Polypeptides having cellobiohydrolase activity and polynucleotides encoding same

Patent metadata

Field	Value
Publication number	US-9957493-B2
Application number	US-201615015854-A
Country	US
Kind code	B2
Filing date	Feb 4, 2016
Priority date	Nov 6, 2009
Publication date	May 1, 2018
Grant date	May 1, 2018

Abstract

The present invention relates to isolated polypeptides having cellobiohydrolase 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.

First claim

What is claimed is:

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, and wherein the polypeptide having cellobiohydrolase activity is selected from the group consisting of:
(a) a polypeptide comprising an amino acid sequence having at least 95% 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 the full-length complement of the mature polypeptide coding sequence of SEQ ID NO: 1, or the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 1, wherein the 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.; and
(c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% identity to the mature polypeptide coding sequence of SEQ ID NO: 1.

2. A method of producing a polypeptide having cellobiohydrolase activity, comprising: (a) cultivating a host cell comprising the nucleic acid construct of claim 1 under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide.

3. A method of producing a polypeptide having cellobiohydrolase activity, comprising: (a) cultivating a transgenic plant or a plant cell comprising the nucleic acid construct of claim 1 under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide.

4. A transgenic plant, plant part or plant cell transformed with the nucleic acid construct of claim 1 .

5. A method for degrading or converting a cellulosic material, comprising:
treating the cellulosic material with an enzyme composition comprising a polypeptide having cellobiohydrolase activity, wherein the polypeptide having cellobiohydrolase activity is selected from the group consisting of:
(a) a polypeptide comprising an amino acid sequence having at least 95% 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 the full-length complement of the mature polypeptide coding sequence of SEQ ID NO: 1, or the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 1, wherein the 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.; and
(c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% % identity to the mature polypeptide coding sequence of SEQ ID NO: 1.

6. The method of claim 5 , further comprising recovering the degraded or converted cellulosic material.

7. A method for producing a fermentation product, comprising:
(a) saccharifying a cellulosic material with an enzyme composition comprising a polypeptide having cellobiohydrolase activity, wherein the polypeptide having cellobiohydrolase activity is selected from the group consisting of:
(i) a polypeptide comprising an amino acid sequence having at least 95% 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 the full-length complement of the mature polypeptide coding sequence of SEQ ID NO: 1, or the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 1, wherein the 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.; and
(iii) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% identity to the mature polypeptide coding sequence of SEQ ID NO: 1;
(b) fermenting the saccharified cellulosic material with one or more fermenting microorganisms to produce the fermentation product; and
(c) recovering the fermentation product from the fermentation.

8. A method of fermenting a cellulosic material, comprising: fermenting the cellulosic material with one or more fermenting microorganisms, wherein the cellulosic material is saccharified with an enzyme composition comprising a polypeptide having cellobiohydrolase activity, wherein the polypeptide having cellobiohydrolase activity is selected from the group consisting of:
(a) a polypeptide comprising an amino acid sequence having at least 95% 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 the full-length complement of the mature polypeptide coding sequence of SEQ ID NO: 1, or the genomic DNA sequence of the mature polypeptide coding sequence of SEQ ID NO: 1, wherein the 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.; and
(c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% identity to the mature polypeptide coding sequence of SEQ ID NO: 1.

9. The method of claim 8 , wherein the fermenting of the cellulosic material produces a fermentation product.

10. The method of claim 9 , further comprising recovering the fermentation product from the fermentation.

11. The method of claim 9 , wherein the fermentation product is an alcohol, an organic acid, a ketone, an amino acid, or a gas.

12. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity comprises an amino acid sequence having at least 95% identity to the mature polypeptide of SEQ ID NO: 2.

13. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity comprises the mature polypeptide of SEQ ID NO: 2.

14. The nucleic acid construct of claim 1 , wherein the polypeptide having cellobiohydrolase activity is encoded by the polynucleotide contained in plasmid pMStr199 which is contained in E. coli DSM 23379.

15. The method of claim 5 , wherein the polypeptide having cellobiohydrolase activity comprises an amino acid sequence having at least 95% identity to the mature polypeptide of SEQ ID NO: 2.

16. The method of claim 5 , wherein the polypeptide having cellobiohydrolase activity comprises the mature polypeptide of SEQ ID NO: 2.

17. The method of claim 7 , wherein the polypeptide having cellobiohydrolase activity comprises an amino acid sequence having at least 95% identity to the mature polypeptide of SEQ ID NO: 2.

18. The method of claim 7 , wherein the polypeptide having cellobiohydrolase activity comprises the mature polypeptide of SEQ ID NO: 2.

19. The method of claim 8 , wherein the polypeptide having cellobiohydrolase activity comprises an amino acid sequence having at least 95% identity to the mature polypeptide of SEQ ID NO: 2.

20. The method of

Assignees

• Novozymes Inc
• Novozymes As

Inventors

• Vlasenko Elena
• Mcbrayer Brett
• Stringer Mary
• Schnorr Kirk

Classifications

• C12P19/12

  Disaccharides · CPC title

• C12N15/1137

  against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138) · CPC title

• C12Y302/01091

  Cellulose 1,4-beta-cellobiosidase (3.2.1.91) · CPC title

• C12N9/2437 Primary

  Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150) · CPC title

• C12P19/02

  Monosaccharides (2-ketogulonic acid C12P7/60) · CPC title