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 for producing and using the polypeptides.

1 . An isolated protease having an amino acid sequence which has at least 95% identity with amino acids 2 to 148 of SEQ ID NO: 5. 
     
     
         2 . The polypeptide of  claim 1 , comprising the amino acid sequence of SEQ ID NO: 5. 
     
     
         3 . An isolated polynucleotide comprising a nucleotide sequence which encodes the polypeptide of  claim 1 . 
     
     
         4 . A method for producing a mutant of a parent cell, which comprises disrupting or deleting a nucleotide sequence encoding the polypeptide of  claim 1 , which results in the mutant producing less of the polypeptide than the parent cell. 
     
     
         5 . A mutant cell produced by the method of  claim 4 . 
     
     
         6 . The mutant cell of  claim 5 , which further comprises a gene encoding a native or heterologous protein. 
     
     
         7 . A method for producing a protein product comprising a polypeptide comprising two or more domains of which one domain is carbohydrate binding module, wherein the method comprises the steps of:
 c) fermentation of a cell having reduced expression of the polypeptide according to  claim 1 , which cell produces said polypeptide comprising two or more domains, and   d) recovering the product from fermentation broth.   
     
     
         8 . A method for producing a protein product essentially free of the protease activity of the polypeptide according to  claim 1 , wherein the method comprises the steps of:
 d) fermentation of a cell expressing a polypeptide according to  claim 1  as well as a protein product of interest,   e) adding agent capable of inhibiting protease activity of a polypeptide according to the invention to the fermentation broth before, during or after the fermentation has been completed, and   f) recovering product of interest from fermentation broth.   
     
     
         9 . A method for producing a protein product essentially free of the protease activity of the polypeptide according to  claim 1 , wherein the method comprises the steps of:
 d) cultivating a cell under conditions permitting expression of said protein product,   e) subjecting the culture broth to combined pH and temperature treatment, and   f) recovering the product from culture broth.   
     
     
         10 . A method according to  claim 8 , wherein the protease activity is removed by at least 60%.