Polypeptides Having Cellulolytic Enhancing Activity And Polynucleotides Encoding Same

Abstract

The present invention relates to isolated polypeptides having cellulolytic enhancing activity, catalytic domains, cellulose binding domains and polynucleotides encoding the polypeptides, catalytic domains or cellulose binding domains. 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, catalytic domains or cellulose binding domains.

1 . An isolated polypeptide having cellulolytic enhancing activity, selected from the group consisting of:
 (a) a polypeptide having at least 60% sequence identity to the mature polypeptide of SEQ ID NO: 6, at least 65% sequence identity to the mature polypeptide of SEQ ID NO: 8, at least 75% sequence identity to the mature polypeptide of SEQ ID NO: 2, or at least 80% sequence identity to the mature polypeptide of SEQ ID NO: 4;   (b) a polypeptide encoded by a polynucleotide that hybridizes under at least high stringency conditions with the mature polypeptide coding sequence of SEQ ID NO: 1 or the cDNA sequence thereof, the mature polypeptide coding sequence of SEQ ID NO: 3 or the cDNA sequence thereof, the mature polypeptide coding sequence of SEQ ID NO: 5 or the cDNA sequence thereof, or the mature polypeptide coding sequence of SEQ ID NO: 7; or the full-length complement thereof;   (c) a polypeptide encoded by a polynucleotide having at least 60% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 5 or the cDNA sequence thereof, at least 65% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 7, at least 75% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or the cDNA sequence thereof, or at least 80% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 3 or the cDNA sequence thereof;   (d) a variant of the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8 comprising a substitution, deletion, and/or insertion at one or more positions; and   (e) a fragment of the polypeptide of (a), (b), (c), or (d) that has cellulolytic enhancing activity.   
     
     
         2 . The polypeptide of  claim 1 , comprising or consisting of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8 or the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8. 
     
     
         3 . An isolated polypeptide comprising a catalytic domain selected from the group consisting of:
 (a) a catalytic domain having at least 80% sequence identity to amino acids 20 to 245 of SEQ ID NO: 2;   (b) a catalytic domain encoded by a polynucleotide that hybridizes under at least very high stringency conditions with nucleotides 58 to 1122 of SEQ ID NO: 1; the cDNA sequence thereof; or the full-length complement thereof;   (c) a catalytic domain encoded by a polynucleotide having at least 80% sequence identity to nucleotides 58 to 1122 of SEQ ID NO: 1 or the cDNA sequence thereof;   (d) a variant of amino acids 20 to 245 of SEQ ID NO: 2 comprising a substitution, deletion, and/or insertion at one or more positions; and   (e) a fragment of the catalytic domain of (a), (b), (c), or (d) that has cellulolytic enhancing activity.   
     
     
         4 . An isolated polypeptide comprising a cellulose binding domain operably linked to a catalytic domain, wherein the binding domain is selected from the group consisting of:
 (a) a cellulose binding domain having at least 80% sequence identity to amino acids 284 to 316 of SEQ ID NO: 2;   (b) a cellulose binding domain encoded by a polynucleotide that hybridizes under at least very high stringency conditions with nucleotides 1237 to 1458 of SEQ ID NO: 1; the cDNA sequence thereof; or the full-length complement thereof;   (c) a cellulose binding domain encoded by a polynucleotide having at least 80% sequence identity to nucleotides 1237 to 1458 of SEQ ID NO: 1 or the cDNA sequence thereof;   (d) a variant of amino acids 284 to 316 of SEQ ID NO: 2 comprising a substitution, deletion, and/or insertion at one or more positions; and   (e) a fragment of the cellulose binding domain of (a), (b), (c), or (d) that has binding activity.   
     
     
         5 . A composition comprising the polypeptide of  claim 1 . 
     
     
         6 . An isolated polynucleotide encoding the polypeptide of  claim 1 . 
     
     
         7 . A recombinant host cell comprising the polynucleotide of  claim 14  operably linked to one or more control sequences that direct the production of the polypeptide. 
     
     
         8 . A method of producing the polypeptide of  claim 1 , comprising: cultivating a cell, which in its wild-type form produces the polypeptide, under conditions conducive for production of the polypeptide. 
     
     
         9 . A method of producing a polypeptide having cellulolytic enhancing activity, comprising: cultivating the host cell of  claim 7  under conditions conducive for production of the polypeptide. 
     
     
         10 . A transgenic plant, plant part or plant cell transformed with a polynucleotide encoding the polypeptide of  claim 1 . 
     
     
         11 . A method of producing a polypeptide having cellulolytic enhancing activity, comprising: cultivating the transgenic plant or plant cell of  claim 10  under conditions conducive for production of the polypeptide. 
     
     
         12 . A method of producing a mutant of a parent cell, comprising inactivating a polynucleotide encoding the polypeptide of  claim 1 , which results in the mutant producing less of the polypeptide than the parent cell. 
     
     
         13 . A double-stranded inhibitory RNA (dsRNA) molecule comprising a subsequence of the polynucleotide of  claim 6 , wherein optionally the dsRNA is an siRNA or an miRNA molecule. 
     
     
         14 . An isolated polynucleotide encoding a signal peptide comprising or consisting of amino acids 1 to 19 of SEQ ID NO: 2, amino acids 1 to 19 of SEQ ID NO: 4, amino acids 1 to 19 of SEQ ID NO: 6, or amino acids 1 to 16 of SEQ ID NO: 8. 
     
     
         15 . A method of producing a protein, comprising: cultivating a recombinant host cell comprising a gene encoding a protein operably linked to the polynucleotide of  claim 14 , wherein the gene is foreign to the polynucleotide encoding the signal peptide, under conditions conducive for production of the protein. 
     
     
         16 . A process for degrading a cellulosic material, comprising: treating the cellulosic material with an enzyme composition in the presence of the polypeptide having cellulolytic enhancing activity of  claim 1 . 
     
     
         17 . A process for producing a fermentation product, comprising:
 (a) saccharifying a cellulosic material with an enzyme composition in the presence of the polypeptide having cellulolytic enhancing activity of  claim 1 ;   (b) fermenting the saccharified cellulosic material with one or more fermenting microorganisms to produce the fermentation product; and   (c) recovering the fermentation product from the fermentation.   
     
     
         18 . A process of fermenting a cellulosic material, comprising: fermenting the cellulosic material with one or more fermenting microorganisms, wherein the cellulosic material is saccharified with an enzyme composition in the presence of the polypeptide having cellulolytic enhancing activity of  claim 1 . 
     
     
         19 . The process of  claim 18 , wherein the fermenting of the cellulosic material produces a fermentation product. 
     
     
         20 . A whole broth formulation or cell culture composition comprising the polypeptide of  claim 1 .