Polypeptides having cellulolytic enhancing activity and polynucleotides encoding same

Patent metadata

Field	Value
Publication number	US-10006012-B2
Application number	US-201414518841-A
Country	US
Kind code	B2
Filing date	Oct 20, 2014
Priority date	Sep 17, 2009
Publication date	Jun 26, 2018
Grant date	Jun 26, 2018

Abstract

The present invention relates to isolated polypeptides having cellulolytic enhancing 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 DNA construct or recombinant expression vector comprising an isolated polynucleotide comprising a nucleotide sequence that encodes a GH61 polypeptide having cellulolytic enhancing activity, wherein the isolated polynucleotide is operably linked to one or more heterologous control sequences that direct the production of the GH61 polypeptide in an expression host, and wherein the GH61 polypeptide having cellulolytic enhancing activity is selected from the group consisting of:
(a) a GH61 polypeptide having at least 95% sequence identity to amino acids 18 to 227 of SEQ ID NO: 6; and
(b) a GH61 polypeptide encoded by a polynucleotide having at least 99% sequence identity to nucleotides 52 to 795 of SEQ ID NO: 5, or a GH61 polypeptide encoded by the cDNA of nucleotides 52 to 795 of SEQ ID NO: 5.

2. The DNA construct or recombinant expression vector of claim 1 , wherein the GH61 polypeptide has at least 96% sequence identity to amino acids 18 to 227 of SEQ ID NO: 6.

3. The DNA construct or recombinant expression vector of claim 1 , wherein the GH61 polypeptide has at least 97% sequence identity to amino acids 18 to 227 of SEQ ID NO: 6.

4. The DNA construct or recombinant expression vector of claim 1 , wherein the GH61 polypeptide has at least 98% sequence identity to amino acids 18 to 227 of SEQ ID NO: 6.

5. The DNA construct or recombinant expression vector of claim 1 , wherein the GH61 polypeptide has at least 99% sequence identity to amino acids 18 to 227 of SEQ ID NO: 6.

6. The DNA construct or recombinant expression vector of claim 1 , wherein the GH61 polypeptide comprises amino acids 18 to 227 of SEQ ID NO: 6.

7. The DNA construct or recombinant expression vector of claim 1 , wherein the GH61 polypeptide consists of amino acids 18 to 227 of SEQ ID NO: 6.

8. The DNA construct or recombinant expression vector of claim 1 , wherein the GH61 polypeptide is encoded by a polynucleotide comprising nucleotides 52 to 795 of SEQ ID NO: 5, or the cDNA of nucleotides 52 to 795 of SEQ ID NO: 5.

9. The DNA construct or recombinant expression vector of claim 1 , wherein the GH61 polypeptide is encoded by a polynucleotide consisting of nucleotides 52 to 795 of SEQ ID NO: 5, or the cDNA of nucleotides 52 to 795 of SEQ ID NO: 5.

10. The DNA construct or recombinant expression vector of claim 1 , wherein the GH61 polypeptide is encoded by nucleotides 52 to 795 of SEQ ID NO: 5 contained in pSMai218 which is contained in E. coli NRRL B-50303 deposited with the Northern Regional Research Center of the Agriculture Research Service Patent Culture Collection.

11. A recombinant host cell comprising the DNA construct or recombinant expression vector of claim 1 .

12. A method of producing a GH61 polypeptide having cellulolytic enhancing activity, said method comprising:
(a) cultivating the recombinant host cell of claim 11 under conditions conducive for production of the GH61 polypeptide; and
(b) recovering the GH61 polypeptide.

13. A DNA construct comprising a nucleic acid encoding a signal peptide comprising amino acids 1 to 17 of SEQ ID NO: 6, wherein the nucleic acid is operably linked to a heterologous nucleic acid encoding a protein.

14. A recombinant host cell comprising a nucleic acid encoding a protein, wherein the nucleic acid is operably linked to an isolated polynucleotide encoding a signal peptide comprising amino acids 1 to 17 of SEQ ID NO: 6, wherein the nucleic acid is foreign to the polynucleotide encoding the signal peptide.

15. A method of producing a protein, said method comprising:
(a) cultivating the recombinant host cell of claim 14 under conditions conducive for production of the protein; and
(b) recovering the protein.

16. A transgenic plant, plant part or plant cell transformed with a polynucleotide encoding a GH61 polypeptide having cellulolytic enhancing activity, wherein the GH61 polypeptide comprises a signal peptide directing the GH61 polypeptide into the secretory pathway, and wherein the GH61 polypeptide having cellulolytic enhancing activity is selected from the group consisting of:
(a) a GH61 polypeptide having at least 95% sequence identity to amino acids 18 to 227 of SEQ ID NO: 6;
(b) a GH61 polypeptide encoded by a polynucleotide having at least 99% sequence identity to nucleotides 52 to 795 of SEQ ID NO: 5, or a GH61 polypeptide encoded by the cDNA of nucleotides 52 to 795 of SEQ ID NO: 5;
(c) a GH61 polypeptide comprising amino acids 18 to 227 of SEQ ID NO: 6.

17. A method of producing a GH61 polypeptide having cellulolytic enhancing activity, said method comprising:
(a) cultivating the transgenic plant or plant cell of claim 16 under conditions conducive for production of the GH61 polypeptide; and
(b) recovering the GH61 polypeptide.

18. A method for degrading a cellulosic material comprising:
(i) treating the cellulosic material with an enzyme composition comprising a GH61 having cellulolytic enhancing activity, and
(ii) recovering the degraded cellulosic material;

wherein the GH61 polypeptide having cellulolytic enhancing activity is selected from the group consisting of:
(a) a GH61 polypeptide having at least 95% sequence identity to amino acids 18 to 227 of SEQ ID NO: 6; and
(b) a GH61 polypeptide encoded by a polynucleotide having at least 99% sequence identity to nucleotides 52 to 795 of SEQ ID NO: 5, or a GH61 polypeptide encoded by the cDNA of nucleotides 52 to 795 of SEQ ID NO: 5.

19. A method for producing a fermentation product, said method comprising:
(i) saccharifying a cellulosic material with an enzyme composition comprising a GH61 polypeptide having cellulolytic enhancing activity;
(ii) fermenting the saccharified cellulosic material with one or more fermenting microorganisms to produce the fermentation product; and
(ii) recovering the fermentation product from the fermentation;

wherein the GH61 polypeptide having cellulolytic enhancing activity is selected from the group consisting of:
(a) a GH61 polypeptide having at least 95% sequence identity to amino acids 18 to 227 of SEQ ID NO: 6; and
(b) a GH61 polypeptide encoded by a polynucleotide having at least 99% sequence identity to nucleotides 52 to 795 of SEQ ID NO: 5, or a GH61 polypeptide encoded by the cDNA of nucleotides 52 to 795 of SEQ ID NO: 5.

20. The method of claim 19 , wherein steps (i) and (ii) are performed simultaneously.

21. A method of fermenting a cellulosic material comprising:
(i) fermenting the cellulosic material with one or more fermenting microorganisms, wherein the cellulosic material is saccharified with an enzyme composition comprising a GH61 polypeptide having cellulolytic enhancing activity, wherein the fermenting of the cellulosic material produces a fermentation product, and
(ii) recovering the fermentation product from the fermentation;

wherein the GH61 polypeptide having cellulolytic enhancing activity is selected from the group consisting of:
(a) a GH61 polypeptide having at least 95% sequence identity to amino acids 18 to 227 of SEQ ID NO: 6; and
(b) a GH61 polypeptide encoded by a polynucleotide having at least 99% sequence identity to nucleotides 52 to 795 of SEQ ID NO: 5, or a GH61 polypeptide encoded by the cDNA of nucleotides 52 to 795 of SEQ ID NO: 5.

22. The method of claim 18 , wherein the GH61 polypeptide comprises amino acids 18 to 227 of SEQ ID NO: 6.

23. The method of claim 19 , wherein the GH61 polypeptide comprises amino acids 18 to 227 of SEQ ID NO:

Assignees

• Novozymes Inc

Inventors

• Maiyuran Suchindra
• Kramer Randall
• Harris Paul

Classifications

• C12P19/02

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

• C11D3/38645

  containing cellulase · CPC title

• C12P19/14

  produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase} · CPC title

• C12Y302/01

  Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1) · CPC title

• C12N9/2437 Primary

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