Subtilases

Abstract

The present invention relates to novel JP170 like subtilases from wild-type bacteria, hybrids thereof and to methods of construction and production of these proteases. Further, the present invention relates to use of the claimed subtilases in detergents, such as a laundry or an automatic dishwashing detergent.

1 . A subtilase enzyme which
 a) has an amino acid sequence which is at least 95% identical with SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12 or SEQ ID NO: 14 or   b) is encoded by a nucleotide sequence obtainable from a deposited strain selected from the group consisting of DSM16714, DSM16715, DSM16719, DSM16722, DSM16712, DSM16713 and DSM16716.   
     
     
         2 . An isolated polypeptide having subtilase activity, which polypeptide is selected from the group consisting of:
 a) a polypeptide which is encoded by a nucleic acid sequence which is at least 85% identical with SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11 or SEQ ID NO: 13,   b) a polypeptide comprising a segment, which segment is encoded by a subsequence of the DNA sequence set forth in SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11 or SEQ ID NO: 13, wherein said subsequence consists of the nucleic acids starting from one of positions 1-210 and ending with one of positions 600-1260, and   c) a polypeptide which is encoded by a nucleic acid sequence which is capable of hybridizing under medium/high stringency conditions with (i) the nucleic acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11 or SEQ ID NO: 13, (ii) its complementary strand, or (iii) a subsequence of (i) or (ii).   
     
     
         3 . A nucleic acid sequence which is contained on a plasmid in the deposited strain DSM16712, DSM16713, DSM16714, DSM16715, DSM16716, DSM16719 or DSM16722. 
     
     
         4 . A nucleic acid sequence as shown in SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11 or SEQ ID NO: 13. 
     
     
         5 . A nucleic acid construct comprising the nucleic acid sequence of  claim 4  operably linked to one or more control sequences capable of directing the expression of the polypeptide in a suitable expression host. 
     
     
         6 . A recombinant expression vector comprising the nucleic acid construct of  claim 5 , a promoter, and transcriptional and translational stop signals. 
     
     
         7 . The vector according to  claim 6 , further comprising a selectable marker. 
     
     
         8 . A recombinant host cell comprising the nucleic acid construct of  claim 5 . 
     
     
         9 . The cell according to  claim 8 , wherein the nucleic acid construct is contained on a vector. 
     
     
         10 . The cell according to  claim 9 , wherein the nucleic acid construct is integrated into the host cell genome. 
     
     
         11 . The cell according to  claim 10 , wherein the nucleic acid sequence encodes an amino acid sequence set forth in SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12 or SEQ ID NO: 14. 
     
     
         12 . The cell according to  claim 11 , wherein the nucleic acid sequence is set forth in SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11 or SEQ ID NO: 13. 
     
     
         13 . A method for producing the polypeptide of  claim 1  comprising
 (a) cultivating a  Bacillus  strain to produce a supernatant comprising the polypeptide; and 
 (b) recovering the polypeptide. 
 
     
     
         14 . A method for producing the polypeptide of  claim 1  comprising
 (a) cultivating a host cell comprising a nucleic acid construct comprising a nucleic acid sequence encoding the polypeptide under conditions conducive to expression of the polypeptide; and 
 (b) recovering the polypeptide. 
 
     
     
         15 . A core sequence of the subtilase of  claim 1 , which core sequence consists of the amino acids starting from one of positions 1-70 and ending with one of position 200-420 of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12 or SEQ ID NO: 14. 
     
     
         16 . The core sequence of  claim 15 , which has 96% identity with the amino acids in position 49-391 of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12 or SEQ ID NO: 14. 
     
     
         17 . (canceled) 
     
     
         18 . A hybrid subtilase comprising a core sequence of  claim 15 . 
     
     
         19 . (canceled) 
     
     
         20 . A detergent composition comprising a subtilase of  claim 1 . 
     
     
         21 . The detergent composition of  claim 20 , which is a laundry detergent or an automatic dishwashing detergent.