# US 8,772,012 B2 · Xylose isomerase and use thereof

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- Number: US 8,772,012 B2
- Document ID: US8772012B2
- Status: Granted
- Kind: B2
- Application number: 13/518,251
- Filed: 2010-12-22
- Published: 2013-04-18
- Granted: 2014-07-08
- Earliest claimed: 2009-12-22
- Assignees: Kabushiki Kaisha Toyota Chuo Kenkyusho; Riken
- Applicants: Satoshi Katahira; Kenro Tokuhiro; Nobuhiko Muramoto; Shigeharu Moriya; Moriya Ohkuma; Yoshina Takahashi
- Inventors: Satoshi Katahira; Kenro Tokuhiro; Nobuhiko Muramoto; Haruo Takahashi; Shigeharu Moriya; Moriya Ohkuma
- Web page: https://patentgrep.com/patent/US8772012B2

## CPC

- C12N 9/90: Isomerases (5.)
- C12N 9/92: Glucose isomerase {(5.3.1.5; 5.3.1.9; 5.3.1.18)}
- C12P 7/10: substrate containing cellulosic material
- C12Y 503/01005: Xylose isomerase (5.3.1.5)
- 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

It is an object of the disclosure of the present description to provide an eukaryotic cell having xylose utilization ability. The disclosure of the present description provides a novel eukaryotic cell having xylose utilization ability by transforming a yeast or other eukaryotic cell using DNA that codes a xylose isomerase from a termite protist.

## Claims

### Claim 1

A nucleic acid selected from any of the following nucleic acids:
a nucleic acid comprising a nucleotide sequence having at least 85% sequence identity with any of the full-length cDNA sequences of SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13 and the full-length nucleotide sequences complementary thereto; and
a cDNA coding for a polypeptide having xylose isomerase activity comprising an amino acid sequence having at least 85% sequence identity with any of the full-length amino acid sequences of SEQ ID NOS: 2, 4, 6, 8, 10, 12, and 14.

### Claim 2 (depends on 1)

The nucleic acid of claim 1, wherein the nucleic acid comprises a nucleotide sequence having at least 85% sequence identity with any of the full-length cDNA sequences of SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13 and the full-length nucleotide sequences complementary thereto.

### Claim 3 (depends on 1)

The nucleic acid of claim 1, wherein the nucleic acid comprises any of the full-length cDNA sequences of SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13 and the full-length nucleotide sequences complementary thereto.

### Claim 4 (depends on 1)

The nucleic acid of claim 1, wherein the nucleic acid is a cDNA coding for a polypeptide having xylose isomerase activity comprising an amino acid sequence having at least 85% sequence identity with any of the full-length amino acid sequences of SEQ ID NOS: 2, 4, 6, 8, 10, 12, and 14.

### Claim 5 (depends on 1)

The nucleic acid of claim 1, wherein the nucleic acid is a cDNA coding for a polypeptide having xylose isomerase activity comprising any of the full-length amino acid sequences of SEQ ID NOS: 2, 4, 6, 8, 10, 12, and 14.

### Claim 6 (depends on 2)

An eukaryotic expression vector comprising the nucleic acid of claim 2.

### Claim 7 (depends on 4)

An eukaryotic expression vector comprising the nucleic acid of claim 4.

### Claim 8 (depends on 6)

An eukaryotic cell that has been transformed with the eukaryotic expression vector of claim 6.

### Claim 9 (depends on 7)

An eukaryotic cell that has been transformed with the eukaryotic expression vector of claim 7.

### Claim 10 (depends on 8)

The eukaryotic cell of claim 8, wherein the eukaryotic cell is a yeast.

### Claim 11 (depends on 9)

The eukaryotic cell of claim 9, wherein the eukaryotic cell is a yeast.

### Claim 12 (depends on 8)

The eukaryotic cell of claim 8, wherein the eukaryotic cell secretorily produces a cellulase.

### Claim 13 (depends on 9)

The eukaryotic cell of claim 9, wherein the eukaryotic cell secretorily produces a cellulase.

### Claim 14 (depends on 8)

The eukaryotic cell of claim 8, wherein the eukaryotic cell produces one or more enzymes included in an enzyme group of an arabinose metabolic pathway.

### Claim 15 (depends on 9)

The eukaryotic cell of claim 9, wherein the eukaryotic cell produces one or more enzymes included in an enzyme group of an arabinose metabolic pathway.

### Claim 16 (depends on 8)

The eukaryotic cell of claim 8, wherein the eukaryotic cell is able to convert xylose to any selected from the group consisting of ethanol, lactic acid, acetic acid, 1,3-propane-diole, propanol, butanol, succinic acid, ethylene, glycerol, farnesol, geranylgeraniol and squalene.

### Claim 17 (depends on 9)

The eukaryotic cell of claim 9, wherein the eukaryotic cell is able to convert xylose to any selected from the group consisting of ethanol, lactic acid, acetic acid, 1,3-propane-diole, propanol, butanol, succinic acid, ethylene, glycerol, farnesol, geranylgeraniol and squalene.

### Claim 18 (depends on 6)

A method of transforming an eukaryotic cell to express an exogenous xylose isomerase, comprising transforming the eukaryotic cell with the eukaryotic expression vector of claim 6.

### Claim 19 (depends on 7)

A method of transforming an eukaryotic cell to express an exogenous xylose isomerase, comprising transforming the eukaryotic cell with the eukaryotic expression vector of claim 7.