# US 8,771,993 B1 · Polypeptides having endoglucanse activity and polynucleotides encoding same

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- Number: US 8,771,993 B1
- Document ID: US8771993B1
- Status: Granted
- Kind: B1
- Application number: 13/764,912
- Filed: 2013-02-12
- Granted: 2014-07-08
- Earliest claimed: 2013-02-12
- Assignees: Novozymes A/S; Novozymes, Inc.
- Applicants: Novozymes A/S; Novozymes Inc.
- Inventors: Nikolaj Spodsberg
- Web page: https://patentgrep.com/patent/US8771993B1

## CPC

- C12P 19/14: produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase}
- C12N 1/14: Fungi (culture of mushrooms A01G18/00; as new plants A01H15/00); Culture media therefor
- C12N 1/20: Bacteria; Culture media therefor
- C12N 9/2437: {Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)}
- C12P 7/10: substrate containing cellulosic material
- C12P 19/02: Monosaccharides (2-ketogulonic acid C12P7/60)
- C12Y 302/01004: Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
- Y02E 50/10: Biofuels, e.g. bio-diesel
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## Abstract

The present invention relates to isolated polypeptides having endoglucanase 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.

## Claims

### Claim 1

An isolated polypeptide, which has at least 98% sequence identity to the sequence of amino acids 22-419 of SEQ ID NO: 2 or is a fragment of the sequence of amino acids 22-419 of SEQ ID NO: 2, wherein the polypeptide has endoglucanase activity.

### Claim 2 (depends on 1)

The polypeptide of claim 1, having at least 98% sequence identity to the sequence of amino acids 22-419 of SEQ ID NO: 2.

### Claim 3 (depends on 1)

The polypeptide of claim 1, having at least 99% sequence identity to the sequence of amino acids 22-419 of SEQ ID NO: 2.

### Claim 4 (depends on 1)

The polypeptide of claim 1, which is a variant of the sequence of amino acids 22-419 of SEQ ID NO: 2, comprising a substitution, deletion, and/or insertion of one or more amino acids.

### Claim 5 (depends on 1)

The polypeptide of claim 1, comprising the sequence of amino acids 22-419 of SEQ ID NO: 2.

### Claim 6 (depends on 1)

The polypeptide of claim 1, which is encoded by a polynucleotide that hybridizes with the mature polypeptide coding sequence of SEQ ID NO: 1 or the cDNA sequence thereof under high stringency conditions.

### Claim 7 (depends on 1)

The polypeptide of claim 1, which is a fragment of the sequence of amino acids 22-419 of SEQ ID NO: 2.

### Claim 8 (depends on 1)

The polypeptide of claim 1, further comprising a cellulose binding domain.

### Claim 9 (depends on 1)

A composition comprising a polypeptide of claim 1 and one or more enzymes selected from the group consisting of a cellulase, a GH61 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.

### Claim 10 (depends on 1)

A process for degrading a cellulosic material, comprising: treating the cellulosic material with a polypeptide of claim 1, a cellobiohydrolase, a beta-glucosidase, and a GH61 polypeptide having cellulolytic enhancing activity.

### Claim 11 (depends on 1)

A process for producing a fermentation product, comprising:
(a) saccharifying a cellulosic material with a polypeptide of claim 1, a cellobiohydrolase, a beta-glucosidase, and a GH61 polypeptide having cellulolytic enhancing activity; and
(b) fermenting the saccharified cellulosic material with one or more fermenting microorganisms to produce the fermentation product.

### Claim 12 (depends on 11)

The process of claim 11, further comprising recovering the fermentation product.

### Claim 13

An isolated polypeptide comprising a catalytic domain, which has at least 99% sequence identity to the sequence of amino acids 92-419 of SEQ ID NO: 2 or is a fragment of the sequence of amino acids 92-419 of SEQ ID NO: 2, wherein the catalytic domain has endoglucanase activity.

### Claim 14 (depends on 13)

The polypeptide of claim 13, further comprising a cellulose binding domain.

### Claim 15 (depends on 13)

A composition comprising the polypeptide of claim 13 and one or more enzymes selected from the group consisting of a cellulase, a GH61 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.

### Claim 16 (depends on 13)

A process for degrading a cellulosic material, comprising: treating the cellulosic material with a polypeptide of claim 13, a cellobiohydrolase, a beta-glucosidase, and a GH61 polypeptide having cellulolytic enhancing activity.

### Claim 17 (depends on 13)

A process for producing a fermentation product, comprising:
(a) saccharifying a cellulosic material with a polypeptide of claim 13, a cellobiohydrolase, a beta-glucosidase, and a GH61 polypeptide having cellulolytic enhancing activity; and
(b) fermenting the saccharified cellulosic material with one or more fermenting microorganisms to produce the fermentation product.

### Claim 18 (depends on 17)

The process of claim 17, further comprising recovering the fermentation product.