Polypeptides having xylanase activity and polynucleotides encoding same
View Patent ↗The present invention relates to polypeptides having xylanase activity, catalytic domains, and carbohydrate binding domains, and polynucleotides encoding the polypeptides, catalytic domains, and carbohydrate binding domains. The present invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides, catalytic domains, and carbohydrate binding domains.
1. A process for degrading or converting a cellulosic or xylan-containing material, comprising: treating the cellulosic or xylan-containing material with an enzyme composition in the presence of a polypeptide having xylanase activity selected from the group consisting of:
(a) a polypeptide having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2;
(b) a polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the cDNA sequence thereof, or (iii) the full-length complement of (i) or (ii), 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 2X SSC, 0.2% SDS at 70° C.;
(c) a polypeptide encoded by a polynucleotide having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or the cDNA sequence thereof;
(d) a polypeptide comprising SEQ ID NO: 2 or the mature polypeptide thereof; and
(e) a fragment of the polypeptide of (a), (b), (c), or (d), that has xylanase activity.
2. A process for producing a fermentation product, comprising:
(a) saccharifying a cellulosic or xylan-containing material with an enzyme composition in the presence of a polypeptide having xylanase activity selected from the group consisting of:
(i) a polypeptide having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2;
(ii) a polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the cDNA sequence thereof, or (iii) the full-length complement of (i) or (ii), 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 2X SSC, 0.2% SDS at 70° C.;
(iii) a polypeptide encoded by a polynucleotide having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or the cDNA sequence thereof;
(iv) a polypeptide comprising SEQ ID NO: 2, or the mature polypeptide thereof; and
(v) a fragment of the polypeptide of (i), (ii), (iii), or (iv), that has xylanase activity;
(b) fermenting the saccharified cellulosic or xylan-containing material with one or more fermenting microorganisms to produce the fermentation product; and
(c) recovering the fermentation product from the fermentation.
3. A process of fermenting a cellulosic or xylan-containing material, comprising: fermenting the cellulosic or xylan-containing material with one or more fermenting microorganisms, wherein the cellulosic or xylan-containing material is saccharified with an enzyme composition in the presence of a polypeptide having xylanase activity selected from the group consisting of:
(a) a polypeptide having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2;
(b) a polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the cDNA sequence thereof, or (iii) the full-length complement of (i) or (ii), 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 2X SSC, 0.2% SDS at 70° C.;
(c) a polypeptide encoded by a polynucleotide having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or the cDNA sequence thereof;
(d) a polypeptide comprising SEQ ID NO: 2 or the mature polypeptide thereof; and
(e) a fragment of the polypeptide of (a), (b), (c), or (d), that has xylanase activity.
4. A nucleic acid construct comprising a polynucleotide encoding a polypeptide having xylanase activity, wherein the polynucleotide is operably linked to one or more heterologous control sequences that direct the production of the polypeptide, and wherein the polypeptide having xylanase activity is selected from the group consisting of:
(a) a polypeptide having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2;
(b) a polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the cDNA sequence thereof, or (iii) the full-length complement of (i) or (ii), 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 2X SSC, 0.2% SDS at 70° C.;
(c) a polypeptide encoded by a polynucleotide having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or the cDNA sequence thereof;
(d) a polypeptide comprising SEQ ID NO: 2 or the mature polypeptide thereof; and
(e) a fragment of the polypeptide of (a), (b), (c), or (d), that has xylanase activity.
5. A recombinant host cell comprising the nucleic acid construct of claim 4 .
6. A method of producing a polypeptide having xylanase activity, comprising:
(a) cultivating the recombinant host cell of claim 5 under conditions conducive for production of the polypeptide; and optionally
(b) recovering the polypeptide.
7. A transgenic plant, plant part or plant cell transformed with a polynucleotide encoding a polypeptide having xylanase activity, wherein the polypeptide having xylanase activity is selected from the group consisting of:
(a) a polypeptide having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2;
(b) a polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the cDNA sequence thereof, or (iii) the full-length complement of (i) or (ii), 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 2X SSC, 0.2% SDS at 70° C.;
(c) a polypeptide encoded by a polynucleotide having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or the cDNA sequence thereof;
(d) a polypeptide comprising SEQ ID NO: 2 or the mature polypeptide thereof; and
(e) a fragment of the polypeptide of (a), (b), (c), or (d), that has xylanase activity.
8. A method of producing a polypeptide having xylanase activity, comprising:
(a) cultivating the transgenic plant or plant cell of claim 7 under conditions conducive for production of the polypeptide; and optionally
(b) recovering the polypeptide.
9. A method of producing a mutant of a parent cell, comprising inactivating a polynucleotide encoding a polypeptide having xylanase activity, which results in the mutant producing less of the polypeptide than the parent cell, wherein the polypeptide having xylanase activity is selected from the group consisting of:
(a) a polypeptide having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2;
(b) a polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the cDNA sequence thereof, or (iii) the full-length complement of (i) or (ii), 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 2X SSC, 0.2% SDS at 70° C.;
(c) a polypeptide encoded by a polynucleotide having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or the cDNA sequence thereof;
(d) a polypeptide comprising SEQ ID NO: 2 or the mature polypeptide thereof; and
(e) a fragment of the polypeptide of (a), (b), (c), or (d), that has xylanase activity.
10. A method of producing a protein, comprising:
(a) cultivating a recombinant host cell comprising a gene encoding a protein operably linked to a polynucleotide encoding a signal peptide comprising or consisting of amino acids 1 to 22 of SEQ ID NO: 2, wherein the gene is foreign to the polynucleotide encoding the signal peptide, under conditions conducive for production of the protein; and optionally
(b) recovering the protein.
11. The process of claim 1 , wherein the cellulosic or xylan-containing material is pretreated.
12. The process of claim 1 , wherein the enzyme composition comprises one or more enzymes selected from the group consisting of a cellulase, a polypeptide having cellulolytic enhancing activity, a hemicellulase, an esterase, an expansin, a laccase, a ligninolytic enzyme, a pectinase, a peroxidase, a protease, and a swollenin.
13. The process of claim 1 , further comprising recovering the degraded or converted cellulosic or xylan-containing material.
14. The process of claim 1 , wherein the degraded or converted cellulosic or xylan-containing material is a sugar.
15. The process of claim 14 , wherein the sugar is selected from the group consisting of glucose, xylose, mannose, galactose, and arabinose.
16. The process of claim 1 , wherein the polypeptide having xylanase activity comprises the mature polypeptide of SEQ ID NO: 2.
17. The process of claim 2 , wherein the cellulosic or xylan-containing material is pretreated.
18. The process of claim 2 , wherein the enzyme composition comprises one or more enzymes selected from the group consisting of a cellulase, a polypeptide having cellulolytic enhancing activity, a hemicellulase, an esterase, an expansin, a laccase, a ligninolytic enzyme, a pectinase, a peroxidase, a protease, and a swollenin.
19. The process of claim 2 , wherein steps (a) and (b) are performed simultaneously in a simultaneous saccharification and fermentation.
20. The process of claim 2 , wherein the fermentation product is an alcohol, an alkane, a cycloalkane, an alkene, an amino acid, a gas, isoprene, a ketone, an organic acid, or polyketide.
21. The process of claim 2 , wherein the polypeptide having xylanase activity comprises the mature polypeptide of SEQ ID NO: 2.
22. The process of claim 3 , wherein the cellulosic or xylan-containing material is pretreated.
23. The process of claim 3 , wherein the enzyme composition comprises one or more enzymes selected from the group consisting of a cellulase, a polypeptide having cellulolytic enhancing activity, a hemicellulase, an esterase, an expansin, a laccase, a ligninolytic enzyme, a pectinase, a peroxidase, a protease, and a swollenin.
24. The process of claim 3 , wherein the fermenting of the cellulosic or xylan-containing material produces a fermentation product.
25. The process of claim 24 , further comprising recovering the fermentation product from the fermentation.
26. The process of claim 24 , wherein the fermentation product is an alcohol, an alkane, a cycloalkane, an alkene, an amino acid, a gas, isoprene, a ketone, an organic acid, or polyketide.
27. The process of claim 3 , wherein the polypeptide having xylanase activity comprises the mature polypeptide of SEQ ID NO: 2.
28. A recombinant host cell comprising a nucleic acid construct comprising a polynucleotide encoding a polypeptide having xylanase activity, wherein the polynucleotide is operably linked to one or more control sequences that direct the production of the polypeptide, wherein the polypeptide having xylanase activity is heterologous to the recombinant host cell, and wherein the polypeptide having xylanase activity is selected from the group consisting of:
(a) a polypeptide having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2;
(b) a polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the cDNA sequence thereof, or (iii) the full-length complement of (i) or (ii), 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 2X SSC, 0.2% SDS at 70° C.;
(c) a polypeptide encoded by a polynucleotide having at least 95% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 or the cDNA sequence thereof;
(d) a polypeptide comprising SEQ ID NO: 2 or the mature polypeptide thereof; and
(e) a fragment of the polypeptide of (a), (b), (c), or (d), that has xylanase activity.
29. A method of producing a polypeptide having xylanase activity, comprising:
(a) cultivating the recombinant host cell of claim 28 under conditions conducive for production of the polypeptide; and optionally
(b) recovering the polypeptide.
30. The process of claim 1 , wherein the polypeptide having xylanase activity has at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2.
31. The process of claim 1 , wherein the polypeptide having xylanase activity has at least 96% sequence identity to the mature polypeptide of SEQ ID NO: 2.
32. The process of claim 1 , wherein the polypeptide having xylanase activity has at least 97% sequence identity to the mature polypeptide of SEQ ID NO: 2.
33. The process of claim 1 , wherein the polypeptide having xylanase activity has at least 98% sequence identity to the mature polypeptide of SEQ ID NO: 2.
34. The process of claim 1 , wherein the polypeptide having xylanase activity has at least 99% sequence identity to the mature polypeptide of SEQ ID NO: 2.
35. The process of claim 2 , wherein the polypeptide having xylanase activity has at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2.
36. The process of claim 2 , wherein the polypeptide having xylanase activity has at least 96% sequence identity to the mature polypeptide of SEQ ID NO: 2.
37. The process of claim 2 , wherein the polypeptide having xylanase activity has at least 97% sequence identity to the mature polypeptide of SEQ ID NO: 2.
38. The process of claim 2 , wherein the polypeptide having xylanase activity has at least 98% sequence identity to the mature polypeptide of SEQ ID NO: 2.
39. The process of claim 2 , wherein the polypeptide having xylanase activity has at least 99% sequence identity to the mature polypeptide of SEQ ID NO: 2.
40. The process of claim 3 , wherein the polypeptide having xylanase activity has at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2.
41. The process of claim 3 , wherein the polypeptide having xylanase activity has at least 96% sequence identity to the mature polypeptide of SEQ ID NO: 2.
42. The process of claim 3 , wherein the polypeptide having xylanase activity has at least 97% sequence identity to the mature polypeptide of SEQ ID NO: 2.
43. The process of claim 3 , wherein the polypeptide having xylanase activity has at least 98% sequence identity to the mature polypeptide of SEQ ID NO: 2.
44. The process of claim 3 , wherein the polypeptide having xylanase activity has at least 99% sequence identity to the mature polypeptide of SEQ ID NO: 2.
45. The nucleic acid construct of claim 5 , wherein the polypeptide having xylanase activity has at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2.
46. The nucleic acid construct of claim 5 , wherein the polypeptide having xylanase activity has at least 96% sequence identity to the mature polypeptide of SEQ ID NO: 2.
47. The nucleic acid construct of claim 5 , wherein the polypeptide having xylanase activity has at least 97% sequence identity to the mature polypeptide of SEQ ID NO: 2.
48. The nucleic acid construct of claim 5 , wherein the polypeptide having xylanase activity has at least 98% sequence identity to the mature polypeptide of SEQ ID NO: 2.
49. The nucleic acid construct of claim 5 , wherein the polypeptide having xylanase activity has at least 99% sequence identity to the mature polypeptide of SEQ ID NO: 2.
50. The nucleic acid construct of claim 5 , wherein the polypeptide having xylanase activity comprises the mature polypeptide of SEQ ID NO: 2.
51. The recombinant host cell of claim 28 , wherein the polypeptide having xylanase activity has at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2.
52. The recombinant host cell of claim 28 , wherein the polypeptide having xylanase activity has at least 96% sequence identity to the mature polypeptide of SEQ ID NO: 2.
53. The recombinant host cell of claim 28 , wherein the polypeptide having xylanase activity has at least 97% sequence identity to the mature polypeptide of SEQ ID NO: 2.
54. The recombinant host cell of claim 28 , wherein the polypeptide having xylanase activity has at least 98% sequence identity to the mature polypeptide of SEQ ID NO: 2.
55. The recombinant host cell of claim 28 , wherein the polypeptide having xylanase activity has at least 99% sequence identity to the mature polypeptide of SEQ ID NO: 2.
56. The recombinant host cell of claim 28 , wherein the polypeptide having xylanase activity comprises the mature polypeptide of SEQ ID NO: 2.