JP2012055286A - New yeast and method for producing food fermented by the yeast - Google Patents
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サッカロマイセス・セレビシエ（Saccharomyces cerevisiae）酵母及びチーズホエー等の食品原料をその酵母で発酵処理した新規の食品の製造法に関するものである。 The present invention relates to a method for producing a novel food obtained by fermenting food materials such as Saccharomyces cerevisiae yeast and cheese whey with the yeast.
食品原料を発酵させる時（パン、酒類、乳製品、味噌、醤油等の製造）にフレーバーを産生する酵母はあるが、チーズホエー用としてフレーバー産生、糖の発酵性、継代培養時の安定性で、満足いく酵母がない。食品原料の１つとして、チーズホエーが十分利用されておらず、これを価値あるものに再生するべく、これが持つ匂いの改善に、先ず取組んだ。
チーズホエーとは哺乳類の乳を酸（乳酸発酵を含む）または凝乳酵素等で主にたんぱく質を凝固させ、その固形物（カード）を除去して得られる液体部分である。通常はチーズ製造の際の副産物として大量に排出される。
There are yeasts that produce flavor when fermenting food ingredients (production of bread, liquor, dairy products, miso, soy sauce, etc.), but for cheese whey, flavor production, sugar fermentability, and stability during subculture There is no satisfactory yeast. As one of the raw materials for food, cheese whey is not fully utilized, so we first tried to improve the odor of this product in order to regenerate it into something valuable.
Cheese whey is a liquid part obtained by coagulating protein in mammal milk mainly with acid (including lactic acid fermentation) or curdling enzyme, and removing the solid matter (curd). It is usually discharged in large quantities as a by-product during cheese manufacture.
このようなホエーは独特の不快臭があり、飲用として適さない。そこで、微生物発酵を用いた改善法として次のような方法が知られている。
（１）下記特許文献１に示されるように、乳性原料を酸素含有ガスの強制供給下でケフィア菌により発酵させ、たんぱく質等の沈殿物を除去し、次いでフィルター除菌または加熱殺菌する乳酸菌飲料の製造法。
（２）特許文献２に示されるように、ホエーを乳酸発酵及び／又は酸を添加し、糖類の濃度が２０〜３５％重量％となるように添加し、かつタンニンを添加し、ｐＨ２．８〜４．５に調製した溶液を酵母発酵してなる酸性乳清発酵製品。
（３）特許文献３に示されるように、ホエーを酵母発酵させることにより良好な風味を有する発酵風味付与剤の製造法。
Such whey has a unique unpleasant odor and is not suitable for drinking. Therefore, the following method is known as an improved method using microbial fermentation.
(1) As shown in Patent Document 1 below, a lactic acid bacteria beverage that ferments a milky raw material with kefir bacteria under forced supply of oxygen-containing gas, removes precipitates such as proteins, and then filters or sterilizes by heating Manufacturing method.
(2) As shown in Patent Document 2, whey is added with lactic acid fermentation and / or acid so that the sugar concentration is 20 to 35% by weight, tannin is added, and pH is 2.8. An acidic whey fermentation product obtained by yeast fermentation of a solution prepared to ˜4.5.
(3) As shown in Patent Document 3, a method for producing a fermented flavor imparting agent having a good flavor by yeast fermentation of whey.
上記のようにホエーを乳酸菌及び／又は酵母による発酵により発酵させることで、発酵風味を付与する試みが行われている。しかし、ホエーが持つ独特の臭みの除去については（１）ではたんぱく質を沈殿させて除去することで行われており、ホエーが本来持つ栄養成分、乳酸菌体が失われている。また、新たな副産物が生じることになる。（２）は低ｐＨ、高糖濃度、タンニン添加の条件で酵母発酵によって生じる香気成分をコントロールしているが、発酵日数がかかり過ぎてチーズ工房での製造としては現実的ではない。また、高糖濃度であることから、そのまま飲料として用いるには適さない。（３）についても、酵母で発酵風味を増強してはいるが、そのままではホエーの不快臭除去に至らず、沈殿物除去工程を含めた発酵風味付与剤としての利用にとどまっている。
これらのように、ホエーをその含有成分を保持したまま、添加剤を加えたり特別な製造工程を加えずにストレート飲料として現実的に活用できる技術は見当たらない。
本発明ではチーズホエーに含まれる乳由来成分、菌体成分を除去することなしに最適な酵母発酵および加工条件を確立することで、チーズホエーが本来持つ不快臭の改善を行い、良好な風味のホエー乳酸菌飲料の製造法及びその製造法から製造される乳酸食品を完成させるに至った。
Attempts have been made to impart a fermentation flavor by fermenting whey by fermentation with lactic acid bacteria and / or yeast as described above. However, the unique odor removal of whey is carried out by precipitating and removing the protein in (1), and the whey's inherent nutrients and lactic acid bacteria are lost. In addition, a new by-product is generated. (2) controls aroma components produced by yeast fermentation under the conditions of low pH, high sugar concentration, and tannin addition, but it takes too many days of fermentation and is not practical for production in a cheese workshop. Further, since it has a high sugar concentration, it is not suitable for use as it is as a beverage. Regarding (3), the fermentation flavor is enhanced with yeast, but as it is, it does not lead to removal of unpleasant odor of whey, and it is only used as a fermentation flavor imparting agent including a precipitate removing step.
As described above, there is no technology that can be practically used as a straight beverage without adding an additive or adding a special manufacturing process while retaining its components.
In the present invention, it is possible to improve the unpleasant odor inherent in cheese whey by establishing optimal yeast fermentation and processing conditions without removing milk-derived components and cell components contained in cheese whey, and having a good flavor. It came to complete the manufacturing method of a whey lactic acid bacteria drink, and the lactic acid food manufactured from the manufacturing method.
牛乳、ヤギ乳、羊乳等哺乳類の乳のチーズ製造時に排出されるホエーは一部粉末化して利用されているが、近年、増加傾向にある小規模チーズ工房では一部の農家で家畜に飲ませている以外は利用されておらず、廃棄処理しているのが現状である。チーズホエーは乳のミネラルや乳酸菌による代謝物等、健康維持に有効とされる成分が残存しており、ヨーロッパでは飲料として活用されている。しかし、チーズホエーには独特のホエー臭があることから日本人には受け入れられてこなかった経緯がある。これらのことから、小規模チーズ工房における排水処理および環境への負荷を軽減させ、なおかつ、複雑な工程をとらずにホエー臭を除去可能な発酵食品の製造法の確立が課題であった。 Whey discharged during the production of cheese from mammals such as cow's milk, goat milk, and sheep milk is partly powdered and used. It is not used except for the current status, and is currently being disposed of. In cheese whey, components that are effective for maintaining health such as milk minerals and metabolites by lactic acid bacteria remain, and are used as beverages in Europe. However, because cheese whey has a unique whey odor, it has not been accepted by Japanese people. From these things, the establishment of the manufacturing method of the fermented food which can reduce the waste water treatment in a small-scale cheese factory and an environment, and can remove a whey odor without taking a complicated process was a subject.
本発明は数多くある酵母（Saccharomyces cerevisiae）から、香気成分が多く、ホエー臭をマスキングする酵母を鋭意選抜した結果、北海道十勝地域で分離された天然酵母（Saccharomyces cerevisiae）TENSAI株（FERM P-21891）に辿り着いた。このTENSAI株（FERM P-21891）は甜菜糖製造工程から分離されたものである。この酵母はチーズホエーの発酵において、適切な発酵および加熱処理をすることにより、良好な発酵風味を付与すると同時に不快臭を低減させる効果があることを見出したものである。 The present invention is a natural yeast (Saccharomyces cerevisiae) TENSAI strain (FERM P-21891) isolated in the Tokachi region of Hokkaido as a result of intensive selection of yeasts that have a large aroma component and mask whey odor from a number of yeasts (Saccharomyces cerevisiae). I arrived at. This TENSAI strain (FERM P-21891) was isolated from the beet sugar manufacturing process. The yeast has been found to have an effect of reducing unpleasant odor while imparting a good fermentation flavor by appropriate fermentation and heat treatment in cheese whey fermentation.
また、本発明は上記製造法で得られた、ホエーを主原料とするもので風味増強添加物に頼らない、酵母発酵風味が付与されたストレートタイプの発酵食品である。 Moreover, this invention is the straight type fermented food provided with the yeast fermentation flavor obtained by the said manufacturing method, using whey as the main raw material and not relying on a flavor enhancement additive.
すなわち、本発明は、下記のとおりである。
［１］サッカロマイセス・セレビシエ（Saccharomyces cerevisiae）TENSAI株（FERM P-21891）。
［２］チーズ製造で排出されるチーズホエーに糖類を添加し、直ちに１次殺菌後、サッカロマイセス・セレビシエ（Saccharomyces cerevisiae）TENSAI株（FERM P-21891）を用いて発酵させることを特徴とする、発酵食品の製造法。
［３］更に、２次殺菌後、糖類、果物風味等を添加した後、３次殺菌として、容器充填後、密閉容器中で殺菌することを特徴とする、［２］に記載の発酵食品の製造法。
That is, the present invention is as follows.
[1] Saccharomyces cerevisiae TENSAI strain (FERM P-21891).
[2] Fermentation characterized by adding saccharides to cheese whey discharged from cheese manufacture, immediately after first sterilization, and then fermenting with Saccharomyces cerevisiae TENSAI strain (FERM P-21891) Food manufacturing method.
[3] The fermented food according to [2], further comprising, after secondary sterilization, adding saccharides, fruit flavor, etc., and then sterilizing in a closed container after filling the container as the tertiary sterilization. Manufacturing method.
従来、小規模チーズ工房で排出されるチーズホエーは、食品としては利用されておらず、排水として捨てられていたのがほとんどである。このため有用成分の利用及び環境の面から問題であった。このホエーを食品として利用可能にしたものである。ホエーの持つ本来の成分を保持し、利用した酵母、乳酸菌の成分も含有しており、且つホエー臭を除去したものであり、小規模チーズ工房でも簡単に製造できる発酵食品である。更に、本発明の酵母は風味を付与した様々な発酵食品を提供することができる。 Conventionally, cheese whey discharged from small-scale cheese workshops has not been used as food, and is mostly discarded as wastewater. For this reason, it was a problem from the viewpoint of utilization of useful components and environment. This whey can be used as food. This is a fermented food that retains the original components of whey, contains yeast and lactic acid bacteria components, removes whey odor, and can be easily manufactured even in a small cheese factory. Furthermore, the yeast of the present invention can provide various fermented foods with a flavor.
本発明におけるチーズホエーの不快臭とは、チーズ製造工程の乳酸発酵の際、生成する独特の不快臭気が、チーズホエーに残存するものを指し、マスキングとは、本発明の酵母で発酵処理する際、発生するイソアミルアルコール、カプロン酸エチル等の香気成分を産生し、アセトインに代表されるチーズホエーの不快臭を隠すことをいう。 The unpleasant odor of cheese whey in the present invention refers to a unique unpleasant odor that is generated in cheese whey during the lactic acid fermentation in the cheese manufacturing process, and masking refers to when fermenting with the yeast of the present invention. This refers to producing aroma components such as isoamyl alcohol and ethyl caproate that are generated, and hiding the unpleasant odor of cheese whey represented by acetoin.
また、新規な天然酵母サッカロマイセス・セレビシエ（Saccharomyces cerevisiae）とは、甜菜糖製造工程より選抜した酵母であり、公知の手法により分離されたものであり、チーズホエーの不快臭をマスキングする効果のあるものを指す。 The new natural yeast Saccharomyces cerevisiae is a yeast selected from the beet sugar manufacturing process and has been isolated by known methods and has the effect of masking the unpleasant odor of cheese whey. Point to.
以下に、本発明者らが、本発明のサッカロマイセス・セレビシエに属する新規酵母、すなわち、TENSAI株（FERM P-21891）を見出すに至った経緯を説明する。 Below, the background of the present inventors finding a novel yeast belonging to the Saccharomyces cerevisiae of the present invention, that is, the TENSAI strain (FERM P-21891) will be described.
本発明の試験を行うにあたり、北海道十勝地域の野生酵母を採取し、最終的に継代培養時に安定して増殖可能な１２種類の酵母を単離した。継代培養はPDA培地（バレイショ浸出液２０％、ブドウ糖２％、カンテン１．５％）を用いた。 In conducting the test of the present invention, wild yeasts were collected from the Tokachi area of Hokkaido, and finally 12 types of yeasts that could be stably propagated during subculture were isolated. For subculture, PDA medium (potato exudate 20%, glucose 2%, agar 1.5%) was used.
チーズホエーを発酵するのに要求される項目はショ糖の発酵性が良いこと、乳糖を資化しないこと、耐酸性（ｐＨ３．０における生育）があること、良い香りの産生能があることが挙げられる。 Items required for fermenting cheese whey are good sucrose fermentability, not assimilating lactose, acid resistance (growth at pH 3.0), good scent production ability Can be mentioned.
これらの酵母について発酵性試験をYPD培地（酵母エキス１％、ペプトン２％、ブドウ糖２％）を基本とした改変培地で行った。すなわち、糖の発酵性試験についてはブドウ糖に代えてショ糖または乳糖を用いた。また、ｐＨを２、３、４、５、６にした場合の生育について調べた。 For these yeasts, fermentability tests were performed on a modified medium based on YPD medium (yeast extract 1%, peptone 2%, glucose 2%). That is, for the sugar fermentability test, sucrose or lactose was used instead of glucose. Further, the growth when the pH was adjusted to 2, 3, 4, 5, 6 was examined.
まず、各酵母をPDA培地で３０℃、２日間培養し、得られた菌体をリン酸緩衝生理食塩水でOD600nm：１になるように懸濁した。各酵母液１００μｌについて糖をブドウ糖、ショ糖、乳糖の３種類に分けて調製したYPD培地、ｐＨを２、３、４、５、６で調製したYPD培地各１０ｍｌに接種した。 First, each yeast was cultured in a PDA medium at 30 ° C. for 2 days, and the obtained bacterial cells were suspended in phosphate buffered saline so as to have an OD 600 nm of 1. 100 μl of each yeast solution was inoculated into 10 ml each of YPD medium prepared by dividing sugar into three types of glucose, sucrose, and lactose, and YPD medium prepared with pH 2, 3, 4, 5 and 6.
その結果、３０℃、２４時間培養でショ糖発酵性が良く、芳香性が優れている株として、本発明に用いるサッカロマイセス・セレビシエ（Saccharomyces cerevisiae）TENSAI株を選択した。同株は乳糖の資化性が無く、耐酸性（ｐＨ３．０で生育可）があることから、ホエー発酵飲料に用いる株として適していた。 As a result, the Saccharomyces cerevisiae TENSAI strain used in the present invention was selected as a strain having good sucrose fermentability and excellent aromaticity when cultured at 30 ° C. for 24 hours. This strain was suitable for use as a whey fermented beverage because it lacks lactose assimilation and has acid resistance (can grow at pH 3.0).
選択した株は26S rDNA-D1/D2 塩基配列が子嚢菌系酵母の一種であるSaccharomyces cerevisiae 基準株に対して100％の相同率を示し、Saccharomyces cerevisiaeに帰属することが確認された。 The selected strain showed 100% homology with the Saccharomyces cerevisiae reference strain, whose 26S rDNA-D1 / D2 nucleotide sequence was a kind of ascomycetous yeast, and was confirmed to belong to Saccharomyces cerevisiae.
ホエー液をベースに本発明に使用した酵母の優位性について調べた。比較は酵母発酵しない対照区、本発明で用いるSaccharomyces cerevisiae TENSAI株（FERM P-21891）、Saccharomyces cerevisiae 標準株（ATCC2601）、醸造協会の清酒用７０１号、焼酎１号（SH-4）、ワイン１号（OC-2）で行った。なお、上述の市販酵母はいずれもサッカロマイセス・セレビシエに属するものである。 Based on the whey solution, the superiority of the yeast used in the present invention was examined. For comparison, the control group without yeast fermentation, Saccharomyces cerevisiae TENSAI strain (FERM P-21891), Saccharomyces cerevisiae standard strain (ATCC2601) used in the present invention, sake 701, shochu 1 (SH-4), wine 1 (OC-2). All the above-mentioned commercially available yeasts belong to Saccharomyces cerevisiae.
本発明の殺菌処理は、１次及び３次は、常圧又は加圧の加熱殺菌が好ましい。発酵食品の香気成分を保持し、かつ、チーズホエーが持つ不快成分をマスキングする条件であればよい。そして、ホエー本来の成分及び発酵菌体の成分が変性することなく保持することが好ましい。２次殺菌は、本来の殺菌が目的と、アルコールを除去する目的がある。このため減圧又は開放常圧殺菌が適当である。 In the sterilization treatment of the present invention, primary and tertiary sterilization is preferably performed under normal pressure or pressurized heat. What is necessary is just the conditions which hold | maintain the aromatic component of fermented food, and mask the unpleasant component which cheese whey has. And it is preferable to hold | maintain the original component of whey and the component of a fermentation microbial cell, without denaturing. Secondary sterilization has the purpose of original sterilization and the purpose of removing alcohol. For this reason, decompression or open atmospheric sterilization is appropriate.
本発酵食品は、飲料、冷菓、食品添加物、食品原料及び食品等に利用される。ゼリーやアイスクリーム等の原料の成分にもなりうる。また、これらの発酵食品に必要に応じて添加剤を添加することができる。添加剤として、増粘剤、食物繊維、香料、ビタミン類等を添加してもよい。 The fermented food is used for beverages, frozen desserts, food additives, food raw materials, foods, and the like. It can also be a component of raw materials such as jelly and ice cream. Moreover, an additive can be added to these fermented foods as needed. As additives, thickeners, dietary fibers, fragrances, vitamins and the like may be added.
また、本発明の新規酵母は、上記にあげた発酵食品に限定されるものではない。酒類、パン、味噌、醤油等、酵母を使用する食品に使用することができる。 Further, the novel yeast of the present invention is not limited to the fermented foods listed above. It can be used for foods that use yeast, such as alcoholic beverages, bread, miso, and soy sauce.
以下に、本発明を実施例等により詳細に説明するが、本発明の技術的範囲が当該実施例により限定されるものではない。 Hereinafter, the present invention will be described in detail with reference to examples and the like, but the technical scope of the present invention is not limited to the examples.
［実施例１］原料のチーズホエーの製造法
チーズ工房で実際のチーズ製造工程で排出されたホエーの一例として下記に示す。
例）ラクレットチーズホエー
スターターとして乳酸菌Lactococcus 属使用
その成分は、
乳酸菌数：１０８／ｍｌ〜１０９／ｍｌレベル
たんぱく質：１％程度、脂質：１％程度、乳糖：５％程度
[Example 1] Production method of raw material cheese whey An example of whey discharged in an actual cheese production process in a cheese workshop is shown below.
Example) Raclette cheese whey Starter use genus Lactococcus Lactococcus
Number of lactic acid bacteria: 10 8 / ml to 10 9 / ml level Protein: about 1%, lipid: about 1%, lactose: about 5%
［実施例２］発酵飲料の製造工程の一例
製造工程を図１に示す。
[Example 2] An example of the production process of a fermented beverage is shown in FIG.
原料の乳としては、一般的に流通している牛乳が好ましい。発酵飲料の製造工程の一例を図１に記述する。加熱殺菌条件として、チーズホエーに糖添加後の１次加熱殺菌は、乳酸菌の死滅及び雑菌の死滅が目的であり、常圧加熱の場合は、液温が７０〜９０℃、１０〜２０分であり、好ましくは８５℃、１０分保持が必要である。酵母スターターの添加量はホエーに対して、菌数１×１０４〜１×１０５/ｍｌが適量であり、その後の酵母スターター添加による発酵は、２５〜３０℃で２０〜２４時間程度の培養が好ましい。また、その後の味付けとしての糖添加後の２次加熱殺菌の場合、解放容器中で７０〜８５℃、２〜２０分、好ましくは８０℃、２分が適当である。また、減圧殺菌の場合、０．０２〜０．１MPa 、７５℃、１５分以上が適当である。これは、酵母の殺菌とフレーバーに影響のあるアルコール残量を出来るだけ少なくする条件であればよい。例えばアルコール残量が０．１〜０．３％程度に抑えることが必要である。また、使用する糖として、単糖類、二等類、三等類、オリゴ糖、及びこれ等の混合物が使用される。糖類の味付け量は、好みにより決めるので限定されるものではない。更に、糖類にプラスした味付けの成分として、この発酵飲料にプルーン、ハスカップ、ブルーベリー、ラズベリー、イチゴ味の果汁等を添加することもできる。一方、３次加熱殺菌は、発酵飲料のフレーバーの飛散を最小限に食い止めるため、密閉容器中で行うことが条件となる。発酵飲料の液温が８５℃に到達すれば殺菌上その目的は達するが、余り品温が高すぎても、時間が長すぎても品質が変性する恐れがあるので、８０〜９５℃、７〜１０分程度保持すればよい。 As the raw material milk, generally distributed milk is preferable. An example of the manufacturing process of a fermented drink is described in FIG. As the heat sterilization conditions, the primary heat sterilization after adding sugar to cheese whey is aimed at the killing of lactic acid bacteria and the killing of miscellaneous bacteria. Yes, preferably holding at 85 ° C. for 10 minutes. The appropriate amount of yeast starter to be added is 1 × 10 4 to 1 × 10 5 / ml of bacteria relative to whey, and subsequent fermentation by addition of yeast starter is performed at 25 to 30 ° C. for about 20 to 24 hours. Is preferred. In addition, in the case of secondary heat sterilization after addition of sugar as a seasoning, 70 to 85 ° C. and 2 to 20 minutes, preferably 80 ° C. and 2 minutes are appropriate in an open container. In the case of sterilization under reduced pressure, 0.02 to 0.1 MPa, 75 ° C., and 15 minutes or more are appropriate. This should just be the conditions which reduce the residual amount of alcohol which has influence on the disinfection and flavor of yeast as much as possible. For example, it is necessary to suppress the remaining amount of alcohol to about 0.1 to 0.3%. In addition, monosaccharide, second class, third class, oligosaccharide, and a mixture thereof are used as the sugar to be used. The amount of saccharide seasoning is not limited because it is determined by preference. Furthermore, as a seasoning component added to sugars, prunes, lotus cups, blueberries, raspberries, strawberry-flavored juices, and the like can be added to the fermented beverage. On the other hand, the third heat sterilization is performed in a closed container in order to prevent the flavor of the fermented beverage from being scattered to a minimum. If the liquid temperature of the fermented beverage reaches 85 ° C., the purpose of sterilization will be reached, but the quality may be denatured even if the product temperature is too high or too long, so 80-95 ° C., 7 What is necessary is just to hold | maintain for about 10 minutes.
［実施例３］
各酵母をPDA培地に接種して３０℃、２日間培養し、得られた菌体をリン酸緩衝生理食塩水でOD600nm：１になるように懸濁した。各酵母液１ｍｌについて、８５℃、１０分間加熱殺菌した１％ショ糖を含有するホエー液１００ｍｌに添加し、３０℃、２２時間発酵させた。発酵後、２ｍｌをサンプル瓶に採取し、８５℃、１０分間殺菌し、４℃に冷却した。
[Example 3]
Each yeast was inoculated into PDA medium and cultured at 30 ° C. for 2 days, and the obtained bacterial cells were suspended in phosphate buffered saline so as to have an OD 600 nm of 1. 1 ml of each yeast solution was added to 100 ml of whey solution containing 1% sucrose that had been sterilized by heating at 85 ° C. for 10 minutes, and fermented at 30 ° C. for 22 hours. After fermentation, 2 ml was collected in a sample bottle, sterilized at 85 ° C. for 10 minutes, and cooled to 4 ° C.
Saccharomyces cerevisiae TENSAI株（FERM P-21891）を用いたホエー発酵液の揮発成分をGC-MS分析した。揮発成分（フレーバー）の分析は、GC-MSによる一般的な方法で行った。発酵ホエー液のサンプル１００μｌをSPME用対応４ｍｌ容バイアル管に入れ、３０℃、１０分間保持した。ヘッドスペースの揮発成分を７５μｍ Carboxen/PDMSファイバー（SUPELCO）に１分間吸着し、島津ガスクロマトグラフ質量分析計GCMS-QP2010（カラム：TC−WAX ６０ｍ×０．２５ｍｍ ｉ.ｄ.膜厚０．２５μｍ）にて分析した。得られたピークの中でブランデー香の成分としても知られるイソアミルアルコール、日本酒の香気成分として知られるカプロン酸エチルを芳香性成分の指標として、発酵バター臭として知られるアセトインがホエー不快臭の指標として認められた。ＴＥＮＳＡＩ酵母で発酵された飲料は、他の供試酵母から発酵される飲料より酢酸エチル、イソアミルアルコール、カプロン酸の含量が高く、また、アセトインの含量は少なかった。それぞれの指標は実際に嗅いだホエー液のフレーバーの傾向と一致していた。それぞれの酵母株で発酵させたホエー液の官能評価試験を行った。官能試験のパネラー人数を１，2回それぞれ１０名で行った。その結果を表１に示した。 The volatile components of the whey fermentation broth using Saccharomyces cerevisiae TENSAI strain (FERM P-21891) were analyzed by GC-MS. Analysis of volatile components (flavors) was performed by a general method by GC-MS. A sample of fermentation whey solution (100 μl) was placed in a 4 ml vial tube for SPME and kept at 30 ° C. for 10 minutes. The volatile components in the head space are adsorbed on a 75 μm Carboxen / PDMS fiber (SUPELCO) for 1 minute, and the Shimadzu gas chromatograph mass spectrometer GCMS-QP2010 (column: TC-WAX 60 m × 0.25 mm id, film thickness 0.25 μm) Analyzed in Among the obtained peaks, isoamyl alcohol, which is also known as a component of brandy fragrance, ethyl caproate, which is known as an aroma component of sake, is used as an indicator of aromatic components, and acetoin, known as fermentation butter odor, is used as an indicator of unpleasant odor of whey. Admitted. Beverages fermented with TENSAI yeast had a higher content of ethyl acetate, isoamyl alcohol and caproic acid, and a lower content of acetoin than beverages fermented from other test yeasts. Each index agreed with the tendency of the flavor of whey solution actually smelled. The sensory evaluation test of the whey solution fermented with each yeast strain was performed. The number of panelists for the sensory test was 1 and 2 times, 10 people each. The results are shown in Table 1.
GC-MS分析した結果及び表１に示した官能試験結果より、Saccharomyces cerevisiae TENSAI株（FERM P-21891）で発酵させたホエーは最も芳香性があり、また、良い香りの香気成分として知られるイソアミルアルコール、カプロン酸エチルのピーク強度が高く、不快臭の指標としたアセトインのピーク強度が低く、ホエー飲料を製造するにあたって、TENSAI株（FERM P-21891）が風味改善をもたらす特徴がある酵母であることが確認された。 From the results of GC-MS analysis and the sensory test results shown in Table 1, whey fermented with Saccharomyces cerevisiae TENSAI strain (FERM P-21891) is the most aromatic and also known as isoamyl, a fragrant component with good fragrance. Alcohol and ethyl caproate have high peak intensity, acetoin peak intensity is low as an indicator of unpleasant odor, TENSAI strain (FERM P-21891) is a yeast that has the characteristics of improving flavor when producing whey beverage It was confirmed.
この新規なホエー発酵飲料用酵母Saccharomyces cerevisiae TENSAI株は２０１０年３月８日に、独立行政法人 産業技術総合研究所 特許生物寄託センターにFERM P-21891として寄託されている。当該Saccharomyces cerevisiae TENSAI株（FERM P-21891）の菌体学的性質を以下に示す。 This new strain of yeast for fermentation of whey, Saccharomyces cerevisiae TENSAI, was deposited on March 8, 2010 as FERM P-21891 at the Patent Organism Depositary, National Institute of Advanced Industrial Science and Technology. The mycological properties of the Saccharomyces cerevisiae TENSAI strain (FERM P-21891) are shown below.
巨視的観察（コロニー観察）
YM寒天平板培地上で２５℃、３日間の培養において、コロニーの性状は、全縁、円錐状、平滑、輝光性、バター様、湿性、クリーム色を示した。
Macroscopic observation (colony observation)
In the culture at 25 ° C. for 3 days on the YM agar plate medium, the colony properties showed full edge, conical shape, smoothness, brightness, butter-like, wetness, and cream color.
微視的観察（形態性状観察）
YM寒天平板培地上で２５℃、３日間の培養において、栄養細胞は亜球形から広楕円形であり、増殖は多極出芽による。また、同培養６日目において、子嚢に４個の亜球形の子嚢胞子が形成することが認められた。
Microscopic observation (morphological observation)
In the culture on YM agar plate medium at 25 ° C. for 3 days, the vegetative cells are subspherical to wide-elliptical, and the growth is by multipolar budding. In addition, on the 6th day of the same culture, it was observed that four subspherical ascospores were formed in the ascomy.
サッカロマイセス・セルビシエ（Saccharomyces cerevisiae）TENSAI株（FERM P-21891）の生理性状試験結果を表２に示す。
［実施例３］
乳酸飲料の製造方法の一例を下記する。
チーズホエー５０ｋｇを乳用パステライザーに入れ、重量に対して１％のショ糖（０．５ｋｇ）を添加し、攪拌しながら８５℃、１０分間加熱殺菌を行う。３０℃まで冷却した後、酵母スターターを添加する。
酵母スターターは（Saccharomyces cerevisiae）TENSAI株（FERM P-21891）をポテトデキストロース培地にて継代培養していたものを１白金時、滅菌した１％ショ糖を含有したホエー液１００ｍｌに加え、３０℃で２０〜２４時間発酵させたものを５０ｍｌ使用する。酵母スターターを添加した後、２５〜３０℃で２２時間発酵処理を行う。その後、ホエーに対して５％のショ糖（２．５ｋｇ）を添加して７５℃、１５分間またはそれと同等以上の加熱殺菌を行う。次に１０℃以下まで冷却し、容器に充填する。充填したホエー発酵飲料は８５℃、１０分間（中心温度到達８０℃以上）の３次殺菌を行う。１０℃以下に冷却する。製品分類としてはチーズホエーを主原料とした乳酸菌飲料（殺菌済み）となる。
[Example 3]
An example of a method for producing a lactic acid beverage is described below.
50 kg of cheese whey is placed in a dairy pasteurizer, 1% sucrose (0.5 kg) is added to the weight, and sterilized by heating at 85 ° C. for 10 minutes while stirring. After cooling to 30 ° C., a yeast starter is added.
The yeast starter (Saccharomyces cerevisiae) TENSAI strain (FERM P-21891) subcultured in potato dextrose medium was added to 100 ml of whey solution containing 1% sucrose sterilized at 1 platinum, and 30 ° C. Use 50 ml of fermented for 20-24 hours. After adding the yeast starter, fermentation treatment is performed at 25 to 30 ° C. for 22 hours. Thereafter, 5% sucrose (2.5 kg) is added to whey and sterilized by heating at 75 ° C. for 15 minutes or more. Next, it is cooled to 10 ° C. or lower and filled into a container. The filled whey fermented beverage is subjected to tertiary sterilization at 85 ° C. for 10 minutes (center temperature reaching 80 ° C. or higher). Cool to below 10 ° C. The product classification is lactic acid bacteria beverage (sterilized) with cheese whey as the main ingredient.
密閉容器充填後３次殺菌（中心温度到達８０℃以上）した発酵飲料の１０℃における保存試験を実施した。３９日保存後の一般性菌数、大腸菌群、真菌数とも陰性であり、官能試験に於いてもフレーバー、味とも良好であった。 A storage test was conducted at 10 ° C. for fermented beverages that were tertiary sterilized (filled with a central temperature of 80 ° C. or higher) after filling the sealed container. The general bacterial count, coliform group, and fungal count after 39 days of storage were negative, and the flavor and taste were also good in the sensory test.
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| WO2018056424A1 (en) * | 2016-09-26 | 2018-03-29 | 株式会社明治 | Fermented milk having rich flavor and method for producing same |
| WO2018056427A1 (en) * | 2016-09-26 | 2018-03-29 | 株式会社明治 | Fermented milk rich in flavor, and method for producing same |
| JP2018050523A (en) * | 2016-09-28 | 2018-04-05 | 株式会社明治 | Method for producing whey preparation |
| KR20200020684A (en) | 2017-06-26 | 2020-02-26 | 가부시키가이샤 야쿠르트 혼샤 | Aromatic Microbial Culture |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20170103789A (en) | 2015-01-19 | 2017-09-13 | 가부시키가이샤 야쿠르트 혼샤 | Culture product of microorganism belonging to the genus wickerhamomyces |
| US10485255B2 (en) | 2015-01-19 | 2019-11-26 | Kabushiki Kaisha Yakult Honsha | Culture product of microorganism belonging to the genus Wickerhamomyces |
| WO2018056424A1 (en) * | 2016-09-26 | 2018-03-29 | 株式会社明治 | Fermented milk having rich flavor and method for producing same |
| WO2018056427A1 (en) * | 2016-09-26 | 2018-03-29 | 株式会社明治 | Fermented milk rich in flavor, and method for producing same |
| JP2018050523A (en) * | 2016-09-28 | 2018-04-05 | 株式会社明治 | Method for producing whey preparation |
| KR20200020684A (en) | 2017-06-26 | 2020-02-26 | 가부시키가이샤 야쿠르트 혼샤 | Aromatic Microbial Culture |
| US11248205B2 (en) | 2017-06-26 | 2022-02-15 | Kabushiki Kaisha Yakult Honsha | Aromatic culture product of microorganism |
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