# US 8,771,994 B1 · Polypeptides having cellobiohydrolase activity and polynucleotides encoding same

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- Number: US 8,771,994 B1
- Document ID: US8771994B1
- Status: Granted
- Kind: B1
- Application number: 13/765,018
- Filed: 2013-02-12
- Granted: 2014-07-08
- Earliest claimed: 2013-02-12
- Assignees: Novozymes, Inc.
- Applicants: Novozymes A/S
- Inventors: Nikolaj Spodsberg
- Web page: https://patentgrep.com/patent/US8771994B1

## CPC

- C12N 9/2437: {Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention relates to isolated polypeptides having cellobiohydrolase 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 95% sequence identity to the sequence of amino acids 21 to 446 of SEQ ID NO: 2 or is a fragment of the sequence of amino acids 21 to 446 of SEQ ID NO: 2, wherein the polypeptide has cellobiohydrolase activity.

### Claim 2 (depends on 1)

The polypeptide of claim 1, which is a variant of the sequence of amino acids 21 to 446 of SEQ ID NO: 2, comprising a substitution, deletion, and/or insertion of one or more amino acids.

### Claim 3 (depends on 1)

The polypeptide of claim 1, comprising or consisting of the sequence of SEQ ID NO: 2.

### Claim 4 (depends on 1)

The polypeptide of claim 1, comprising or consisting of the sequence of amino acids 21 to 446 of SEQ ID NO: 2.

### Claim 5 (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 6 (depends on 1)

The polypeptide of claim 1, which is a fragment of the sequence of amino acids 21 to 446 of SEQ ID NO: 2.

### Claim 7 (depends on 1)

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

### Claim 8 (depends on 1)

A composition comprising the 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 expansin, an esterase, a laccase, a ligninolytic enzyme, a pectinase, a peroxidase, a protease, and a swollenin.

### Claim 9 (depends on 1)

A method for degrading or converting a cellulosic material, comprising: treating the cellulosic material with a polypeptide of claim 1, an endoglucanase, a beta-glucosidase, and a GH61 polypeptide having cellulolytic enhancing activity.

### Claim 10 (depends on 9)

The method of claim 9, wherein the cellulosic material is pretreated.

### Claim 11 (depends on 9)

The method of claim 9, further comprising recovering the degraded cellulosic material.

### Claim 12 (depends on 1)

A method for producing a fermentation product, comprising:
(a) saccharifying a cellulosic material with a polypeptide of claim 1, an endoglucanase, a beta-glucosidase, and a GH61 polypeptide having cellulolytic enhancing activity;
(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.

### Claim 13 (depends on 12)

The method of claim 12, wherein the cellulosic material is pretreated.

### Claim 14 (depends on 12)

The method of claim 12, wherein steps (a) and (b) are performed simultaneously in a simultaneous saccharification and fermentation.

### Claim 15 (depends on 12)

The method of claim 12, wherein the fermentation product is an alcohol, an organic acid, a ketone, an amino acid, or a gas.

### Claim 16

An isolated polypeptide comprising a catalytic domain, which has at least 95% sequence identity to the sequence of amino acids 92 to 446 of SEQ ID NO: 2 or is a fragment of the sequence of amino acids 92 to 446 of SEQ ID NO: 2, wherein the catalytic domain has cellobiohydrolase activity.

### Claim 17 (depends on 16)

The polypeptide of claim 16, wherein the catalytic domain has the sequence of amino acids 92 to 446 of SEQ ID NO: 2.

### Claim 18 (depends on 16)

A composition comprising a polypeptide of claim 16 and one or more enzymes selected from the group consisting of a cellulase, a GH61 polypeptide having cellulolytic enhancing activity, a hemicellulase, an expansin, an esterase, a laccase, a ligninolytic enzyme, a pectinase, a peroxidase, a protease, and a swollenin.

### Claim 19 (depends on 16)

A method for degrading or converting a cellulosic material, comprising: treating the cellulosic material with a polypeptide of claim 16, an endoglucanase, a beta-glucosidase, and a GH61 polypeptide having cellulolytic enhancing activity.

### Claim 20 (depends on 16)

A method for producing a fermentation product, comprising:
(a) saccharifying a cellulosic material with a polypeptide of claim 16, an endoglucanase, a beta-glucosidase, and a GH61 polypeptide having cellulolytic enhancing activity;
(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.