Polypeptides having cellulolytic enhancing activity and nucleic acids 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 for producing and using the polypeptides.
1. An isolated recombinant host cell transformed with a nucleic acid construct comprising a polynucleotide encoding a GH61 polypeptide having cellulolytic enhancing activity, wherein the polynucleotide is operably linked to one or more control sequences that direct the production of the polypeptide, wherein the GH61 polypeptide is heterologous to the recombinant host cell, and wherein the GH61 polypeptide having cellulolytic enhancing activity is selected from the group consisting of:
(a) a GH61 polypeptide comprising an amino acid sequence having at least 90% sequence identity to amino acids 19 to 226 of SEQ ID NO: 8;
(b) a GH61 polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions with the full-length complement of nucleotides 55 to 678 of the polynucleotide of SEQ ID NO: 7, 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 GH61 polypeptide encoded by a polynucleotide having at least 90% sequence identity to nucleotides 55 to 678 of the polynucleotide of SEQ ID NO: 7.
2. The recombinant host cell of claim 1 , wherein the GH61 polypeptide comprises an amino acid sequence having at least 90% sequence identity to amino acids 19 to 226 of SEQ ID NO: 8.
3. The recombinant host cell of claim 1 , wherein the GH61 polypeptide comprises an amino acid sequence having at least 95% sequence identity to amino acids 19 to 226 of SEQ ID NO: 8.
4. The recombinant host cell of claim 1 , wherein the GH61 polypeptide comprises an amino acid sequence having at least 97% sequence identity to amino acids 19 to 226 of SEQ ID NO: 8.
5. The recombinant host cell of claim 1 , wherein the GH61 polypeptide comprises an amino acid sequence having at least 98% sequence identity to amino acids 19 to 226 of SEQ ID NO: 8.
6. The recombinant host cell of claim 1 , wherein the GH61 polypeptide comprises an amino acid sequence having at least 99% sequence identity to amino acids 19 to 226 of SEQ ID NO: 8.
7. The recombinant host cell of claim 1 , wherein the GH61 polypeptide comprises SEQ ID NO: 8 or amino acids 19 to 226 of SEQ ID NO: 8.
8. The recombinant host cell of claim 1 , wherein the GH61 polypeptide is encoded by a polynucleotide that hybridizes under high stringency conditions with the full-length complement of nucleotides 55 to 678 of the polynucleotide of SEQ ID NO: 7, 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.
9. The recombinant host cell of claim 1 , wherein the GH61 polypeptide is encoded by a polynucleotide that hybridizes under very high stringency conditions with the full-length complement of nucleotides 55 to 678 of the polynucleotide of SEQ ID NO: 7, 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.
10. The recombinant host cell of claim 1 , wherein the GH61 polypeptide is encoded by the polynucleotide contained in plasmid pTter61E which is contained in E. coli NRRL B-30814.
11. A method for producing a GH61 polypeptide having cellulolytic enhancing activity, comprising (a) cultivating the recombinant host cell of claim 1 under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide.
12. A nucleic acid construct comprising a polynucleotide encoding a GH61 polypeptide having cellulolytic enhancing activity, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct the production of the polypeptide, and wherein the GH61 polypeptide having cellulolytic enhancing activity is selected from the group consisting of:
(a) a GH61 polypeptide comprising an amino acid sequence having at least 90% sequence identity to amino acids 19 to 226 of SEQ ID NO: 8;
(b) a GH61 polypeptide encoded by a polynucleotide that hybridizes under high stringency conditions with the full-length complement of nucleotides 55 to 678 of the polynucleotide of SEQ ID NO: 7, 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 GH61 polypeptide encoded by a polynucleotide having at least 90% sequence identity to nucleotides 55 to 678 of the polynucleotide of SEQ ID NO: 7.
13. The nucleic acid construct of claim 12 , wherein the GH61 polypeptide comprises SEQ ID NO: 8 or amino acids 19 to 226 of SEQ ID NO: 8.
14. A recombinant host cell transformed with the nucleic acid construct of claim 12 .
15. A method for producing a GH61 polypeptide having cellulolytic enhancing activity, comprising (a) cultivating a recombinant host cell comprising the nucleic acid construct of claim 12 under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide.
16. The nucleic acid construct of claim 12 , wherein the GH61 polypeptide comprises an amino acid sequence having at least 90% sequence identity to amino acids 19 to 226 of SEQ ID NO: 8.
17. The nucleic acid construct of claim 12 , wherein the GH61 polypeptide comprises an amino acid sequence having at least 95% sequence identity to amino acids 19 to 226 of SEQ ID NO: 8.
18. The nucleic acid construct of claim 12 , wherein the GH61 polypeptide comprises an amino acid sequence having at least 97% sequence identity to amino acids 19 to 226 of SEQ ID NO: 8.
19. The nucleic acid construct of claim 12 , wherein the GH61 polypeptide comprises an amino acid sequence having at least 98% sequence identity to amino acids 19 to 226 of SEQ ID NO: 8.
20. The nucleic acid construct of claim 12 , wherein the GH61 polypeptide comprises an amino acid sequence having at least 99% sequence identity to amino acids 19 to 226 of SEQ ID NO: 8.
21. The nucleic acid construct of claim 12 , wherein the GH61 polypeptide is encoded by a polynucleotide that hybridizes under high stringency conditions with the full-length complement of nucleotides 55 to 678 of the polynucleotide of SEQ ID NO: 7, 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.
22. The nucleic acid construct of claim 12 , wherein the GH61 polypeptide is encoded by a polynucleotide that hybridizes under very high stringency conditions with the full-length complement of nucleotides 55 to 678 of the polynucleotide of SEQ ID NO: 7, 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.
23. The nucleic acid construct of claim 12 , wherein the GH61 polypeptide is encoded by the polynucleotide contained in plasmid pTter61E which is contained in E. coli NRRL B-30814.