JP4300157B2 - Chilled beverage and method for producing the same - Google Patents
Chilled beverage and method for producing the same Download PDFInfo
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本発明は野菜汁及び／又は果実汁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、野菜汁及び／又は果実汁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 vegetable juice and / or fruit juice and a method for producing the chilled beverage. More specifically, it is a chilled beverage comprising a beverage composition based on vegetable juice and / or fruit juice, wherein the lactic acid bacteria are present in a state of viable bacteria and cooled to a condition where lactic acid fermentation is not performed. The present invention relates to a chilled beverage and a manufacturing method thereof.
野菜／果実飲料とは、野菜／果実（野菜汁、果実汁等）を主要原料とする飲料の総称で、原料野菜／果実の種類、野菜汁、果実汁等の含有量の違いなどにより、いろいろな種類の製品となり市場で見ることができる。上述したように野菜／果実飲料は野菜汁、果実汁を主成分とするので、ショ糖、ブドウ糖、果糖などの糖質、有機酸類に加えて、野菜汁、果実汁に由来するアミノ酸、無機質などの栄養成分が適度の濃度で含まれており、このために、ある種の微生物とって格好の培地としての性質を持っていることになる。これらの微生物の繁殖の結果、製品の風味を変えたり、外観にも変化を与えたり、時には沈でん物を発生させたりする。これら、微生物の増殖を防ぎ、風味等を維持した状態での保存を可能にするために、飲料の加工途上で加熱による殺菌を実施することになるのであるが、加熱殺菌により、野菜汁、果実汁の変色、並びに加熱臭の発生などの香りの劣化が生じ、製品の風味を変え、外観にも変化を与えることになる。 Vegetable / fruit drink is a general term for beverages that use vegetables / fruits (vegetable juice, fruit juice, etc.) as the main ingredient, and there are various types depending on the type of raw vegetables / fruit, content of vegetable juice, fruit juice, etc. Can be seen in the market. As described above, vegetable / fruit drinks are mainly made from vegetable juice and fruit juice. Therefore, in addition to sugars such as sucrose, glucose, and fructose, organic acids, amino acids derived from vegetable juice and fruit juice, inorganic substances, etc. Is contained in a moderate concentration, and for this reason, it has properties as a suitable medium for certain microorganisms. As a result of the propagation of these microorganisms, the flavor of the product is changed, the appearance is changed, and sometimes sediment is generated. In order to prevent the growth of these microorganisms and enable preservation in a state where the flavor is maintained, sterilization by heating is carried out during the processing of beverages. Degradation of the scent such as discoloration of the juice and generation of heated odor occurs, changing the flavor of the product and changing the appearance.
これに対し、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。一方、特開２００１−２５２０１２には、長期間のチルド保管後も、所望の生菌数、ｐＨおよび風味を保持する乳酸飲料およびその製造法が、特開２００１−０５６２３１には、中性付近のｐＨで好ましい風味を与える素材（コーヒー、メロン等）により風味付けされた調味乳発酵食品が開示されている。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、若しくは野菜／果実（野菜汁、果実汁等）を極めて強い殺菌条件にて処理を行っているため、野菜／果実素材本来の味・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・色合い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 heating for disinfection and hold | maintained the original taste, fragrance, and color 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）野菜汁及び／又は果実汁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、該飲料組成物を下記の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。
条件：前記飲料組成物がｐＨ４．０未満である場合は、６５℃で０．１７分〜６５℃
で１０分に相当する殺菌処理、また、前記飲料組成物がｐＨ４．０以上４．６未満の場合は、６５℃で０．１７分〜７５℃で１分に相当する殺菌処理、更に、前記飲料組成物がｐＨ４．６以上７．０以下の場合は、６５℃で０．１７分〜１２１．１℃で０．３分に相当する殺菌処理。
（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 method for producing a chilled beverage comprising a beverage composition based on vegetable juice and / or fruit juice, wherein the concentration of lactic acid produced by lactic acid fermentation is less than 0.1% (lactic acid equivalent: W / V%). The beverage composition is sterilized under the following conditions, and after cooling to 5 to 40 ° C., the number of viable lactic acid bacteria is in the range of 1 × 10 6 to 1 × 10 9 cells / ml. In addition, a method for producing a chilled beverage characterized by cooling to a condition that does not cause lactic acid fermentation.
Conditions: When the beverage composition has a pH of less than 4.0, it is 0.17 minutes to 65 ° C at 65 ° C.
Sterilization treatment corresponding to 10 minutes, and when the beverage composition has a pH of 4.0 or more and less than 4.6, sterilization treatment corresponding to 1 minute at 65 ° C. for 0.17 minutes to 75 ° C., When the beverage composition has a pH of 4.6 to 7.0, a sterilization treatment corresponding to 0.17 minutes at 65 ° C to 0.3 minutes at 121.1 ° C.
(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・色合い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, aroma, and color of the material by reducing the heat for sterilization.
＜１＞本発明の野菜汁及び／又は果実汁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 the vegetable juice and / or fruit juice of the present invention (hereinafter also referred to as the present chilled beverage).
＜１−１＞本発明チルド飲料に使用する野菜汁及び／又は果実汁（以下、飲料原料ともいう）
本発明チルド飲料に使用する飲料原料としては、グレープフルーツ、オレンジ、リンゴ、ブドウ、イチゴ、パイナップル、キウイ、グアバ、マンゴー、アセロラ、ブルーベリー、ザクロ、桃、洋なし、パパイヤ、メロン、スイカ、バナナ、イチジク等の果実汁、及び／又はトマト、赤ピーマン、ニンジン、キャベツ、ほうれん草、ケール、プチヴェール（商標：ケールと芽キャベツの交配種）等の野菜汁が挙げられる。また、当該飲料原料は、その野菜汁及び／又は果実汁のｐＨ範囲により、ｐＨ４．０未満のグループである、グレープフルーツ、オレンジ、リンゴ、ブドウ、イチゴ、パイナップル、キウイ、グアバ、マ
ンゴー、アセロラ、ブルーベリー、ザクロ等、ｐＨ４．０以上ｐＨ４．６未満のグループであるトマト、赤ピーマン、桃、洋なし、パパイヤ等、ｐＨ４．６以上ｐＨ７．０以下のグループであるニンジン、キャベツ、ほうれん草、ケール、プチヴェール（商標：ケールと芽キャベツの交配種）、メロン、スイカ、バナナ、イチジク等に分類することも可能である。
<1-1> Vegetable juice and / or fruit juice used for the chilled beverage of the present invention (hereinafter also referred to as beverage ingredients)
The beverage ingredients used in the chilled beverage of the present invention include grapefruit, orange, apple, grape, strawberry, pineapple, kiwi, guava, mango, acerola, blueberry, pomegranate, peach, pear, papaya, melon, watermelon, banana, fig. And / or vegetable juices such as tomato, red pepper, carrot, cabbage, spinach, kale, and petit veil (trademark: hybrid of kale and brussels sprouts). Moreover, the said drink raw material is the group below pH 4.0 according to the pH range of the vegetable juice and / or fruit juice, grapefruit, orange, apple, grape, strawberry, pineapple, kiwi, guava, mango, acerola, blueberry Such as pomegranate, pomegranate, tomato, red pepper, peach, pear, papaya, etc., carrot, cabbage, spinach, kale, petit It is also possible to classify into veil (trademark: hybrid of kale and brussels sprouts), melon, watermelon, banana, fig and the like.
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が、飲料原料がストレート換算で５０％以上１００％以下で含まれることが好ましく、９０％以上１００％以下で含まれることが更に好ましい。なお、ストレート換算で飲料原料１００％という値は、濃縮液を用いれば可能である。従って、本飲料原料はストレート液であっても濃縮液であってもかまわない。ここでいう、ストレート液とは、非濃縮液のことを意味し、濃縮液とは、特に濃縮の方法に限定されることはないが、原料（果物や野菜）から絞ったそのままの果汁、野菜汁（ストレート果汁、ストレート野菜汁）を、逆浸透濃縮や前進凍結濃縮のように低加熱濃縮したものであることが好ましい。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、「％」は質量／体積（Ｗ／Ｖ）％を示す。
本発明の野菜汁及び／又は果実汁をベースとする飲料組成からなるチルド飲料において、“野菜汁及び／又は果実汁をベースとする”とは、当該野菜汁及び／又は果実汁を主要原料としていることを意味する。
Although there is no restriction | limiting in particular in the ratio with respect to the whole chilled drink of the drink raw material used for this invention, It is preferable that a drink raw material is contained in 50% or more and 100% or less in straight conversion, and it is further contained in 90% or more and 100% or less. preferable. In addition, the value of 100% of beverage ingredients in straight conversion is possible if a concentrated liquid is used. Therefore, the beverage ingredient may be a straight liquid or a concentrated liquid. The straight liquid here means a non-concentrated liquid, and the concentrated liquid is not particularly limited to the concentration method, but it is a fruit juice or vegetable that is squeezed from the raw material (fruit or vegetable). It is preferable that juice (straight fruit juice, straight vegetable juice) is obtained by concentrating with low heat like reverse osmosis concentration or forward freeze concentration. In addition, the beverage ingredients can be used alone or in combination of two or more. In the present specification, unless otherwise specified, “%” indicates mass / volume (W / V)%.
In the chilled beverage having a beverage composition based on vegetable juice and / or fruit juice of the present invention, “based on vegetable juice and / or fruit juice” means that the vegetable juice and / or fruit juice is the main ingredient. Means that
＜２＞本発明に利用する乳酸菌
本発明のチルド飲料は、野菜汁及び／又は果実汁をベースとする飲料組成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 vegetable juice and / or fruit juice, and after sterilizing the beverage composition, It is a chilled beverage characterized by being added in a state and cooled to a condition where lactic acid fermentation is not performed. 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. From the fact that there is little influence on taste, fragrance, color tone and simple preparation without the process of collecting bacteria is possible.
The method of adding the lactic acid bacteria culture solution obtained by the method of 4) to this invention chilled drink as it is is preferable. 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、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、色合い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, a beverage composition based on a 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, aroma and color of the material.
＜３−１＞殺菌
加熱殺菌は、野菜汁、果実汁の着色、並びに加熱臭を発生させ、製品の風味を変え、外観にも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、色合い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、飲料原料をベースとする飲料組成物のｐＨによって変化させることが好ましく、例えば、飲料原料をベースとする飲料組成物が４．０未満の場合は、６５℃で０．１７分〜６５℃で１０分に相当する殺菌条件で殺菌処理する。また、飲料原料をベースとする飲料組成物がｐＨ４．０以上４．６未満の場合は、６５℃で０．１７分〜７５℃で１分に相当する殺菌条件で殺菌処理する。さらに、飲料原料をベースとする飲料組成物がｐＨ４．６以上７．０以下の場合は、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。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
本発明の野菜汁及び／又は果実汁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、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 vegetable juice and / or fruit juice of the present invention, the beverage composition may optionally include other various additives. Is possible. 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 liquor obtained by the above method was subjected to centrifuge (H-
103N) and centrifuged at 3,000 rpm for 15 minutes, and 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＞
オレンジ濃縮果汁（カーギル社製 ＲＩ ６５％）を蒸留水で希釈した、オレンジ果汁（ＲＩ １２％）（株式会社アタゴ製 アッベ式屈折計 ＮＡＲ−３Ｔ）、ｐＨ３．８（堀場製作所製 ｐＨメーター Ｆ−１２）を６５℃で１０分間殺菌した。この加熱殺菌したオレンジ果汁を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。一方、比較例１は実施例２と同様に調製したオレンジ果汁（ＲＩ １２％）（株式会社アタゴ製 アッベ式屈折計 ＮＡＲ−３Ｔ）、ｐＨ３．８（堀場製作所製 ｐＨメーター Ｆ−１２）を１０５℃で１分間殺菌した。この加熱殺菌したオレンジ果汁を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。本実施例２の製品を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および酸分（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、容器に充填直後の色調評価（Ｌ値、ａ値、ｂ値，ａ／ｂ値）および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>
Orange juice (RI 12%) (Atago Co., Ltd. Abbe refractometer NAR-3T), pH 3.8 (Horiba, Ltd., pH meter F-), diluted with distilled water, concentrated orange juice (RI 65%, manufactured by Cargill) 12) was sterilized at 65 ° C. for 10 minutes. After cooling this heat-sterilized orange juice 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 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, Comparative Example 1 was prepared in the same manner as in Example 2 with orange juice (RI 12%) (Abago Abbe Refractometer NAR-3T), pH 3.8 (Horiba Seisakusho pH Meter F-12) 105. Sterilized at ° C for 1 minute. After cooling this heat-sterilized orange juice 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 this 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. Proliferation confirmation was performed. For the products of Example 2 and Comparative Example 1 , the color tone evaluation (L value, a value, b value, a / b value) immediately after filling the container and the sensory evaluation of the product stored chilled for 14 days were performed.
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。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。
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. 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.
色調評価（Ｌ値、ａ値、ｂ値、ａ／ｂ値）は以下の方法で実施した。すなわち、実施例
２および比較例１とも、容器に充填後０日目のサンプルのＬ値、ａ値、ｂ値を分光測色型測色計（ＣＭ−３５００ｄ：ミノルタ社製）にて、本機器の取扱説明書の記載に従い計測した。
Color tone evaluation (L value, a value, b value, a / b value) was carried out by the following method. That is, in both Example 2 and Comparative Example 1, the L value, a value, and b value of the sample on the 0th day after filling the container were measured with a spectrocolorimetric colorimeter (CM-3500d: manufactured by Minolta). Measurements were made according to the instructions in the equipment instruction manual.
官能評価は、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。 The sensory evaluation compares the beverages of the above example and the beverages of the comparative examples at two points. The sensory evaluators of 20 men and 20 women (40 people in total) select which one is preferable, and the beverages of each example The number of people who chose to be preferred and the risk factor as its significance test were determined. The risk rate means the probability of making the first type of error in the statistical hypothesis test.
実施例３： 洗浄した完熟トマトを破砕し、直ちに８５℃達温まで加熱して酵素失活した後、開口径２ｍｍΦのパルパーで搾汁したトマトジュース ＲＩ ６％（株式会社アタゴ製 アッベ式屈折計 ＮＡＲ−３Ｔ）、ｐＨ４．４（堀場製作所製 ｐＨメーター Ｆ−１２）を７５℃で１分間殺菌した。この加熱殺菌したトマトジュースを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と同様に搾汁したトマトジュース
ＲＩ ６％（株式会社アタゴ製 アッベ式屈折計 ＮＡＲ−３Ｔ）、ｐＨ４．４（堀場製作所製 ｐＨメーター Ｆ−１２）を１２１．１℃で４５秒殺菌した。この加熱殺菌したトマトジュースを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、比較例１と同様に実施した。
Example 3: The washed ripe tomato was crushed and immediately heated to 85 ° C to inactivate the enzyme, then tomato juice RI 6% squeezed with a pulper having an opening diameter of 2 mmΦ (Abbe refractometer manufactured by Atago Co., Ltd.) NAR-3T), pH 4.4 (pH meter F-12 manufactured by Horiba, Ltd.) was sterilized at 75 ° C. for 1 minute. After cooling this heat-sterilized tomato juice 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 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, tomato juice RI 6% (Abago Abbe refractometer NAR-3T manufactured by Atago Co., Ltd.), pH 4.4 (Horiba, Ltd. pH meter F-12) 121.1 ° C. was squeezed in the same manner as Example 3. For 45 seconds. After cooling this heat-sterilized tomato juice 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. The product of Comparative Example 2 was prepared by cooling. 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４： 洗浄したメロンを破砕し、直ちに９０℃達温まで加熱して酵素失活した後、開口径１．５ｍｍΦのパルパー及び開口径０．５ｍｍのフィニッシャーで搾汁したメロン果汁、ＲＩ １２％（株式会社アタゴ製 アッベ式屈折計 ＮＡＲ−３Ｔ）、ｐＨ５．１（堀場製作所製 ｐＨメーター Ｆ−１２）を１２１．１℃で１０秒間殺菌した。この加熱殺菌したメロン果汁を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３は実施例４と同様に搾汁したメロン果汁、ＲＩ １２％（株式会社アタゴ製 アッベ式屈折計 ＮＡＲ−３Ｔ）、ｐＨ５．１（堀場製作所製 ｐＨメーター Ｆ−１２）を１２１．１℃で３．１分間殺菌した。この加熱殺菌したメロンジュースを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３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、比較例１と同様に実施した。 Example 4: The washed melon was crushed, immediately heated to 90 ° C. and heated to inactivate the enzyme, and then squeezed with a pulper having an opening diameter of 1.5 mmΦ and a finisher having an opening diameter of 0.5 mm, RI 12 % (Abbe refractometer NAR-3T manufactured by Atago Co., Ltd.), pH 5.1 (pH meter F-12 manufactured by Horiba, Ltd.) was sterilized at 121.1 ° C. for 10 seconds. After cooling this heat-sterilized melon juice 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 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 3, melon juice squeezed in the same manner as in Example 4, RI 12% (Abago Abbe refractometer NAR-3T, manufactured by Atago Co., Ltd.), pH 5.1 (Horiba, Ltd., pH meter F-12) 121.1 Sterilized at ° C for 3.1 minutes. After cooling the heat-sterilized melon juice 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 3 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 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２〜４および比較例１〜３の結果＞
実施例２〜４および比較例１〜３の結果を表１、表２、表３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、実施例４の乳酸菌の生菌数と保存日数の関係を図５に酸分と保存日数の関係を図６に示す。図２，４，６に示されるように、実施例２，実施例３、実施例４のいずれの温度（５℃、１０℃）での保存においても、酸分の上昇は０．０５％（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。また、表３の色調評価からは、実施例２に対し比較例１は暗く、赤褐色がかっていることが、実施例３に対し比較例２は暗く、茶褐色がかっていることが、実施例４に対し比較例３は暗く、黄褐色がかっていることが示された。これらの結果より、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、色合いを保持できることが示された。
<Results of Examples 2 to 4 and Comparative Examples 1 to 3>
The results of Examples 2 to 4 and Comparative Examples 1 to 3 are summarized in Table 1, Table 2, and Table 3. Regarding the acidity increase, the acid content on the 0th day after filling and the 14th day after filling (lactic acid equivalent, W / V%) is compared, and the acidity rise is 0.1% (lactic acid equivalent: 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. FIG. 4 shows the relationship between the acid content and the number of storage days, FIG. 5 shows the relationship between the number of viable bacteria and the storage days of Example 4, and FIG. 6 shows the relationship between the acid content and the storage days. As shown in FIGS. 2, 4, and 6, the increase in acid content is 0.05% when stored at any temperature (5 ° C., 10 ° C.) in Examples 2, 3 and 4 (5 ° C., 10 ° C.). Lactic acid conversion, W / V%) or less, indicating that lactic acid fermentation was not performed. Moreover, from the color tone evaluation of Table 3, it was found in Example 4 that Comparative Example 1 was darker and reddish brown than Example 2, and that Comparative Example 2 was darker and brownish than Example 3. On the other hand, Comparative Example 3 was dark and shown to be tan. From these results, the chilled beverage of the present invention is sterilized under reduced sterilization strength, and after adding lactic acid bacteria, it is cooled and stored (refrigerated) to a condition where lactic acid fermentation is not performed, so that the contaminating bacteria grow in the beverage. It was shown that the original taste, fragrance and color of the material can be retained.
＜試験例１〜１２＞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と同様に搾汁したオレンジ果汁（ＲＩ １２％、ｐＨ３．８）を１０５℃で１分間殺菌した。この加熱殺菌したオレンジ果汁を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４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４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 1-12> Growth Inhibitory Activity of Contaminated Bacteria Added by Lactic Acid Bacteria Test Examples 1-4:
Orange juice (RI 12%, pH 3.8) squeezed in the same manner as in Example 2 was sterilized at 105 ° C. for 1 minute. After cooling this heat-sterilized orange juice to 10 ° C., the transparent milk fermented liquid prepared in Example 1 <1> had a viable number of lactic acid bacteria of 1.0 × 10 6 cells / ml and the number of contaminating bacteria shown in Table 4. Test samples 1 and 3 were prepared by adding 1.0 × 10 2 pieces / 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 2 and 4 were prepared by adding only the contaminating bacteria shown in Table 4 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と同様に搾汁したトマトジュース（ＲＩ ６％、ｐＨ４．４）を１２１．１℃で４５秒間殺菌した。この加熱殺菌したトマトジュースを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４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４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:
Tomato juice (RI 6%, pH 4.4) squeezed in the same manner as in Example 3 was sterilized at 121.1 ° C. for 45 seconds. After the heat-sterilized tomato juice 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 4 was 1. It added so that it might become 0x10 < 2 > piece / ml, and after filling a container, it cooled to 5-10 degreeC, and prepared Test Example 5 and 7 sample. 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 4 to 1.0 × 10 2 cells / ml in the 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.
試験例９〜１２：
通常の方法で搾汁したメロン果汁（ＲＩ １２％、ｐＨ５．１）を１２１．１℃で３．１分間殺菌した。この加熱殺菌したメロン果汁を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４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９および１１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１０および１２試料は、試験例９および１１試料において、表４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 9-12:
Melon juice (RI 12%, pH 5.1) squeezed by a normal method was sterilized at 121.1 ° C. for 3.1 minutes. The heat-sterilized melon juice was cooled to 10 ° C., and the transparent milk fermented liquid prepared in Example 1 <1> had a viable lactic acid bacteria count of 1.0 × 10 6 cells / ml and the contaminating bacteria count shown in Table 4 was 1. It added so that it might become 0x10 < 2 > piece / ml, and after filling a container, it cooled to 5-10 degreeC, and prepared the test example 9 and 11 sample. Thereafter, changes in the numbers of lactic acid bacteria and contaminating bacteria were observed for 14 days. Test examples 10 and 12 were prepared by adding only the contaminating bacteria shown in Table 4 to 1.0 × 10 2 / ml in test examples 9 and 11 samples . 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１〜１２において使用する汚染菌を表４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、１２１℃、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（普通寒天培地（日水製薬（株）製）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 4 shows the contaminating bacteria used in Test Examples 1-12. 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, ordinary containing Bacillus cereus 1501 strain (distributed from the Japan Canning Association) (soil extract (suspending 200 g of field soil in 1000 ml of distilled water, heating at 121 ° C. for 4 hours, and filtering the supernatant)) Using an agar medium (ordinary agar medium (manufactured by Nissui Pharmaceutical Co., Ltd.) 35 g, the above-mentioned soil extract 250 ml, distilled water 750 ml mixed and sterilized at 121 ° C. for 15 minutes), the cells were cultured at 35 ° C. for 1 week. It was prepared by suspending it in physiological saline.
結果を図７〜図１２に示す。試験例１、３、５，７，９，１１においては、汚染菌が調製直後から１４日目まで殆ど増殖しなかった。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２，４，６，８，１０，１２においては、試験例２、４は試験例１，３とほぼ同様に汚染菌数が減少したが，試験例６，８，１０，１２は、試験例５，７，９，１１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１０〜１５０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, 7, 9, and 11, the contaminating bacteria hardly grew from immediately after preparation until the 14th day. On the other hand, in Test Examples 2, 4, 6, 8, 10, and 12 in which only the number of contaminating bacteria was added to be 1.0 × 10 2 / ml, Test Examples 2 and 4 were almost the same as Test Examples 1 and 3. In contrast to Test Examples 5, 7, 9, and 11, in Test Examples 6, 8, 10, and 12, the number of contaminated bacteria began to increase immediately after preparation, and 14 days after preparation. The eye increased to about 10 to 150 times that at the time of preparation. 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.
Claims (4)
条件：前記飲料組成物がｐＨ４．０未満である場合は、６５℃で０．１７分〜６５℃で１０分に相当する殺菌処理、また、前記飲料組成物がｐＨ４．０以上４．６未満の場合は、６５℃で０．１７分〜７５℃で１分に相当する殺菌処理、更に、前記飲料組成物がｐＨ４．６以上７．０以下の場合は、６５℃で０．１７分〜１２１．１℃で０．３分に相当する殺菌処理。 A method for producing a chilled beverage comprising a beverage composition based on vegetable juice and / or fruit juice, wherein the concentration of lactic acid produced by lactic acid fermentation is less than 0.1% (lactic acid equivalent: W / V%) , After sterilizing the beverage composition under the following conditions, after cooling to 5 to 40 ° C., lactic acid bacteria are added so that the viable cell count is in the range of 1 × 10 6 to 1 × 10 9 cells / ml, and lactic acid A method for producing a chilled beverage characterized by cooling to a condition that does not allow fermentation.
Conditions: When the beverage composition has a pH of less than 4.0, a sterilization treatment corresponding to 0.17 minutes at 65 ° C to 10 minutes at 65 ° C, and the beverage composition has a pH of 4.0 or more and less than 4.6. In the case of sterilization treatment corresponding to 1 minute at 65 ° C. for 0.17 minutes to 75 ° C., and further when the beverage composition has a pH of 4.6 or more and 7.0 or less, it is 0.17 minutes at 65 ° C. Sterilization treatment corresponding to 0.3 minutes at 121.1 ° C.
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