JP6582275B2 - Caproic acid low-producing yeast - Google Patents
Caproic acid low-producing yeast Download PDFInfo
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本発明は、カプロン酸生成の少ない酵母に関する。さらに詳しくは、カプロン酸エチル生成能は高いがカプロン酸生成能は低く且つアルコール耐性を持つ酵母、及びその酵母を用いた酒類の製造方法に関する。 The present invention relates to a yeast with low caproic acid production. More specifically, the present invention relates to a yeast that has a high ability to produce ethyl caproate but has a low ability to produce caproic acid and has alcohol resistance, and a method for producing alcoholic beverages using the yeast.
近年、醸造酒の多様化が進み、様々な商品の開発が進んでいる。例えば、吟醸香の高い清酒の開発である。清酒においては、代表的な吟醸香として果物様の香りに形容されるカプロン酸エチルや酢酸イソアミル等が知られている。以前はこれらの吟醸香を生成させるには「吟醸造り」という高精白（精米歩合６０％以下）の原料米を用いて低温発酵させる方法が取られていた。しかしこのような方法では安定した吟醸香を生成させることは困難であった。このため「吟醸造り」に取って変わる方法として、吟醸香を高生成する酵母を選抜・分離して清酒を製造する方法が試みられている。 In recent years, diversification of brewed sake has progressed, and various products have been developed. For example, the development of sake with a high ginjo aroma. In refined sake, ethyl caproate, isoamyl acetate, and the like, which are described as fruit-like fragrances, are known as typical ginjo aromas. In the past, to produce these ginjo incense, a method of low-temperature fermentation using raw rice of “Ginjo-zukuri”, which is highly refined (milled rice ratio 60% or less), was used. However, it was difficult to produce a stable ginjo aroma by such a method. For this reason, as a method that replaces “Ginjo-zukuri”, a method of producing sake by selecting and separating yeasts that produce ginjo aroma highly has been attempted.
代表的な吟醸香であるカプロン酸エチルの高生成酵母に関しては、以下の方法が開発され、既に実用化が成されている。ひとつは、カプロン酸エチルの材料となるカプロン酸の合成に関わる脂肪酸合成酵素について、その阻害剤セルレニンの耐性を指標として、耐性が高いものを選抜・分離する方法である（特許文献１）。また、カプロン酸には酵母の生育を阻止する作用があり一定濃度のカプロン酸を含む培地ではカプロン酸高生産酵母は増殖できなくなるが、このカプロン酸感受性を指標として感受性が高い株を選抜・分離する方法である（特許文献２及び３）。さらに、生成されたカプロン酸エチルは、同成分に対して特異性の高いエステラーゼにより分解されることから、このエステラーゼ活性を低下させた酵母の分離が試みられている。具体的には、分解により毒性のあるフルオロエタノールが生じるカプロン酸２−フルオロエチルに対する耐性を指標にして、耐性の高い酵母の取得が試みられている（特許文献４）。
これらの方法で、実際にカプロン酸エチル高生成酵母が分離・選抜され、その一部は清酒製造に使用されている。
The following method has been developed and put into practical use for a high-producing yeast of ethyl caproate, which is a typical ginjo aroma. One is a method of selecting and separating a fatty acid synthase involved in the synthesis of caproic acid, which is a material of ethyl caproate, with high resistance using the resistance of its inhibitor cerulenin as an index (Patent Document 1). In addition, caproic acid has the effect of inhibiting the growth of yeast, and in a medium containing a certain concentration of caproic acid, yeast that produces high caproic acid cannot grow. (Patent Documents 2 and 3). Furthermore, since the produced ethyl caproate is decomposed by esterase having high specificity for the same component, attempts have been made to isolate yeast having reduced esterase activity. Specifically, attempts have been made to obtain highly resistant yeast using resistance to 2-fluoroethyl caproate, which produces toxic fluoroethanol by decomposition, as an index (Patent Document 4).
By these methods, ethyl caproate high-producing yeast is actually separated and selected, and a part thereof is used for sake production.
ところで、代表的な吟醸香とされるカプロン酸エチルは、リンゴ様の華やかな香りであるが、その材料となるカプロン酸はヤギの体臭様に例えられる不快臭であり、清酒の官能評価においては脂肪酸臭として敬遠される。この代表的な吟醸香であるカプロン酸エチルを酵母が生成するには２つの経路があり、一つはアルコールアシルトランスフェラーゼによりカプロイルＣｏＡとエタノールから生成される経路、もう一つはエステラーゼによりカプロン酸とエタノールから生成される経路である。両経路から生成されるカプロン酸エチルは、カプロイルＣｏＡとカプロン酸の濃度が、それぞれ律速となっていることが明らかにされている（非特許文献２）。 By the way, ethyl caproate, which is a typical ginjo aroma, has a gorgeous apple-like scent, but caproic acid, which is the material, has an unpleasant odor similar to the body odor of goats. It is shunned as a fatty acid odor. There are two pathways for yeast to produce this typical ginjo aroma, ethyl caproate, one is produced from caproyl CoA and ethanol by alcohol acyltransferase, and the other is produced by esterase and caproic acid. It is a pathway generated from ethanol. It has been clarified that the caproyl CoA and caproic acid concentrations of ethyl caproate produced from both pathways are rate limiting (Non-patent Document 2).
カプロン酸エチルの生成反応は、カプロイルＣｏＡやカプロン酸の濃度が律速なので、カプロン酸エチルを多く生成する酵母はカプロイルＣｏＡやカプロン酸も多く生成すると考えられ、カプロン酸エチル高生成酵母により醸造された製成酒は、概して不快臭とされるカプロン酸の含有量も多いという問題がある。実際、清酒中において、カプロン酸濃度とカプロン酸エチル濃度には強い正の相関関係が認められ、清酒中のカプロン酸エチル濃度はカプロン酸濃度に依存することが報告されている（非特許文献１）。
このカプロン酸エチルの材料であるカプロン酸の生成能が高いことや、カプロン酸エチル分解能が低いことに着目してカプロン酸エチル高生成酵母の分離を行った例は存在する（特許文献２、３及び４）。しかし、カプロン酸エチル低分解性と考えられるカプロン酸２−フルオロエチル耐性を示し、尚且つカプロン酸の生成量が少ない酵母の取得及びこれを利用した清酒の製造方法は知られていない。
Since the caproyl CoA and caproic acid concentrations are rate-limiting in the production reaction of ethyl caproate, yeast that produces a large amount of ethyl caproate is thought to produce a large amount of caproyl CoA and caproic acid. Seisei has a problem that it contains a large amount of caproic acid, which is generally regarded as an unpleasant odor. In fact, in sake, a strong positive correlation was observed between caproic acid concentration and ethyl caproate concentration, and it was reported that the ethyl caproate concentration in sake depends on the caproic acid concentration (Non-patent Document 1). ).
There are examples in which ethyl caproate high-producing yeasts were separated by paying attention to the high ability to produce caproic acid, which is a material of this ethyl caproate, and low ability of ethyl caproate (Patent Documents 2 and 3). And 4). However, it has not been known how to obtain yeast that exhibits resistance to 2-fluoroethyl caproate, which is considered to have a low degradability of ethyl caproate, and that produces a small amount of caproic acid, and a method for producing sake using this.
本発明の目的は、カプロン酸エチルの生成能は高いが、カプロン酸の生成能は低い酵母を提供することである。本発明の他の目的は、醸造酒の香味成分の変化をできるかぎり少なくすべく、上記酵母のアルコール耐性株を取得することである。本発明のさらに他の目的は当該酵母を使用してカプロン酸エチルの濃度は高いが、カプロン酸濃度は低い、香気成分豊かで不快臭の少ない清酒を提供することである。 An object of the present invention is to provide a yeast having a high ability to produce ethyl caproate but a low ability to produce caproic acid. Another object of the present invention is to obtain an alcohol-tolerant strain of the yeast so as to minimize the change in flavor components of brewed sake. Still another object of the present invention is to provide sake using the yeast, which has a high concentration of ethyl caproate but a low concentration of caproic acid, which is rich in aroma components and has little unpleasant odor.
本発明者は、上記目的を達成するために種々検討の結果、出願人が所有するカプロン酸エチル高生成酵母株を親株として変異を加え、この中から、カプロン酸エチル分解活性とカプロン酸濃度に着目して、酵母を選抜・分離した。さらに、この酵母のアルコール耐性株を取得した。加えて、この酵母を用いて清酒を製造することで本発明に到達した。すなわち本発明は以下のとおりである。 As a result of various studies to achieve the above object, the present inventor added a mutation as a parent strain to an ethyl caproate high-producing yeast strain owned by the applicant, and from these, the caproate ethyl degradation activity and caproic acid concentration were changed. Paying attention, yeast was selected and separated. Furthermore, an alcohol-resistant strain of this yeast was obtained. In addition, the present invention has been achieved by producing sake using this yeast. That is, the present invention is as follows.
１．受託番号がＮＩＴＥ Ｐ―０２１３９である酵母Ｂ２−１−２株
２．前記１の酵母で発酵する工程を含む加工食品の製造方法
1. Yeast B2-1-2 strain whose accession number is NITE P-021392. Process for producing processed food comprising the step of fermenting with said yeast of 1
カプロン酸エチル高生成酵母は、その材料であり不快臭とされるカプロン酸も多く生成すると考えられる。しかし、本発明の酵母は、突然変異を起こさせる前の親株の酵母と比較しカプロン酸エチルの高生成能は維持しながら、カプロン酸生成が少ない。すなわち、通常、清酒中において、カプロン酸濃度とカプロン酸エチル濃度には強い正の相関関係が認められるが（非特許文献１）、本発明の酵母を使用すれば、カプロン酸エチルの高濃度を維持したまま、カプロン酸濃度は低い清酒を製造することができる。アルコール耐性を有することともあわせて、香気成分豊かで不快臭の少ない清酒を提供できる。 It is considered that ethyl caproate high-producing yeast produces a large amount of caproic acid, which is a material and has an unpleasant odor. However, the yeast of the present invention produces less caproic acid while maintaining the high ability to produce ethyl caproate compared to the parental yeast before the mutation. That is, usually, in sake, a strong positive correlation is found between caproic acid concentration and ethyl caproate concentration (Non-patent Document 1). However, if the yeast of the present invention is used, a high concentration of ethyl caproate is increased. While maintaining this, sake with a low caproic acid concentration can be produced. Along with having alcohol resistance, it is possible to provide sake with abundant aroma components and less unpleasant odor.
以下に本発明を詳細に説明する。
本発明の酵母は、カプロン酸エチルの生成能は高いが、カプロン酸生成能は低い。この酵母を使用して、常法により清酒の仕込試験を行うと、代表的な吟醸香であるカプロン酸エチル濃度が４．０ｐｐｍ以上、不快臭である遊離カプロン酸が２５．４ｐｐｍ以下の製成酒を得ることができる。また、本発明の酵母はアルコール耐性を持つので、酵母の自己消化による香気成分の変化が少ない。これらのことから、本酵母を用いれば香気成分の豊かな製成酒を製造することができる。
The present invention is described in detail below.
The yeast of the present invention has a high ability to produce ethyl caproate, but has a low ability to produce caproic acid. Using this yeast, when a sake brewing test was conducted by a conventional method, a typical ginjo incense concentration of ethyl caproate of 4.0 ppm or higher and an unpleasant odor of free caproic acid of 25.4 ppm or lower was produced. You can get sake. Moreover, since the yeast of this invention has alcohol tolerance, the change of the aroma component by yeast self-digestion is few. From these things, if this yeast is used, a refined liquor rich in aroma components can be produced.
さらに、本発明の酵母は、発酵速度にも優れており、変異処理や薬剤耐性処理を行ったにもかかわらず、親株と同等の発酵速度を有している。例えば、総米５００ｇの清酒製造試験を行えば、醪日数２７日目に親株と同等のアルコール度数で搾ることが可能である。 Furthermore, the yeast of this invention is excellent also in the fermentation rate, and has the fermentation rate equivalent to a parent strain | stump | stock despite having performed the mutation process and the chemical | medical agent resistance process. For example, if a sake production test of 500 g of total rice is performed, it can be squeezed with the same alcohol content as the parent strain on the 27th day of the roast.
本発明の酵母は、２倍体から直接分離した２倍体の酵母である。この２倍体の酵母は、１倍体や１倍体を接合した２倍体の酵母と違い、劣性遺伝の形質が発現する頻度が低いので、取得するのが難しい。しかし変異処理の強弱を工夫することにより、例えばＵＶの波長や照射時間、エチルメタンスルフォネート濃度を調整したりすることで、取得することができる。また、この酵母で製造した生成酒は、変異処理や薬剤耐性処理を行ったにもかかわらず、１倍体や接合された２倍体では実現できないような、親株とほぼ同等の発酵力を有し、清酒に必要なアルコール、酸、アミノ酸、香気成分等もほぼ親株と同等の生産量である。
本発明の酵母を用いて生産する加工食品は特に制限されないが、例えば、醸造酒、蒸留酒、それらを混合した酒、味噌、醤油、パンが挙げられる。醸造酒、蒸留酒、それらを混合した酒には、果実や穀物を原料としたものが含まれる。さらに、これらの酒類には、日本酒、ビール、発泡酒、ワイン、老酒、リンゴ酒等が含まれる。
The yeast of the present invention is a diploid yeast that is directly separated from the diploid. This diploid yeast is difficult to obtain because it has a low frequency of recessive inheritance, unlike a haploid or a diploid yeast conjugated with a haploid. However, it can be obtained by devising the strength of the mutation treatment, for example, by adjusting the wavelength of UV, the irradiation time, and the concentration of ethyl methanesulfonate. In addition, the sake produced with this yeast has almost the same fermentative power as the parent strain, which cannot be realized with haploids or conjugated diploids, despite mutation treatment and drug resistance treatment. And the alcohol, acids, amino acids, fragrance components, etc. necessary for sake are almost the same as the parent strain.
The processed food produced using the yeast of the present invention is not particularly limited, and examples thereof include brewed liquor, distilled liquor, liquor mixed with them, miso, soy sauce, and bread. Brewed liquor, distilled liquor, and liquor mixed with them include those made from fruits and grains. Furthermore, these liquors include sake, beer, sparkling wine, wine, old sake, apple wine and the like.
本発明の酵母は以下のようにして取得することができる。
カプロン酸エチルを多く生成する株を親株とし、突然変異を起こさせる。親株は、系列が異なるものを複数選ぶのが好ましい。突然変異させる方法は、特に制限されるわけではないが、例えば、エチルメタンスルフォネート処理したり、ＵＶ処理することが挙げられる。特許文献４の方法に従って、突然変異させた酵母から、カプロン酸エチル高生成株を選抜・分離する。すなわち、得られた突然変異株を、親株が生育できない濃度のカプロン酸２−フルオロエチルを含む培地で培養し、生育できたものを分離する。
The yeast of the present invention can be obtained as follows.
A strain that produces a large amount of ethyl caproate is used as a parent strain to cause mutation. It is preferable to select a plurality of parent strains having different series. The method of mutating is not particularly limited, and examples thereof include ethyl methanesulfonate treatment and UV treatment. According to the method of Patent Document 4, an ethyl caproate high-producing strain is selected and isolated from the mutated yeast. That is, the obtained mutant strain is cultured in a medium containing 2-fluoroethyl caproate at a concentration at which the parent strain cannot grow, and the grown strain is separated.
分離した酵母についてそれぞれ、発酵試験を行う。例えば、この酵母を麹エキス培地で常法により培養する。得られた上清のカプロン酸濃度、一般成分（日本酒度、アルコール、酸度、アミノ酸度）、及び香気成分を分析し、これらを総合的に判断して、酵母を選抜する。 A fermentation test is performed for each separated yeast. For example, this yeast is cultured by a conventional method in a koji extract medium. The obtained supernatant is analyzed for caproic acid concentration, general components (sake degree, alcohol, acidity, amino acid degree), and aroma components, and these are comprehensively judged to select yeast.
選抜した酵母にエタノール溶液を加えて培養し、得られた株をアルコール耐性株とする。このアルコール耐性株の発酵試験を行い、二酸化炭素の減少量から仮定した発酵速度、カプロン酸濃度、一般成分（日本酒度、アルコール、酸度、アミノ酸度）及び香気成分の分析結果から総合的に判断して酵母をさらに選抜する。 The selected yeast is added with an ethanol solution and cultured, and the resulting strain is designated as an alcohol-resistant strain. This alcohol-resistant strain is fermented and comprehensively judged from the analysis results of fermentation rate, caproic acid concentration, general components (sake degree, alcohol, acidity, amino acid degree) and aroma components assumed from the amount of carbon dioxide reduction. Select yeast further.
この選抜された酵母について実際に小スケールの仕込試験を行う。仕込試験は、常法により行う。仕込試験により得られた生成酒のカプロン酸濃度、一般成分（日本酒度、アルコール、酸度、アミノ酸度）及び香気成分を分析し、さらにはきき酒による官能評価を行い、最終的に酵母を選抜する。 The selected yeast is actually subjected to a small-scale preparation test. The preparation test is conducted by a conventional method. The caproic acid concentration, general components (sake degree, alcohol, acidity, amino acid degree) and aroma components of the produced liquor obtained by the preparation test are analyzed, and further sensory evaluation is performed with sake, and finally yeast is selected.
この酵母で発酵することで、香気成分豊かで不快臭の少ない加工食品を生産することができる。 By fermenting with this yeast, it is possible to produce processed foods that are rich in aroma components and have less unpleasant odor.
以下に、本発明を実施例で説明する。 Hereinafter, the present invention will be described with reference to examples.
実施例１
［カプロン酸エチルを多く生成し且つカプロン酸生成が少ないアルコール耐性酵母の選抜］
（１）カプロン酸２−フルオロエチル耐性株の分離
出願人がこれまでに育種分離したカプロン酸エチル高生成株を親株として用いた。各２ｍｌの麹エキス培地（ボーメ６．５）で一晩培養後の親株（３系列作成）を集菌、洗浄した。これらの親株をそれぞれ１．エチルメタンスルフォネート処理、２．ＵＶ処理、３．変異処理なし、の操作に供した。次いで、親株が生育できない濃度（１０００ｐｐｍ）のカプロン酸２−フルオロエチルを含むＹＮＢ培地（０．６７重量％イーストナイトロジェンベース、２重量％寒天、５重量％エタノール）に塗抹した後、３０℃で５日間培養し、生育したコロニー（２５０株以上）を分離して、カプロン酸２−フルオロエチル耐性株とした。
Example 1
[Selection of alcohol-resistant yeast that produces a large amount of ethyl caproate and a small amount of caproic acid]
(1) Isolation of 2-fluoroethyl caproate-resistant strain The high caproate-producing strain that the applicant has so far bred and isolated was used as a parent strain. The parent strain (prepared in 3 series) after overnight culture in 2 ml of each koji extract medium (Baume 6.5) was collected and washed. Each of these parent strains is 1. 1. Ethyl methane sulfonate treatment 2. UV treatment; It was subjected to an operation without mutation treatment. Then, after smearing on a YNB medium (0.67 wt% yeast nitrogen base, 2 wt% agar, 5 wt% ethanol) containing 2-fluoroethyl caproate at a concentration (1000 ppm) at which the parent strain cannot grow, After culturing for 5 days, the grown colonies (250 strains or more) were isolated and made into 2-fluoroethyl caproate resistant strains.
（２）カプロン酸生成に着目した発酵試験
上記のカプロン酸２−フルオロエチル耐性株を用いて発酵試験を行った。発酵試験は以下のように実施した。すなわち、前述の酵母を麹エキス培地に植菌した後、３０℃で３日間静置培養した。次に８ｇのアルコール脱水麹と２０ｍｌの麹エキスを混合したものに上記の培養液をそれぞれ１ｍｌずつ添加し、１３℃で１４日間静置した。発酵試験の終了後、遠心分離（０℃、５０００ｒｐｍ、１０分間）を行い、得られた上清のカプロン酸濃度、一般成分（日本酒度、アルコール、酸度、アミノ酸度）および香気成分を分析した。親株と比較してカプロン酸エチル濃度を維持しつつカプロン酸濃度が低下しているものを中心に総合的に判断して酵母を選抜した。
(2) Fermentation test focusing on caproic acid production Fermentation test was performed using the above-mentioned 2-fluoroethyl-resistant caproic acid strain. The fermentation test was performed as follows. That is, after inoculating the yeast described above in a koji extract medium, the yeast was statically cultured at 30 ° C. for 3 days. Next, 1 ml of the above culture solution was added to a mixture of 8 g of alcohol-dehydrated koji and 20 ml of koji extract, and the mixture was allowed to stand at 13 ° C. for 14 days. After completion of the fermentation test, centrifugation (0 ° C., 5000 rpm, 10 minutes) was performed, and the obtained supernatant was analyzed for caproic acid concentration, general components (sake degree, alcohol, acidity, amino acid degree) and aroma components. Yeasts were selected based on a comprehensive judgment centered on the decrease in caproic acid concentration while maintaining the ethyl caproate concentration compared to the parent strain.
（３）アルコール耐性株の分離
上記の方法で選抜した酵母からエタノール耐性株を分離した。すなわち、前述の酵母を麹エキス培地に植菌した後、３０℃で３日間静置培養した。培養後に遠心分離（０℃、５０００ｒｐｍ、１０分間）して、得られた菌体にエタノール溶液（２０重量％エタノール、１重量％グルコース、０．１Ｍ酢酸緩衝液、ｐＨ４．２）を加え、軽く懸濁した後に１５℃で７日間培養した。次いで、培養液を滅菌水にて適宜希釈した後、ＹＰＤ培地（１重量％イーストエキス、２重量％ポリペプトン、２重量％グルコース、２重量％寒天）に塗抹して、３０℃で３日間静置培養した。この操作を２回繰り返して合計３回行い、得られた株をアルコール耐性株とした。
(3) Isolation of alcohol-resistant strain An ethanol-resistant strain was isolated from the yeast selected by the above method. That is, after inoculating the yeast described above in a koji extract medium, the yeast was statically cultured at 30 ° C. for 3 days. After incubation, the mixture was centrifuged (0 ° C., 5000 rpm, 10 minutes), and an ethanol solution (20 wt% ethanol, 1 wt% glucose, 0.1 M acetate buffer, pH 4.2) was added to the obtained cells, and lightly added. After suspension, the cells were cultured at 15 ° C. for 7 days. Next, after appropriately diluting the culture solution with sterilized water, it is smeared on YPD medium (1 wt% yeast extract, 2 wt% polypeptone, 2 wt% glucose, 2 wt% agar) and allowed to stand at 30 ° C for 3 days. Cultured. This operation was repeated twice for a total of 3 times, and the resulting strain was designated as an alcohol resistant strain.
（４）発酵速度に着目した発酵試験
上記の酵母を用いて発酵試験を行った。試験後、遠心分離（０℃、５０００ｒｐｍ、１０分間）を行い、得られた上清のカプロン酸濃度、一般成分（日本酒度、アルコール、酸度、アミノ酸度）および香気成分を分析した。発酵による二酸化炭素の減少量を発酵速度と仮定して、試験開始から容器を含めた重量測定を毎日実施した。各種分析および発酵速度の結果から総合的に判断して１８株を下記の総米５００ｇの仕込試験に供した。
(4) Fermentation test paying attention to fermentation rate Fermentation test was done using said yeast. After the test, centrifugation (0 ° C., 5000 rpm, 10 minutes) was performed, and the obtained supernatant was analyzed for caproic acid concentration, general components (sake degree, alcohol, acidity, amino acid degree) and aroma components. Assuming that the decrease in carbon dioxide due to fermentation was the fermentation rate, gravimetric measurement including the container was performed daily from the start of the test. Judging from the results of various analyzes and fermentation rates, 18 strains were subjected to the following 500 g total rice preparation test.
実施例２
［カプロン酸エチルを多く生成し且つカプロン酸生成が少ないアルコール耐性酵母を用いた仕込試験］
掛米および麹米に精米歩合４０％の秋田酒こまち、麹菌は出願人が開発したＮ５４Ｇを使用して、３段仕込みにて総米５００ｇの仕込試験を行った。仕込配合を表１に示す。品温は、添仕込１３℃、仲仕込９℃、留仕込６℃として、最高温度（１０℃）に達するまで１日あたり０．７〜０．８℃ずつ上昇させた。もろみの管理は主としてＢＭＤ値（留め後日数×ボーメ度）にて行い、定期的にサンプリングを実施した。最終的に遠心分離（０℃、３０００ｒｐｍ、５０分間）により上槽した。得られた製成酒の一般、香気成分および遊離カプロン酸濃度を分析し、きき酒による官能評価を行った（それぞれの結果を表２〜４に示す）。なお、官能評価は４人の醸造試験場職員が５点法により実施した（親株を３．０とした相対評価、点数が低いほど評価が高い）。
Example 2
[Preparation test using alcohol-resistant yeast that produces a large amount of ethyl caproate and a small amount of caproic acid]
A rice cake ratio of 40% for Akita Sake Komachi, and Koji molds were tested in a three-stage process for 500g of rice in total using a rice milling ratio of 40%. The charging composition is shown in Table 1. The product temperature was increased by 0.7 to 0.8 ° C. per day until the maximum temperature (10 ° C.) was reached, with an additive charge of 13 ° C., an intermediate charge of 9 ° C., and a distillate charge of 6 ° C. Moromi was managed mainly by BMD value (days after retention x Baume degree) and periodically sampled. Finally, the upper tank was obtained by centrifugation (0 ° C., 3000 rpm, 50 minutes). General, aroma components, and free caproic acid concentrations of the resulting sake were analyzed, and sensory evaluation was performed using suki-shu (each result is shown in Tables 2 to 4). In addition, sensory evaluation was carried out by four brewing laboratory staff by the five-point method (relative evaluation with the parent stock being 3.0, the lower the score, the higher the evaluation).
以上の方法を用いて得られた結果から、総合的に判断して、酵母を最終的に選抜し、親株と同等のカプロン酸エチルを生成し、尚且つカプロン酸の生成量が少ないアルコール耐性株Ｂ２−１−２株を取得した。Ｂ２−１−２株は親株と比較してカプロン酸エチルの生成量が同等であったが、カプロン酸生成量が約５３％減少していた（表３、図１）。 From the results obtained by using the above method, a comprehensive judgment is made, and the yeast is finally selected to produce an ethyl caproate equivalent to the parent strain, and the alcohol-resistant strain with a small amount of caproic acid produced. B2-1-2 strain was acquired. Although the amount of ethyl caproate produced in the B2-1-2 strain was equivalent to that of the parent strain, the amount of caproic acid produced was reduced by about 53% (Table 3, FIG. 1).
なお、このＢ２−１−２株は、「ＵＴ−１」として、独立行政法人製品評価技術基盤機構 特許微生物寄託センターに寄託されている。
ＵＴ−１：受託番号ＮＩＴＥ Ｐ―０２１３９
This B2-1-2 strain has been deposited as “UT-1” at the Patent Microorganism Deposit Center, National Institute of Technology and Evaluation.
UT-1: Accession number NITE P-02139
本発明の酵母は、主要な吟醸香の一つであるカプロン酸エチルは多く生成するが、不快臭とされているカプロン酸生成が少ない。さらに、醸造酒の香味成分の変化を出来る限り少なくできるアルコール耐性を有する。これらより、本発明の酵母は加工食品、特に清酒の製造に有用である。
The yeast of the present invention produces a large amount of ethyl caproate, which is one of the main ginjo aromas, but produces less caproic acid, which is considered an unpleasant odor. Furthermore, it has an alcohol resistance capable of minimizing changes in flavor components of brewed sake. From these, the yeast of this invention is useful for manufacture of processed food, especially refined sake.
Claims (2)
A method for producing a processed food comprising the step of fermenting with the yeast of claim 1
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