Polypeptides having xylanase activity and polynucleotides thereof

Patent metadata

Field	Value
Publication number	US-9657283-B2
Application number	US-201514668599-A
Country	US
Kind code	B2
Filing date	Mar 25, 2015
Priority date	Feb 12, 2004
Publication date	May 23, 2017
Grant date	May 23, 2017

Abstract

The present invention relates to isolated polypeptides having xylanase 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.

First claim

What is claimed is:

1. A nucleic acid construct comprising an isolated polynucleotide encoding a polypeptide having xylanase activity operably linked to one or more heterologous control sequences that direct production of the polypeptide in a recombinant expression host, wherein the polynucleotide is selected from the group consisting of:
(a) a polynucleotide encoding a polypeptide having xylanase activity comprising an amino acid sequence having at least 95% identity to amino acids 20 to 323 of SEQ ID NO: 4;
(b) a polynucleotide encoding a polypeptide having xylanase activity which hybridizes under at least high stringency conditions with (i) nucleotides 58 to 1400 of SEQ ID NO: 3, (ii) the cDNA sequence of nucleotides 58 to 1400 of SEQ ID NO: 3, or (iii) the full-length complement of (i) or (ii), 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 in 2×SSC, 0.2% SDS at 65° C.;
(c) a polynucleotide encoding a polypeptide having xylanase activity comprising a polynucleotide sequence having at least 95% identity to nucleotides 58 to 1400 of SEQ ID NO: 3;
(d) a polynucleotide encoding a polypeptide having xylanase activity comprising amino acids 20 to 323 of SEQ ID NO: 4; and
(e) a polynucleotide encoding a polypeptide having xylanase activity comprising nucleotides 58 to 1400 of SEQ ID NO: 3.

2. The nucleic acid construct of claim 1 , wherein the polynucleotide encodes a polypeptide having xylanase activity comprising an amino acid sequence having at least 95% identity to amino acids 20 to 323 of SEQ ID NO: 4.

3. The nucleic acid construct of claim 1 , wherein the polynucleotide encodes a polypeptide having xylanase activity comprising an amino acid sequence having at least 97% identity to amino acids 20 to 323 of SEQ ID NO: 4.

4. The nucleic acid construct of claim 1 , wherein the polynucleotide encodes a polypeptide having xylanase activity comprising the amino acid sequence of SEQ ID NO: 4, or a fragment thereof having xylanase activity.

5. The nucleic acid construct of claim 1 , wherein the polynucleotide encodes a polypeptide having xylanase activity comprising the amino acid sequence of SEQ ID NO: 4.

6. The nucleic acid construct of claim 1 , wherein the polynucleotide encodes a polypeptide having xylanase activity comprising amino acids 20 to 323 of SEQ ID NO: 4.

7. The nucleic acid construct of claim 1 , wherein the polynucleotide hybridizes under at least high stringency conditions with (i) nucleotides 58 to 1400 of SEQ ID NO: 3, (ii) the cDNA sequence of nucleotides 58 to 1400 of SEQ ID NO: 3, or (iii) the full-length complement of (i) or (ii), 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 in 2×SSC, 0.2% SDS at 65° C.

8. The nucleic acid construct of claim 7 , wherein the polynucleotide hybridizes under at least very high stringency conditions with (i) nucleotides 58 to 1400 of SEQ ID NO: 3, (ii) the cDNA sequence of nucleotides 58 to 1400 of SEQ ID NO: 3, 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 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.

9. The nucleic acid construct of claim 1 , wherein the polynucleotide comprises a polynucleotide sequence having at least 95% identity to nucleotides 58 to 1400 of SEQ ID NO: 3.

10. The nucleic acid construct of claim 9 , wherein the polynucleotide comprises a polynucleotide sequence having at least 97% identity to nucleotides 58 to 1400 of SEQ ID NO: 3.

11. The nucleic acid construct of claim 1 , wherein the polynucleotide comprises SEQ ID NO: 3 or nucleotides 58 to 1400 of SEQ ID NO: 3.

12. The nucleic acid construct of claim 1 , wherein the polynucleotide comprises the polynucleotide sequence contained in plasmid pJLin162 which is contained in E. coli NRRL B-30702.

13. A recombinant expression vector comprising the nucleic acid construct of claim 1 .

14. An isolated recombinant host cell comprising the nucleic acid construct of claim 1 .

15. A method for producing a polypeptide having xylanase activity comprising: (a) cultivating the recombinant host cell of claim 14 under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide.

16. The nucleic acid construct of claim 1 , wherein the polynucleotide is obtained by (a) hybridizing a population of DNA under at least high stringency conditions with (i) nucleotides 58 to 1400 of SEQ ID NO: 3, (ii) the cDNA sequence of nucleotides 58 to 1400 of SEQ ID NO: 3, or (iii) the full-length complementary strand of (i) or (ii); and (b) isolating the hybridizing polynucleotide, which encodes a polypeptide having xylanase activity, 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 in 2×SSC, 0.2% SDS at 65° C.

17. The nucleic acid construct of claim 16 , wherein the polynucleotide is obtained by (a) hybridizing a population of DNA under very high stringency conditions with (i) nucleotides 58 to 1400 of SEQ ID NO: 3, (ii) the cDNA sequence of nucleotides 58 to 1400 of SEQ ID NO: 3, or (iii) the full-length complementary strand of (i) or (ii); and (b) isolating the hybridizing polynucleotide, which encodes a polypeptide having xylanase activity, 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.

18. A transgenic plant, plant part or plant cell, which has been transformed with the nucleic acid construct of claim 1 .

19. A method for producing a polypeptide having xylanase activity, comprising: (a) cultivating the transgenic plant or the plant cell of claim 18 under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide.

20. The nucleic acid construct of claim 1 , wherein the polynucleotide encodes a polypeptide having xylanase activity comprising an amino acid sequence having at least 98% identity to amino acids 20 to 323 of SEQ ID NO: 4.

21. The nucleic acid construct of claim 1 , wherein the polynucleotide encodes a polypeptide having xylanase activity comprising an amino acid sequence having at least 99% identity to amino acids 20 to 323 of SEQ ID NO: 4.

22. An isolated recombinant host cell transformed with a nucleic acid construct comprising an isolated polynucleotide encoding a polypeptide having xylanase activity operably linked to one or more control sequences that direct production of the polypeptide in the recombinant host cell, wherein the polypeptide having xylanase activity is heterologous to the recombinant host cell and wherein the polynucleotide is selected from the group consisting of:
(a) a polynucleotide encoding a polypeptide having xylanase activity comprising an amino acid sequence having at least 95% identity to amino acids 20 to

Assignees

• Novozymes Inc

Inventors

• Yaver Debbie
• Harris Paul
• Otani Suzanne
• Lin Janine
• Ding Hanshu

Classifications

• C12N9/248

  Xylanases · CPC title

• C12N9/2482 Primary

  Endo-1,4-beta-xylanase (3.2.1.8) · CPC title

• C12Y302/01008

  Endo-1,4-beta-xylanase (3.2.1.8) · CPC title