WO1996020272A1 - Novel aromatic yeast strains - Google Patents

Novel aromatic yeast strains Download PDF

Info

Publication number
WO1996020272A1
WO1996020272A1 PCT/JP1995/002632 JP9502632W WO9620272A1 WO 1996020272 A1 WO1996020272 A1 WO 1996020272A1 JP 9502632 W JP9502632 W JP 9502632W WO 9620272 A1 WO9620272 A1 WO 9620272A1
Authority
WO
WIPO (PCT)
Prior art keywords
yeast
strain
strains
sake
ethyl
Prior art date
Application number
PCT/JP1995/002632
Other languages
French (fr)
Japanese (ja)
Inventor
Naotaka Kurose
Tadao Asano
Nobutsugu Hiraoka
Tadaki Shigeno
Tadanori Yano
Original Assignee
Takara Shuzo Co., Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Takara Shuzo Co., Ltd. filed Critical Takara Shuzo Co., Ltd.
Priority to AU43146/96A priority Critical patent/AU4314696A/en
Priority to KR1019970704352A priority patent/KR987001032A/en
Priority to JP52035396A priority patent/JP3544987B2/en
Publication of WO1996020272A1 publication Critical patent/WO1996020272A1/en

Links

Classifications

    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
    • A21D8/00—Methods for preparing or baking dough
    • A21D8/02—Methods for preparing dough; Treating dough prior to baking
    • A21D8/04—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes
    • A21D8/047—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes with yeasts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C12N1/18—Baker's yeast; Brewer's yeast
    • C12N1/185—Saccharomyces isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
    • C12C11/00—Fermentation processes for beer
    • C12C11/02—Pitching yeast
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
    • C12G1/00—Preparation of wine or sparkling wine
    • C12G1/02—Preparation of must from grapes; Must treatment and fermentation
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
    • C12G3/00—Preparation of other alcoholic beverages
    • C12G3/02—Preparation of other alcoholic beverages by fermentation
    • C12G3/021—Preparation of other alcoholic beverages by fermentation of botanical family Poaceae, e.g. wheat, millet, sorghum, barley, rye, or corn
    • C12G3/022—Preparation of other alcoholic beverages by fermentation of botanical family Poaceae, e.g. wheat, millet, sorghum, barley, rye, or corn of botanical genus Oryza, e.g. rice
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi
    • C12R2001/85—Saccharomyces

Definitions

  • the present invention relates to a novel yeast having a high aromaticity and a method for producing alcoholic beverages and foods rich in an aromaticity by the yeast.
  • the present invention relates to a method for easily producing a yeast which produces a large amount of carboxylate and / or isoamyl acetate among the aroma components. Relates to a method for producing aromatic liquors and foods using selected yeasts.
  • the present inventors have obtained a new aromatic sake yeast strain which is bred by crossing between haploids derived from Japan Brewing Association No. 71 and Japan Brewing Association No. 91 (Japanese Unexamined Patent Publication No. 5-3 1 7 0 3 4).
  • they have found a simple and high-frequency method for producing haploid strains of sake yeast (Japanese Patent Application No. 5-3177035).
  • it has been desired to provide a yeast in which ethyl ethyl cabronate and Z or isoamyl acetate are more highly produced by the hybridization treatment.
  • liquors other than sake have similar problems in the production of shochu, and also in the production of foods, in order to produce fragrant shochu. It was the same.
  • Ginjo sake which has a rich aroma component
  • it is difficult to produce a large amount of Ginjo sake in a stable manner because the cost of raw materials is high and the production period is long. I did.
  • sake rich in ginjo-scent when brewing ordinary sake using low-polished rice.
  • the purpose of the present invention is to solve the above-mentioned problems of the prior art, and to produce sake with a high Ginjo aroma or a new Ginjo aroma with ordinary sake. For that purpose. And to provide a novel yeast having a high ability to produce methyl and ethyl or ethyl beet carboxylate, and to use the yeast to stably and largely produce a liquor rich in ginjo aroma from low-polished rice. Another object of the present invention is to provide a method for producing a food, and a method for producing a food rich in ginjo aroma using the yeast.
  • the first invention of the present invention is directed to a clonal breed by crossing cells of haploids from the same source of yeast belonging to the genus Saccharomyces.
  • the second invention relates to a novel aromatic yeast strain that produces high levels of ethyl ether and / or isoamyl acetate.
  • the second invention relates to alcoholic beverages and foods rich in aroma components characterized by using the yeast. Related to manufacturing method.
  • the present inventors crossed haploid strains derived from the same source of yeast belonging to the genus Saccharomyces, thereby obtaining ethyl cabronate and / or isoacetate.
  • the yeast belonging to the genus Saccharomyces in the present invention includes sake yeast, shochu yeast, Wien yeast, Shelley yeast, brewer's yeast, Wisky yeast, and alky yeast. Any yeast belonging to the genus Saccharomyces such as call yeast and pan yeast can be used, and yeast used for sake brewing is, for example, pure culture at the Honshu Brewing Association. Yeast No. 6, Yeast No. 7, Association No. 9 Yeast, Kyokai No. 10 Yeast, Kyokai No. 11 Yeast, Kyokai No. 12 Yeast, Kyokai No. 13 Yeast, Kyokai No. 13 Yeast, Kyokai No. 13 Yeast, Kyokai No.
  • shochu yeast for example, Kyosho Shochu No.2 Yeast, Kagoshima test yeast, Miyazaki test yeast, Awamori No. 1 yeast, and the like. Mutated strains, hybrid strains, acclimatized strains, cell fusion strains, artificial mutant strains such as transformants by brasmid, etc., and even wild-type strains belonging to the genus Saccharomyces All yeasts to which they belong are included.
  • the haploid strains of the same origin according to the present invention are defined as the a type of a haploid strain isolated from the same strain of yeast having a diploid or higher ploidy.
  • the term refers to a strain and an ⁇ -type strain, and includes, for example, haploid strains derived from the Association No. 7'01 yeast and haploid strains derived from the Association No. 91 yeast.
  • At least one of the haploid strains to be hybridized in the present invention is a yeast belonging to the genus Saccharomyces, which is resistant to 18 ⁇ % ethanol. It is easy and preferable to select and hybridize a haploid strain derived from the same origin of E. coli, but the haploid strain derived from the same origin of the yeast shown in Saccharomyces You may use one that has become familiar with Nor and has become resistant to 18 ⁇ ⁇ ⁇ % ethanol. Furthermore, it is possible to cross haploid strains derived from the same origin of yeast belonging to the genus Saccharomyces cerevisiae, which is not resistant to 18 ⁇ ⁇ ⁇ % ethanol. It is.
  • the diploid hybrid strains obtained here include sake yeast, shochu yeast, Wien yeast, brewery yeast, brewer's yeast, Wisky yeast, alcohol yeast, pan yeast and the like. In both cases, a large amount of ethyl carboxylate and / or iso-isoamyl acetate is produced.
  • sake yeast is not limited to the production of sake, but sake yeast can be used for the production of shochu and the like other than sake. The same applies to other yeasts.
  • the method for producing sake, shochu, wine, beer, whiskey, fragrance liquid, pan, etc. is not particularly limited, but may be produced according to a general method. it can .
  • a haploid strain of the Japan Brewing Association No. 71 (hereinafter abbreviated as K-701) strain
  • the mating type a: 3 strains and the mating type ⁇ : 3 strains were respectively transformed into YPD liquid medium (yeast extract lw / v%, peptone 2 wZv%, glucose 2 wZv%). In 5 ml, 30. C.
  • the cells were cultured with shaking for 24 hours.
  • Culture solution of K-701 strain a-type haploid strain 0.1 ⁇ m1 and culture solution of K-701 strain ⁇ -type haploid strain 0.1 m1 to 1 m1 YPD liquid medium
  • the cells were cultured at 30 ° C. for 24 hours. This culture solution is diluted appropriately with sterile water, applied to a solid YPD plate medium, cultured at 30 ° C for 48 hours, and colonies having a diameter of more than 2 mm are crossed (diploid).
  • Strain ).
  • the rice used was rice with a polishing rate of 77% (w / w) of alpha rice (manufactured by Seven Rice Industries, Ltd.). Zeni was manufactured using 72% (w / w) of white rice. Yeast was added also contain 1 X 1 0 9 pieces in 5 ml. The fermentation temperature was constant at 15 ° C. After the distilling, the filtrate of the moromi mash was analyzed for low-boiling aroma components using gas chromatography, and as a result, all hybrids were found to have the parent strain (K-171). It was found that 1.7 to 6 times the amount of ethyl ether bromide and 2.4 to 3.9 times the amount of isoamyl acetate were formed and regenerated.
  • the above-mentioned high production of ethyl bromide and isoamyl acetate indicates that the hybrid strain is a novel yeast different from the K-701 strain.
  • the five strains according to the present invention ⁇ 8 — 30 — 1, 8 — 30 — 2, 9 — 30 — 3, 10 — 30 — 2, and 10 — 41 — 3 ⁇ Is a hybrid of a haploid strain derived from the K7011 strain, and its bacteriological properties are shown below.
  • a medium for test for assimilation of carbon compounds from Wickerham (manufactured by Difco) is dispensed into a test tube into a durham tube, and the 5 strains are inoculated. After culturing for a day, the presence or absence of the generation of carbon dioxide was observed.
  • the nitrate was potassium nitrate, and the growth was observed by the Okizano-graft method using a medium for test for assimilation of carbon compounds from Wickerham (manufactured by DFCO). Was.
  • the morphological and biochemical results indicate that the five yeast strains of the present invention are yeast belonging to Saccharomyces cerevisiae. . Also, 1 over ⁇ La double down medium, 3 5 e growth in C is negative, and whether we and the small specification write have you in the test have also a recognized form of high-foam this sake, the 5 strains K This indicates that it is a hybrid of 701 strains. 6. Resistance to drugs
  • the haploid strains of ⁇ -701 strains can be isolated from each other.
  • a highly aroma-producing yeast which can be used for ordinary sake brewing and which can produce ethyl carboxylate and isoamyl acetate at a high level was provided.
  • the Awako-no-Yamaha for sake and the Nihon Brewery Association No. 91 (hereinafter abbreviated as 91-001) strain can also be used.
  • the five strains ⁇ 3—112, 3—2—2, 3—411, 411, 411 ⁇ according to the present invention are derived from about 901 strains. Although it is a hybrid of a diploid strain, its bacteriological properties are shown below. (Mycological properties)
  • Sporulation medium (acetic mosquito re U arm 2 w / v%, Group courses 0. 0 5 w / V% , agar 2 w, v%) was Ma ⁇ nutrient 3 0 e C, 5 days, microscopic Observed at.
  • Wickerham's medium for carbon compound assimilation test (manufactured by Difco) is dispensed into a durham tube into a test tube, inoculated with the five strains, and incubated at 30 ° C. After culturing for a day, the presence or absence of the generation of carbon dioxide was observed.
  • the nitrate was potassium nitrate, and the growth was observed by the Okizano-graft method using a medium for test for assimilation of carbon compounds from Wickerham (manufactured by Difco).
  • the morphological and biochemical results indicate that the five yeast strains of the present invention are yeasts belonging to Saccharomyces 13 mis.
  • the formation of high foam was not observed in the small-batch test of sake, indicating that the five strains were hybrids of the K191 strain.
  • the haploid strains of K-901 strains are crossed with each other, whereby ethyl cabronate and / or ethyl acetate can be obtained.
  • Yeast-producing yeasts that produce high amounts of sodium and can be used for ordinary sake brewing were provided.
  • the representative strains 10 — 4 1 — 3 and 3 — 2 — 2 are designated as Saccharomyces cerevisiae 10 — 41 13 and iaccharomyces cerevisiae ⁇ -2-2, respectively.
  • Saccharomyces cerevisiae 10-13 was transferred to the International Depositary on October 25, 1994 and then to the International Depositary on January 11, 1995. Also, Saccharomyces cerevisiae 3 — 2 — 2 was deposited internationally on January 11, 1995.
  • the method for producing sake, shochu and other alcoholic beverages of the present invention is characterized by using these yeast strains, and the brewing method is not particularly limited. Similarly, the method of producing food is not particularly limited.
  • Sake was produced from the five mixed strains derived from the K-701 strain with the blending formula shown in Table 1.
  • the rice used was rice with a polishing rate of 77% (w / w) of ⁇ rice (manufactured by Seven Rice Industry Co., Ltd.). ⁇ was manufactured using 72% (w / w) of white rice.
  • the yeast contained 1 ⁇ 10 9 cells in 5 ml. The fermentation temperature was kept constant at 15.
  • As the control strain the parent strain K-701 was used, and a representative sake yeast, Japan Brewing Association No. 91 (hereinafter abbreviated as K-901) was used.
  • Table 6 shows the analysis results of the upper tank liquid. Table 6 Results of ffi solution analysis on small sake preparation Control strain Hybrid strain
  • the sensory test was performed by a three-point method (1: good, 2: normal, 3: bad), and expressed as the average value of 10 panelists.
  • Shochu was produced with the blending formula shown in Table 7 for two hybrid strains derived from the K-701 strain.
  • K-701 strain which is frequently used for rice shochu, was used.
  • the rice used was low-grade rice with a polishing rate of 70% (w / w). ⁇ was manufactured using 72% (w / w) of white rice.
  • the yeast contained 2 ⁇ 10 8 cells in 5 ml.
  • Svitase M Naagase Seikagaku Corporation] was used as the enzyme preparation.
  • the fermentation temperature was constant at 20 ° C.
  • the mash on the 14th day after distillation was distilled under reduced pressure at a decompression degree of 700 mm Hg, and alcohol was distilled off to a distillation alcohol content of 20 vZv%.
  • the water split to V / V% was analyzed. Table 8 shows the results. Table 8 Results of analysis of rice baked S decompression distillate
  • the sensory test was performed by a three-point method (1: good, 2: normal, 3: bad), and expressed as the average value of 10 panelists.
  • rice shochu produced using 10 to 30 strains or 10 to 4 13 strains had a content of ethyl bromide, ethyl bromide and isoamyl acetate. But more than K-701 strains In particular, it became a new type of shochu with a light and fruity fragrance.
  • Sake was produced from five hybrid strains derived from the K-901 strain using the blending recipe shown in Table 1.
  • the rice used was polished rice (manufactured by Seven Rice Industries, Ltd.) with a polishing rate of 77% (w / w). Satoshi was manufactured using 72% (w / w) of milled rice.
  • Yeast was added at a concentration of 1 ⁇ 10 9 in 5 ml. Fermentation temperature was Tsu line in 1 5 e C- constant.
  • As a control strain a parent strain K-901 and a representative sake yeast K-701 were used. Table 9 shows the analysis results of the upper tank liquid.
  • the content was higher than that of the K-901 strain, a strong fragrance was recognized, and a good organoleptic evaluation was obtained.
  • sake manufactured using 3-1 2 strains had a higher isoamyl acetate content than K-901 strains, and was manufactured using 4 4-1 3 strains.
  • the resulting sake has a higher cabonic acid content than that of the K-901 strain, and has a different organoleptic flavor compared to the sake produced using each of the K-901 strains. It has a quality of liquor.
  • Ethyl cabronate and Z or iso-isoamyl acetate bred by crossing haploid strains of the same origin of yeast belonging to the genus Saccharomyces according to the present invention By using high-yield new yeast, it is not necessary to use high-purity rice and low-temperature long-term ginjo brewing, but also to use ordinary sake or shochu with low-cost rice.
  • the production of aromatic liquor, shochu and other liquors, which is rich in fragrant esters mainly consisting of ethyl butyl bromide and diisoamyl acetate is low cost. This makes it possible to achieve stable operations in a short period of time. Same for food production As described above, by using the novel yeast according to the present invention, it is possible to produce a food rich in aroma components.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Mycology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Botany (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Medicinal Chemistry (AREA)
  • Food Science & Technology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Alcoholic Beverages (AREA)

Abstract

Novel yeasts having a high isoamyl acetate and/or ethyl caproate productivity and a process for producing alcoholic drinks or foods having rich ginjo-flavors with the use of the yeasts. Novel aromatic yeast strains having been bred by hybridizing haploids of yeasts which belong to the genus Saccharomyces and have the same origin with each other and having a high ethyl caproate and/or isoamyl acetate productivity. The use of these yeasts makes it possible to produce sake, shochu, wine, beer, whisky, flavoring liquids, bread, etc.

Description

明 細 書  Specification
芳香性酵母新菌株  New aromatic yeast strain
技術分野 Technical field
本発明 は、 芳香性の高い新規酵母及び該酵母に よ る 芳香 性 に富ん だ酒類、 食品の製造方法に 関す る 。  The present invention relates to a novel yeast having a high aromaticity and a method for producing alcoholic beverages and foods rich in an aromaticity by the yeast.
更 に詳細に は、 本発明 は、 芳香成分の う ち カ ブロ ン酸ェ チ ル及び /又は酢酸ィ ソ ァ ミ ルを多 く 生成す る 酵母を容易 に造成す る 方法に 関 し 、 更に は選択 し た酵母を用 いて 、 香 気の豊か な酒類、 食品を製造す る 方法に関す る 。  More specifically, the present invention relates to a method for easily producing a yeast which produces a large amount of carboxylate and / or isoamyl acetate among the aroma components. Relates to a method for producing aromatic liquors and foods using selected yeasts.
背景技術 Background art
酒類、 特に清酒の製造 に おいて 、 フ ルーテ ィ な吟醸香成 分の含量の高い清酒を製造す る に は、 低精白米を使用 し 、 In the production of alcoholic beverages, especially sake, in order to produce a high-quality sake with a high content of ginjo aroma, low-polished rice is used.
1 5 "C前後で発酵さ せ る 普通酒仕込で は困難であ る 。 従来 、 吟醸香 に富ん だ清酒を得 る ため に は高度精白米を使用 し 、 低温発酵を行 っ て い た。 し か し なが ら 、 こ の方法で は時 間 と コ ス ト がかか る 。 It is difficult to prepare regular sake that is fermented at around 15 "C. Conventionally, low-temperature fermentation was performed using highly polished rice to obtain sake rich in ginjo aroma. However, this method is time-consuming and costly.
現状で は 、 低精 白米の加圧蒸 しや リ パーゼ処理等が行わ れ る よ う に な っ たが、 低精白米の普通酒仕込か ら吟醸香に 富ん だ清酒を製造す る こ と は、 困難であ る こ と よ り 、 酵母 の育種 と い う 観点か らの技術開発に依存す る 方向が進め ら れ、 こ れ ま で に 、 カ ブロ ン酸ェチル高生産酵母の育種に関 し て は 、 セル レニ ン耐性を用 い る 方法 （特開昭 6 3 — 3 0 9 1 7 5 号） が公開 さ れて い る 。 ま た、 酢酸イ ソ ア ミ ル高 生産酵母の育種 に 関 し て は、 5 ' , 5 ' , 5 ' 一 ト リ フ ル オ ロ ー D , L — 口 イ シ ン耐性を用 い る 方法 （特開昭 6 2 — 6 6 6 9 号） 、 4 一 ァザ ー D L — 口 イ シ ン耐性を用 レ、 る 方 法 （特閧平 1 一 2 5 7 4 2 3 号） が公開さ れて い る 。 更に 、 カ ブロ ン酸ェチル及び 又は酢酸イ ソ ア ミ ル高生産酵母 に 関 し て は 、 固形色素平板培地上で異な る 色調を示す株を 選択す る 方法 （特開平 2 - 1 3 3 6 8 号） が公開さ れて い る 。 上記の いずれの方法 も 、 酵母菌株を変異処理す る こ と が好ま し い と さ れ、 交雑処理について は十分な検討がなさ れて い なか っ た。 At present, low-grade rice is steamed under pressure and lipase treated.However, the production of refined sake with a rich ginjo flavor from low-grade rice ordinary sake is started. However, it has been difficult to rely on the development of technology from the viewpoint of breeding yeast, and so far, breeding of yeast with high production of ethyl ethyl cabronate has been promoted. With respect to this, a method using cerulenin resistance (Japanese Patent Application Laid-Open No. 63-309175) has been disclosed. In addition, regarding the breeding of yeasts producing high isoamyl acetate, 5 ′, 5 ′, 5 ′ trifluoride D, L—method using oral insulin resistance (Japanese Unexamined Patent Publication (Kokai) No. 62-66669), 41-user DL — mouth The law has been disclosed. Furthermore, for yeasts that produce high levels of ethyl acetyl bromide and / or isoamyl acetate, a method for selecting strains showing different color tones on a solid dye plate medium (Japanese Patent Laid-Open No. No. 8) has been published. In any of the above methods, it is said that it is preferable to perform mutation treatment on the yeast strain, and no sufficient study has been made on the hybridization treatment.
—方、 本発明者 ら は、 日本醸造協会 7 0 1 号 と 日本醸造 協会 9 0 1 号由来一倍体同士の交雑に よ り 育種さ れる 芳香 性清酒酵母新菌株を取得 し た （特開平 5 - 3 1 7 0 3 4 号 ) 。 ま た 、 清酒酵母の一倍体株を簡便に 、 かつ、 高頻度に 製造す る 方法を見出 し た （特閧平 5 - 3 1 7 0 3 5 号） 。 し か し な が ら 、 よ り カ ブロ ン酸ェチル及び Z又は酢酸イ ソ ァ ミ ルが交雑処理に よ り 高生成さ れる 酵母の提供が望ま れ て い た 。  On the other hand, the present inventors have obtained a new aromatic sake yeast strain which is bred by crossing between haploids derived from Japan Brewing Association No. 71 and Japan Brewing Association No. 91 (Japanese Unexamined Patent Publication No. 5-3 1 7 0 3 4). In addition, they have found a simple and high-frequency method for producing haploid strains of sake yeast (Japanese Patent Application No. 5-3177035). However, it has been desired to provide a yeast in which ethyl ethyl cabronate and Z or isoamyl acetate are more highly produced by the hybridization treatment.
ま た 、 清酒以外の酒類で は、 焼酎の製造に おいて も 、 香 り の良い焼酎を製造す る ため に は、 同様の問題点を有 し 、 更 に は食品の製造にお いて も 、 同様であ っ た。  In addition, liquors other than sake have similar problems in the production of shochu, and also in the production of foods, in order to produce fragrant shochu. It was the same.
香気成分の豊かな吟醸酒の場合、 原料 コ ス ト が高 く 、 ま た製造 日 数 も長 く かか る こ と か ら 、 安定に大量の吟醸酒を 製造す る こ と は困難であ っ た。 ま た、 低精白米を用 い る 普 通酒仕込で は、 吟醸香に富ん だ清酒を製造す る こ と は更に 困難で あ っ た。  In the case of Ginjo sake, which has a rich aroma component, it is difficult to produce a large amount of Ginjo sake in a stable manner because the cost of raw materials is high and the production period is long. I did. In addition, it was even more difficult to produce sake rich in ginjo-scent when brewing ordinary sake using low-polished rice.
本発明 の 目 的 は、 前記の従来技術の問題点を解決すべ く 、 普通酒仕込で吟醸香の高い、 あ る い は新規な吟醸香を有 す る 清酒 を製造す る こ と に あ り 、 そ の た め に酢酸イ ソ ア ミ ル及びノ又はカ ブ口 ン酸ェチルの生成能の高い新規酵母菌 を提供す る こ と 、 及び該酵母を用 いて低精白米か ら 、 安定 に かつ大置に 、 吟醸香の豊かな酒類を製造す る方法、 更に は該酵母を用 いて吟醸香の豊かな食品を製造す る 方法を提 供す る こ と に あ る 。 The purpose of the present invention is to solve the above-mentioned problems of the prior art, and to produce sake with a high Ginjo aroma or a new Ginjo aroma with ordinary sake. For that purpose. And to provide a novel yeast having a high ability to produce methyl and ethyl or ethyl beet carboxylate, and to use the yeast to stably and largely produce a liquor rich in ginjo aroma from low-polished rice. Another object of the present invention is to provide a method for producing a food, and a method for producing a food rich in ginjo aroma using the yeast.
発明 の開示 DISCLOSURE OF THE INVENTION
本発明 を概説すれば、 本発明の第 1 の発明 は、 サ ッ カ ロ ミ セ ス属 に属す る酵母の同一起源由来の一倍体同士の細胞 の交雑 に よ り 育種さ れる カ ブロ ン酸ェチル及び 又は酢酸 イ ソ ア ミ ル高生産の芳香性酵母新菌株に関 し 、 第 2 の発明 は 、 当該酵母を用 い る こ と を特徴 と す る香気成分の豊かな 酒類、 食品の製造方法に関す る 。  In summary of the present invention, the first invention of the present invention is directed to a clonal breed by crossing cells of haploids from the same source of yeast belonging to the genus Saccharomyces. The second invention relates to a novel aromatic yeast strain that produces high levels of ethyl ether and / or isoamyl acetate. The second invention relates to alcoholic beverages and foods rich in aroma components characterized by using the yeast. Related to manufacturing method.
本発明者 ら は、 サ ッ カ ロ ミ セス属 に属す る 酵母の同一起 源由来の一倍体株同士を交雑さ せ る こ と に よ り 、 カ ブロ ン 酸ェチル及び /又は酢酸ィ ソ ァ ミ ル高生産株を造成す る こ と に成功 し 、 本発明 を完成さ せ た。  The present inventors crossed haploid strains derived from the same source of yeast belonging to the genus Saccharomyces, thereby obtaining ethyl cabronate and / or isoacetate. We succeeded in creating a high-amylase producing strain and completed the present invention.
本発明 に おけ る サ 、ソ カ ロ ミ セス属 に属す る 酵母 と は、 清 酒酵母、 焼酎酵母、 ワ イ ン酵母、 シ ェ リ ー酵母、 ビール酵 母、 ウ ィ ス キー酵母、 ア ル コ ール酵母、 及びパ ン酵母等で サ ッ カ ロ ミ セ ス属 に属す る 酵母であればいずれで も よ く 、 清酒醸造 に使用 さ れる 酵母では例え ば'曰本醸造協会で純粋 培養 し て 頒布さ れて い る 協会 6 号酵母、 協会 7 号酵母、 協 会 9 号酵母、 協会 1 0 号酵母、 協会 1 1 号酵母、 協会 1 2 号酵母、 協会 1 3 号酵母、 協会 1 4 号酵母、 協会 6 0 1 号 酵母、 協会 7 0 1 号酵母、 協会 9 0 1 号酵母、 協会 1 0 0 1 号酵母等が挙げ られ、 焼酎酵母で は例 え ば協会焼酎 2 号 酵母、 鹿児島ェ試酵母、 宮崎ェ試酵母、 泡盛 1 号酵母等が 挙げ られ る 。 変異処理株、 交雑株、 馴養株、 細胞融合株及 びブラ ス ミ ド 等に よ る 形質転換株等の人工変異株、 更に は 野生株であ っ て も 、 サ ッ カ ロ ミ セス属 に属す る 酵母はすべ て 含む 。 The yeast belonging to the genus Saccharomyces in the present invention includes sake yeast, shochu yeast, Wien yeast, Shelley yeast, brewer's yeast, Wisky yeast, and alky yeast. Any yeast belonging to the genus Saccharomyces such as call yeast and pan yeast can be used, and yeast used for sake brewing is, for example, pure culture at the Honshu Brewing Association. Yeast No. 6, Yeast No. 7, Association No. 9 Yeast, Kyokai No. 10 Yeast, Kyokai No. 11 Yeast, Kyokai No. 12 Yeast, Kyokai No. 13 Yeast, Kyokai No. 13 Yeast, Kyokai No. 1 Yeast No.4, Kyokai No.601 Yeast, Kyokai No.701 Yeast, Kyokai No.901 Yeast, Kyokai No.101 Yeast, etc.For shochu yeast, for example, Kyosho Shochu No.2 Yeast, Kagoshima test yeast, Miyazaki test yeast, Awamori No. 1 yeast, and the like. Mutated strains, hybrid strains, acclimatized strains, cell fusion strains, artificial mutant strains such as transformants by brasmid, etc., and even wild-type strains belonging to the genus Saccharomyces All yeasts to which they belong are included.
本発明 で い う 同一起源由来の一倍体株同士 と は、 二倍体 あ る い は それ以上の倍数性を有す る 同一株の酵母か ら分離 さ れ た一倍体株の a 型株 と α 型株の こ と を指 し 、 例え ば協 会 7 ' 0 1 号酵母由来の一倍体株同士、 協会 9 0 1 号酵母由 来の一倍体株同士が挙げ られる 。  The haploid strains of the same origin according to the present invention are defined as the a type of a haploid strain isolated from the same strain of yeast having a diploid or higher ploidy. The term refers to a strain and an α-type strain, and includes, for example, haploid strains derived from the Association No. 7'01 yeast and haploid strains derived from the Association No. 91 yeast.
本発明 に お いて交雑さ せ る 一倍体株は、 少な く と も一方 が、 1 8 ν Ζ ν %エ タ ノ ール耐性を有す る サ ッ カ ロ ミ セス 属 に属 す る酵母の同一起源由来の一倍体株を選択 し て交雑 さ せ る の が容易で好ま し いが、 サ ッ カ ロ ミ セス厲に厲す る 酵母の 同一起源由来の一倍体株をエ タ ノ ールで馴養 し て 1 8 ν Ζ ν %エ タ ノ ールに耐性を示す よ う に な つ た も の を用 い て も よ い。 更に 、 1 8 ν Ζ ν %エ タ ノ ール耐性を有 し な ぃサ ッ 力 口 ミ セ ス属 に属す る酵母の同一起源由来の一倍体 株同士 を交雑さ せ る こ と も可能であ る 。  At least one of the haploid strains to be hybridized in the present invention is a yeast belonging to the genus Saccharomyces, which is resistant to 18 νΖν% ethanol. It is easy and preferable to select and hybridize a haploid strain derived from the same origin of E. coli, but the haploid strain derived from the same origin of the yeast shown in Saccharomyces You may use one that has become familiar with Nor and has become resistant to 18 ν Ζ ν% ethanol. Furthermore, it is possible to cross haploid strains derived from the same origin of yeast belonging to the genus Saccharomyces cerevisiae, which is not resistant to 18 ν Ζ ν% ethanol. It is.
こ こ に得 られる交雑二倍体株は、 清酒酵母、 焼酎酵母、 ワ イ ン酵母、 シ ヱ リ ー酵母、 ビール酵母、 ウ ィ ス キー酵母 、 ア ル コ ール酵母、 及びパ ン酵母等のいずれで も 、 カ ブ口 ン酸ェチル及び /又は酢酸ィ ソ ァ ミ ルを多 く 生成す る ので The diploid hybrid strains obtained here include sake yeast, shochu yeast, Wien yeast, brewery yeast, brewer's yeast, Wisky yeast, alcohol yeast, pan yeast and the like. In both cases, a large amount of ethyl carboxylate and / or iso-isoamyl acetate is produced.
、 こ れ ら の酵母を用 いて清酒、 焼酎、 ワ イ ン 、 ビール、 ゥ イ ス キー等の酒類、 又はパ ン等の食品を製造すれば、 香気 の豊か な製品を製造す る こ と がで き る 。 更 に 、 こ れ らの酵 母 を用 い て芳香性の香気液を製造す る こ と も可能であ る 。 な お 、 清酒酵母は清酒の製造に限定さ れる も のでは な く 、 清酒酵母 を清酒以外の焼酎等の製造に使用 す る こ と がで き る 。 他の酵母について も 同様であ る 。 If these yeasts are used to produce alcoholic beverages such as sake, shochu, wine, beer, and whisky, or food products such as bread, it is possible to produce products with a rich aroma. it can . Furthermore, these yeasts It is also possible to produce aromatic fragrances using the mother. Incidentally, sake yeast is not limited to the production of sake, but sake yeast can be used for the production of shochu and the like other than sake. The same applies to other yeasts.
清酒、 焼酎、 ワ イ ン 、 ビール、 ウ ィ ス キー、 香気液、 パ ン 等の製造方法は特に 限定す る も のでは な く 、 一般的な方 法 に従 っ て製造す る こ と がで き る 。  The method for producing sake, shochu, wine, beer, whiskey, fragrance liquid, pan, etc. is not particularly limited, but may be produced according to a general method. it can .
以下 に本発明の酵母の取得方法の 1 例を示す。  Hereinafter, an example of the method for obtaining the yeast of the present invention will be described.
(一倍体酵母の交雑方法）  (Method of crossing haploid yeast)
1 8 v Z v %エ タ ノ ール耐性を示す清酒用泡な し酵母、 日 本醸造協会 7 0 1 号 （以下、 K 一 7 0 1 と 略述す る ） 株 の一倍体株 （接合型 a ： 3 株、 接合型 α ： 3 株） を 、 そ れ ぞれ Y P D液体培地 （酵母エキス l w / v %、 ペプ ト ン 2 w Z v % 、 グル コ ース 2 w Z v % ) 5 m l で、 3 0 。Cに て 2 4 時間振 と う 培養 し た。 K 一 7 0 1 株の a型一倍体株の 培養液 0 . l m 1 と K 一 7 0 1 株の α 型一倍体株の培養液 0 . 1 m 1 を 1 m 1 の Y P D液体培地に力□え 、 3 0 °Cに て 2 4 時間静置培養 し た。 こ の培養液を滅菌水で適当 に希釈 し て 、 固形 Y P D平板培地に塗布 し 、 3 0 で に て 4 8 時間 培養 し 、 直径が 2 m m を越え る コ ロ ニー を交雑株 （二倍体 株 ） と し て分離 し た。  18v Zv% ethanol resistant foamless yeast for sake, a haploid strain of the Japan Brewing Association No. 71 (hereinafter abbreviated as K-701) strain The mating type a: 3 strains and the mating type α: 3 strains were respectively transformed into YPD liquid medium (yeast extract lw / v%, peptone 2 wZv%, glucose 2 wZv%). In 5 ml, 30. C. The cells were cultured with shaking for 24 hours. Culture solution of K-701 strain a-type haploid strain 0.1 μm1 and culture solution of K-701 strain α-type haploid strain 0.1 m1 to 1 m1 YPD liquid medium The cells were cultured at 30 ° C. for 24 hours. This culture solution is diluted appropriately with sterile water, applied to a solid YPD plate medium, cultured at 30 ° C for 48 hours, and colonies having a diameter of more than 2 mm are crossed (diploid). Strain).
(芳香性の高い交雑株の分離方法）  (Method of isolating highly aromatic hybrids)
9 種 3 株ずつ、 合計 2 7株の交雑二倍体株を 1 8 v Z v %濃度のエ タ ノ ールを含む麴汁培地 （ ブ リ ッ ク ス度 1 . 8 、 p H 4 . 4 ) に接種 し 、 1 0 °Cで 1 0 日 間静置 し た後、 メ チ レ ン ブルー に よ る 染色率が K 一 7 0 1 株よ り も少な い 株を 5 株選択 し 、 8 - 3 0 - 1 株、 8 — 3 0 - 2 株、 9 一 3 0 - 3 株、 1 0 - 3 0 — 2 株、 及び 1 0 - 4 1 一 3株 と し た。 A total of 27 hybrid diploid strains, each consisting of 3 strains of 9 species, were cultured in a broth medium containing ethanol at a concentration of 18 vZv% (Bricks degree 1.8, pH4. 4) After inoculation at 10 ° C for 10 days, the staining rate with methylene blue is lower than that of the K-701 strain. Select 5 shares, and select 8-30-1 shares, 8-30-2 shares, 9-30-3 shares, 10-30-2 shares, and 10-41-13 shares did.
こ れ ら選択さ れた菌株を表 1 に示す仕込配合で清酒の小 仕込試験を行つ た。 表 1 仕 込 合 初 添 仲 添 留 添 合 S十 総 米 (g) 33 60 107 200  A small brewing test of sake was performed on the selected strains using the brewing formulations shown in Table 1. Table 1 Initial mashing mashing mashing mashing S-ten rice (g) 33 60 107 200
掛 米 (g) 21 47 91 159  Rice (g) 21 47 91 159
米 (g) 12 13 16 41 乳 酸 (m 1 ) 0.23 0.23 汲 水 (m 1 ) 51 85 168 304  Rice (g) 12 13 16 41 Lactic acid (m 1) 0.23 0.23 Pumped water (m 1) 51 85 168 304
酵 母 (m 1 ) 5 5  Yeast mother (m 1) 5 5
掛米は精米歩合 7 7 % ( w / w ) の α 米 [ セ ブ ン ラ イ ス 工業 （株） 製】 を使用 し た。 錢は、 精米歩合 7 2 % ( w / w ) の 白米を用 いて製造 し た。 酵母は 5 m l 中に 1 X 1 0 9 個含む も の を添加 し た。 発酵温度は 1 5 °C—定で行っ た 。 留後 1 8 曰 目 の醪の ろ液について 、 ガス ク ロ マ ト グラ フ ィ ー を用 いて低沸点香気成分の分析を行っ た結果、 いずれ の交雑株 も親株 （ K 一 7 0 1 ) の 1 . 7 〜 6 倍のカ ブロ ン 酸ェチル及び 2 . 4〜 3 . 9 倍の酢酸イ ソ ア ミ ルを生成 し て レヽ る こ と を認め た。 The rice used was rice with a polishing rate of 77% (w / w) of alpha rice (manufactured by Seven Rice Industries, Ltd.). Zeni was manufactured using 72% (w / w) of white rice. Yeast was added also contain 1 X 1 0 9 pieces in 5 ml. The fermentation temperature was constant at 15 ° C. After the distilling, the filtrate of the moromi mash was analyzed for low-boiling aroma components using gas chromatography, and as a result, all hybrids were found to have the parent strain (K-171). It was found that 1.7 to 6 times the amount of ethyl ether bromide and 2.4 to 3.9 times the amount of isoamyl acetate were formed and regenerated.
以上のカ ブロ ン酸ェチル及び酢酸ィ ソ ァ ミ ル高生産 と い う 形質は 、 当該交雑株が K - 7 0 1 株 と は異な る 新規酵母 菌で あ る こ と を示す も のであ る 。 The above-mentioned high production of ethyl bromide and isoamyl acetate The phenotype indicates that the hybrid strain is a novel yeast different from the K-701 strain.
上記の よ う に 、 本発明 に よ る 5 菌株 { 8 — 3 0 — 1 、 8 - 3 0 - 2 , 9 - 3 0 - 3 , 1 0 - 3 0 - 2 、 1 0 - 4 1 - 3 } は 、 K 一 7 0 1 株由来一倍体株の交雑株であ る が、 そ の菌学的性質を以下に示す。  As described above, the five strains according to the present invention {8 — 30 — 1, 8 — 30 — 2, 9 — 30 — 3, 10 — 30 — 2, and 10 — 41 — 3 } Is a hybrid of a haploid strain derived from the K7011 strain, and its bacteriological properties are shown below.
(菌学的性質）  (Bacteriological properties)
1 . 形態学的性質  1. Morphological properties
Y P D 培地で 3 0 で、 2 日 間培養 し た後、 顕微鏡で観察 し た 。  After culturing at 30 in YPD medium for 2 days, the cells were observed under a microscope.
a ) 形 ： 卵円形  a) Shape: oval
b ) 大 き さ ： 長さ 4 . 7 〜 7 . 9 μ ηι、 幅 3 . 8〜 5 . 5 μ m b) Size: Length 4.7 to 7.9 μηι, width 3.8 to 5.5 μm
2 . 胞子形成 ： 有 り  2. Sporulation: Yes
胞子形成用培地 （酢酸カ リ ウ ム 2 w Z v % 、 グルコ ース 0 . 0 5 w / V % , 寒天 2 \^ ％ ) で 3 0 T：、 5 日 間培 養 し 、 顕微鏡で観察 し た。  Culture in a medium for sporulation (2% Zw% of potassium acetate, 0.05% w / v of glucose, 2% of agar) at 30 T: Cultured for 5 days and observed under a microscope did.
3 . 増殖の形態 ： 出芽  3. Form of growth: budding
4 . 生化学的観察  4. Biochemical observations
a ) 糖の発酵性 a) Sugar fermentability
ウ ィ ッ カ ー八ム の炭素化合物同化試験用培地 （ デ ィ フ コ 社製 ） を ダー ラ ム管入 り 試験管に分注 し て 、 当該 5 菌株を 接種 し 、 3 0 -Cで 7 日 間培養 し て 、 そ の炭酸ガス発生の有 無を観察 し た。  A medium for test for assimilation of carbon compounds from Wickerham (manufactured by Difco) is dispensed into a test tube into a durham tube, and the 5 strains are inoculated. After culturing for a day, the presence or absence of the generation of carbon dioxide was observed.
グル コ ー ス （ + ) ガ ラ ク ト ー ス （ + ) ス ク ロ ー ス （ + ) マル ト ー ス （ + ) ラ ク 卜 一 ス （ 一 ） メ リ ビオ ー ス （ 一 ） ラ フ イ ノ ース （ + ) Glucose (+) Galactose (+) Scroose (+) Maltose (+) Lactose (1) Melibiose (1) Raffinose (+)
b ) 糖の資化性 b) Utilization of sugar
ウ ィ ッ カ ーハ ム の炭素化合物同化試験用培地 （ デ ィ フ コ 社製 ） を用 いて 、 ォ キザノ グラ フ 法に よ り 、 3 0 °C、 1 4 曰 後の生育を観察 し た。  Using Wickerham's medium for carbon compound assimilation test (manufactured by Difco), growth was observed at 14 ° C and 14 ° C by the Okizanograph method. .
グル コ ー ス （ + ) ガ ラ ク 卜 一 ス （ + ) ス ク ロ ース （ + ) マル ト ー ス （ + ) ラ ク 卜 ース （ 一 ）  Glucose (+) Garcose (+) Cross (+) Maltose (+) Lactose (1)
c ) 硝酸塩の 同化性 ： （ 一 ） c) Nitrate assimilation: (I)
硝酸塩は硝酸カ リ ウ ム と し 、 ウ ィ ッ カ ーハ ム の炭素化合 物同化試験用培地 （デ ィ フ コ 社製） を用 いて 、 ォ キザノ グ ラ フ 法 に よ り 生育を観察 し た。  The nitrate was potassium nitrate, and the growth was observed by the Okizano-graft method using a medium for test for assimilation of carbon compounds from Wickerham (manufactured by DFCO). Was.
d ) T T C 染色性 ： 赤 d) TTC staining: red
e ) (3 — ァ ラ ニ ン培地、 3 5 。C、 3 日 間培養での生育 ： （ 一 ) e) (3—Aranine medium, 35. C, growth in 3 day culture: (1)
5 . 高泡の形成  5. Formation of high bubbles
清酒の小仕込を行っ た と こ ろ 、 高泡の形成は観察さ れな か っ た 。  During the small brewing of sake, the formation of high bubbles was not observed.
以上、 形態学的、 生化学的結果は、 本発明酵母 5 菌株が サ ッ カ ロ ミ セ ス · セ レ ビ シェ ( Saccharomyces cerevisiae ) に属す る 酵母菌であ る こ と を示す も のであ る 。 ま た、 1 ー ァ ラ ニ ン培地、 3 5 eCでの生育が陰性、 かつ清酒の小仕 込試験 に お いて高泡の形成 も認め られな い こ と か ら 、 当該 5 菌株 は K 一 7 0 1 株の交雑株であ る こ と を示す も のであ る 。 6 . 薬剤 に対す る 耐性 As described above, the morphological and biochemical results indicate that the five yeast strains of the present invention are yeast belonging to Saccharomyces cerevisiae. . Also, 1 over § La double down medium, 3 5 e growth in C is negative, and whether we and the small specification write have you in the test have also a recognized form of high-foam this sake, the 5 strains K This indicates that it is a hybrid of 701 strains. 6. Resistance to drugs
そ れぞれの薬剤を含む S D 培地を用 いて 、 3 0 eCで 3 曰 間培養 し た。 そ の結果を表 2 に示す。 表 2 And have use the SD medium its containing drugs respectively, were cultured for 3曰at 3 0 e C. The results are shown in Table 2. Table 2
Figure imgf000011_0001
a ) 5 ' , 5 ' , 5 ' 一 ト リ フ ルオ ロ ー D , L —口 イ シ ン耐性
Figure imgf000011_0001
a) 5 ', 5', 5 'trifluoro D, L-mouth resistant
b ) セ ル レニ ン耐性  b) Cellulenine resistance
c ) 4 一 ァザ ー D L —口 イ シ ン耐性  c) 4 OTHER D L — mouth resistant
d ) ク ロ ト リ マ ゾール耐性  d) Clotrimazole resistance
7 . カ ブロ ン酸に対す る感受性  7. Sensitivity to cabronic acid
カ ブロ ン酸を含む S D 培地を用 いて 、 3 0。Cで 3 日 間培 養 し た。 そ の結果を表 3 に示す。 3 カ ブロ ン酸 30 using SD medium containing carboxylic acid. C cultivated for 3 days. The results are shown in Table 3. 3 Cavonic acid
菌 株  Strain
0 . 0 3 V / V %  0.03 V / V%
K - 7 0 1 + K-7 0 1 +
8 - 3 0 - 1  8-3 0-1
8 - 3 0 - 2  8-3 0-2
9 - 3 0 - 3 +  9-3 0-3 +
1 0 - 3 0 - 2  1 0-3 0-2
1 0 - 4 1 - 3 + か く し て 、 本発明 に よ り 、 Κ — 7 0 1 株の一倍体株、 好 ま し く はエ タ ノ ール耐性を示す一倍体株同士を交雑さ せ る こ と に よ っ て 、 カ ブロ ン酸ェチル及び酢酸イ ソ ア ミ ルを高 生成す る 、 普通酒仕込に使用可能な香気高生産酵母が提供 さ れ た 。  Thus, according to the present invention, the haploid strains of Κ-701 strains, preferably haploid strains exhibiting ethanol resistance, can be isolated from each other. By the hybridization, a highly aroma-producing yeast which can be used for ordinary sake brewing and which can produce ethyl carboxylate and isoamyl acetate at a high level was provided.
前記 Κ 一 7 0 1 株 と 同様の手法に よ り 、 清酒用泡な し酵 母、 日 本醸造協会 9 0 1 号 （以下、 Κ 一 9 0 1 と 略述す る ) 株 に つ いて も 、 カ ブロ ン酸ェチル及びノ又は酢酸イ ソ ァ ミ ル高生産の交雑二倍体株を 5 株選択 し 、 3 - 1 - 2 株、 3 - 2 - 2 株、 3 - 4 一 1 株、 4 一 1 一 1 株、 及び 4 一 4 一 3 株 と し た。  According to the same method as that of the above-mentioned 70-1 strain, the Awako-no-Yamaha for sake and the Nihon Brewery Association No. 91 (hereinafter abbreviated as 91-001) strain can also be used. , 5 selected hybrid diploid strains that produce high amounts of ethyl ethyl and bromo or isoamyl acetate, 3-1 -2 strain, 3-2-2 strain, 3-4 1 strain, 411 and 413 shares.
本発明 に よ る 5 菌株 { 3 — 1 一 2 、 3 — 2 — 2 、 3 - 4 一 1 、 4 一 1 一 1 、 4 一 4 一 3 } は、 Κ 一 9 0 1 株由来一 倍体株の交雑株であ る が、 その菌学的性質を以下に示す。 ( 菌学的性質） The five strains {3—112, 3—2—2, 3—411, 411, 411} according to the present invention are derived from about 901 strains. Although it is a hybrid of a diploid strain, its bacteriological properties are shown below. (Mycological properties)
1 . 形態学的性質  1. Morphological properties
Y P D 培地で 3 0 eC、 2 日 間培養 し た後、 顕微鏡で観察 し た 。 After cultured for 3 0 e C, 2 days in YPD medium and observed under a microscope.
a ) 形 ： 卵円形  a) Shape: oval
b ) 大 き さ ： 長さ 4 . 7 〜 7 . 9 i m 、 幅 3 . 8 〜 5 . 5 μ m  b) Size: Length 4.7 to 7.9 im, width 3.8 to 5.5 μm
2 . 胞子形成 ： 有 り  2. Sporulation: Yes
胞子形成用培地 （酢酸カ リ ウ ム 2 w / v % 、 グル コ ー ス 0 . 0 5 w / V % , 寒天 2 w , v % ) で 3 0 eC、 5 日 間培 養 し 、 顕微鏡で観察 し た。 Sporulation medium (acetic mosquito re U arm 2 w / v%, Group courses 0. 0 5 w / V% , agar 2 w, v%) was Ma培nutrient 3 0 e C, 5 days, microscopic Observed at.
3 . 増殖の形態 ： 出芽  3. Form of growth: budding
4 . 生化学的観察  4. Biochemical observations
a ) 糖の発酵性 a) Sugar fermentability
ウ ィ ッ カ ーハ ム の炭素化合物同化試験用培地 （ デ ィ フ コ 社製 ） を ダー ラ ム管入 り 試験管に分注 し て 、 当該 5 菌株を 接種 し 、 3 0 °Cで 7 日 間培養 し て 、 そ の炭酸ガス発生の有 無を観察 し た。  Wickerham's medium for carbon compound assimilation test (manufactured by Difco) is dispensed into a durham tube into a test tube, inoculated with the five strains, and incubated at 30 ° C. After culturing for a day, the presence or absence of the generation of carbon dioxide was observed.
グル コ ー ス （ + ) ガ ラ ク ト ー ス （ + ) ス ク ロ ー ス （ + ) マ ル ト ース （ + ) ラ ク ト ー ス （ 一 ） メ リ ビ オ ー ス （ 一 ） ラ フ イ ノ 一 ス （ + )  Glucose (+) galactose (+) sucrose (+) maltose (+) lactose (1) meliviose (1) la Finance (+)
b ) 糖の資化性 b) Utilization of sugar
ウ ィ ッ カ ーノ、 ム の炭素化合物同化試験用培地 （ デ ィ フ コ 社製 ） を 用 いて 、 ォ キザ ノ グラ フ 法に よ り 、 3 0 °C、 1 4 曰 後の生育を観察 し た。 Using a medium for test of carbon compound assimilation of Wickano and Mu (Difco) at 30 ° C and 14 ° C by the Oxanograph method. He observed the subsequent growth.
グル コ — ス （ + ) ガラ ク 卜 ース ( + ) ス ク d ース （ + ) マル ト ース ( + ) ラ ク 卜 ース （ 一 ）  Glucose (+) galactose (+) sugar dose (+) maltose (+) lactose (1)
C ) 硝酸塩の同化性 ： （ 一 ）  C) Nitrate assimilation: (I)
硝酸塩は硝酸カ リ ウ ム と し 、 ウ ィ ッ カ ーハ ム の炭素化合 物同化試験用培地 （ デ ィ フ コ 社製） を用 いて 、 ォ キザノ グ ラ フ 法 に よ り 生育を観察 し た  The nitrate was potassium nitrate, and the growth was observed by the Okizano-graft method using a medium for test for assimilation of carbon compounds from Wickerham (manufactured by Difco). Was
d ) T T C染色性 ： 赤 d) TTC staining: red
e ) 13 一 ァ ラ ニ ン培地、 3 5 °Cでの生育 ： （ + ) e) Growth in 13-aranin medium at 35 ° C: (+)
5 . 高泡の形成 5. Formation of high bubbles
酒の小仕込を行っ た と こ ろ 、 高泡の形成は観察さ れな か つ た 。  During the small brewing of sake, the formation of high bubbles was not observed.
以上、 形態学的、 生化学的結果は、 本発明酵母 5 菌株が サ ッ 力 13 ミ セ ス ♦ セ レ ピ シェ に属す る 酵母菌であ る こ と を 示す も の であ る 。 ま た、 清酒の小仕込試験に おいて高泡の 形成 も 認め られな レヽ こ と か ら 、 当該 5 菌株は K 一 9 0 1 株 の交雑株であ る こ と を示す も のであ る 。  As described above, the morphological and biochemical results indicate that the five yeast strains of the present invention are yeasts belonging to Saccharomyces 13 mis. In addition, the formation of high foam was not observed in the small-batch test of sake, indicating that the five strains were hybrids of the K191 strain.
6 . 薬剤 に対す る 耐性 6. Resistance to drugs
そ れぞれの薬剤を含む S D 培地を用 いて 、 3 0 °Cで 3 曰 間培養 し た。 そ の結果を表 4 に示す。 表 4 The cells were cultured at 30 ° C for 3 hours using SD medium containing each drug. The results are shown in Table 4. Table 4
Figure imgf000015_0001
Figure imgf000015_0001
ン耐性 Resistance
b ) セ ル レ ニ ン耐性  b) Cellulenine resistance
c ) 4 一 ァザ ー D L —口 イ シ ン耐性  c) 4 OTHER D L — mouth resistant
d ) ク ロ ト リ マ ゾール耐性  d) Clotrimazole resistance
7 . カ ブロ ン酸に対す る感受性 7. Sensitivity to cabronic acid
カ ブロ ン酸を含む S D 培地を用 いて 、 3 0 eCで 3 日 間培 養 し た 。 そ の結果を表 5 に示す。 5 カ ブ口 ン酸 And you have use the SD medium containing mosquito Bro phosphate, 3 days was Ma培nourishment at 3 0 e C. Table 5 shows the results. 5 Turnip acid
休  Rest
0 . 0 3 V / V %  0.03 V / V%
κ 一 9 0 1 +  κ 1 9 0 1 +
3 一 1 一 2 + 3 1 1 1 2 +
一 一 +  One one +
3 一 4 一 1 +  3 1 4 1 1 +
4 一 1 一 1 +  4 1 1 1 1 +
4 一 4 一 3 + か く し て 、 本発明 に よ り 、 K — 9 0 1 株の一倍体株同士 を交雑さ せ る こ と に よ っ て 、 カ ブロ ン酸ェチル及び 又は 酢酸イ ソ ア ミ ルを高生成す る 、 普通酒仕込に使用可能な香 気高生産酵母が提供さ れた。  4 14 13 + Thus, according to the present invention, the haploid strains of K-901 strains are crossed with each other, whereby ethyl cabronate and / or ethyl acetate can be obtained. Yeast-producing yeasts that produce high amounts of sodium and can be used for ordinary sake brewing were provided.
代表的 な菌株であ る 1 0 — 4 1 — 3 株、 3 — 2 — 2 株は 、 そ れぞれ Saccharomyces cerevisiae 1 0 — 4 1 一 3 、 iaccharomyces cerevisiae ό ― 2 ― 2 と 表示 し 、 通産省 工業技術院生命工学工業技術研究所に 、 そ れぞれ F E R Μ The representative strains 10 — 4 1 — 3 and 3 — 2 — 2 are designated as Saccharomyces cerevisiae 10 — 41 13 and iaccharomyces cerevisiae ό-2-2, respectively. The Institute of Biotechnology and Industrial Technology, the National Institute of Advanced Industrial Science and Technology (FER)
B P — 5 3 3 2 、 F E R M B P — 5 3 3 1 と し て寄託 し て あ る 。 な お 、 Saccharomyces cerevisiae 1 0 — 4 1 一 3 は平成 6 年 1 0 月 2 5 曰 に寄託、 そ の後平成 7 年 1 2 月 1 1 曰 に 国際寄託に移管さ れた。 ま た 、 Saccharomyces c e r e v i s i a e 3 — 2 — 2 は平成 7 年 1 2 月 1 1 曰 に 国際寄 託さ れ た 。 They have been deposited as BP-533 32 and FERMBP-5331. Saccharomyces cerevisiae 10-13 was transferred to the International Depositary on October 25, 1994 and then to the International Depositary on January 11, 1995. Also, Saccharomyces cerevisiae 3 — 2 — 2 was deposited internationally on January 11, 1995.
2 株取得で き たが、 こ れ ら に 限定す る も ので は ない。 本発明 の清酒、 焼酎及びそ の他の酒類の製造方法は、 こ れ ら の酵母菌株を用 い る こ と を特徴 と し 、 醸造方法は特に 限定す る も のでは な い。 ま た、 食品の製造方法に おいて も 同様 に製造方法は特に限定す る も ので はな い。  Two shares were acquired, but not limited to these. The method for producing sake, shochu and other alcoholic beverages of the present invention is characterized by using these yeast strains, and the brewing method is not particularly limited. Similarly, the method of producing food is not particularly limited.
発明 を実施す る ための最良の形態 BEST MODE FOR CARRYING OUT THE INVENTION
次 に 、 本発明の菌株を用 い た酒類製造の具体例を挙げて Next, specific examples of alcoholic beverage production using the strain of the present invention will be given.
、 本発明 を更に具体的 に説明 す る が、 本発明 は こ れ らの実 施例 に 限定さ れな い。 However, the present invention will be described more specifically, but the present invention is not limited to these embodiments.
実施例 1 Example 1
K 一 7 0 1 株由来交雑株 5 株につ いて 、 表 1 に示す仕込 配合で清酒の製造を行っ た。 掛米は精米歩合 7 7 % ( w / w ) の α 米 [ セブン ラ イ ス工業 （株） 製〕 を使用 し た。 麴 は 、 精米歩合 7 2 % ( w / w ) の 白米を用 いて製造 し た。 酵母は 5 m 1 中 に 1 X 1 0 9 個含む も の を添加 し た。 発酵 温度 は 1 5 で一定で行っ た。 対照株 と し て親株の K 一 7 0 1 株、 及び代表的な清酒酵母であ る 日本醸造協会 9 0 1 号 (以下 K - 9 0 1 と 略述す る ） を用 い た。 上槽液の分析結 果 を表 6 に示す。 表 6 淸酒小仕込上 ffi液分析結果 対照株 交雑株 Sake was produced from the five mixed strains derived from the K-701 strain with the blending formula shown in Table 1. The rice used was rice with a polishing rate of 77% (w / w) of α rice (manufactured by Seven Rice Industry Co., Ltd.).麴 was manufactured using 72% (w / w) of white rice. The yeast contained 1 × 10 9 cells in 5 ml. The fermentation temperature was kept constant at 15. As the control strain, the parent strain K-701 was used, and a representative sake yeast, Japan Brewing Association No. 91 (hereinafter abbreviated as K-901) was used. Table 6 shows the analysis results of the upper tank liquid. Table 6 Results of ffi solution analysis on small sake preparation Control strain Hybrid strain
K-701 K-901 8-30-1 8-30-2 9-30-3 10-30- 10-41-K-701 K-901 8-30-1 8-30-2 9-30-3 10-30- 10-41-
2 3 アルコ-ル (v/vX) 19.0 19.8 18.4 18.2 18.0 17.8 18.8 日本酒度 ±0 +4.0 -7.0 -8.0 -8.0 -10.0 -2.02 3 Alcohol (v / vX) 19.0 19.8 18.4 18.2 18.0 17.8 18.8 Sake degree ± 0 +4.0 -7.0 -8.0 -8.0 -10.0 -2.0
P H 4.27 4.31 4.34 4.33 4.34 4.34 4.39P H 4.27 4.31 4.34 4.33 4.34 4.34 4.39
^ χ» rtir 、 ^ χ »rtir,
3.25 3.25 3.25 3. 30 3. 20 3.00 2.70 ノ 8S (ml j 1.85 1.95 2.25 2.15 2.20 2.40 2.30  3.25 3.25 3.25 3.30 3.20 3.00 2.70 no 8S (ml j 1.85 1.95 2.25 2.15 2.20 2.40 2.30
«¾ 7 ppn^ «¾ 7 ppn ^
fx Γ f ァ 1* 19.2 20.3 19.9 20.1 19.3 14.5 21.9 酢酸 Iチル 87.6 97.3 161.3 Η8.1 1 0.0 119.1 153.5 n-フ 'ΠΛ'ノ-ル 223.2 170.8 223.7 214.1 208.4 125.9 170.3 ノ、 、  fx Γ f a 1 * 19.2 20.3 19.9 20.1 19.3 14.5 21.9 I-yl acetate 87.6 97.3 161.3 Η8.1 1 0.0 119.1 153.5
4.8 4.2 18.5 15.6 15.2 16.6 13.2 イソフ タノール 108.7 93.2 87.5 84.2 84.4 91.5 117.2 カフ ·αン ¾ιチル 0.7 0.7 1.2 1.4 1. 4.2 1.9 イソァミル 7ルコ-ル 238.1 251.0 240.0 237.4 237.8 237.0 274.4 能 検 査  4.8 4.2 18.5 15.6 15.2 16.6 13.2 isophthalanol 108.7 93.2 87.5 84.2 84.4 91.5 117.2
2.2 2.1 1.4 1.5 1.5 1.4 1.3 2.2 2.1 1.4 1.5 1.5 1.4 1.3
(芳香性） ―. (Aromatic) ―.
96/20272 96/20272
官能検査は 3点法 （ 1 ： 良、 2 ： 普通、 3 ： 悪） で行い 、 パネ ラ ー 1 0 名 の平均値で表 し た。  The sensory test was performed by a three-point method (1: good, 2: normal, 3: bad), and expressed as the average value of 10 panelists.
実施例 2 Example 2
K 一 7 0 1 株由来交雑株 2 株について 、 表 7 に示す仕込 配合で焼酎の製造を行っ た。 対照株 と し て米焼酎用 に よ く 用 い られ る K 一 7 0 1 株を使用 し た。  Shochu was produced with the blending formula shown in Table 7 for two hybrid strains derived from the K-701 strain. As a control strain, K-701 strain, which is frequently used for rice shochu, was used.
Figure imgf000019_0001
掛米は精米歩合 7 0 % ( w / w ) の低品位米を使用 し た 。 麴は 、 精米歩合 7 2 % ( w / w ) の 白米を用 いて製造 し た。 酵母は 5 m 1 中に 2 X 1 0 8 個含む も の を添加 し た。 酵素剤は ス ビ タ ーゼ M [ナガセ生化学工業 （株） 製〕 を使 用 し た。 発酵温度は 2 0 °C—定で行っ た。 留後 1 4 日 目 の 醪を減圧度一 7 0 0 m m H g で減圧蒸留 し 、 留出 アル コ ー ル分 2 0 v Z v % ま での垂ロ を ア ルコ ール 2 5 . 0 V / V % に割水 し た も の を分析 し た。 そ の結果を表 8 に示す。 表 8 米焼 S滅圧蒸留液分析結果
Figure imgf000019_0001
The rice used was low-grade rice with a polishing rate of 70% (w / w).麴 was manufactured using 72% (w / w) of white rice. The yeast contained 2 × 10 8 cells in 5 ml. Svitase M [Nagase Seikagaku Corporation] was used as the enzyme preparation. The fermentation temperature was constant at 20 ° C. The mash on the 14th day after distillation was distilled under reduced pressure at a decompression degree of 700 mm Hg, and alcohol was distilled off to a distillation alcohol content of 20 vZv%. The water split to V / V% was analyzed. Table 8 shows the results. Table 8 Results of analysis of rice baked S decompression distillate
Figure imgf000020_0001
官能検査は 3 点法 （ 1 ： 良、 2 ： 普通、 3 ： 悪） で行い 、 パネ ラ ー 1 0 名 の平均値で表 し た。
Figure imgf000020_0001
The sensory test was performed by a three-point method (1: good, 2: normal, 3: bad), and expressed as the average value of 10 panelists.
こ の結果、 1 0 — 3 0 — 2 株又は 1 0 — 4 1 一 3 株を用 いて 製造 し た米焼酎は、 吟醸香であ る カ ブロ ン酸ェチル及 び酢酸イ ソ ア ミ ル含量が K 一 7 0 1 株に比べて 多 く 、 官能 的 に も 軽快で フ ルーテ ィ な香 り が強 く 認め られた新 し い タ イ ブの焼酎 と な っ た。 As a result, rice shochu produced using 10 to 30 strains or 10 to 4 13 strains had a content of ethyl bromide, ethyl bromide and isoamyl acetate. But more than K-701 strains In particular, it became a new type of shochu with a light and fruity fragrance.
以上の結果は 、 8 - 3 0 - 1 株、 8 — 3 0 - 2 株、 9 一 The above results are as follows: 8-30-1 shares, 8-30-2 shares, 9-1 shares
3 0 - 3 株、 1 0 — 3 0 - 2 株、 及び 1 0 - 4 1 一 3 株の 交雑株 5 株が、 K 一 7 0 1 株に ない有用 な性質を もつ、 す なわ ち香気エステルを多 く 生成す る 新規酵母菌であ る こ と を示す も のであ る 。 Five hybrids of 30-3, 10-3 0-2, and 10-4 13 strains have useful properties not found in the K7-101 strain, that is, aromatic esters. This indicates that it is a novel yeast that produces a large amount of E. coli.
実施例 3 Example 3
K 一 9 0 1 株由来交雑株 5 株につ いて 、 表 1 に示す仕込 配合で清酒の製造を行 っ た。 掛米は精米歩合 7 7 % ( w / w ) の α 米 [ セ ブン ラ イ ス工業 （株） 製 ] を使用 し た。 飽 は 、 精米歩合 7 2 % ( w / w ) の 白米を用 いて製造 し た。 酵母は 5 m 1 中 に 1 X 1 0 9 個含む も の を添加 し た。 発酵 温度は 1 5 eC—定で行 っ た。 対照株 と し て親株の K 一 9 0 1 株、 及び代表的 な清酒酵母であ る K 一 7 0 1 株を用 い た 。 上槽液の分析結果を表 9 に示す。 Sake was produced from five hybrid strains derived from the K-901 strain using the blending recipe shown in Table 1. The rice used was polished rice (manufactured by Seven Rice Industries, Ltd.) with a polishing rate of 77% (w / w). Satoshi was manufactured using 72% (w / w) of milled rice. Yeast was added at a concentration of 1 × 10 9 in 5 ml. Fermentation temperature was Tsu line in 1 5 e C- constant. As a control strain, a parent strain K-901 and a representative sake yeast K-701 were used. Table 9 shows the analysis results of the upper tank liquid.
表 9 清酒小仕込上槽液分析結果 対照株 交雑株 Table 9 Results of analysis of sake liquor in the top liquor of control sake
K-701 -901 3-1-2 3-2-2 3-4-1 4-1-1 4-4-3 アルコ-ル (v/vX) 19.5 19.8 19.3 19.8 19.4 19.5 19.2 日本酒度 + 3.0 +5.0 + 1.5 +1.5 +1.5 -1.0 -0.5K-701 -901 3-1-2 3-2-2 3-4-1 4-1-1 4-4-3 alcohol (v / vX) 19.5 19.8 19.3 19.8 19.4 19.5 19.2 Sake degree + 3.0 + 5.0 + 1.5 +1.5 +1.5 -1.0 -0.5
P H 4.13 4.22 4.17 4.22 4.21 4.06 4.08 滴定酸度（Bl) 2.75 2.85 2.85 2.75 2.70 3.10 3.10PH 4.13 4.22 4.17 4.22 4.21 4.06 4.08 Titrate (Bl) 2.75 2.85 2.85 2.75 2.70 3.10 3.10
7ミノ酸度 (,1) 1.95 2.10 Z.15 2.15 2.20 1.95 1.95 低沸点香気 7 Mino acidity (, 1) 1.95 2.10 Z.15 2.15 2.20 1.95 1.95 Low boiling odor
成分 （PPB) Ingredient (PPB)
ァ tトァ ΛΓヒト' 18.6 18. 15.3 14.3 13.8 11.9 12.8 酢黢 If 120.0 104.5 134.5 145.9 120.0 119.4 123.0 n-フ' Wノ-ル 166.1 136.0 140.7 138.9 126.0 103.5 82.0 齚酸イソァミル 8.6 6.3 10.8 14.9 14.3 10.4 6.3 イソフ'タノ-ル 95.9 98.1 102.2 109.5 103.0 98.4 108.2 カフ 'ロン酸 ifル 0.9 0.7 0.7 1.3 1.2 1.4 1.2 イソアミ 11アルコ- 236.0 242.0 218.5 224.4 212.1 203.5 208.5 官 能 検 査  T ト ΛΓ human 1 18.6 18. 15.3 14.3 13.8 11.9 12.8 Vinegar 12 If 120.0 104.5 134.5 145.9 120.0 119.4 123.0 'Tanol 95.9 98.1 102.2 109.5 103.0 98.4 108.2 Cuff' ifonic acid 0.9 0.7 0.7 1.3 1.2 1.4 1.2 Isoamiric 11 alcohol-236.0 242.0 218.5 224.4 212.1 203.5 208.5 Public inspection
2.1 2.0 1.8 1.3 1.4 1.7 1.8 2.1 2.0 1.8 1.3 1.4 1.7 1.8
(芳香性） 官能検査は 3 点法 （ 1 ： 良、 2 ： 普通、 3 ： 悪） で行い 、 パネ ラ ー 1 0 名 の平均値で表 し た。 (Aromatic) The sensory test was performed by a three-point method (1: good, 2: normal, 3: bad), and expressed as the average value of 10 panelists.
こ の結果、 3 — 2 - 2 株、 3 — 4 - 1 株、 4 一 1 一 1 株 を用 いて製造 し た清酒は、 吟醸香であ る カ ブロ ン酸ェチル 及び酢酸イ ソ ア ミ ル含量が K 一 9 0 1 株に比べて 多 く 、 フ ルーテ ィ な香 り が強 く 認め られ、 官能的に も良好な評価が 得 ら れた 。 ま た、 3 — 1 一 2 株を用 いて製造 し た清酒は、 酢酸イ ソ ア ミ ル含量が K — 9 0 1 株に比べて 多 く な り 、 4 一 4 一 3 株を用 いて製造 し た清酒は、 カ ブロ ン酸含量が K 一 9 0 1 株 に比べて 多 く な り 、 それぞれ K 一 9 0 1 株を用 い て 製造 し た清酒に比べて官能的 に も異な る 香気を有す る 酒質 と な っ た。  As a result, the sake produced using the 3-2, 2-4, 3-4-1, and 4-11 shares produced the ethyl savory, ethyl bromide and isoamyl acetate. The content was higher than that of the K-901 strain, a strong fragrance was recognized, and a good organoleptic evaluation was obtained. In addition, sake manufactured using 3-1 2 strains had a higher isoamyl acetate content than K-901 strains, and was manufactured using 4 4-1 3 strains. The resulting sake has a higher cabonic acid content than that of the K-901 strain, and has a different organoleptic flavor compared to the sake produced using each of the K-901 strains. It has a quality of liquor.
以上の結果は、 3 - 1 一 2 株、 3 - 2 - 2 株、 3 — 4 - 1 株、 4 — 1 一 1 株、 及び 4 — 4 一 3 株の交雑株 5 株が、 K 一 9 0 1 株 に な い有用 な性質を も つ、 す なわ ち香気エス テ ルを 多 く 生成す る 新規酵母菌であ る こ と を示す も のであ る 。  The above results show that 5 crosses of 3-1 1 2, 3-2-2, 3-4-1, 4-1 1 1 and 4-4 1 3 It indicates that it is a novel yeast that has useful properties not found in the 01 strain and that produces a large amount of aroma esters.
本発明 に よ る サ ッ カ ロ ミ セス属 に属す る 酵母の同一起源 由来の一倍体株同士の交雑 に よ り 育種さ れる カ ブロ ン酸ェ チ ル及び Z又は酢酸ィ ソ ァ ミ ル高生産の新規酵母菌を使用 す る こ と に よ り 、 高精 白米、 低温長期仕込の吟醸仕込を行 わ な く と も 、 普通酒仕込あ る いは低価格米に よ る 焼酎仕込 に お いて カ ブロ ン酸ェチル及びノ又は酢酸ィ ソ ァ ミ ルを主 と す る 香気エ ス テ ルの豊か な、 芳香性に富ん だ清酒、 焼酎 及びそ の他の酒類の製造が、 低 コ ス ト で 、 し か も短期間で 安定 し て 行 う こ と が可能 と な る 。 食品の製造に お いて も 同 様 に 、 本発明 に よ る新規酵母菌を用 い る こ と に よ り 、 香気 成分の豊かな食品を製造す る こ と が可能 と な る 。 Ethyl cabronate and Z or iso-isoamyl acetate bred by crossing haploid strains of the same origin of yeast belonging to the genus Saccharomyces according to the present invention By using high-yield new yeast, it is not necessary to use high-purity rice and low-temperature long-term ginjo brewing, but also to use ordinary sake or shochu with low-cost rice. In addition, the production of aromatic liquor, shochu and other liquors, which is rich in fragrant esters mainly consisting of ethyl butyl bromide and diisoamyl acetate, is low cost. This makes it possible to achieve stable operations in a short period of time. Same for food production As described above, by using the novel yeast according to the present invention, it is possible to produce a food rich in aroma components.

Claims

請 求 の 範 囲 The scope of the claims
1 . サ ッ カ ロ ミ セ ス属 に属す る酵母の同一起源由来の一 倍体株同士の交雑 に よ り 育種さ れる カ ブロ ン酸ェチル及び 又は酢酸ィ ソ ァ ミ ル高生産の芳香性酵母新菌株。  1. High fragrance of high production of ethyl etyl and / or isoamyl acetate bred by crossing of haploid strains of the same origin of yeast belonging to the genus Saccharomyces New yeast strain.
2 . 該新菌株が、 カ ブロ ン酸ェチル及び酢酸イ ソ ア ミ ル 髙生産であ る 請求項 1 記載の芳香性酵母新菌株。  2. The new aromatic yeast strain according to claim 1, wherein the new strain is produced by ethyl ethyl carboxylate and isoamyl acetate.
3 . 該新菌株が、 1 8 V ノ V %エ タ ノ ール耐性を示す請 求項 1 に記載の芳香性酵母新菌株。  3. The new aromatic yeast strain according to claim 1, wherein the new strain exhibits 18 V / V% ethanol resistance.
4 . 請求項 1 に記載の酵母新菌株を用 い る こ と を特徴 と す る 酒類、 食品の製造方法。  4. A method for producing alcoholic beverages and foods, which comprises using the novel yeast strain according to claim 1.
PCT/JP1995/002632 1994-12-26 1995-12-22 Novel aromatic yeast strains WO1996020272A1 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
AU43146/96A AU4314696A (en) 1994-12-26 1995-12-22 Novel aromatic yeast strains
KR1019970704352A KR987001032A (en) 1994-12-26 1995-12-22 Novel Aromatic Yeast Strains
JP52035396A JP3544987B2 (en) 1994-12-26 1995-12-22 New aromatic yeast strain

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP6/336638 1994-12-26
JP33663894 1994-12-26

Publications (1)

Publication Number Publication Date
WO1996020272A1 true WO1996020272A1 (en) 1996-07-04

Family

ID=18301239

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP1995/002632 WO1996020272A1 (en) 1994-12-26 1995-12-22 Novel aromatic yeast strains

Country Status (4)

Country Link
JP (1) JP3544987B2 (en)
KR (1) KR987001032A (en)
AU (1) AU4314696A (en)
WO (1) WO1996020272A1 (en)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2002253211A (en) * 2001-02-28 2002-09-10 Gekkeikan Sake Co Ltd High-flavor component productive yeast
KR100448516B1 (en) * 2002-04-04 2004-09-13 주식회사 진로 Saccharomyces cerevisiae jy-209 and process for preparing alcoholic liquors using same
KR100733071B1 (en) * 2006-11-21 2007-06-28 주식회사 두산 Saccharomyces cerevisiae t084 increasing aromatic compounds in alcoholic liquor and method for preparing alcoholic liquor using the same
JP2009219381A (en) * 2008-03-13 2009-10-01 Kirin Holdings Co Ltd New wine yeast obtained by hybridizing shochu yeast with wine yeast and method for creating the same
JP6119881B1 (en) * 2016-01-07 2017-04-26 菊正宗酒造株式会社 Yeast having high ability to produce ethyl caproate and method for producing fermented product using the yeast
JP2017121251A (en) * 2017-03-10 2017-07-13 菊正宗酒造株式会社 Yeast with high production capacity of ethyl caproate, and method of producing fermentation product using that yeast

Citations (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS626669A (en) * 1985-07-03 1987-01-13 Ookura Syuzo Kk Method of cultivating variant yeast
JPS63309175A (en) * 1987-06-09 1988-12-16 Getsukeikan Kk Culture of yeast variant
JPH01257423A (en) * 1988-02-16 1989-10-13 Kyowa Hakko Kogyo Co Ltd Variant yeast and use thereof
JPH03108476A (en) * 1989-09-21 1991-05-08 Satsuma Syuzo Kk Yeast fused cell strain and alcoholic beverage
JPH0491782A (en) * 1990-08-03 1992-03-25 Kyowa Hakko Kogyo Co Ltd Preparation of food and drink
JPH05317034A (en) * 1992-02-27 1993-12-03 Takara Shuzo Co Ltd New strain of aromatic sake yeast
JPH06169748A (en) * 1992-09-30 1994-06-21 Hakutsuru Shuzo Kk Production of refined sake using yeast having low decomposition of ethyl caproate, method for breeding yeast having low decomposition of ethyl caproate and its yeast strain
JPH06169749A (en) * 1992-12-08 1994-06-21 Fukushima Pref Gov Method for culturing variant yeast
JPH06169747A (en) * 1992-09-30 1994-06-21 Hakutsuru Shuzo Kk Method for breeding yeast having low decomposition of isoamyl acetate, its yeast strain and production of refined sake using the same
JPH0751053A (en) * 1993-08-11 1995-02-28 Kumamoto Pref Gov Yeast for brewing shochu and method for producing shochu with the yeast
JPH0779766A (en) * 1993-09-17 1995-03-28 Takara Shuzo Co Ltd Novel yeast and its use
JPH07147972A (en) * 1993-11-29 1995-06-13 Asahi Chem Ind Co Ltd New saccharomyces cerevisiae and use thereof
JPH07203951A (en) * 1994-01-14 1995-08-08 Mercian Corp Yeast variant strain and production of liquors using the same

Patent Citations (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS626669A (en) * 1985-07-03 1987-01-13 Ookura Syuzo Kk Method of cultivating variant yeast
JPS63309175A (en) * 1987-06-09 1988-12-16 Getsukeikan Kk Culture of yeast variant
JPH01257423A (en) * 1988-02-16 1989-10-13 Kyowa Hakko Kogyo Co Ltd Variant yeast and use thereof
JPH03108476A (en) * 1989-09-21 1991-05-08 Satsuma Syuzo Kk Yeast fused cell strain and alcoholic beverage
JPH0491782A (en) * 1990-08-03 1992-03-25 Kyowa Hakko Kogyo Co Ltd Preparation of food and drink
JPH05317034A (en) * 1992-02-27 1993-12-03 Takara Shuzo Co Ltd New strain of aromatic sake yeast
JPH06169748A (en) * 1992-09-30 1994-06-21 Hakutsuru Shuzo Kk Production of refined sake using yeast having low decomposition of ethyl caproate, method for breeding yeast having low decomposition of ethyl caproate and its yeast strain
JPH06169747A (en) * 1992-09-30 1994-06-21 Hakutsuru Shuzo Kk Method for breeding yeast having low decomposition of isoamyl acetate, its yeast strain and production of refined sake using the same
JPH06169749A (en) * 1992-12-08 1994-06-21 Fukushima Pref Gov Method for culturing variant yeast
JPH0751053A (en) * 1993-08-11 1995-02-28 Kumamoto Pref Gov Yeast for brewing shochu and method for producing shochu with the yeast
JPH0779766A (en) * 1993-09-17 1995-03-28 Takara Shuzo Co Ltd Novel yeast and its use
JPH07147972A (en) * 1993-11-29 1995-06-13 Asahi Chem Ind Co Ltd New saccharomyces cerevisiae and use thereof
JPH07203951A (en) * 1994-01-14 1995-08-08 Mercian Corp Yeast variant strain and production of liquors using the same

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
J. FERMENT. BIOENG., (1992), Vol. 73, No. 6, HIRATA et al., "Breeding of a Useful Strain of Sake Yeast for Ginjo-Sake Brewing", p. 497-499. *
J. FERMENT. BIOENG., (1994), Vol. 77, No. 4, KISHIMOTO, "Fermentation Chatacteristics of Hybrids Between the Cryophilic Wine Yeast Saccharomyces Bayanus and the Mesophilic Wine Yeast Saccharomyces Cerevisiae", p. 432-435. *
J. FERMENT. BIOENG., (1994), Vol. 77, No. 4, SHINOHARA et al., "Selection and Hybridization of Wine Yeasts for Improved Winemaking Properties: Fermentation Rate and Aroma Productivity", p. 428-431. *
JOURNAL OF FERMENTATION SOCIETY, (1989), Vol. 67, No. 3, YANAGIUCHI, KIYOWAKA, WAKAI, "Separation of Sake Yeast with a High Accumulation Power of Isoamyl Acetate", p. 159-165. *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2002253211A (en) * 2001-02-28 2002-09-10 Gekkeikan Sake Co Ltd High-flavor component productive yeast
KR100448516B1 (en) * 2002-04-04 2004-09-13 주식회사 진로 Saccharomyces cerevisiae jy-209 and process for preparing alcoholic liquors using same
KR100733071B1 (en) * 2006-11-21 2007-06-28 주식회사 두산 Saccharomyces cerevisiae t084 increasing aromatic compounds in alcoholic liquor and method for preparing alcoholic liquor using the same
JP2009219381A (en) * 2008-03-13 2009-10-01 Kirin Holdings Co Ltd New wine yeast obtained by hybridizing shochu yeast with wine yeast and method for creating the same
JP6119881B1 (en) * 2016-01-07 2017-04-26 菊正宗酒造株式会社 Yeast having high ability to produce ethyl caproate and method for producing fermented product using the yeast
JP2017121204A (en) * 2016-01-07 2017-07-13 菊正宗酒造株式会社 Yeast with high production capacity of ethyl caproate, and method of producing fermentation product using that yeast
JP2017121251A (en) * 2017-03-10 2017-07-13 菊正宗酒造株式会社 Yeast with high production capacity of ethyl caproate, and method of producing fermentation product using that yeast

Also Published As

Publication number Publication date
JP3544987B2 (en) 2004-07-21
KR987001032A (en) 1998-04-30
AU4314696A (en) 1996-07-19

Similar Documents

Publication Publication Date Title
CN101962603B (en) Process for brewing high-degree aromatic grape spirit
JP6400690B2 (en) Method for preparing fermented beverages and beverages thus produced
Bhat et al. An overview on the biological production of vinegar
CN108504480B (en) Production method of beer distilled liquor and beer distilled liquor obtained by production method
CN108753530B (en) Production method of hop whisky and obtained hop whisky
CN102994362A (en) Method for fermenting apple vinegar
Lentz et al. Genetic and physiological characterization of yeast isolated from ripe fruit and analysis of fermentation and brewing potential
CN101889073A (en) Method of producing distilled spirit
CN110564563B (en) Low-alcohol fermentation type aerated rice wine and brewing method thereof
CN101748025A (en) Compound lemon wine and preparation method thereof
JP2007037436A (en) Method for producing liquor
CN114540138A (en) Method for producing novel fruity acid beer by double-yeast mixed fermentation of wort
WO1996020272A1 (en) Novel aromatic yeast strains
US20090238919A1 (en) Method of manufacturing vinegar with high extract content
WO2017214673A1 (en) A yeast strain and uses thereof
JP5965795B2 (en) Distilled liquor with a high content of 4-vinyl guaiacol and method for producing the same, distilled liquor obtained using the distilled liquor with a high content of vanillin
JPS63309175A (en) Culture of yeast variant
CN108823015B (en) Production method of wheat-flavor distilled liquor and wheat-flavor distilled liquor obtained by production method
JP2003116523A (en) Yeast isolated from cherry blossom, method for obtaining the same, and method for producing sake or other food and beverage using the same
CN1145949A (en) Second fermentation process of naturally fermented condensed wine
JP5965796B2 (en) Distilled liquor with a high content of 4-vinyl guaiacol and method for producing the same, distilled liquor obtained using the distilled liquor with a high content of vanillin
JPH06133703A (en) Production of food or drink
JP3090786B2 (en) Vinegar production method
JP3260919B2 (en) Food and beverage manufacturing method
JP3932321B2 (en) Aroma-producing yeast, food and drink using the same, and method for producing the same

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU CN JP KR US

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
ENP Entry into the national phase

Ref country code: US

Ref document number: 1997 849552

Date of ref document: 19970602

Kind code of ref document: A

Format of ref document f/p: F

WWE Wipo information: entry into national phase

Ref document number: 1019970704352

Country of ref document: KR

WWP Wipo information: published in national office

Ref document number: 1019970704352

Country of ref document: KR

WWW Wipo information: withdrawn in national office

Ref document number: 1019970704352

Country of ref document: KR