# US 8,569,033 B2 · Surfactant tolerant cellulase and method for modification thereof

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- Number: US 8,569,033 B2
- Document ID: US8569033B2
- Status: Granted
- Kind: B2
- Application number: 10/582,002
- Filed: 2004-12-07
- Published: 2007-05-03
- Granted: 2013-10-29
- Earliest claimed: 2003-12-08
- Assignees: Meiji Seika Pharma Co., Ltd.
- Applicants: Manabu Watanabe; Koji Yanai; Yumiko Tsuyuki
- Inventors: Manabu Watanabe; Koji Yanai; Yumiko Tsuyuki
- Web page: https://patentgrep.com/patent/US8569033B2

## CPC

- C12N 9/2437: {Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)}
- 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)
- C12Y: ENZYMES

## Abstract

A method for suppressing a reduction in an endoglucanase activity in the presence of a surfactant, characterized by modifying a protein having the endoglucanase activity in which the N-terminus is an amino acid other than pyroglutamic acid, to a protein having the N-terminus of pyroglutamic acid, is disclosed. Further, a modified protein having an endoglucanase activity wherein the N-terminal amino acid is converted into pyroglutamic acid by an amino acid modification, a polynucleotide encoding the protein, an expression vector comprising the polynucleotide, a host cell transformed with the expression vector, and a process for producing the protein by cultivating the host cell, are disclosed.

## Claims

### Claim 1

An isolated protein selected from the group consisting of:
(a) a protein comprising the amino acid sequence of SEQ ID NO: 2, 4, 38, or 40, wherein the N-terminal amino acid is pyroglutamic acid; and
(b) a protein comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 2, 4, 38, or 40, and having an endoglucanase activity, wherein the N-terminal amino acid is pyroglutamic acid.

### Claim 2 (depends on 1)

A protein produced by the steps comprising:
cultivating a host cell transformed with an expression vector comprising a polynucleotide encoding the protein according to claim 1, and recovering the protein from the host cell or culture obtained by the cultivation.

### Claim 3 (depends on 1)

A method for suppressing a reduction in an endoglucanase activity in the presence of a surfactant, characterized by modifying a protein having the endoglucanase activity in which the N-terminus is an amino acid other than pyroglutamic acid, to a protein according to claim 1.

### Claim 4 (depends on 3)

The method according to claim 3, wherein the modification is carried out by adding pyroglutamic acid or an amino acid convertible into pyroglutamic acid, or a peptide having the N-terminus of pyroglutamic acid or an amino acid convertible into pyroglutamic acid, to the N-terminus of the protein having the endoglucanase activity in which the N-terminus is an amino acid other than pyroglutamic acid.

### Claim 5 (depends on 3)

The method according to claim 3, wherein the modification is carried out by substituting pyroglutamic acid or an amino acid convertible into pyroglutamic acid, or a peptide having the N-terminus of pyroglutamic acid or an amino acid convertible into pyroglutamic acid, for the N-terminal amino acid or an N-terminal region of the protein having the endoglucanase activity in which the N-terminus is an amino acid other than pyroglutamic acid.

### Claim 6 (depends on 3)

The method according to claim 3, wherein the protein having the endoglucanase activity in which the N-terminus is an amino acid other than pyroglutamic acid is a cellulase belonging to family 45.

### Claim 7

The method according to 4, wherein the protein having the endoglucanase activity in which the N-terminus is an amino acid other than pyroglutamic acid is a cellulase belonging to family 45.

### Claim 8 (depends on 5)

The method according to claim 5, wherein the protein having the endoglucanase activity in which the N-terminus is an amino acid other than pyroglutamic acid is a cellulase belonging to family 45.