# US 8,541,213 B2 · Recombinant yeast and branched alcohol production method using recombinant yeast

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- Number: US 8,541,213 B2
- Document ID: US8541213B2
- Status: Granted
- Kind: B2
- Application number: 13/619,386
- Filed: 2012-09-14
- Published: 2013-01-10
- Granted: 2013-09-24
- Earliest claimed: 2009-06-04
- Assignees: Toyota Jidosha Kabushiki Kaisha
- Applicants: Satoshi Yoneda; Masayoshi Muramatsu; Shusei Obata; Ai Inoue
- Inventors: Satoshi Yoneda; Masayoshi Muramatsu; Shusei Obata; Ai Inoue
- Web page: https://patentgrep.com/patent/US8541213B2

## CPC

- C12N 9/1241: {Nucleotidyltransferases (2.7.7)}
- C12N 9/0006: {acting on CH-OH groups as donors (1.1)}
- C12N 15/52: Genes encoding for enzymes or proenzymes
- C12P 7/04: acyclic
- 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)}

## Abstract

This invention provides a recombinant yeast that can produce branched alcohol appropriate for automotive fuel and the like and a branched alcohol production method whereby branched alcohol can be produced at low cost with the use of the recombinant yeast. A recombinant yeast in which a hydroxymethyl glutaryl-CoA reductase gene has been expressed to a high degree and the ADP-ribose pyrophosphatase gene and/or the yhfR gene are introduced so as to be expressed therein is provided.

## Claims

### Claim 1

A method of production of saturated branched alcohol, comprising:
culturing in culture medium a recombinant yeast in which the HMG1 gene of Saccharomyces cerevisiae has been expressed to a high degree and the ADP-ribose pyrophosphatase gene and/or the yhfR gene are introduced so as to be expressed therein, wherein the ADP-ribose pyrophosphatase gene encodes a protein having the amino acid sequence of SEQ ID NO: 2 or a protein having an amino acid sequence with a 90% or more similarity to the amino acid sequence of SEQ ID NO: 2, and wherein the yhfR gene encodes a protein having the amino acid sequence of SEQ ID NO: 4 or a protein having an amino acid sequence with a 90% or more similarity to the amino acid sequence of SEQ ID NO: 4; and
collecting the branched alcohol from the medium.

### Claim 2 (depends on 1)

The method of production of saturated branched alcohol according to claim 1, wherein the ADP-ribose pyrophosphatase gene in the recombinant yeast is the nudF gene of Bacillus subtilis.

### Claim 3 (depends on 1)

The method of production of saturated branched alcohol according to claim 1, wherein the yhfR gene in the recombinant yeast is the yhfR gene of Bacillus subtilis.

### Claim 4 (depends on 1)

The method of production of saturated branched alcohol according to claim 1, wherein the recombinant yeast is a yeast having a mevalonic acid pathway.

### Claim 5 (depends on 1)

The method of production of saturated branched alcohol according to claim 1, wherein the recombinant yeast is a yeast of the genus Saccharomyces.

### Claim 6 (depends on 5)

The method of production of saturated branched alcohol according to claim 5, wherein the recombinant yeast is Saccharomyces cerevisiae.

### Claim 7 (depends on 1)

The method of production of saturated branched alcohol according to claim 1, wherein 2-methyl-1-butanol and 3-methyl-1-butanol is collected as the branched alcohol.