Polypeptides having protease activity and polynucleotides encoding same
The present invention relates to isolated polypeptides having protease activity, animal feed additives comprising said polypeptides improving the nutritional value of an animal feed by adding the protease. invention also relates to polynucleotides encoding the polypeptides, nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing the polypeptides.
1 . A nucleic acid construct or expression vector comprising a polynucleotide encoding a polypeptide having protease activity operably linked to one or more control sequences that direct the production of the polypeptide in an expression host,
wherein the polypeptide has at least 98% sequence identity to amino acids 26-425 of SEQ ID NO: 1 or wherein the polypeptide is a fragment of amino acids 26-425, wherein the polypeptide optionally further comprises an N-terminal and/or C-terminal His-tag and/or HO-tag and/or the polypeptide optionally further comprises an N-terminal and/or C-terminal extension of up to 10 amino acids.
2 . A recombinant host cell comprising a polynucleotide encoding a polypeptide having protease activity operably linked to one or more control sequences that direct the production of the polypeptide,
wherein the polypeptide has at least 98% sequence identity to amino acids 26-425 of SEQ ID NO: 1 or wherein the polypeptide is a fragment of amino acids 26-425, wherein the polypeptide optionally further comprises an N-terminal and/or C-terminal His-tag and/or HO-tag and/or the polypeptide optionally further comprises an N-terminal and/or C-terminal extension of up to 10 amino acids.
3 . The recombinant host cell of claim 2 , wherein the polypeptide has at least 98% sequence identity to amino acids 26-425 of SEQ ID NO: 1.
4 . The recombinant host cell of claim 2 , wherein the polypeptide has at least 99% sequence identity to amino acids 26-425 of SEQ ID NO: 1.
5 . The recombinant host cell of claim 2 , wherein the polypeptide has at least 99.5% sequence identity to amino acids 26-425 of SEQ ID NO: 1.
6 . The recombinant host cell of claim 2 , wherein the polypeptide comprises amino acids 26-425 of SEQ ID NO: 1.
7 . The recombinant host cell of claim 2 , wherein the polypeptide is a fragment of amino acids 26-425 of SEQ ID NO: 1, wherein the fragment has protease activity.
8 . The recombinant host cell of claim 2 , wherein the polypeptide further comprises an N-terminal and/or C-terminal His-tag and/or HQ-tag.
9 . The recombinant host cell of claim 2 , wherein the polypeptide further comprises an N-terminal and/or C-terminal extension of up to 10 amino acids.
10 . The recombinant host cell of claim 2 , which is a yeast recombinant host cell selected from the group consisting of Candida, Hansenula, Kluyveromyces, Pichia, Saccharomyces, Schizosaccharomyces , and Yarrowia cells.
11 . The recombinant host cell of claim 2 , which is a filamentous fungal recombinant host cell selected from the group consisting of Acremonium, Aspergillus, Aureobasidium, Bjerkandera, Ceriporiopsis, Chrysosporium, Coprinus, Coriolus, Cryptococcus, Filibasidium, Fusarium, Humicola, Magnaporthe, Mucor, Myceliophthora, Neocallimastix, Neurospora, Paecilomyces, Penicillium, Phanerochaete, Phlebia, Piromyces, Pleurotus, Schizophyllum, Talaromyces, Thermoascus, Thielavia, Tolypocladium, Trametes , and Trichoderma cells.
12 . The recombinant host cell of claim 2 , which is a Gram-positive cell selected from the group consisting of Bacillus, Clostridium, Enterococcus, Geobacillus, Lactobacillus, Lactococcus, Oceanobacillus, Staphylococcus, Streptococcus , or Streptomyces cells, or a Gram-negative bacteria selected from the group consisting of Campylobacter, E. coli, Flavobacterium, Fusobacterium, Helicobacter, Ilyobacter, Neisseria, Pseudomonas, Salmonella , and Ureaplasma cells.
13 . A method of producing a polypeptide having protease activity, comprising cultivating a recombinant host cell of claim 12 under conditions conducive for production of the polypeptide.
14 . The method of claim 13 , further comprising recovering the polypeptide.
15 . The recombinant host cell of claim 2 , which is selected from the group consisting of Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillus brevis, Bacillus circulans, Bacillus clausii, Bacillus coagulans, Bacillus firmus, Bacillus lautus, Bacillus lentus, Bacillus licheniformis, Bacillus megaterium, Bacillus pumilus, Bacillus stearothermophilus, Bacillus subtilis, Bacillus thuringiensis, Streptococcus equisimilis, Streptococcus pyogenes, Streptococcus uberis , and Streptococcus equi subsp. Zooepidemicus, Streptomyces achromogenes, Streptomyces avermitilis, Streptomyces coelicolor, Streptomyces griseus , and Streptomyces lividans cells.
16 . A method of producing a polypeptide having protease activity, comprising cultivating a recombinant host cell of claim 15 under conditions conducive for production of the polypeptide.
17 . The method of claim 16 , further comprising recovering the polypeptide.