Polypeptides Having Protease Activity and Polynucleotides Encoding Same

Abstract

The present invention relates to isolated polypeptides having protease activity and isolated 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.

1 - 21 . (canceled) 
     
     
         22 . An isolated polypeptide having protease activity, selected from the group consisting of:
 (a) a polypeptide having at least 60% sequence identity to the mature polypeptide of SEQ ID NO: 2 or 4;   (b) a polypeptide encoded by a polynucleotide that hybridizes under medium stringency conditions, medium-high stringency conditions, high stringency conditions, or very high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1 or 3, (ii) the genomic DNA sequence comprising the mature polypeptide coding sequence of SEQ ID NO: 1 or 3, or (iii) the full-length complementary strand of (i) or (ii);   (c) a polypeptide encoded by a polynucleotide having at least 60% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or 3;   (d) a variant comprising a substitution, deletion, and/or insertion of one or more (several) amino acids of the mature polypeptide of SEQ ID NO: 2 or 4;   (e) a fragment of a polypeptide of (a), (b), (c), or (d) that has protease activity; and   (f) a polypeptide having protease activity and further comprising the motif:   
       
         
           
                 
               
                   (SEQ ID NO: 13) 
                 
                   His Gly Thr (Xaa) 6  Ala Ser Tyr Gly Ser Val Ser Gly 
                 
             
                
                
               
            
           
         
         wherein Xaa is any natural amino acid and (Xaa) 6  mean 6 consecutive amino acids, where each amino acid may be any natural amino acid; 
         or the corresponding motif comprising one or two substitutions among the last 8 amino acids in the motif, such as a motif selected among SEQ ID NO: 14 to SEQ ID NO: 49. 
       
     
     
         23 . The polypeptide of  claim 22 , having at least 80% sequence identity to the mature polypeptide of SEQ ID NO: 2 or 4. 
     
     
         24 . The polypeptide of  claim 22 , having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 2 or 4. 
     
     
         25 . The polypeptide of  claim 22 , comprising or consisting of SEQ ID NO: 2 or 4. 
     
     
         26 . The polypeptide of  claim 22 , comprising or consisting of the mature polypeptide of SEQ ID NO: 2 or 4. 
     
     
         27 . The polypeptide of  claim 26 , wherein the mature polypeptide is amino acids 144 to 418 of SEQ ID NO: 2 or amino acids 1 to 275 of SEQ ID NO: 4. 
     
     
         28 . The polypeptide of  claim 22 , comprising a S8 serin protease domain. 
     
     
         29 . A polypeptide having at least 60% sequence identity to the mature polypeptide of SEQ ID NO: 2, 4 or 6, wherein said polypeptide comprises the motif His Gly Thr (Xaa) 6  Ala Ser Tyr Gly Ser Val Ser Gly (SEQ ID NO: 13). 
     
     
         30 . The polypeptide of  claim 22 , wherein the protease has high detergency performance at low temperatures and the stability is not affected by the presence of a strong chelator. 
     
     
         31 . An isolated polypeptide comprising a catalytic domain selected from the group consisting of:
 (a) a catalytic domain having at least 60% sequence identity to amino acids 143 to 418 of SEQ ID NO: 2   (b) a catalytic domain encoded by a polynucleotide that hybridizes under medium-high stringency conditions with (i) nucleotides 711 to 1535 of SEQ ID NO: 1, (ii) cDNA sequence comprising the catalytic domain of SEQ ID NO: 1, or (iii) the full-length complementary strand of (i) or (ii);   (c) a catalytic domain encoded by a polynucleotide having at least 60% sequence identity to the catalytic domain of SEQ ID NO: 1;   (d) a variant of a catalytic domain comprising a substitution, deletion, and/or insertion of one or more (several) amino acids of the catalytic domain of SEQ ID NO: 2; and   (e) a fragment of a catalytic domain of (a), (b), (c) or (d) that has protease activity.   
     
     
         32 . A composition comprising the polypeptide of  claim 22 . 
     
     
         33 . The composition of  claim 32 , which is a detergent composition for laundry or automatic dish washing. 
     
     
         34 . The composition of  claim 33 , further comprising one of more additional enzymes selected from the group consisting of a protease, an amylase, a lipase, a cutinase, a cellulase, an endoglucanase, a xyloglucanase, a pectinase, a pectin lyase, a xanthanase, a peroxidase, a haloperoxygenase, a catalase, a mannanase, or any mixture thereof. 
     
     
         35 . The composition of  claim 32 , comprising a surfactant. 
     
     
         36 . An isolated polynucleotide encoding the polypeptide of  claim 22 . 
     
     
         37 . A nucleic acid construct or expression vector comprising the polynucleotide of  claim 36  operably linked to one or more (several) control sequences that direct the production of the polypeptide in an expression host. 
     
     
         38 . A recombinant host cell comprising the polynucleotide of  claim 36  operably linked to one or more control sequences that direct the production of the polypeptide. 
     
     
         39 . A method of producing the polypeptide of  claim 22 , comprising:
 (a) cultivating a cell, which in its wild-type form produces the polypeptide, under conditions conducive for production of the polypeptide; and   (b) recovering the polypeptide.   
     
     
         40 . A method of producing a polypeptide, comprising:
 (a) cultivating a host cell of  claim 38  under conditions conducive for production of the polypeptide; and   (b) recovering the polypeptide.