Polypeptides Having Alpha-Glucuronidase Activity And Polynucleotides Encoding Same

Abstract

The present invention relates to isolated polypeptides having alpha-glucuronidase activity, catalytic domains and polynucleotides encoding the polypeptides, 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 - 18 . (canceled) 
     
     
         19 . A nucleic acid construct comprising a polynucleotide encoding a polypeptide having alpha-glucuronidase activity, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct the production of the polypeptide in an expression host, and wherein the polypeptide having alpha-glucuronidase activity is selected from the group consisting of:
 (a) a polypeptide having at least 80% sequence identity to the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 8;   (b) a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO:1, (ii) the cDNA sequence thereof, or (iii) the full-length complement of (i) or (ii), wherein 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.; and   (c) a polypeptide encoded by a polynucleotide having at least 80% sequence identity to the mature polypeptide coding sequence of SEQ ID NO:1 or the cDNA sequence thereof.   
     
     
         20 . The nucleic acid construct of  claim 19 , wherein the polypeptide has at least 80% sequence identity to the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 8. 
     
     
         21 . The nucleic acid construct of  claim 19 , wherein the polypeptide has at least 85% sequence identity to the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 8. 
     
     
         22 . The nucleic acid construct of  claim 19 , wherein the polypeptide has at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 8. 
     
     
         23 . The nucleic acid construct of  claim 19 , wherein the polypeptide has at least 91% sequence identity to the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 8. 
     
     
         24 . The nucleic acid construct of  claim 19 , wherein the polypeptide has at least 92% sequence identity to the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 8. 
     
     
         25 . The nucleic acid construct of  claim 19 , wherein the polypeptide has at least 93% sequence identity to the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 8. 
     
     
         26 . The nucleic acid construct of  claim 19 , wherein the polypeptide has at least 94% sequence identity to the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 8. 
     
     
         27 . The nucleic acid construct of  claim 19 , wherein the polypeptide has at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 8. 
     
     
         28 . The nucleic acid construct of  claim 19 , wherein the polypeptide has at least 96% sequence identity to the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 8. 
     
     
         29 . The nucleic acid construct of  claim 19 , wherein the polypeptide has at least 97% sequence identity to the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 8. 
     
     
         30 . The nucleic acid construct of  claim 19 , wherein the polypeptide has at least 98% sequence identity to the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 8. 
     
     
         31 . The nucleic acid construct of  claim 19 , wherein the polypeptide has at least 99% sequence identity to the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 8. 
     
     
         32 . The nucleic acid construct of  claim 19 , wherein the polypeptide comprises SEQ ID NO: 6 or SEQ ID NO: 8. 
     
     
         33 . The nucleic acid construct of  claim 19 , wherein the polypeptide consists of SEQ ID NO: 6 or SEQ ID NO: 8. 
     
     
         25 . The nucleic acid construct of  claim 19 , wherein the polypeptide comprises the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 8. 
     
     
         26 . The nucleic acid construct of  claim 19 , wherein the polypeptide consists of the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 8. 
     
     
         27 . The nucleic acid construct of  claim 19 , wherein the polypeptide is encoded by a polynucleotide that hybridizes under high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 5 or the mature polypeptide coding sequence of SEQ ID NO: 7, (ii) the cDNA sequence thereof, or (iii) the full-length complement of (i) or (ii), wherein 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. 
     
     
         28 . The nucleic acid construct of  claim 19 , wherein the polypeptide is encoded by a polynucleotide that hybridizes under very high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 5 or the mature polypeptide coding sequence of SEQ ID NO: 7, (ii) the cDNA sequence thereof, or (iii) the full-length complement of (i) or (ii), wherein 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. 
     
     
         29 . The nucleic acid construct of  claim 19 , wherein the polypeptide is encoded by a polynucleotide having at least 80% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 5 or the mature polypeptide coding sequence of SEQ ID NO: 7. or the cDNA sequence thereof. 
     
     
         30 . The nucleic acid construct of  claim 19 , wherein the polypeptide is encoded by a polynucleotide having at least 85% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 5 or the mature polypeptide coding sequence of SEQ ID NO: 7, or the cDNA sequence thereof. 
     
     
         31 . The nucleic acid construct of  claim 19 , wherein the polypeptide is encoded by a polynucleotide having at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 5 or the mature polypeptide coding sequence of SEQ ID NO: 7, or the cDNA sequence thereof. 
     
     
         32 . The nucleic acid construct of  claim 19 , wherein the polypeptide is encoded by a polynucleotide having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 5 or the mature polypeptide coding sequence of SEQ ID NO: 7, or the cDNA sequence thereof. 
     
     
         33 . The nucleic acid construct of  claim 19 , wherein the polypeptide is a fragment of the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 8, which has alpha-glucuronidase activity. 
     
     
         34 . An expression vector comprising the nucleic acid construct of  claim 19 . 
     
     
         35 . An isolated recombinant host cell transformed with the nucleic acid construct of  claim 19 . 
     
     
         36 . A method of producing a polypeptide having alpha-glucuronidase activity, comprising:
 (a) cultivating the recombinant host cell of  claim 35  under conditions conducive for production of the polypeptide; and   (b) recovering the polypeptide.   
     
     
         37 . An isolated recombinant host cell transformed with a nucleic acid construct comprising a polynucleotide encoding a polypeptide having alpha-glucuronidase activity, wherein the polynucleotide is operably linked to one or more control sequences that direct the production of the polypeptide in an expression host, and wherein the polypeptide having alpha-glucuronidase activity is selected from the group consisting of:
 (a) a polypeptide having at least 80% sequence identity to the mature polypeptide of SEQ ID NO: 6 or the mature polypeptide of SEQ ID NO: 8;   (b) a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO:1, (ii) the cDNA sequence thereof, or (iii) the full-length complement of (i) or (ii), wherein 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.; and   (c) a polypeptide encoded by a polynucleotide having at least 80% sequence identity to the mature polypeptide coding sequence of SEQ ID NO:1 or the cDNA sequence thereof.   
     
     
         38 . A method of producing a polypeptide having alpha-glucuronidase activity, comprising:
 (a) cultivating the recombinant host cell of  claim 37  under conditions conducive for production of the polypeptide; and   (b) recovering the polypeptide.