# US 11,299,757 B2 · Mutant gene associated with improvement in ethanol productivity via ethanol fermentation and method for producing ethanol using the same

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- Number: US 11,299,757 B2
- Document ID: US11299757B2
- Status: Granted
- Kind: B2
- Application number: 16/761,676
- Filed: 2018-11-05
- Published: 2021-06-17
- Granted: 2022-04-12
- Earliest claimed: 2017-11-06
- Assignees: Toyota Jidosha Kabushiki Kaisha
- Applicants: Toyota Jidosha Kabushiki Kaisha
- Inventors: Junji Ito; Toru Onishi; Nobuki Tada; Rie Hirao
- Web page: https://patentgrep.com/patent/US11299757B2

## CPC

- C12P 7/06: Ethanol, i.e. non-beverage
- C07K 14/39: from yeasts
- C12N 9/12: transferring phosphorus containing groups, e.g. kinases (2.7)
- Y02E 50/10: Biofuels, e.g. bio-diesel
- 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)}
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION

## Abstract

This invention is intended to improve the ethanol fermentation ability of a yeast strain having xylose-metabolizing ability with the use of a mutant gene encoding a mutant protein comprising a consensus sequence comprising a substitution of amino acid in the 30th position in SEQ ID NO: 1, amino acid in the 43rd position in SEQ ID NO: 4, and amino acid in the 31st position in SEQ ID NO: 7 with other amino acid residues.

## Claims

### Claim 1

A mutant gene encoding a mutant CDC123 (cell division cycle 123) protein, wherein said mutant CDC123 protein comprises:
(a) the amino acid sequence of SEQ ID NO: 3; or
(b) an amino acid sequence having 95% or higher identity to the amino acid sequence of SEQ ID NO: 3 in which an amino acid residue corresponding to the 324th position from the N terminus of the amino acid sequence of SEQ ID NO: 3 is cysteine.

### Claim 2 (depends on 1)

A mutant yeast strain having xylose-metabolizing ability, which comprises the mutant gene according to claim 1.

### Claim 3 (depends on 2)

A method for producing ethanol comprising a step of culturing the mutant yeast strain according to claim 2 in a xylose-containing medium and performing ethanol fermentation.

### Claim 4 (depends on 3)

The method for producing ethanol according to claim 3, wherein the medium contains cellulose and the ethanol fermentation proceeds simultaneously at least with the cellulose saccharification.