JP4296133B2 - Chilled beverage and method for producing the same - Google Patents
Chilled beverage and method for producing the same Download PDFInfo
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Description
本発明はコーヒー、紅茶、日本茶、麦茶、ウーロン茶、ハーブ茶またはフレーバードウォーター等の中性域で良好な風味を有する飲料原料をベースとする飲料組成物からなるチルド飲料およびその製造方法に関する。より具体的には、コーヒー、紅茶、日本茶、麦茶、ウーロン茶、ハーブ茶またはフレーバードウォーター等の中性域で良好な風味を有する飲料原料をベースとする飲料組成物からなるチルド飲料であって、乳酸菌が生菌の状態で存在し、且つ乳酸発酵しない条件まで冷却することを特徴とするチルド飲料およびその製造方法に関する。 The present invention relates to a chilled beverage comprising a beverage composition based on a beverage ingredient having a good flavor in a neutral region such as coffee, black tea, Japanese tea, barley tea, oolong tea, herbal tea or flavored water, and a method for producing the same. More specifically, a chilled beverage comprising a beverage composition based on a beverage ingredient having a good flavor in a neutral range such as coffee, black tea, Japanese tea, barley tea, oolong tea, herbal tea or flavored water, The present invention relates to a chilled beverage and a method for producing the chilled beverage, wherein the lactic acid bacteria are cooled to a condition in which lactic acid bacteria exist in a live state and do not undergo lactic acid fermentation.
近年の消費者は嗜好が多様化している。中でも風味についての感覚が鋭敏となっている。従って、原料の風味がより好ましい飲料の開発が望まれているが、従来法により製造された飲料では、一般的な加熱殺菌により、加熱臭が発生するなど風味の劣化が避けられない状況にある。また、加熱殺菌条件の軽減を目的として、クエン酸等の有機酸を添加することにより飲料のｐＨを低下させた後に殺菌する方法も実施されているが、その有機酸等の酸味により風味が損なわれるという結果となっている。 Consumers in recent years have diversified preferences. Above all, the sense of flavor is sensitive. Therefore, development of beverages with more preferable flavors of raw materials is desired, but beverages produced by conventional methods are in a situation in which deterioration of the flavor is unavoidable due to generation of heated odor due to general heat sterilization. . In addition, for the purpose of reducing heat sterilization conditions, a method of sterilization after lowering the pH of a beverage by adding an organic acid such as citric acid has been implemented, but the flavor is impaired by the acidity of the organic acid or the like. As a result.
一方、飲料の製造過程において、乳酸菌を利用し、風味を保持する試みも見られる。例えば、特開昭５０−０１８６５３には、長期保存可能な野菜ジュースの製法を目的に、特開昭５７−１３８３７０には野菜ジュース本来の風味を損なうことなくこの加熱臭を除去する目的で、野菜ジュースに乳酸菌を添加し乳酸発酵させた後に低温殺菌をする野菜ジュースの製造法が開示されている。また、特開昭５９−０９８６７２には、果汁の乳酸発酵液からなる乳酸菌飲料が、特開昭６０−２５１８６７には、広範囲の果実の果汁から高濃度に乳酸菌を含む乳酸菌飲料を製造するための果汁の処理に特徴を有する乳酸菌飲料の製造法が開示されている。しかしこれらはいずれも、その製造過程において乳酸発酵を実施しているか、若しくは飲料原料を強い殺菌条件にて処理を行っているため、素材本来の味・香り・色合いを保持した飲料とはなっていない。
本発明の課題は、殺菌のための加熱を軽減し、素材本来の味・香りを保持したチルド飲料を提供することにある。 The subject of this invention is providing the chilled drink which reduced the heat | fever for disinfection and hold | maintained the original taste and aroma of a raw material.
本発明によれば、下記要旨の発明が提供される。
（１）中性域で良好な風味を有する飲料原料をベースとする飲料組成物からなる、乳酸発酵による生成乳酸の濃度が０．１％（乳酸換算：Ｗ／Ｖ％）未満のチルド飲料の製造方法であって、該飲料組成物を６５℃で０．１７分〜１２１．１℃で０．３分に相当する殺菌条件で殺菌処理した後、５〜４０℃に冷却してから乳酸菌をその生菌数が１×１０6〜１×１０9個／ｍｌの範囲内となるよう加え、乳酸発酵しない条件まで冷却することを特徴とするチルド飲料の製造方法。
（２）前記乳酸菌がラクトバチルス属のものである、（１）に記載の製造方法。
（３）前記乳酸菌がラクトバチルス・カゼイである、（２）に記載の製造方法。
（４）前記乳酸菌の生菌数が１×１０7〜１×１０8個／ｍｌの範囲にある、（１）〜（３）のいずれかに記載の製造方法。
According to the present invention, the following inventions are provided.
(1) A chilled beverage having a concentration of lactic acid produced by lactic acid fermentation of less than 0.1% (in terms of lactic acid: W / V%), comprising a beverage composition based on a beverage ingredient having a good flavor in the neutral range. In the production method, the beverage composition is sterilized at 65 ° C. under sterilization conditions corresponding to 0.17 minutes to 121.1 ° C. for 0.3 minutes, and then cooled to 5 to 40 ° C. A method for producing a chilled beverage, wherein the number of viable bacteria is added so as to be within a range of 1 × 10 6 to 1 × 10 9 cells / ml, and the mixture is cooled to a condition where lactic acid fermentation is not performed.
(2) The production method according to (1) , wherein the lactic acid bacterium is of the genus Lactobacillus.
(3) The production method according to (2) , wherein the lactic acid bacterium is Lactobacillus casei.
(4) The production method according to any one of (1) to (3) , wherein the number of viable bacteria of the lactic acid bacteria is in the range of 1 × 10 7 to 1 × 10 8 cells / ml.
本発明の効果は、殺菌のための加熱が軽減されたことにより、素材本来の味・香りを保持したチルド飲料が提供できることにある。 The effect of the present invention is to provide a chilled beverage that retains the original taste and aroma of the material by reducing the heating for sterilization.
＜１＞本発明の中性域で良好な風味を有する飲料原料をベースとする飲料組成物からなるチルド飲料（以下、本発明チルド飲料ともいう） <1> A chilled beverage comprising a beverage composition based on a beverage material having a good flavor in the neutral range of the present invention (hereinafter also referred to as the chilled beverage of the present invention).
＜１−１＞本発明チルド飲料に使用する中性域で良好な風味を有する飲料原料（以下、単に飲料原料ともいう）
本発明チルド飲料に使用する中性域で良好な風味を有する飲料原料としては、コーヒー、紅茶、日本茶、麦茶、ウーロン茶、ハーブ茶（薬草茶）またはドクダミ茶、杜中茶、オリーブ茶、グアバ茶、枇杷茶、柿の葉茶、桑の葉茶等の健康茶、香料をナチュラルウォーター等の水に添加したフレーバードウォーター等の中性域で良好な風味を有するものが挙げられる。また、当該飲料原料のｐＨ範囲は、概ねｐＨ４．６〜７．０を示す。
<1-1> Beverage materials having a good flavor in the neutral range used in the chilled beverage of the present invention (hereinafter also simply referred to as beverage materials)
Beverage materials having a good flavor in the neutral range used in the chilled beverages of the present invention include coffee, tea, Japanese tea, barley tea, oolong tea, herbal tea (medicine tea) or dokudami tea, chuchu tea, olive tea, guava Examples include healthy teas such as tea, persimmon tea, persimmon leaf tea, and mulberry leaf tea, and neutral flavors such as flavored water obtained by adding a fragrance to water such as natural water. Moreover, the pH range of the said beverage raw material shows pH 4.6-7.0 in general.
本発明に用いる飲料原料のチルド飲料全体に対する割合は特に制限はない。また、飲料原料は１種単独でも使用でき、２種以上の併用も可能である。なお、本明細書で、特に指示がない限りにおいて「％」は質量／体積（Ｗ／Ｖ）％を示す。
本発明の、中性域で良好な風味を有する飲料原料をベースとする飲料組成からなるチルド飲料において、“中性域で良好な風味を有する飲料原料をベースとする”とは、当該コーヒー、紅茶、日本茶、麦茶、ウーロン茶、ハーブ茶、フレーバードウォーター等の中性域で良好な風味を有する飲料原料を主要原料としていることを意味する。
The ratio with respect to the whole chilled drink of the drink raw material used for this invention does not have a restriction | limiting in particular. In addition, the beverage ingredients can be used alone or in combination of two or more. In this specification, “%” indicates mass / volume (W / V)% unless otherwise specified.
In the chilled beverage comprising a beverage composition based on a beverage ingredient having a good flavor in the neutral range of the present invention, "based on a beverage ingredient having a good flavor in the neutral range" means the coffee, This means that beverage ingredients having a good flavor in the neutral range such as black tea, Japanese tea, barley tea, oolong tea, herbal tea, and flavored water are used as main ingredients.
＜２＞本発明に利用する乳酸菌
本発明のチルド飲料は、上記飲料原料をベースとする飲料組成物からなるチルド飲料であって、該飲料組成物を殺菌後、乳酸菌を生菌の状態で添加し、且つ乳酸発酵しない条件まで冷却することを特徴とするチルド飲料である。本発明のチルド飲料に添加する乳酸菌としては、発酵食品などに広く利用されている乳酸菌を用いることが可能である。例えば、ラクトバチルス属（Lactobacillus）、ラクトコッカス属（Lactococcus）、ペディオコッカス属（Pediococcus）、ストレプトコッカス属（Streptococcus）、エノコッカス属（Oenococcus）、ロイコノストック属（Leuconostoc）、ビフィドバクテリウム属（Bifidobacterium）が挙げられる。これら乳酸菌のうちラクトバチルス属が好ましく、摂取した場
合、生きて腸に届くプロバイオティクス菌といわれるラクトバチルス・カゼイ（Lactobacillus casei）が特に好ましい。さらに具体的には、平成４年１月１０日に通商産業省工業技術院微生物工業技術研究所（現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）に微工研菌寄第１２７０４号（ＦＥＲＭ Ｐ−１２７０４）として寄託されているラクトバチルス・カゼイ ＡＳＴ−８（ＦＥＲＭ Ｐ−１２７０４）が更に好ましい。
<2> Lactic Acid Bacteria Used in the Present Invention The chilled beverage of the present invention is a chilled beverage comprising a beverage composition based on the above beverage ingredients, and after sterilizing the beverage composition, the lactic acid bacteria are added in the state of viable bacteria And it is a chilled drink characterized by cooling to the conditions which do not lactic-acid-ferment. As lactic acid bacteria added to the chilled beverage of the present invention, lactic acid bacteria widely used for fermented foods and the like can be used. For example, Lactobacillus, Lactococcus, Pediococcus, Streptococcus, Oenococcus, Leuconostoc, Bifidobacterium ( Bifidobacterium). Among these lactic acid bacteria, the genus Lactobacillus is preferable, and Lactobacillus casei, which is said to be a probiotic bacterium that reaches the intestines when ingested, is particularly preferable. More specifically, on January 10, 1992, the Ministry of International Trade and Industry, Institute of Industrial Science, Microbial Industrial Technology Research Institute (currently the National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center) More preferred is Lactobacillus casei AST-8 (FERM P-12704) deposited as FERM P-12704).
＜２−１＞本発明に利用する乳酸菌の本発明チルド飲料への添加方法
本発明のチルド飲料は、添加された乳酸菌が生菌の状態で存在していることを特徴とする。本発明に利用する乳酸菌の本発明チルド飲料への添加形態であるが、例えば、（１）：全粉乳、脱脂粉乳、生乳等の乳培地で培養して得られる乳酸菌培養液をそのまま本発明チルド飲料へ添加する方法。（２）：上記培養液から乳酸菌を遠心分離や膜分離等で集菌した集菌物を本発明チルド飲料へ添加する方法。（３）：上記培養液を乾燥させた培養液乾燥物または上記集菌物を乾燥させた乾燥菌体を本発明チルド飲料へ添加する方法。（４）：全粉乳、脱脂粉乳、生乳等の乳をプロテアーゼ処理した透明乳培地（以下、プロテアーゼ透明乳培地ともいう）で培養して得られる乳酸菌培養液（以下、透明乳発酵液ともいう）をそのまま本発明チルド飲料へ添加する方法。（５）：（４）で得られる培養液から乳酸菌を遠心分離や膜分離等で集菌した集菌物を本発明チルド飲料へ添加する方法。（６）：（４）で得られる培養液を乾燥させた培養液乾燥物または（５）で得られる集菌物を乾燥させた乾燥菌体を本発明チルド飲料へ添加する方法。が挙げられる。味、香り、色調への影響も少ないこと及び集菌する工程を有しない簡便な調製が可能なことより、上記（４）の方法で得られる乳酸菌培養液をそのまま本発明チルド飲料へ添加する方法が好ましい。ここで、上記乳酸菌の培地、培養方法等において、格別の制限はなく、公知の乳酸菌用培地、培養条件で培養することが可能であり、また乳酸菌の集菌方法、培養液または集菌物の乾燥法においても格別の制限はなく公知の方法を用いることが可能である。
<2-1> Method for Adding Lactic Acid Bacteria Utilized in the Present Invention to the Chilled Beverage of the Present Invention The chilled beverage of the present invention is characterized in that the added lactic acid bacterium is present in a viable state. The lactic acid bacteria used in the present invention are added to the chilled beverage of the present invention. For example, (1): a lactic acid bacterium culture solution obtained by culturing in a milk medium such as whole milk powder, skim milk powder, raw milk, etc. A method of adding to beverages. (2): A method of adding a collected product obtained by collecting lactic acid bacteria from the culture solution by centrifugation or membrane separation to the chilled beverage of the present invention. (3): A method of adding a dried culture solution obtained by drying the above-mentioned culture solution or a dried bacterial cell obtained by drying the above-mentioned collected bacteria to the chilled beverage of the present invention. (4): Lactic acid bacteria culture medium (hereinafter also referred to as transparent milk fermentation liquid) obtained by culturing milk such as whole milk powder, skim milk powder, and raw milk in a protease-treated transparent milk medium (hereinafter also referred to as protease transparent milk medium). Is added to the chilled beverage of the present invention as it is. (5): A method of adding a collected product obtained by collecting lactic acid bacteria from the culture solution obtained in (4) by centrifugation or membrane separation to the chilled beverage of the present invention. (6): A method of adding a dried culture solution obtained by drying the culture solution obtained in (4) or a dried bacterial cell obtained by drying the collected product obtained in (5) to the chilled beverage of the present invention. Is mentioned. A method of adding the lactic acid bacteria culture solution obtained by the method (4) as it is to the chilled beverage of the present invention as it is because it has little influence on the taste, aroma, and color and can be easily prepared without the step of collecting bacteria. Is preferred. Here, the lactic acid bacteria culture medium, culture method, etc. are not particularly limited, and can be cultured under known lactic acid bacteria culture media and culture conditions. There is no particular limitation on the drying method, and a known method can be used.
＜２−２＞本発明チルド飲料へ添加する乳酸菌の生菌数および添加直後の本発明チルド飲料に含まれる乳酸菌の生菌数
本発明チルド飲料へ添加する乳酸菌の生菌数は、１×１０７〜１×１０１１個／mlの範囲にあれば好ましく、１×１０９〜１×１０１１個／mlの範囲にあればさらに好ましい。
また、添加直後の本発明チルド飲料に含まれる乳酸菌の生菌数は、１×１０６〜１×１０９個／mlの範囲にあれば好ましく、１×１０７〜１×１０８個／mlの範囲であればさらに好ましい。尚、生菌数の測定は、例えば、乳及び乳製品の成分規格に関する省令の乳酸菌数の測定法により測定することが可能である。
<2-2> Viable count of lactic acid bacteria added to the chilled beverage of the present invention and viable count of lactic acid bacteria contained in the chilled beverage of the present invention immediately after addition It is preferably in the range of 7 to 1 × 10 11 pieces / ml, and more preferably in the range of 1 × 10 9 to 1 × 10 11 pieces / ml.
The viable count of lactic acid bacteria contained in the chilled beverage of the present invention immediately after addition is preferably in the range of 1 × 10 6 to 1 × 10 9 cells / ml, and 1 × 10 7 to 1 × 10 8 cells / ml. If it is the range, it is still more preferable. The number of viable bacteria can be measured by, for example, the method for measuring the number of lactic acid bacteria according to the ministerial ordinance relating to the component specifications of milk and dairy products.
＜３＞本発明のチルド飲料の製造方法
本発明のチルド飲料の製造方法は、上記飲料原料をベースとする飲料組成物を殺菌後冷却し、乳酸菌を上記方法にて添加する。その後、完成した飲料を容器等に充填、密封し、乳酸発酵しない条件まで冷却して、保存するというものである。すなわち、本製造方法は、飲料原料をベースとする飲料組成物を、殺菌の強度を減じた条件で殺菌後冷却し、乳酸菌を添加、乳酸発酵しない条件下まで冷却し保存（冷蔵）することで、飲料中での汚染菌の増殖を防ぎ且つ素材本来の味、香りを保持したチルド飲料を製造するというものである。
<3> Method for Producing a Chilled Beverage of the Present Invention In the method for producing a chilled beverage of the present invention, the beverage composition based on the beverage raw material is sterilized and then cooled, and lactic acid bacteria are added by the above method. Thereafter, the finished beverage is filled in a container or the like, sealed, cooled to a condition that does not undergo lactic acid fermentation, and stored. That is, the present production method is to cool a beverage composition based on a beverage ingredient after sterilization under reduced sterilization strength, add lactic acid bacteria, and cool and store (refrigerate) until no lactic acid fermentation occurs. The production of a chilled beverage that prevents the growth of contaminating bacteria in the beverage and retains the original taste and aroma of the material.
＜３−１＞殺菌
加熱殺菌は、飲料原料をベースとする飲料組成物本来の味を損ない、さらに加熱臭を発生させ、製品の風味に変化を与えることになる。本発明のチルド飲料は、素材本来の味、香りを保持した状態での提供を目的とすることより、本発明における飲料原料をベースとする飲料組成物の殺菌条件は、６５℃で０．１７分〜１２１．１℃で０．３分に相当する殺菌条件で殺菌処理するものである。ここで、「相当する殺菌条件」とは、殺菌温度がより低温であっても、また、より高温であってもよく、温度と時間で表される殺菌の強度
が同等である殺菌条件をいう。殺菌方法は、格別の制限はなく、バッチ殺菌法、連続殺菌法等の公知の方法を用いることが可能である。
なお、相当する殺菌条件へは次式で変換可能である。
The corresponding sterilization conditions can be converted by the following equation.
＜３−２＞殺菌後の冷却
殺菌後は、添加する乳酸菌が死滅しない程度、例えば５℃〜４０℃程度まで冷却することが好ましい。また、冷却方法は、格別の制限はなく、プレート冷却器等の公知の冷却方法を用いることが可能である。
<3-2> Cooling after sterilization After sterilization, it is preferable that the lactic acid bacteria to be added are cooled to such an extent that they do not die, for example, about 5 to 40 ° C. The cooling method is not particularly limited, and a known cooling method such as a plate cooler can be used.
＜３−３＞乳酸菌添加
＜２−１＞に記載の方法にて乳酸菌を殺菌冷却後の飲料原料をベースとする飲料組成物に添加する。また、添加する乳酸菌の生菌数は、１×１０７〜１×１０１１個／mlの範囲にあれば好ましく、１×１０９〜１×１０１１個／mlの範囲にあればさらに好ましい。さらに、添加直後の本発明チルド飲料に含まれる乳酸菌の生菌数は、１×１０６〜１×１０９個／mlの範囲にあれば好ましく、１×１０７〜１×１０８個／mlの範囲であればさらに好ましい。
<3-3> Addition of lactic acid bacteria Lactic acid bacteria are added to the beverage composition based on the beverage raw material after sterilization cooling by the method described in <2-1>. The number of viable lactic acid bacteria added is preferably in the range of 1 × 10 7 to 1 × 10 11 cells / ml, and more preferably in the range of 1 × 10 9 to 1 × 10 11 cells / ml. Furthermore, the viable count of lactic acid bacteria contained in the chilled beverage of the present invention immediately after addition is preferably in the range of 1 × 10 6 to 1 × 10 9 cells / ml, and 1 × 10 7 to 1 × 10 8 cells / ml. If it is the range, it is still more preferable.
＜３−４＞充填、密封、保存
乳酸菌を上記方法にて添加後、容器等に充填、密封し、更に乳酸発酵をしない条件下まで冷却して保存する。飲料の充填、密封には格別の制限はなく、公知の方法を用いることが可能である。また、本発明チルド飲料を保存する期間は、汚染菌の増殖を防ぎ、風味を維持するために、乳酸発酵をしない条件下まで冷却しておくことが好ましい。ここで、「乳酸発酵をしない条件」とは、乳酸発酵を全くしないか、したとしても通常の保存期間、例えば２週間で、乳酸発酵による生成乳酸の濃度が０．１％（乳酸換算：Ｗ／Ｖ％）未満となる条件をいう。具体的には０℃〜１５℃の温度が好ましい。雑菌汚染防止効果の観点を加味すると、５℃〜１０℃の温度がより好ましい。また、保存期間としては、添加乳酸菌数が維持され、汚染菌の抑制が可能である期間は保存が可能である。具体的には、本発明のチルド飲料は２週間程度の保存が可能である。一方、保存方法は温度が上記の条件に維持される限りにおいては、格別の制限はなく、公知の方法を用いることが可能である。
<3-4> Filling, sealing, and storage After adding lactic acid bacteria by the above method, the container is filled and sealed, and further cooled and stored under conditions that do not cause lactic acid fermentation. There are no particular restrictions on the filling and sealing of beverages, and known methods can be used. Moreover, it is preferable to cool to the conditions which do not carry out lactic acid fermentation during the period which preserve | saves this invention chilled drink, in order to prevent the proliferation of a contamination microbe and to maintain flavor. Here, the “condition of not performing lactic acid fermentation” means that the concentration of lactic acid produced by lactic acid fermentation is 0.1% (lactic acid conversion: W) in a normal storage period, for example, 2 weeks, even if lactic acid fermentation is not performed at all. / V%). Specifically, a temperature of 0 ° C. to 15 ° C. is preferable. In view of the effect of preventing contamination with various bacteria, a temperature of 5 ° C to 10 ° C is more preferable. Moreover, as a preservation | save period, preservation | save is possible for the period when the number of addition lactic acid bacteria is maintained and contamination bacteria can be suppressed. Specifically, the chilled beverage of the present invention can be stored for about 2 weeks. On the other hand, the storage method is not particularly limited as long as the temperature is maintained at the above-described conditions, and a known method can be used.
＜４＞各種添加剤の添加
本発明の上記飲料原料をベースとする飲料組成物からなるチルド飲料において、該飲料組成物は、上記飲料原料のみで構成されていても良く、その他の各種添加剤等を任意に添加することも可能である。ここで利用できる添加剤としては、飲料に添加配合できることが知られている各種のものが挙げられる。例えば、ハーブ、ショウガ、シナモン等の香辛料、オレンジフレーバー、リンゴフレーバー等の香料、シュークロース、グルコース、フラクトース等の糖類、キシリトール、エリスリトール、ソルビトール等の糖アルコール類、乳果オリゴ糖、大豆オリゴ糖、フラクトオリゴ糖、ガラクトオリゴ糖、キシロオリゴ糖等の各種オリゴ糖類、ステビア、甘茶抽出物、甘草抽出物等の甘味料、ポリデキストロース等の食物繊維類、ビタミンＣ等の各種ビタミン類、イソロイシン、バリン等のアミノ酸類、炭酸カルシウム、塩化第二鉄等の無機塩類、水、液糖等の水溶液を例示することができる。これらの配合量は特に限定されるものではないが、通常０．１％〜５％（Ｗ／Ｖ）程度の範囲から選ぶことが可能である。
<4> Addition of various additives In the chilled beverage comprising the beverage composition based on the beverage raw material of the present invention, the beverage composition may be composed only of the beverage raw material, and other various additives. It is also possible to add etc. arbitrarily. Examples of the additive that can be used here include various types that are known to be added and blended in beverages. For example, spices such as herbs, ginger and cinnamon, flavors such as orange flavor and apple flavor, sugars such as sucrose, glucose and fructose, sugar alcohols such as xylitol, erythritol and sorbitol, whey oligosaccharides, soybean oligosaccharides, Various oligosaccharides such as fructooligosaccharide, galactooligosaccharide and xylo-oligosaccharide, sweeteners such as stevia, candy tea extract and licorice extract, dietary fibers such as polydextrose, various vitamins such as vitamin C, amino acids such as isoleucine and valine , Inorganic salts such as calcium carbonate and ferric chloride, and aqueous solutions such as water and liquid sugar. These blending amounts are not particularly limited, but can usually be selected from a range of about 0.1% to 5% (W / V).
以下、本発明を実施例により具体的に詳述するが、本発明はこれらに限定されない。 EXAMPLES Hereinafter, although an Example demonstrates this invention concretely in detail, this invention is not limited to these.
＜実施例１＞本発明に利用する乳酸菌の調製
本発明に利用する乳酸菌は、以下のように調製した。
＜１＞透明乳発酵液の調整法
脱脂粉乳８％水溶液（Ｗ／Ｖ）を炭酸カリウムでｐＨ８．０に調製した。この調製液に、プロテアーゼＳ「アマノ」Ｇ（天野エンザイム株式会社製）を０．０５％（Ｗ／Ｖ）添加し、７０℃で４時間反応させる。反応終了後、９５℃で２０分加熱して酵素失活及び殺菌を行った。このプロテアーゼ透明乳培地にラクトバチルス・カゼイ ＡＳＴ−８（ＦＥＲＭ Ｐ−１２７０４）を１ｘ１０７個／mｌとなるように接種し、30℃で48時間発酵させた。得られた透明乳酸発酵液の生菌数は１．５ｘ１０９個／mｌであった。尚、生菌数の測定は乳等省令の乳酸菌数測定法に準じ、Ｂ．Ｃ．Ｐ．加プレートカウント培地を用いて測定した。
＜２＞集菌物の調整法
上記の方法により得られた透明乳酸発酵液を、遠心分離機（株式会社コクサイ製 Ｈ−１０３Ｎ）を用いて、３，０００ｒｐｍ １５分の条件で遠心分離し、沈殿した菌体を回収し、集菌物を得た。なお、得られた集菌物の生菌数は、２．０×１０１０個／ｇであった。
＜３＞乾燥菌体の調整法
上記の方法により得られた透明乳酸発酵液を液体窒素で急速凍結した。急速凍結した透明乳酸菌発酵液を凍結乾燥器（LABCONCO社製ＦＤ−１０）を用いて凍結乾燥させ、乾燥菌体を得た。なお、得られた集菌物の生菌数は、１．０×１０１１個／ｇであった。
<Example 1> Preparation of lactic acid bacteria used in the present invention The lactic acid bacteria used in the present invention were prepared as follows.
<1> Preparation Method of Transparent Milk Fermentation Solution A skim milk powder 8% aqueous solution (W / V) was adjusted to pH 8.0 with potassium carbonate. To this preparation solution, 0.05% (W / V) of protease S “Amano” G (Amano Enzyme Co., Ltd.) is added and reacted at 70 ° C. for 4 hours. After completion of the reaction, the enzyme was deactivated and sterilized by heating at 95 ° C. for 20 minutes. This protease transparent milk medium was inoculated with Lactobacillus casei AST-8 (FERM P-12704) at 1 × 10 7 cells / ml and fermented at 30 ° C. for 48 hours. The number of viable bacteria in the obtained transparent lactic acid fermentation broth was 1.5 × 10 9 cells / ml. The measurement of the number of viable bacteria is in accordance with the method for measuring the number of lactic acid bacteria according to the Ministerial Ordinance. C. P. Measurement was performed using an additional plate count medium.
<2> Preparation method of bacterial collection The transparent lactic acid fermentation broth obtained by the above method is centrifuged using a centrifuge (H-103N, manufactured by Kokusai Co., Ltd.) under conditions of 3,000 rpm for 15 minutes, The precipitated cells were collected to obtain a collected product. In addition, the number of viable bacteria of the obtained collected product was 2.0 × 10 10 cells / g.
<3> Preparation method of dried microbial cells The transparent lactic acid fermentation broth obtained by the above method was rapidly frozen with liquid nitrogen. The rapidly frozen transparent lactic acid bacteria fermentation broth was freeze-dried using a freeze dryer (LAB-10 manufactured by LABCONCO) to obtain dried cells. In addition, the number of viable bacteria in the obtained collected product was 1.0 × 10 11 cells / g.
＜実施例２〜４および比較例１〜３＞本発明のチルド飲料の製造およびその評価
実施例１で調製した乳酸菌および下記に示す条件で調製した飲料原料を用いて本発明のチルド飲料を製造した。
<Examples 2 to 4 and Comparative Examples 1 to 3> Production of the chilled beverage of the present invention and its evaluation Production of the chilled beverage of the present invention using the lactic acid bacteria prepared in Example 1 and the beverage ingredients prepared under the conditions shown below. did.
＜実施例２＞
通常の方法で抽出した麦茶、ｐＨ６．５（堀場製作所製 ｐＨメーター Ｆ−１２）を１２１．１℃で１０秒間殺菌した。この加熱殺菌した麦茶を１０℃まで冷却し、実施例１＜１＞で調製した透明乳発酵液を無菌的に１％（Ｖ／Ｖ）添加して、容器に充填後、５〜１０℃に冷却して、本発明チルド飲料製品を調製した。添加された透明乳発酵液中の乳酸菌生菌数は１．５ｘ１０9個／mｌであり、調製された製品中の乳酸菌生菌数は１．５ｘ１０7個／mｌであった。一方、比較例１は実施例２と同様に抽出した麦茶、ｐＨ６．５（堀場製作所製 ｐＨメーター Ｆ−１２）を１２１．１℃で４分間殺菌した。この加熱殺菌した麦茶を１０℃まで冷却後、実施例１＜１＞で調製した透明乳発酵液を無菌的に１％（Ｖ／Ｖ）添加して、容器に充填後、５〜１０℃に冷却して、比較例１製品を調製した。添加された透明乳発酵液中の乳酸菌生菌数は１．５ｘ１０9個／mｌであり、調製された製品中の乳酸菌生菌数は１．５ｘ１０7個／mｌであった。実施例２の製品は１４日間チルド保管（５℃、１０℃）し、経時的に製品中の乳酸菌生菌数、酸分（乳酸換算（Ｗ／Ｖ％））を測定し、汚染菌の増殖確認を実施した。また、実施例２および比較例１の製品について、１４日間チルド保管した製品についての官能評価を実施した。
<Example 2>
Barley tea, pH 6.5 (pH meter F-12 manufactured by Horiba, Ltd.) extracted by a normal method was sterilized at 121.1 ° C. for 10 seconds. This heat-sterilized barley tea is cooled to 10 ° C., and 1% (V / V) of the transparent milk fermentation liquid prepared in Example 1 <1> is added aseptically, and after filling the container, the temperature is increased to 5 to 10 ° C. Upon cooling, a chilled beverage product of the present invention was prepared. The number of viable lactic acid bacteria in the added clear milk fermentation broth was 1.5 × 10 9 cells / ml, and the number of live lactic acid bacteria in the prepared product was 1.5 × 10 7 cells / ml. On the other hand, in Comparative Example 1, barley tea, pH 6.5 (pH meter F-12, manufactured by Horiba, Ltd.) extracted in the same manner as in Example 2 was sterilized at 121.1 ° C. for 4 minutes. After cooling this heat-sterilized barley tea to 10 ° C, aseptically adding 1% (V / V) of the transparent milk fermented liquid prepared in Example 1 <1>, filling the container to 5-10 ° C Upon cooling, a Comparative Example 1 product was prepared. The number of viable lactic acid bacteria in the added clear milk fermentation broth was 1.5 × 10 9 cells / ml, and the number of live lactic acid bacteria in the prepared product was 1.5 × 10 7 cells / ml. The product of Example 2 was chilled for 14 days (5 ° C., 10 ° C.), and the number of live lactic acid bacteria and acid content (lactic acid equivalent (W / V%)) in the product was measured over time, and the growth of contaminating bacteria Confirmation was carried out. In addition, for the products of Example 2 and Comparative Example 1 , sensory evaluation was performed on the products that were chilled for 14 days.
透明乳発酵液および製品中の乳酸菌生菌数は実施例１の方法に従って実施した。 The number of viable lactic acid bacteria in the transparent milk fermentation broth and the product was determined according to the method of Example 1.
また、汚染菌の増殖確認は以下の方法に従って実施した。
リステリア菌：Oxford リステリア選択培地を用い、３０℃ ３日間培養後、計測した。Oxford リステリア選択培地：ペプトン２３．０ｇ、デンプン１．０ｇ、塩化ナトリウム５．０ｇ、 クエン酸鉄アンモニウム０．５ｇ、エスクリン１．０ｇ、塩化リチウム１５．０ｇ 寒天１０．０ｇ、シクロヘキシミド０．４ｇ、硫酸コリスチン０．０２ｇ、 塩
酸アクリフラビン０．００５ｇ、セフォテタン０．００２ｇ、フォスフォマイシン０．０１ｇ を蒸留水に溶解し（総量１Ｌ）、１２１．１℃ １５分で殺菌する。
バチルス・セレウス菌：ＮＧＫＧ培地を用い、３５℃ １日間培養後、計測した。ＮＧＫＧ培地：ペプトン１．０ｇ、酵母エキス０．５ｇ、塩化ナトリウム４．０ｇ、グリシン３．０ｇ、 硫酸ポリミキシン５万単位、フェノールレッド０．０２５ｇ、寒天１８．０ｇ を蒸留水に溶解し（総量１Ｌ）（ｐＨ６．８）を１２１．１℃ １５分で殺菌する。
In addition, confirmation of the growth of contaminating bacteria was performed according to the following method.
Listeria monocytogenes: Counted after culturing at 30 ° C. for 3 days using an Oxford Listeria selective medium. Oxford Listeria selective medium: 23.0 g of peptone, 1.0 g of starch, 5.0 g of sodium chloride, 0.5 g of ammonium iron citrate, 1.0 g of esculin, 15.0 g of lithium chloride, 10.0 g of agar, 0.4 g of cycloheximide, sulfuric acid Dissolve 0.02 g colistin, 0.005 g acriflavine hydrochloride, 0.002 g cefotetan, 0.01 g fosfomycin in distilled water (total amount: 1 L), and sterilize at 121.1 ° C. for 15 minutes.
Bacillus cereus: measured using NGKG medium after culturing at 35 ° C. for 1 day. NGKG medium: 1.0 g of peptone, 0.5 g of yeast extract, 4.0 g of sodium chloride, 3.0 g of glycine, 50,000 units of polymyxin sulfate, 0.025 g of phenol red, and 18.0 g of agar are dissolved in distilled water (total amount: 1 L). ) (PH 6.8) is sterilized at 121.1 ° C. for 15 minutes.
製品中の酸分（乳酸換算（Ｗ／Ｖ％）は０．１Ｎ水酸化ナトリウムを用いた中和滴定により測定した。 Acid content in the product (lactic acid equivalent (W / V%) was measured by neutralization titration using 0.1N sodium hydroxide.
官能評価は、１４日間チルド保管された上記実施例の飲料と比較例の飲料とを２点比較し、男性２０名及び女性２０名（合計４０名）の官能評価員に、どちらが好ましいかを選択させ、各例の飲料を好ましいと選択した人数及びその有意検定結果としての危険率を求めた。なお、危険率は、統計的仮説検定で第一種の過誤をおかす確率をいう For sensory evaluation, the beverages of the above-mentioned example stored in chilled storage for 14 days and the beverages of the comparative example are compared at two points, and which of 20 males and 20 females (40 in total) is selected is preferred. Then, the number of persons who selected the beverage of each example as preferable and the risk factor as a result of the significance test were obtained. The risk factor is the probability of making a type I error in the statistical hypothesis test.
実施例３： 通常の方法で抽出したウーロン茶、ｐＨ６．２（堀場製作所製 ｐＨメーター Ｆ−１２）を１２１．１℃で１０秒間殺菌した。この加熱殺菌したウーロン茶を１０℃まで冷却後、実施例１＜１＞で調製した透明乳発酵液を無菌的に１％（Ｖ／Ｖ）添加し、容器に充填後、５〜１０℃に冷却して、本発明チルド飲料製品を調製した。添加された透明乳発酵液中の乳酸菌生菌数は１．５ｘ１０９個／mｌであり、調製された製品中の乳酸菌生菌数は１．５ｘ１０７個／mｌであった。比較例２は実施例３と同様に抽出したウーロン茶、ｐＨ６．２（堀場製作所製 ｐＨメーター Ｆ−１２）を１２１．１℃で４分間殺菌した。この加熱殺菌したウーロン茶を１０℃まで冷却後、実施例１＜１＞で調製した透明乳発酵液を無菌的に１％（Ｖ／Ｖ）添加し、容器に充填後、５〜１０℃に冷却して、比較例２製品を調製した。添加された透明乳発酵液中の乳酸菌生菌数は１．５ｘ１０９個／mｌであり、調製された製品中の乳酸菌生菌数は１．５ｘ１０７個／mｌであった。なお、各種評価は実施例２および比較例１と同様に実施した。
Example 3 Oolong tea, pH 6.2 (pH meter F-12, manufactured by Horiba, Ltd.) extracted by a normal method was sterilized at 121.1 ° C. for 10 seconds. After cooling this heat-sterilized oolong tea to 10 ° C., aseptically adding 1% (V / V) of the transparent milk fermentation liquid prepared in Example 1 <1>, filling the container, and cooling to 5-10 ° C. The chilled beverage product of the present invention was prepared. The number of viable lactic acid bacteria in the added clear milk fermentation broth was 1.5 × 10 9 cells / ml, and the number of viable lactic acid bacteria in the prepared product was 1.5 × 10 7 cells / ml. In Comparative Example 2, oolong tea, pH 6.2 (pH meter F-12, manufactured by Horiba, Ltd.) extracted in the same manner as in Example 3 was sterilized at 121.1 ° C. for 4 minutes. After cooling this heat-sterilized oolong tea to 10 ° C., aseptically adding 1% (V / V) of the transparent milk fermentation liquid prepared in Example 1 <1>, filling the container, and cooling to 5-10 ° C. Then, a product of Comparative Example 2 was prepared. The number of viable lactic acid bacteria in the added clear milk fermentation broth was 1.5 × 10 9 cells / ml, and the number of viable lactic acid bacteria in the prepared product was 1.5 × 10 7 cells / ml. Various evaluations were performed in the same manner as in Example 2 and Comparative Example 1.
＜実施例２〜３および比較例１〜２の結果＞
実施例２〜３および比較例１〜２の結果を表１、表２に纏めた。酸度上昇については、充填後０日目と充填後１４日目の酸分（乳酸換算：Ｗ／Ｖ％）を比較し、酸分の上昇が０．１％（乳酸換算：Ｗ／Ｖ％）未満の場合を「酸度上昇なし」とした。乳酸菌の残存性は、充填後０日目と充填後１４日目の乳酸菌の生菌数を比較し、充填後０日目に対し３０％以上の生菌数を維持している場合を○とした。汚染菌の増殖については、１４日目の汚染菌数が調整時の１０倍未満の場合を「汚染菌の増殖なし」とし、調製時の１０倍以上の場合を「汚染菌の増殖あり」とした。
さらに、実施例２の乳酸菌の生菌数と保存日数の関係を図１に酸分と保存日数の関係を図２に、実施例３の乳酸菌の生菌数と保存日数の関係を図３に酸分と保存日数の関係を図４に示す。図２，４に示されるように、実施例２，実施例３のいずれの温度（５℃、１０℃）での保存においても、酸分の上昇は０．０１％（乳酸換算、Ｗ／Ｖ％）以下であり、乳酸発酵をしていないことが示された。本結果より、本発明チルド飲料は、殺菌強度を減じた条件で殺菌し、乳酸菌を添加後、乳酸発酵しない条件下まで冷却し保存（冷蔵）することで、飲料中での汚染菌の増殖を防ぎ且つ素材本来の味、香りを保持できることが示された。
<Results of Examples 2-3 and Comparative Examples 1-2>
The results of Examples 2-3 and Comparative Examples 1-2 are summarized in Tables 1 and 2. Regarding the increase in acidity, the acid content on the 0th day after filling and the 14th day after filling (lactic acid conversion: W / V%) was compared, and the increase in acidity was 0.1% (lactic acid conversion: W / V%) The case where it was less than “no increase in acidity”. As for the persistence of lactic acid bacteria, the number of viable bacteria of lactic acid bacteria on the 0th day after filling and the 14th day after filling is compared. did. Concerning the growth of contaminated bacteria, if the number of contaminated bacteria on the 14th day is less than 10 times that at the time of adjustment, “no growth of contaminating bacteria” is assumed, and if more than 10 times that at the time of preparation, “the presence of contaminating bacteria is propagated” did.
Further, FIG. 1 shows the relationship between the number of viable bacteria and storage days in Example 2, FIG. 2 shows the relationship between acid content and storage days, and FIG. 3 shows the relationship between the number of live bacteria and storage days in Example 3. The relationship between the acid content and the storage days is shown in FIG. As shown in FIGS. 2 and 4, the increase in acid content was 0.01% (lactic acid equivalent, W / V) even when stored at any temperature (5 ° C., 10 ° C.) in Examples 2 and 3. %) Or less, indicating that no lactic acid fermentation was performed. From this result, the chilled beverage of the present invention is sterilized under conditions with reduced sterilization strength, and after adding lactic acid bacteria, it is cooled and stored (refrigerated) to a condition where lactic acid fermentation is not carried out. It was shown that it can prevent and retain the original taste and aroma of the material.
＜試験例１〜８＞乳酸菌による添加した汚染菌の増殖抑制活性
試験例１〜４：
実施例２と同様に抽出した麦茶（ｐＨ６．５）を１２１．１℃で４分間殺菌した。この加熱殺菌した麦茶を１０℃まで冷却後、実施例１＜１＞で調製した透明乳発酵液を乳酸菌生菌数が１．０ｘ１０6個／mｌおよび表３に示す汚染菌の菌数が１．０ｘ１０2個／mｌとなるように添加し、容器に充填後、５〜１０℃に冷却して、試験例１および３試料を調製した。その後、当該乳酸菌および汚染菌の菌数の変化を１４日間観察した。試験例２および４試料は、試験例１および３試料において、表３に示す汚染菌のみを１．０ｘ１０2個
／mｌとなるように添加したものである。なお、製品中の乳酸菌生菌数および汚染菌の菌数は実施例１、２の方法に従って計測した。
<Test Examples 1 to 8> Growth inhibitory activity of contaminated bacteria added by lactic acid bacteria Test Examples 1 to 4:
Barley tea (pH 6.5) extracted in the same manner as in Example 2 was sterilized at 121.1 ° C. for 4 minutes. After this heat-sterilized barley tea was cooled to 10 ° C., the number of viable lactic acid bacteria in the transparent milk fermentation broth prepared in Example 1 <1> was 1.0 × 10 6 cells / ml and the number of contaminating bacteria shown in Table 3 was 1. The sample was added at a rate of 0.0 × 10 2 / ml, filled into a container, and then cooled to 5 to 10 ° C. to prepare Test Examples 1 and 3 samples. Thereafter, changes in the numbers of lactic acid bacteria and contaminating bacteria were observed for 14 days. Test examples 2 and 4 were prepared by adding only the contaminating bacteria shown in Table 3 to 1.0 × 10 2 / ml in test examples 1 and 3 . The number of viable lactic acid bacteria and the number of contaminating bacteria in the product were measured according to the methods of Examples 1 and 2.
試験例５〜８：
実施例３と同様に抽出したウーロン茶（ｐＨ６．２）を１２１．１℃で４分間殺菌した。この加熱殺菌したウーロン茶を１０℃まで冷却後、実施例１＜１＞で調製した透明乳発酵液を乳酸菌生菌数が１．０ｘ１０6個／mｌおよび表３に示す汚染菌数が１．０ｘ１０2個／mｌとなるように添加し、容器に充填後、５〜１０℃に冷却して、試験例５および７試料を調製した。その後、当該乳酸菌および汚染菌の菌数の変化を１４日間観察した。試験例６および８試料は、試験例５および７試料において、表３に示す汚染菌のみを１．０ｘ１０2個／mｌとなるように添加したものである。なお、製品中の乳酸菌生菌数および汚染菌の菌数は実施例１、２の方法に従って計測した。
Test Examples 5 to 8:
Oolong tea (pH 6.2) extracted in the same manner as in Example 3 was sterilized at 121.1 ° C. for 4 minutes. After the heat-sterilized oolong tea was cooled to 10 ° C., the transparent milk fermented liquid prepared in Example 1 <1> had a live lactic acid bacteria count of 1.0 × 10 6 cells / ml and the contaminated bacteria count shown in Table 3 was 1.0 × 10 6. Test samples 5 and 7 were prepared by adding 2 / ml, filling the container, and cooling to 5 to 10 ° C. Thereafter, changes in the numbers of lactic acid bacteria and contaminating bacteria were observed for 14 days. Test examples 6 and 8 were prepared by adding only the contaminating bacteria shown in Table 3 to 1.0 × 10 2 / ml in test examples 5 and 7 . The number of viable lactic acid bacteria and the number of contaminating bacteria in the product were measured according to the methods of Examples 1 and 2.
＜添加する汚染菌およびのその調製＞
試験例１〜８において使用する汚染菌を表３に示す。また、リステリア・モノサイトゲネス ＡＴＣＣ７６４４は普通寒天培地（日水製薬（株）製）を用い、３０℃で３日間培養したものを生理食塩水に懸濁し調製した。一方、バチルス・セレウス １５０１株（日本缶詰協会から分与）を土壌エキス（畑土２００ｇを１０００ｍｌの蒸留水に懸濁し、１
２１℃、４時間の加熱抽出後、上澄みを濾過したもの）を含む普通寒天培地（日水製薬（株）製、普通寒天培地３５ｇ、上記土壌エキス２５０ｍｌ、蒸留水７５０ｍｌを混合し１２１℃、１５分の殺菌を施したもの）を用い、３５℃で１週間培養したものを生理食塩水に懸濁し調製した。
<Contaminating bacteria to be added and their preparation>
Table 3 shows the contaminants used in Test Examples 1-8. In addition, Listeria monocytogenes ATCC7644 was prepared by suspending a normal agar medium (manufactured by Nissui Pharmaceutical Co., Ltd.) at 30 ° C. for 3 days in physiological saline. On the other hand, Bacillus cereus 1501 strain (distributed from the Japan Canning Association) was suspended in 1000 g of distilled water.
21 ° C., heated and extracted for 4 hours, and filtered the supernatant) A mixture of 35 g of ordinary agar medium (manufactured by Nissui Pharmaceutical Co., Ltd., ordinary agar medium, 250 ml of the above-mentioned soil extract and 750 ml of distilled water) 1) and cultured for 1 week at 35 ° C. and suspended in physiological saline.
結果を図５〜図８に示す。試験例１、３，５，７においては、汚染菌数が調製直後から１４日目まで増殖せず、微減傾向を示した。一方、汚染菌数のみを１．０ｘ１０２個／mｌとなるように添加した試験例２，４、６，８においては、試験例１、３，５，７とは対照的に、汚染菌数が調製直後から増加し始め、調製後１４日目には調製時の約１５〜６０
倍にまで増加した。つまり、試料中の乳酸菌（ラクトバチルス・カゼイ ＡＳＴ−８（ＦＥＲＭ Ｐ−１２７０４））は、汚染菌の増殖を阻害する能力を有することが示された。
The results are shown in FIGS. In Test Examples 1, 3, 5, and 7, the number of contaminating bacteria did not grow from immediately after preparation until the 14th day, and showed a slight decreasing tendency. On the other hand, in Test Examples 2, 4, 6, and 8 in which only the number of contaminating bacteria was added to 1.0 × 10 2 / ml, in contrast to Test Examples 1, 3, 5, and 7, the number of contaminating bacteria Begins to increase immediately after preparation, and on the 14th day after preparation, about 15-60 at the time of preparation.
Doubled. That is, it was shown that the lactic acid bacteria (Lactobacillus casei AST-8 (FERM P-12704)) in the sample have the ability to inhibit the growth of contaminating bacteria.
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| EP2285230A1 (en) * | 2008-04-30 | 2011-02-23 | Nestec S.A. | Compositions for preparing a coffee beverage comprising hydrolysed chlorogenic acid |
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