# US 10,767,170 B2 · High-temperature neutral cellulase as well as coding gene and application thereof

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- Number: US 10,767,170 B2
- Document ID: US10767170B2
- Status: Granted
- Kind: B2
- Application number: 15/534,491
- Filed: 2014-12-09
- Published: 2019-12-19
- Granted: 2020-09-08
- Earliest claimed: 2014-12-09
- Assignees: Feed Research Institute, Chinese Academy of Agricultural Sciences
- Applicants: Feed Research Institute, Chinese Academy of Agricultural Sciences
- Inventors: Bin Yao; Pengjun Shi; Hong Yang; Huoqing Huang; Huiying Luo; Peilong Yang; Yaru Wang; Xiaoyun Su; Yingguo Bai; Xia Shi; Rui Ma; Kun Meng
- Web page: https://patentgrep.com/patent/US10767170B2

## CPC

- C12N 9/2437: {Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)}
- C12N 1/16: Yeasts; Culture media therefor
- C12N 1/185: Saccharomyces isolates
- C12N 15/63: Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12R 2001/85: Saccharomyces
- C12Y 302/01004: Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12Y: ENZYMES

## Abstract

Provided are a fungus-sourced high-temperature neutral Family-45 cullulase as well as a coding gene and application thereof. The cullulase has optimal pH value of 5.5, and optimal temperature of 60° C., has certain enzyme activity in alkaline condition, and has good alkali resistance, maintains about 70% enzyme activity in optimal condition after being processed at 90° C. for 1 hour, maintains about 50% enzyme activity in optimal condition after being processed in boiling water for 1 hour, and can be well applied in c and other fields.

## Claims

### Claim 1

A method of degrading cellulose contained in a raw textile fiber, comprising applying a cellulase to said raw textile fiber, wherein said cellulase has the amino acid as shown in SEQ ID NO: 1 or SEQ ID NO: 2.

### Claim 2 (depends on 1)

The method of claim 1, wherein said cellulase is produced by transcribing an isolated polynucleotide encoding said cellulase.

### Claim 3 (depends on 2)

The method of claim 2, wherein said isolated polynucleotide has a nucleotide sequence as set in forth in SEQ ID NO.4 or SEQ ID NO.5.

### Claim 4 (depends on 3)

The method of claim 3, wherein said isolated polynucleotide is in a recombinant vector.

### Claim 5 (depends on 3)

The method of claim 3, wherein said isolated polynucleotide is introduced into the genome of a recombinant yeast host cell.

### Claim 6 (depends on 5)

The method of claim 5, wherein said recombinant host cell is a Pichia cell.

### Claim 7 (depends on 1)

The method of claim 1, wherein said cellulase has an optimal pH value of 5.5, the optimal temperature of 60° C., maintains about 70% of its optimal activity after incubation at 60° C. for 1 hour, and maintains 50% of its optimal activity condition after incubation in 100° C. water for 1 hour.

### Claim 8

A method of degrading cellulose contained in paper pulp, comprising applying a cellulase to said paper pulp, wherein said cellulase has the amino acid sequence as shown in SEQ ID NO: 1 or SEQ ID NO: 2.

### Claim 9 (depends on 8)

The method of claim 8, wherein said cellulase is produced by transcribing an isolated polynucleotide encoding said cellulase.

### Claim 10 (depends on 9)

The method of claim 9, wherein said isolated polynucleotide has a nucleotide sequence as set in forth in SEQ ID NO.4 or SEQ ID NO.5.

### Claim 11 (depends on 10)

The method of claim 10, wherein said isolated polynucleotide is in a recombinant vector.

### Claim 12 (depends on 10)

The method of claim 10, wherein said isolated polynucleotide is introduced into the genome of a recombinant yeast host cell.

### Claim 13 (depends on 12)

The method of claim 12, wherein said recombinant host cell is a Pichia cell.

### Claim 14 (depends on 8)

The method of claim 8, wherein said cellulase has an optimal pH value of 5.5, the optimal temperature of 60° C., maintains about 70% of its optimal activity after incubation at 60° C. for 1 hour, and maintains 50% of its optimal activity condition after incubation in 100° C. water for 1 hour.