# US 7,465,557 B2 · Method of selecting barley variety, barley β-amylase gene and process for producing malt alcoholic drink

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- Number: US 7,465,557 B2
- Document ID: US7465557B2
- Status: Granted
- Kind: B2
- Application number: 10/490,378
- Filed: 2002-10-02
- Published: 2005-03-10
- Granted: 2008-12-16
- Earliest claimed: 2001-10-04
- Assignees: Sapporo Breweries Limited
- Inventors: Kazuyoshi Takeda; Takafumi Kaneko; Makoto Kihara; Takashi Asakura; Kazutoshi Ito
- Web page: https://patentgrep.com/patent/US7465557B2

## CPC

- C12N 9/2425: {Beta-amylase (3.2.1.2)}
- A01H 1/045: {using molecular markers}
- A01H 5/10: Seeds
- A01H 6/4624: {Hordeum vulgarus [barley]}
- C12P 7/14: Multiple stages of fermentation; Multiple types of microorganisms or re-use of microorganisms
- C12P 19/22: produced by the action of a beta-amylase, e.g. maltose
- C12Q 1/40: involving amylase
- C12Y 302/01002: Beta-amylase (3.2.1.2)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A01H: NEW PLANTS OR {NON-TRANSGENIC} PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- 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)}
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Y: ENZYMES

## Abstract

A method of selecting a barley variety comprising: a heat treatment step of heat-treating a crude enzyme solution extracted from a seed of a barley (e.g., at 57.5° C. for 30 minutes); an activity-determining step of determining the enzyme activity of β-amylase in the crude enzyme solution heat-treated; and a selection step of selecting the barley variety containing a β-amylase having a residual activity of from 85 to 90% as a result of the activity determination.

## Claims

### Claim 1

A method of selecting a barley variety, which comprises:
heating a crude enzyme solution extracted from a seed of a barley;
determining the enzyme activity of a β-amylase in the crude enzyme solution heat-treated; and
selecting a barley variety comprising a β-amylase having a residual activity of from 85 to 90% as a result of the activity determination compared to an unheated control,
determining a nucleotide sequence of a gene encoding the β-amylase of the selected barley variety, and
selecting the barley variety comprising a structural gene of the β-amylase comprising the sequence set forth in SEQ ID NO:1,
wherein said heating of the crude enzyme solution occurs at a predetermined temperature of 55 to 58° C. and for a time of 10 to 60 minutes.