Polypeptides Having Glucuronyl Esterase Activity and Polynucleotides Encoding Same

Abstract

The present invention relates to isolated polypeptides having glucuronyl esterase activity, catalytic domains and polynucleotides encoding the polypeptides or catalytic 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 or catalytic domains.

1 - 15 . (canceled) 
     
     
         16 . A nucleic acid construct comprising a polynucleotide encoding a polypeptide having glucuronyl esterase activity operably linked to one or more heterologous control sequences that direct the expression of the polypeptide in an expression host, wherein the polypeptide having glucuronyl esterase activity is selected from the group consisting of:
 (a) a polypeptide comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of amino acids 1 to 392 of SEQ ID NO: 6;   (b) a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions with the full-length complement of the nucleotide sequence of nucleotides 1 to 2508 of SEQ ID NO: 5, wherein the high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5× SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide, and washing three times each for 15 minutes using 2× SSC, 0.2% SDS at 65° C.;   (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of nucleotides 1 to 2508 of SEQ ID NO: 5; and   (d) a fragment of the amino acid sequence of amino acids 1 to 392 of SEQ ID NO: 6, wherein the fragment has glucuronyl esterase activity.   
     
     
         17 . The nucleic acid construct of  claim 16 , wherein the polypeptide having glucuronyl esterase activity comprises an amino acid sequence that has at least 95 percent sequence identity to the amino acid sequence of amino acids 1 to 392 of SEQ ID NO: 6. 
     
     
         18 . The nucleic acid construct of  claim 16 , wherein the polypeptide having glucuronyl esterase activity comprises the amino acid sequence of SEQ ID NO: 6. 
     
     
         19 . The nucleic acid construct of  claim 16 , wherein the polypeptide having glucuronyl esterase activity comprises the amino acid sequence of amino acids 1 to 392 of SEQ ID NO:6. 
     
     
         20 . An isolated recombinant host cell comprising the nucleic acid construct of  claim 16 . 
     
     
         21 . The isolated recombinant host cell of  claim 20 , wherein the polypeptide having glucuronyl esterase activity comprises an amino acid sequence that has at least 95 percent sequence identity to the amino acid sequence of amino acids 1 to 392 of SEQ ID NO: 6. 
     
     
         22 . The isolated recombinant host cell of  claim 20 , wherein the polypeptide having glucuronyl esterase activity comprises the amino acid sequence of SEQ ID NO: 6. 
     
     
         23 . The isolated recombinant host cell of  claim 20 , wherein the polypeptide having glucuronyl esterase activity comprises the amino acid sequence of amino acids 1 to 392 of SEQ ID NO: 6. 
     
     
         24 . The isolated recombinant host cell of  claim 20 , which is a yeast host cell. 
     
     
         25 . The isolated recombinant host cell of  24 , wherein the yeast host cell is a  Candida, Hansenula, Kluyveromyces, Pichia, Saccharomyces, Schizosaccharomyces,  or  Yarrowia  cell such as a  Kluyveromyces lactis, Saccharomyces carisbergensis, Saccharomyces cerevisiae, Saccharomyces diastaticus, Saccharomyces douglasii, Saccharomyces kluyveri, Saccharomyces norbensis, Saccharomyces oviformis,  or  Yarrowia lipolytica  cell. 
     
     
         26 . The isolated recombinant host cell of  claim 24 , wherein the yeast host cell is a  Saccharomyces  cell. 
     
     
         27 . The isolated recombinant host cell of  claim 24 , wherein the yeast host cell is a  Saccharomyces cerevisiae  cell. 
     
     
         28 . An expression vector comprising the nucleic acid construct of  claim 16 . 
     
     
         29 . The expression vector of  claim 28 , wherein the polypeptide having glucuronyl esterase activity comprises an amino acid sequence that has at least 95 percent sequence identity to the amino acid sequence of amino acids 1 to 392 of SEQ ID NO: 6. 
     
     
         30 . The expression vector of  claim 28 , wherein the polypeptide having glucuronyl esterase activity comprises the amino acid sequence of SEQ ID NO: 6. 
     
     
         31 . The expression vector of  claim 28 , wherein the polypeptide having glucuronyl esterase activity comprises the amino acid sequence of amino acids 1 to 392 of SEQ ID NO:6. 
     
     
         32 . A method of producing a polypeptide having glucuronyl esterase activity, comprising:
 (a) cultivating an isolated recombinant host cell under conditions conductive for production of the polypeptide, wherein the isolated recombinant host cell comprises a nucleic acid construct comprising a polynucleotide encoding a polypeptide having glucuronyl esterase activity operably linked to one or more heterologous control sequences that direct the expression of the polypeptide in an expression host, wherein the polypeptide having glucuronyl esterase activity is selected from the group consisting of:
 (i) a polypeptide comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence of amino acids 1 to 392 of SEQ ID NO: 6; 
 (ii) a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions with the full-length complement of the nucleotide sequence of nucleotides 1 to 2508 of SEQ ID NO: 5, wherein the high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5× SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon sperm DNA, and 50% formamide, and washing three times each for 15 minutes using 2× SSC, 0.2% SDS at 65° C.; 
 (iii) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of nucleotides 1 to 2508 of SEQ ID NO: 5; and 
 (iv) a fragment of the amino acid sequence of amino acids 1 to 392 of SEQ ID NO: 6, wherein the fragment has glucuronyl esterase activity; and 
   (b) recovering the polypeptide.   
     
     
         33 . The expression vector of  claim 32 , wherein the polypeptide having glucuronyl esterase activity comprises an amino acid sequence that has at least 95 percent sequence identity to the amino acid sequence of amino acids 1 to 392 of SEQ ID NO: 6. 
     
     
         34 . The expression vector of  claim 32 , wherein the polypeptide having glucuronyl esterase activity comprises the amino acid sequence of SEQ ID NO: 6. 
     
     
         35 . The expression vector of  claim 32 , wherein the polypeptide having glucuronyl esterase activity comprises the amino acid sequence of amino acids 1 to 392 of SEQ ID NO:6.