Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same
The present invention relates to isolated polypeptides having cellulolytic enhancing activity and isolated polynucleotides encoding the polypeptides. 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.
1. A nucleic acid construct comprising an isolated polynucleotide encoding a polypeptide having cellulolytic enhancing activity selected from the group consisting of (a) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to amino acids 26 to 253 of SEQ ID NO: 2; (b) a polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions with the full-length complement of nucleotides 76 to 832 of SEQ ID NO: 1, wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 mg/ml sheared and denatured salmon sperm DNA, and 50% formamide for 12 to 24 hours, 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 95% identity to nucleotides 76 to 832 of SEQ ID NO: 1; and (d) a fragment of SEQ ID NO: 2 having cellulolytic enhancing 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.
2. The nucleic acid construct of claim 1 , wherein the polypeptide comprises an amino acid sequence having at least 97% identity to amino acids 26 to 253 of SEQ ID NO: 2.
3. The nucleic acid construct of claim 1 , wherein the polypeptide comprises or consists of the amino acid sequence of SEQ ID NO: 2; or a fragment thereof having cellulolytic enhancing activity.
4. The nucleic acid construct of claim 1 , wherein the polypeptide comprises or consists of amino acids 26 to 253 of SEQ ID NO: 2.
5. The nucleic acid construct of claim 1 , wherein the polypeptide is encoded by a polynucleotide that hybridizes under very high stringency conditions with the full-length complement of nucleotides 76 to 832 of SEQ ID NO: 1, wherein very high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 mg/ml sheared and denatured salmon sperm DNA, and 50% formamide for 12 to 24 hours, and washing three times each for 15 minutes using 2×SSC, 0.2% SDS at 70° C.
6. The nucleic acid construct of claim 1 , wherein the polypeptide is encoded by a polynucleotide comprising a nucleotide sequence having at least 97% identity to nucleotides 76 to 832 of SEQ ID NO: 1.
7. The nucleic acid construct of claim 1 , wherein the polypeptide is encoded by nucleotides 76 to 832 of SEQ ID NO: 1.
8. The nucleic acid construct of claim 1 , wherein the polypeptide is encoded by the polynucleotide contained in plasmid pGEM-T-GH61D23Y4 which is contained in E. coli DSM 22882.
9. An expression vector comprising the nucleic acid construct of claim 1 .
10. A recombinant host cell comprising the nucleic acid construct of claim 1 .
11. A method of producing a polypeptide having cellulolytic enhancing activity, said method comprising:
(a) cultivating the recombinant host cell of claim 10 under conditions conducive for production of the polypeptide; and
(b) recovering the polypeptide.
12. A transgenic plant, plant part or plant cell transformed with the nucleic acid construct of claim 1 .
13. A method of producing a polypeptide having cellulolytic enhancing activity, said method comprising:
(a) cultivating the transgenic plant or plant cell of claim 12 under conditions conducive for production of the polypeptide; and
(b) recovering the polypeptide.