# US 9,752,135 B2 · Thermostable β-glucosidase

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- Number: US 9,752,135 B2
- Document ID: US9752135B2
- Status: Granted
- Kind: B2
- Application number: 14/850,732
- Filed: 2015-09-10
- Published: 2016-03-17
- Granted: 2017-09-05
- Earliest claimed: 2014-09-11
- Assignees: Honda Motor Co., Ltd.
- Applicants: Honda Motor Co., Ltd.
- Inventors: Yoshitsugu Hirose; Takahiro Gunji; Asuka Yamaguchi; Migiwa Suda; Jiro Okuma; Yasuhiro Kondo; Tomohiko Kato; Daisuke Shibata
- Web page: https://patentgrep.com/patent/US9752135B2

## CPC

- C12N 9/2445: {Beta-glucosidase (3.2.1.21)}
- C12N 9/2482: {Endo-1,4-beta-xylanase (3.2.1.8)}
- C12P 19/02: Monosaccharides (2-ketogulonic acid C12P7/60)
- C12P 19/14: produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase}
- C12Y 302/01008: Endo-1,4-beta-xylanase (3.2.1.8)
- C12Y 302/01021: Beta-glucosidase (3.2.1.21)
- C12P 2203/00: Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source
- Y02E 50/10: Biofuels, e.g. bio-diesel
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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)}
- C12Y: ENZYMES
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## Abstract

A thermostable β-glucosidase including a β-glucosidase catalytic domain, the β-glucosidase catalytic domain including: (A) a polypeptide including an amino acid sequence represented by SEQ ID NO: 1; (B) a polypeptide including an amino acid sequence in which at least one amino acid is deleted, substituted, or added in the amino acid sequence represented by SEQ ID NO: 1, and having hydrolytic activity using p-nitrophenyl-β-D-glucopyranoside as a substrate at least under conditions of a temperature of 75° C. and a pH of 7; or (C) a polypeptide including an amino acid sequence having at least 90% sequence identity with the amino acid sequence represented by SEQ ID NO: 1, and having hydrolytic activity using p-nitrophenyl-β-D-glucopyranoside as a substrate at least under conditions of a temperature of 75° C. and a pH of 7.

## Claims

### Claim 1

An isolated recombinant thermostable β-glucosidase comprising:
a polypeptide consisting of the amino acid sequence of SEQ ID NO: 1 and
at least one region selected from the group consisting of a cellulose-binding module, linker domain, a signal peptide and a tag.

### Claim 2 (depends on 1)

The thermostable β-glucosidase according to claim 1, which also has xylanase activity.

### Claim 3 (depends on 1)

A glycoside hydrolase mixture, comprising the thermostable β-glucosidase according to claim 1 and at least one or more types of other glycoside hydrolases.

### Claim 4 (depends on 1)

A method for producing a lignocellulose degradation product, the method comprising producing a lignocellulose degradation product containing a cellulose degradation product by bringing a material composed of lignocellulose containing cellulose into contact with the termostable β-glucosidase according to claim 1.

### Claim 5 (depends on 3)

A method for producing a lignocellulose degradation product, the method comprising producing a lignocellulose degradation product containing a cellulose degradation product by bringing a material composed of lignocellulose containing cellulose into contact with the glycoside hydrolase mixture according to claim 3.