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 21 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 235 to 1491 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 235 to 1491 of SEQ ID NO: 5; and   (d) a fragment of the amino acid sequence of amino acids 21 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% sequence identity to the amino acid sequence of amino acids 21 to 392 of SEQ ID NO: 6. 
     
     
         18 . The nucleic acid construct of  claim 16 , wherein the polypeptide having glucuronyl esterase activity comprises an amino acid sequence that has at least 97% sequence identity to the amino acid sequence of amino acids 21 to 392 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 SEQ ID NO: 6 or the amino acid sequence of amino acids 21 to 392 of SEQ ID NO: 6. 
     
     
         20 . The nucleic acid construct of  claim 16 , wherein the polypeptide having glucuronyl esterase activity consists of the amino acid sequence of SEQ ID NO: 6 or the amino acid sequence of amino acids 21 to 392 of SEQ ID NO: 6. 
     
     
         21 . The nucleic acid construct of  claim 16 , wherein the polypeptide is a fragment of the amino acid sequence of SEQ ID NO: 6, wherein the fragment has glucuronyl esterase activity. 
     
     
         22 . The nucleic acid construct of  claim 16 , wherein the polypeptide having glucuronyl esterase activity is encoded by a polynucleotide that hybridizes under very high stringency conditions with the full-length complement of the nucleotide sequence of nucleotides 235 to 1491 of SEQ ID NO: 5, wherein the very 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 70° C. 
     
     
         23 . An isolated recombinant host cell transformed with the nucleic acid construct of  claim 16 . 
     
     
         24 . A method of producing a polypeptide having glucuronyl esterase activity, comprising:
 (a) cultivating the isolated recombinant host cell of  claim 23  under conditions conducive for production of the polypeptide; and   (b) recovering the polypeptide.   
     
     
         25 . An isolated recombinant host cell transformed with a nucleic acid construct comprising a polynucleotide encoding a polypeptide having glucuronyl esterase activity operably linked to one or more control sequences that direct the expression of the polypeptide in the host cell, 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 21 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 235 to 1491 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 235 to 1491 of SEQ ID NO: 5; and   (d) a fragment of the amino acid sequence of amino acids 21 to 392 of SEQ ID NO: 6, wherein the fragment has glucuronyl esterase activity.   
     
     
         26 . The recombinant host cell of  claim 25 , wherein the polypeptide having glucuronyl esterase activity comprises an amino acid sequence that has at least 95% sequence identity to the amino acid sequence of amino acids 21 to 392 of SEQ ID NO: 6. 
     
     
         27 . The recombinant host cell of  claim 25 , wherein the polypeptide having glucuronyl esterase activity comprises an amino acid sequence that has at least 97% sequence identity to the amino acid sequence of amino acids 21 to 392 of SEQ ID NO: 6. 
     
     
         28 . The recombinant host cell of  claim 25 , wherein the polypeptide having glucuronyl esterase activity comprises the amino acid sequence of SEQ ID NO: 6 or the amino acid sequence of amino acids 21 to 392 of SEQ ID NO: 6. 
     
     
         29 . The recombinant host cell of  claim 25 , wherein the polypeptide having glucuronyl esterase activity consists of the amino acid sequence of SEQ ID NO: 6 or the amino acid sequence of amino acids 21 to 392 of SEQ ID NO: 6. 
     
     
         30 . The recombinant host cell of  claim 25 , wherein the polypeptide is a fragment of the amino acid sequence of SEQ ID NO: 6, wherein the fragment has glucuronyl esterase activity. 
     
     
         31 . The recombinant host cell of  claim 25 , wherein the polypeptide having glucuronyl esterase activity is encoded by a polynucleotide that hybridizes under very high stringency conditions with the full-length complement of the nucleotide sequence of nucleotides 235 to 1491 of SEQ ID NO: 5, wherein the very 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 70° C. 
     
     
         32 . A method of producing a polypeptide having glucuronyl esterase activity, comprising:
 (a) cultivating the isolated recombinant host cell of  claim 22  under conditions conducive for production of the polypeptide; and   (b) recovering the polypeptide.