Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same
View Patent ↗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. An isolated polynucleotide comprising a nucleotide sequence encoding a polypeptide having cellulolytic enhancing activity, selected from the group consisting of:
(a) a polynucleotide encoding a polypeptide having cellulolytic enhancing activity comprising an amino acid sequence having at least 95% identity to amino acids 22 to 332 of SEQ ID NO: 2;
(b) a polynucleotide encoding a polypeptide having cellulolytic enhancing activity that hybridizes under high stringency conditions with the full-length complement of nucleotides 64 to 1018 of SEQ ID NO: 1, wherein high stringency conditions are defined as prehebridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/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 polynucleotide encoding a polypeptide having cellulolytic enhancing activity comprising amino acids 22 to 332 of SEQ ID NO: 2;
(d) a polynucleotide encoding a polypeptide having cellulolytic enhancing activity comprising nucleotides 64 to 1018 of SEQ ID NO: 1; and
(e) a fragment of the polynucleotide of (a), (b), (c), or (d) encoding a polypeptide fragment having cellulolytic enhancing activity.
2. The isolated polynucleotide of claim 1 , wherein the polypeptide having cellulolytic enhancing activity comprises an amino acid sequence having at least 96% sequence identity to amino acids 22 to 332 of SEQ ID NO: 2.
3. The isolated polynucleotide of claim 2 , wherein the polypeptide having cellulolytic enhancing activity comprises an amino acid sequence having at least 97% sequence identity to amino acids 22 to 332 of SEQ ID NO: 2.
4. The isolated polynucleotide of claim 3 , wherein the polypeptide having cellulolytic enhancing activity comprises an amino acid sequence having at least 98% sequence identity to amino acids 22 to 332 of SEQ ID NO: 2.
5. The isolated polynucleotide of claim 4 , wherein the polypeptide having cellulolytic enhancing activity comprises an amino acid sequence having at least 99% sequence identity to amino acids 22 to 332 of SEQ ID NO: 2.
6. The isolated polynucleotide of claim 1 , which hybridizes under very high stringency conditions with the full-length complement of nucleotides 64 to 1018 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 micrograms/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.
7. The isolated polynucleotide of claim 1 , which is contained in plasmid pGEM-T-Ppin7 which is contained in E. coli DSM 22711.
8. A nucleic acid construct or expression vector comprising the polynucleotide of claim 1 operably linked to one or more control sequences that direct the production of the polypeptide in an expression host.
9. A recombinant host cell comprising the polynucleotide of claim 1 operably linked to one or more control sequences that direct the production of the polypeptide.
10. A method of producing a polypeptide having cellulolytic enhancing activity, comprising: (a) cultivating the recombinant host cell of claim 9 under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide.
11. A method of producing a polypeptide having cellulolytic enhancing activity, comprising: (a) cultivating a transgenic plant or a plant cell comprising the polynucleotide of claim 1 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 polynucleotide of claim 1 .