Endoglucanases

Abstract

The present invention relates to enzyme preparations consisting essentially of an enzyme which has cellulytic activity and comprises a first amino acid sequence having the following sequence (SEQ ID NO: 79) Thr Arg Xaa Xaa Asp Cys Cys Xaa Xaa Xaa Cys Xaa 1   2   3   4   5   6   7   8   9   10  11  12 Trp Xaa 13  14 and a second amino acid sequence having the following sequence Trp Cys Cys Xaa Cys (SEQ ID NO: 80) 1   2   3   4   5 wherein, at position 3 of the first sequence, the amino acid is Trp, Tyr or Phe; at position 4 of the first sequence, the amino acid is Trp, Tyr or Phe; at position 8 of the first sequence, the amino acid is Arg, Lys or His; at positions 9, 10, 12 and 14, respectively, of the first sequence, and at position 4 of the second sequence, the amino acid is any of the 20 naturally occurring amino acid residues with the provisos that, in the first amino acid sequence, (i) when the amino residue at position 12 is Ser, then the amino acid residue at position 14 is not Ser, and (ii) when the amino residue at position 12 is Gly, then the amino acid residue at position 14 is not Ala, performs very well in industrial applications such as laundry compositions, for biopolishing of newly manufactured textiles, for providing an abraded look of cellulosic fabric or garment, and for treatment of paper pulp. Further, the invention relates to DNA constructs encoding such enzymes, a method for providing a gene encoding for such enzymes, a method of producing the enzymes, enzyme preparations containing such enzymes, and the use of these enzymes for a number of industrial applications.

1 . An enzyme preparation consisting essentially of an enzyme which has cellulytic activity and comprises a first amino acid sequence having the following sequence 
       
         
           
                 
                 
               
                   (SEQ ID NO: 79) 
                     
                 
                 
                 
               
                   Thr Arg Xaa Xaa Asp Cys Cys Xaa Xaa Xaa Cys Xaa 
                     
                 
                   1   2   3   4   5   6   7   8   9   10  11  12 
                 
                     
                 
                   Trp Xaa 
                 
                   13  14 
                 
             
                
               
            
             
                
                
                
                
                
               
            
           
         
       
       and a second amino acid sequence having the following sequence 
       
         
           
                 
                 
                 
                 
               
                     
                   Trp Cys Cys Xaa Cys 
                   (SEQ ID NO: 80) 
                     
                 
                     
                   1   2   3   4   5 
                 
             
                
                
               
            
           
         
       
       wherein
 (a) the amino acid residue at position 3 of the first sequence is Trp, Tyr or Phe; 
 (b) the amino acid residue at position 4 of the first sequence is Trp, Tyr or Phe; 
 (c) the amino acid residue at position 8 of the first sequence is Arg, Lys or His; and 
 (d) the amino acid residues at positions 9, 10, 12 and 14 of the first sequence and at position 4 of the second sequence are independently any of the 20 naturally occurring amino acid residues, provided that, in the first amino acid sequence, (i) when the amino acid residue at position 12 is Ser, then the amino acid residue at position 14 is not Ser, and (ii) when the amino acid residue at position 12 is Gly, then the amino acid residue at position 14 is not Ala. 
 
     
     
         2 - 10 . (canceled) 
     
     
         11 . An enzyme preparation consisting essentially of an enzyme having cellulytic activity and being obtainable from a strain belonging to Hymenomycetes (Basidiomycota) which enzyme comprises an amino acid sequence selected from the group consisting of the sequences 
       
         
           
                 
                 
                 
                 
               
                     
                   Xaa Thr Arg Xaa Phe Asp Xaa 
                   (SEQ ID NO: 105) 
                     
                 
                     
                   1   2   3   4   5   6   7; 
                 
                     
                     
                 
                     
                   Xaa Thr Arg Xaa Tyr Asp Xaa 
                   (SEQ ID NO: 106) 
                 
                     
                   1   2   3   4   5   6   7; 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   Xaa Thr Arg Xaa Trp Asp Xaa 
                   (SEQ ID NO: 107) 
                 
                     
                   1   2   3   4   5   6   7 
                 
             
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       wherein
 (a) Xaa at position 4 is Trp, Tyr or Phe; and 
 (b) Xaa at positions 1 and 7 is independently any of the 20 naturally occurring amino acid residues. 
 
     
     
         12 - 65 . (canceled) 
     
     
         66 . A DNA construct comprising a DNA sequence encoding an enzyme exhibiting cellulytic activity, which DNA sequence comprises
 (a) the DNA sequence of SEQ ID NO: 1, and/or the DNA sequence obtainable from the plasmid in  Saccharomyces cerevisiae  DSM 9770, or   (b) an analogue of the DNA sequence of SEQ ID NO: 1 or the DNA sequence obtainable from the plasmid in  Saccharomyces cerevisiae  DSM 9770, which
 (i) is homologous, preferably at least 70% homologous, with the DNA sequence of SEQ ID NO: 1 and/or the DNA sequence obtainable from the plasmid in  Saccharomyces cerevisiae  DSM 9770, 
 (ii) hybridizes under the conditions described herein with the same nucleotide probe as the DNA sequence of SEQ ID NO: 1 and/or the DNA sequence obtainable from the plasmid in  Saccharomyces cerevisiae  DSM 9770, 
 (iii) encodes a polypeptide which is homologous preferably at least 65% homologous, with the polypeptide encoded by a DNA sequence comprising the DNA sequence of SEQ ID NO: 1 and/or the DNA sequence obtainable from the plasmid in  Saccharomyces cerevisiae  DSM 9770, 
 (iv) encodes a polypeptide which is immunologically reactive with an antibody raised against the purified endoglucanase encoded by the DNA sequence of SEQ ID NO: 1 or obtainable from the plasmid in  Saccharomyces cerevisiae , DSM 9770. 
   
     
     
         67 . The DNA construct of  claim 66 , in which the DNA sequence is isolated from or produced on the basis of a DNA library of a strain belonging to the family Chaetomiaceae, preferably to the genus  Myceliophthora , in particular a strain of  M. thermophila , especially  M. thermophila , CBS 117.65. 
     
     
         68 - 83 . (canceled) 
     
     
         84 . A recombinant expression vector comprising a DNA construct of  claim 66 . 
     
     
         85 . A cell comprising a DNA construct of  claim 66 . 
     
     
         86 . A cell comprising a recombinant expression vector of  claim 84 . 
     
     
         87 - 88 . (canceled) 
     
     
         89 . A method of producing an enzyme exhibiting endoglucanase activity, comprising culturing a cell of  claim 85  under conditions permitting the production of the enzyme, and recovering the enzyme from the culture. 
     
     
         90 . (canceled) 
     
     
         91 . A method of providing colour clarification of laundry, which method comprising treating the laundry with a soaking, washing or rinsing liquor comprising an enzyme preparation of  claim 1 . 
     
     
         92 - 96 . (canceled) 
     
     
         97 . A laundry composition comprising the enzyme preparation of  claim 1 , and a compound selected from the group consisting of a surfactant, a builder compound, and a fabric softening agent. 
     
     
         98 . The laundry composition of  claim 97 , which further comprises one or more enzymes selected from the group consisting of proteases, amylases, lipases, cellulases, xylanases, peroxidases and laccases. 
     
     
         99 . The composition of  claim 97 , wherein the surfactant is a nonionic, anionic, cationic, zwitterionic, ampholytic or amphoteric surfactant. 
     
     
         100 . The composition of  claim 97 , wherein the fabric softening agent is a cationic or nonionic softening agent, preferably a quaternary ammonium compound, and which optionally further comprises one or more compounds selected from a surfactant an electrolyte, a buffer, an antioxidant and a liquid carrier. 
     
     
         101 - 105 . (canceled)