Glycoside hydrolases from thermophilic fungi

Patent metadata

Field	Value
Publication number	US-9353363-B2
Application number	US-201213884773-A
Country	US
Kind code	B2
Filing date	Jan 26, 2012
Priority date	Jan 26, 2011
Publication date	May 31, 2016
Grant date	May 31, 2016

Abstract

The present invention relates to isolated polypeptides having cellulolytic activity or hemicellulolytic 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.

First claim

The invention 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 having cellobiohydrolase activity in a host cell, and wherein the polypeptide having cellobiohydrolase activity is selected from the group consisting of:
(a) a polypeptide having at least 90% sequence identity to the polypeptide of SEQ ID NO: 4;
(b) a polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with (i) the polypeptide coding sequence of SEQ ID NO: 3, (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 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 polypeptide coding sequence of SEQ ID NO: 3, or the cDNA sequence thereof;
(d) a fragment of the polypeptide of (a), (b), or (c) that has cellobiohydrolase activity;
(e) a polypeptide comprising SEQ ID NO: 4; and
(f) a polypeptide encoded by the polynucleotide of SEQ ID NO: 3 or the cDNA sequence thereof.

2. A recombinant host cell comprising the nucleic acid construct of claim 1 .

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

4. A process for degrading or converting a cellulosic or xylan-containing material, comprising:
(a) treating the cellulosic or xylan-containing material with an enzyme composition comprising a polypeptide having cellobiohydrolase activity at a temperature of about 50° C. to about 80° C., wherein the polypeptide having cellobiohydrolase activity is selected from the group consisting of:
(1) a polypeptide having at least 90% sequence identity to the polypeptide of SEQ ID NO: 4;
(2) a polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with (i) the polypeptide coding sequence of SEQ ID NO: 3, (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 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.;
(3) a polypeptide encoded by a polynucleotide having at least 90% sequence identity to the polypeptide coding sequence of SEQ ID NO: 3, or the cDNA sequence thereof;
(4) a fragment of the polypeptide of (1), (2), or (3) that has cellobiohydrolase activity;
(5) a polypeptide comprising SEQ ID NO: 4; and
(6) a polypeptide encoded by the polynucleotide of SEQ ID NO: 3 or the cDNA sequence thereof.

5. The process of claim 4 , further comprising recovering the degraded or converted cellulosic or xylan-containing material.

6. A process for producing a fermentation product, comprising:
(a) saccharifying a cellulosic or xylan-containing material with an enzyme composition comprising a polypeptide having cellobiohydrolase activity at a temperature of about 50° C. to about 80° C., wherein the polypeptide having cellobiohydrolase activity is selected from the group consisting of:
(1) a polypeptide having at least 90% sequence identity to the polypeptide of SEQ ID NO: 4;
(2) a polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with (i) the polypeptide coding sequence of SEQ ID NO: 3, (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 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.;
(3) a polypeptide encoded by a polynucleotide having at least 90% sequence identity to the polypeptide coding sequence of SEQ ID NO: 3, or the cDNA sequence thereof;
(4) a fragment of the polypeptide of (1), (2), or (3) that has cellobiohydrolase activity
(5) a polypeptide comprising SEQ ID NO: 4; and
(6) a polypeptide encoded by the polynucleotide of SEQ ID NO: 3 or the cDNA sequence thereof;
(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.

7. 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 comprising a polypeptide having cellobiohydrolase activity at a temperature of about 50° C. to about 80° C., wherein the polypeptide having cellobiohydrolase activity is selected from the group consisting of:
(1) a polypeptide having at least 90% sequence identity to the polypeptide of SEQ ID NO: 4;
(2) a polypeptide encoded by a polynucleotide that hybridizes under very high stringency conditions with (i) the polypeptide coding sequence of SEQ ID NO: 3, (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 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.;
(3) a polypeptide encoded by a polynucleotide having at least 90% sequence identity to the polypeptide coding sequence of SEQ ID NO: 3, or the cDNA sequence thereof;
(4) a fragment of the polypeptide of (1), (2), or (3) that has cellobiohydrolase activity;
(5) a polypeptide comprising SEQ ID NO: 4; and
(6) a polypeptide encoded by the polynucleotide of SEQ ID NO: 3 or the cDNA sequence thereof.

8. The nucleic acid construct of claim 1 , wherein the polypeptide has at least 95% sequence identity to the polypeptide of SEQ ID NO: 4.

9. The process of claim 4 , wherein the cellulosic or xylan-containing material is pretreated.

10. The process of claim 4 , wherein the degraded or converted cellulosic or xylan-containing material is a sugar.

11. The process of claim 10 , wherein the sugar is selected from the group consisting of glucose, xylose, mannose, galactose, and arabinose.

12. The process of claim 4 , wherein the polypeptide has at least 95% sequence identity to the polypeptide of SEQ ID NO: 4.

13. The process of claim 4 , wherein the enzyme composition further 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.

14. The process of claim 6 , wherein the cellulosic or xylan-containing material is pretreated.

15. The process of claim 6 , wherein the enzyme composition further 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,

Assignees

• Lange Lene
• Busk Peter K
• Novozymes As

Inventors

• Lange Lene
• Busk Peter K

Classifications

• 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

• C12Y302/01091

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

• C12Y302/01004

  Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase · CPC title

• C12P2201/00

  Pretreatment of cellulosic or lignocellulosic material for subsequent enzymatic treatment or hydrolysis · CPC title

• C12P21/00

  Preparation of peptides or proteins (single cell protein C12N1/00) · CPC title