta, Ceriporiopsis aneirina, Ceriporiopsis caregiea, Ceriporiopsis gilvescens, Ceriporiopsis pannocinta, Ceriporiopsis rivulosa, Ceriporiopsis subrufa, Ceriporiopsis subvermispora, Chrysosporium inops, Chrysosporium keratinophilum, Chrysosporium lucknowense, Chrysosporium merdarium, Chrysosporium pannicola, Chrysosporium queenslandicum, Chrysosporium tropicum, Chrysosporium zonatum, Coprinus cinereus, Coriolus hirsutus, Fusarium bactridioides, Fusarium cerealis, Fusarium crookwellense, Fusarium culmorum, Fusarium graminearum, Fusarium graminum, Fusarium heterosporum, Fusarium negundi, Fusarium oxysporum, Fusarium reticulatum, Fusarium roseum, Fusarium sambucinum, Fusarium sarcochroum, Fusarium sporotrichioides, Fusarium suiphureum, Fusarium torulosum, Fusarium trichothecioides, Fusarium venenatum, Humicola insolens, Humicola lanuginosa, Mucor miehei, Myceliophthora thermophila, Neurospora crassa, Penicillium purpurogenum, Phanerochaete chrysosporium, Phiebia radiata, Pleurotus eryngii, Thielavia terrestris, Trametes villosa, Trametes versicolor, Trichoderma harzianum, Trichoderma Trichoderma longibrachiatum, Trichoderma reesei , and Trichoderma viride recombinant host cells.

21. The method of claim 12 , wherein the recombinant host cell is a recombinant filamentous fungal host cell.

22. The method of claim 21 , wherein the recombinant filamentous fungal host cell is selected from the group consisting of Aspergillus awamori, Aspergillus foetidus, Aspergillus fumigatus, Aspergillus japonicus, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Bjerkandera adusta, Ceriporiopsis aneirina, Ceriporiopsis caregiea, Ceriporiopsis gilvescens, Ceriporiopsis pannocinta, Ceriporiopsis rivulosa, Ceriporiopsis

Assignees

• Novozymes As

Inventors

• Schnorr Kirk Matthew
• Nielsen Jens Erik
• Klausen Mikkel

Classifications

• A61P31/04

  Antibacterial agents · CPC title

• A23K20/174

  Vitamins · CPC title

• C11D3/38636

  containing enzymes other than protease, amylase, lipase, cellulase, oxidase or reductase · CPC title

• A23K20/20

  Inorganic substances, e.g. oligoelements · CPC title

• A23K50/80

  for aquatic animals, e.g. fish, crustaceans or molluscs · CPC title