Polypeptides having cellobiohydrolase activity and polynucleotides encoding same
The present invention relates to isolated polypeptides having cellobiohydrolase activity and 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 a polynucleotide encoding a polypeptide having cellobiohydrolase 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 cellobiohydrolase activity is selected from:
(a) a polypeptide that has at least 95% sequence identity to the sequence of amino acids 19-457 of SEQ ID NO: 4,
(b) a polypeptide that is encoded by a polynucleotide that hybridizes with the full-length complement of SEQ ID NO: 3 or the cDNA sequence thereof under high stringency conditions, wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 μg/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 95% sequence identity to nucleotides 55 to 1479 of SEQ ID NO: 3; or the cDNA sequence thereof.
2. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity has at least 95% sequence identity to the sequence of amino acids 19-457 of SEQ ID NO: 4.
3. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity has at least 96% sequence identity to the sequence of amino acids 19-457 of SEQ ID NO: 4.
4. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity has at least 97% sequence identity to the sequence of amino acids 19-457 of SEQ ID NO: 4.
5. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity has at least 98% sequence identity to the sequence of amino acids 19-457 of SEQ ID NO: 4.
6. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity has at least 99% sequence identity to the sequence of amino acids 19-457 of SEQ ID NO: 4.
7. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity is a variant of the sequence of amino acids 19-457 of SEQ ID NO: 4, comprising a substitution, deletion, and/or insertion of one or more amino acids.
8. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity comprises the sequence of amino acids 19-457 of SEQ ID NO: 4.
9. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity is encoded by a polynucleotide that hybridizes with the mature polypeptide coding sequence of SEQ ID NO: 3 or the cDNA sequence thereof under high stringency conditions, wherein high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 μg/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.
10. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity further comprises a cellulose binding domain.
11. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity is encoded by a polynucleotide that hybridizes with the full-length complement of SEQ ID NO: 3 or the cDNA sequence thereof under very high stringency conditions, wherein very high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5×SSPE, 0.3% SDS, 200 μg/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.
12. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity is encoded by a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to nucleotides 55 to 1479 of SEQ ID NO: 3; or the cDNA sequence thereof.
13. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity is encoded by a polynucleotide comprising a nucleotide sequence having at least 96% sequence identity to nucleotides 55 to 1479 of SEQ ID NO: 3; or the cDNA sequence thereof.
14. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity is encoded by a polynucleotide comprising a nucleotide sequence having at least 97% sequence identity to nucleotides 55 to 1479 of SEQ ID NO: 3; or the cDNA sequence thereof.
15. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity is encoded by a polynucleotide comprising a nucleotide sequence having at least 98% sequence identity to nucleotides 55 to 1479 of SEQ ID NO: 3; or the cDNA sequence thereof.
16. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity is encoded by a polynucleotide comprising a nucleotide sequence having at least 99% sequence identity to nucleotides 55 to 1479 of SEQ ID NO: 3; or the cDNA sequence thereof.
17. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity is encoded by a polynucleotide comprising or consisting of nucleotides 55 to 1479 of SEQ ID NO: 3; or the cDNA sequence thereof.
18. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity is encoded by a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 3.
19. A recombinant expression vector comprising the nucleic acid construct of claim 1 .
20. A recombinant host cell comprising the nucleic acid construct of claim 1 .
21. A transgenic plant, plant part or plant cell transformed with the nucleic acid construct of claim 1 .
22. A method of producing a polypeptide having cellobiohydrolase activity, comprising: (a) cultivating the recombinant host cell of claim 20 under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide.
23. A method of producing a polypeptide having cellobiohydrolase activity, comprising: (a) cultivating the transgenic plant or the plant cell of claim 21 under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide.