Polypeptides having beta-xylosidase activity and polynucleotides encoding same
The present invention relates to isolated polypeptides having beta-xylosidase 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 or expression vector comprising a polynucleotide encoding a polypeptide having beta-xylosidase 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 beta-xylosidase activity is selected from the group consisting of:
(a) a polypeptide having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10;
(b) a polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with the full-length complement of the mature polypeptide coding sequence of SEQ ID NO: 1, SEQ ID NO: 3, or SEQ ID NO: 9, wherein very high stringency conditions are defined as prehybridization and hybridization at 42° C. in 5X 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.;
(c) a polypeptide encoded by a polynucleotide having at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1, SEQ ID NO: 3, or SEQ ID NO: 9, or the cDNA sequence thereof;
(d) a variant of the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10 comprising a substitution, deletion, and/or insertion at one or more positions and having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10; and
(e) a fragment of the polypeptide of (a), (b), (c), or (d) that has beta-xylosidase activity.
2. The nucleic acid construct or expression vector of claim 1 , wherein the polypeptide having beta-xylosidase activity has at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10.
3. The nucleic acid construct or expression vector of claim 1 , wherein the polypeptide having beta-xylosidase activity has at least 96% sequence identity to the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10.
4. The nucleic acid construct or expression vector of claim 1 , wherein the polypeptide having beta-xylosidase activity has at least 97% sequence identity to the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10.
5. The nucleic acid construct or expression vector of claim 1 , wherein the polypeptide having beta-xylosidase activity has at least 98% sequence identity to the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10.
6. The nucleic acid construct or expression vector of claim 1 , wherein the polypeptide having beta-xylosidase activity has at least 99% sequence identity to the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10.
7. The nucleic acid construct or expression vector of claim 1 , wherein the polypeptide having beta-xylosidase activity comprises SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10 or the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10.
8. An isolated recombinant host cell transformed with the nucleic acid construct or expression vector of claim 1 .
9. A method of producing a polypeptide having beta-xylosidase activity, comprising: cultivating the recombinant host cell of claim 8 under conditions conducive for production of the polypeptide, and optionally recovering the polypeptide.
10. A transgenic plant, plant part or plant cell transformed with the nucleic acid construct or expression vector of claim 1 .
11. A method of producing a polypeptide having beta-xylosidase activity, comprising: cultivating the transgenic plant or plant cell of claim 10 under conditions conducive for production of the polypeptide, and optionally recovering the polypeptide.
12. An isolated recombinant host cell transformed with a nucleic acid construct or expression vector comprising a polynucleotide encoding a polypeptide having beta-xylosidase activity, wherein the polynucleotide is operably linked to one or more control sequences that direct the production of the polypeptide in an expression host, wherein the polypeptide having beta-xylosidase activity is heterologous to the recombinant host cell, and wherein the polypeptide having beta-xylosidase activity is selected from the group consisting of:
(a) a polypeptide having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10;
(b) a polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with the full-length complement of the mature polypeptide coding sequence of SEQ ID NO: 1, SEQ ID NO: 3, or SEQ ID NO: 9, 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.;
(c) a polypeptide encoded by a polynucleotide having at least 90% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1, SEQ ID NO: 3, or SEQ ID NO: 9, or the cDNA sequence thereof;
(d) a variant of the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10 comprising a substitution, deletion, and/or insertion at one or more positions and having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10; and
(e) a fragment of the polypeptide of (a), (b), (c), or (d) that has beta-xylosidase activity.
13. The recombinant host cell of claim 12 , wherein the polypeptide having beta-xylosidase activity has at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10.
14. The recombinant host cell of claim 12 , wherein the polypeptide having beta-xylosidase activity has at least 96% sequence identity to the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10.
15. The recombinant host cell of claim 12 , wherein the polypeptide having beta-xylosidase activity has at least 97% sequence identity to the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10.
16. The recombinant host cell of claim 12 , wherein the polypeptide having beta-xylosidase activity has at least 98% sequence identity to the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10.
17. The recombinant host cell of claim 12 , wherein the polypeptide having beta-xylosidase activity has at least 99% sequence identity to the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10.
18. The recombinant host cell of claim 12 , wherein the polypeptide having beta-xylosidase activity comprises SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10 or the mature polypeptide of SEQ ID NO: 2, SEQ ID NO: 4, or SEQ ID NO: 10.
19. A method of producing a polypeptide having beta-xylosidase activity, comprising: cultivating the recombinant host cell of claim 12 under conditions conducive for production of the polypeptide, and optionally recovering the polypeptide.