JP6362845B2 - Beverages containing lactic acid bacteria - Google Patents
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本発明は、保存性の向上した乳酸菌を含有する飲料に関する。具体的に本発明は、保存中における乳酸菌数の減少が抑制された乳酸菌を含有する飲料に関する。 The present invention relates to a beverage containing lactic acid bacteria having improved storage stability. Specifically, the present invention relates to a beverage containing lactic acid bacteria in which a decrease in the number of lactic acid bacteria during storage is suppressed.
人類の歴史上、乳酸菌は、ヨーグルトなどの発酵乳製品、各種漬物類、鮒寿司など、多くの加工飲食品において、味や風味の付与、栄養の強化、食品の保存性改善など様々な目的で用いられてきた。また、乳酸菌は、生きた状態で摂取することで、腸内フローラが改善されると同時に、乳酸菌やその産生物質により人体に有用な効能をもたらすことが知られている。 In the history of mankind, lactic acid bacteria are used for various purposes such as fermented dairy products such as yogurt, various pickles, sushi, and other processed foods and beverages, such as imparting taste and flavor, enhancing nutrition, and improving food storage stability. Has been used. In addition, it is known that lactic acid bacteria, when ingested in a living state, improve intestinal flora, and at the same time bring useful effects to the human body by lactic acid bacteria and their products.
しかしながら、乳酸菌を生きた状態で摂取できる飲食品は賞味期限を気にしなければならない場合がある。すなわち、乳酸菌の中には低ｐＨや冷蔵保存の環境下で経時的に生菌数が減少するものがあり、このような乳酸菌を製品に用いると、保存中に生菌数が製品設計上の下限値を下回ってしまう（例えば、乳酸菌飲料の成分規格上必要とされる乳酸菌数が１．０×１０６／ｍｌ未満となる）。前記のような必要以上の乳酸菌生菌数の減少は、飲食品の賞味期限の設定を短縮しなければならない原因となる。 However, foods and drinks that can be ingested with lactic acid bacteria alive may have to worry about the expiration date. In other words, there are some lactic acid bacteria whose viable cell counts decrease over time under low pH or refrigerated storage conditions. When such lactic acid bacteria are used in products, the viable cell counts may be It falls below the lower limit value (for example, the number of lactic acid bacteria required for component specifications of lactic acid bacteria beverages is less than 1.0 × 10 6 / ml). The above-mentioned decrease in the number of viable lactic acid bacteria more than necessary is a cause of having to shorten the setting of the shelf life of food and drink.
かかる問題を解決するために、（１）製造時の乳酸菌生菌数を多くした状態で製品を出荷する方法、（２）保存中に乳酸菌生菌の生残性を向上する方法、のいずれの方法が利用されている。 In order to solve such a problem, any one of (1) a method of shipping a product in a state where the number of viable lactic acid bacteria at the time of production is increased, and (2) a method of improving the viability of the live lactic acid bacteria during storage The method is being used.
これら方法の内、（１）の方法は、生菌数を長期間一定以上に保つことができるが、製造時及び保存初期に乳酸菌による発酵が通常よりも盛んに行われることで、製品の酸味や発酵臭が強まり、結果として製品の賞味期限が短縮される。 Among these methods, the method (1) can keep the number of viable bacteria above a certain level for a long time, but the fermentation of lactic acid bacteria is more active than usual at the time of production and storage, so that the acidity of the product is increased. As a result, the product expiration date is shortened.
一方、（２）の方法では、さらに、（ア）スクリーニング等による生残性の高い乳酸菌の選抜・育種、（イ）共発酵、（ウ）生残性を低下させる因子の除去、（エ）生残性向上物質の添加、などの方法が利用されている。これらのうち、（ア）〜（ウ）の方法は、生菌の生残性向上に高い効果を示すが、利便性が非常に低い。（エ）の方法は、利便性の面で優れており、添加する物質の例として、エリスリトール（特許文献１）、パーオキシダーゼ（特許文献２）、ドロマイト（特許文献３）、ビタミンＣおよび／またはビタミンＥ（特許文献４）、リン酸カルシウム（特許文献５）、リン脂質（特許文献６）、マンガン（特許文献７）、イヌリン（特許文献８）、低分子化処理ガラクトマンナン（特許文献９）など多数が知られている。しかしながら、生残性向上物質の添加による方法では、製品の味質や風味、物性などに影響を与えることもあるので、添加する物質の選択や添加量を慎重に検討しなければならない。 On the other hand, in the method (2), (a) selection and breeding of lactic acid bacteria having high survival by screening and the like, (b) co-fermentation, (c) removal of factors that reduce survival, (d) Methods such as addition of a survival-enhancing substance are used. Among these, the methods (a) to (c) are highly effective in improving the viability of viable bacteria but are very low in convenience. The method (d) is excellent in terms of convenience, and examples of the substance to be added include erythritol (Patent Document 1), peroxidase (Patent Document 2), dolomite (Patent Document 3), vitamin C and / or Vitamin E (patent document 4), calcium phosphate (patent document 5), phospholipid (patent document 6), manganese (patent document 7), inulin (patent document 8), low molecular weight treated galactomannan (patent document 9) and many others It has been known. However, the method of adding a survival-enhancing substance may affect the quality, flavor, and physical properties of the product, so the selection of the substance to be added and the amount to be added must be carefully considered.
また、乳酸菌やビフィズス菌を多く含む発酵乳飲料に、一定量の乳酸カルシウム等のカルシウム塩を添加することで発酵乳飲料の異臭味をマスキングする方法が知られている（特許文献１０）。さらに、本発明者らは、乳酸菌を含む飲料に乳酸カルシウムを添加することで乳酸菌の増殖を抑制できることも見出している。
しかしながら、乳酸カルシウムが乳酸菌を含む飲料において乳酸菌数の減少も抑制できることはこれまで知られていなかった。
Moreover, the method of masking the off-flavor of a fermented milk drink is known by adding a fixed quantity of calcium salts, such as calcium lactate, to fermented milk drink which contains many lactic acid bacteria and bifidobacteria (patent document 10). Furthermore, the present inventors have also found that the growth of lactic acid bacteria can be suppressed by adding calcium lactate to a beverage containing lactic acid bacteria.
However, it has not been known so far that calcium lactate can suppress a decrease in the number of lactic acid bacteria in a beverage containing lactic acid bacteria.
本発明は、これまで知られていない極めて簡便な方法により、最終製品の味質や風味、物性に影響を与えないで、保存時における乳酸菌生菌数の経時的な減少が抑制された乳酸菌を含有する飲料を提供すること、及び、乳酸菌生菌数の経時的な減少を抑制する方法を提供することを目的とする。 The present invention provides a lactic acid bacterium in which the decrease in the number of viable lactic acid bacteria over time is suppressed without affecting the taste quality, flavor, and physical properties of the final product by a very simple method that has not been known so far. It aims at providing the drink which contains and the method of suppressing the time-dependent decrease of the number of living lactic acid bacteria.
本発明者らは、上記のごとき課題を解決すべく鋭意研究した結果、乳酸菌を含有する飲料に一定量の乳酸カルシウムを含有することにより、味質への影響が極めて軽微であり、さらに長期間保存しても乳酸菌生菌数の減少が効果的に抑制され、結果的に保存性の向上した乳酸菌を含有する飲料が得られることを見出した。 As a result of diligent research to solve the above-described problems, the present inventors have found that a beverage containing lactic acid bacteria contains a certain amount of calcium lactate, so that the influence on taste quality is extremely slight, and for a longer period of time. It was found that even when stored, a decrease in the number of viable lactic acid bacteria was effectively suppressed, and as a result, a beverage containing lactic acid bacteria with improved storage stability was obtained.
すなわち、本発明は、以下の態様を有する乳酸菌を含有する飲料に関する。
（１）乳酸カルシウムを０．１〜３．０質量％含有し、１０℃以下で生菌数が減少する乳酸菌を含有する飲料。
（２）乳酸カルシウムを含有し、該乳酸カルシウムの含有量が０．３６質量％より大きく２．０質量％以下である、（１）に記載の乳酸菌を含有する飲料。
That is, this invention relates to the drink containing the lactic acid bacteria which have the following aspects.
(1) A beverage containing lactic acid bacteria containing 0.1 to 3.0% by mass of calcium lactate and having a viable cell count reduced at 10 ° C or lower.
(2) The beverage containing lactic acid bacteria according to (1), which contains calcium lactate and the content of the calcium lactate is greater than 0.36% by mass and not more than 2.0% by mass.
また、本発明は、以下の態様を有する乳酸菌を含有する飲料における乳酸菌の減少抑制方法に関する。
（３）乳酸カルシウムを添加することによる乳酸菌を含有する飲料における乳酸菌数の減少抑制方法。
（４）乳酸カルシウムを０．１〜３．０質量％添加することによる、（３）に記載の乳酸菌を含有する飲料における乳酸菌数の減少抑制方法。
（５）乳酸カルシウムの添加量が０．３６質量％より大きく２．０質量％以下である、（３）に記載の乳酸菌を含有する飲料における乳酸菌数の減少抑制方法。
（６）前記飲料に含有される乳酸菌が、１０℃以下で生菌数が減少する乳酸菌である、（３）〜（５）のいずれかに記載の乳酸菌を含有する飲料における乳酸菌数の減少抑制方法。
Moreover, this invention relates to the reduction | decrease suppression method of the lactic acid bacteria in the drink containing the lactic acid bacteria which has the following aspects.
(3) A method for suppressing a decrease in the number of lactic acid bacteria in a beverage containing lactic acid bacteria by adding calcium lactate.
(4) The method for suppressing a decrease in the number of lactic acid bacteria in a beverage containing the lactic acid bacteria according to (3), wherein 0.1 to 3.0% by mass of calcium lactate is added.
(5) The method for suppressing a decrease in the number of lactic acid bacteria in a beverage containing lactic acid bacteria according to (3), wherein the addition amount of calcium lactate is greater than 0.36% by mass and 2.0% by mass or less.
(6) Suppression of decrease in the number of lactic acid bacteria in a beverage containing the lactic acid bacteria according to any one of (3) to (5), wherein the lactic acid bacteria contained in the beverage are lactic acid bacteria whose viable cell count is reduced at 10 ° C. or lower. Method.
本発明の乳酸菌を含有する飲料によれば、極めて簡便な方法、かつ、該飲料の味質や風味、物性への影響を極めて軽微に抑えることができる方法により、店頭に商品が陳列しているときや家庭での保管中など、該飲料を保存した際に、乳酸菌生菌数の減少を抑制し、長期間にわたり一定以上の乳酸菌数を保つことができる、乳酸菌を含有する飲料を提供することができる。 According to the beverage containing the lactic acid bacterium of the present invention, products are displayed at the storefront by a very simple method and a method that can suppress the influence on the taste quality, flavor, and physical properties of the beverage. To provide a beverage containing lactic acid bacteria that can suppress the decrease in the number of viable lactic acid bacteria and maintain a certain number of lactic acid bacteria over a long period of time when the beverage is stored, such as during storage at home or at home. Can do.
本発明における「乳酸菌を含有する飲料」とは、乳酸菌の生菌数が１．０×１０６／ｍｌ以上である飲料を示し、乳酸菌が生菌状態で含まれているものを示す。したがって、本発明の乳酸菌を含有する飲料には、乳及び乳製品の成分規格等に関する省令（以下、「乳等省令」という）で定められるはっ酵乳及び乳酸菌飲料を当然包含するものである。 The “beverage containing lactic acid bacteria” in the present invention refers to a beverage in which the number of viable bacteria of lactic acid bacteria is 1.0 × 10 6 / ml or more, and indicates that the lactic acid bacteria are contained in a viable state. Therefore, the beverage containing the lactic acid bacteria of the present invention naturally includes fermented milk and lactic acid bacteria beverages defined by a ministerial ordinance concerning the component specifications of milk and dairy products (hereinafter referred to as “Ministerial Ordinance for Milk”).
なお、本発明における乳酸菌を含有する飲料は、乳成分の有無を問わないが、乳成分を配合する場合、牛乳等の生乳、脱脂粉乳、全脂粉乳、調製粉乳、ホエイパウダー、バターミルクパウダー、クリームパウダー、濃縮乳、生クリーム、無糖練乳、加糖全脂練乳、加糖脱脂練乳、バター、脱脂乳などから選択される１又は２以上を配合することができる。 The beverage containing lactic acid bacteria in the present invention may or may not have milk components, but when milk components are blended, raw milk such as milk, skim milk powder, whole milk powder, prepared milk powder, whey powder, butter milk powder, One or more selected from cream powder, concentrated milk, fresh cream, unsweetened condensed milk, sweetened whole fat condensed milk, sweetened defatted condensed milk, butter, nonfat milk and the like can be blended.
本発明においては、乳酸菌を含有する飲料に乳酸カルシウムが含有されていることを特徴とする。乳酸菌を含有する飲料における乳酸カルシウムの含有量としては、０．１〜３．０質量％が例示され、好ましくは０．３〜２．０質量％、さらに好ましくは０．３６より大きく２．０質量％以下である。すなわち、乳酸カルシウムが前記の含有量となるように、乳酸菌を含有する飲料の製造工程において乳酸カルシウムを添加する。
なお、乳酸カルシウムの含有量は、０．１質量％未満であると乳酸菌の減少抑制効果が不十分であり、０．３質量％以上であると乳酸菌の減少抑制効果が顕著に発揮される。また、乳酸カルシウムの含有量が３．０質量％を超えると乳酸カルシウム特有の苦味や収斂味が目立ってしまう。
In the present invention, the beverage containing lactic acid bacteria contains calcium lactate. As content of the calcium lactate in the drink containing a lactic acid bacterium, 0.1-3.0 mass% is illustrated, Preferably it is 0.3-2.0 mass%, More preferably, it is larger than 0.36 and 2.0. It is below mass%. That is, calcium lactate is added in the production process of a beverage containing lactic acid bacteria so that the calcium lactate has the above content.
In addition, if the content of calcium lactate is less than 0.1% by mass, the effect of suppressing the decrease of lactic acid bacteria is insufficient, and if it is 0.3% by mass or more, the effect of suppressing the decrease of lactic acid bacteria is remarkably exhibited. On the other hand, when the content of calcium lactate exceeds 3.0% by mass, the bitterness and astringent taste peculiar to calcium lactate become conspicuous.
本発明で使用する乳酸菌については、特に制限はなく、発酵飲食品等に用いられる乳酸菌であればいずれも単独又は混合して使用することができる。その具体例として、ラクトバチルス属（Lactobacillus）、ビフィドバクテリウム属（Bifidobacterium）、エンテロコッカス属（Enterococcus）、ラクトコッカス属（Lactococcus）、ペディオコッカス属（Pediococcus）、リューコノストック属（Leuconostoc）、ウィッセラ属（Weissella）などが挙げられる。
また、本発明で使用する乳酸菌として、好ましくは、１０℃以下で生菌数が減少する（静菌状態となる、若しくは死滅する）乳酸菌が挙げられ、例えば、ラクトバチルス・カゼイ（Lactobacillus casei）が挙げられる。
There is no restriction | limiting in particular about the lactic acid bacteria used by this invention, All can be used individually or in mixture, if it is a lactic acid bacterium used for fermented food / beverage products. Specific examples thereof include Lactobacillus, Bifidobacterium, Enterococcus, Lactococcus, Pediococcus, Leuconostoc, Examples include the genus Weissella.
The lactic acid bacterium used in the present invention is preferably a lactic acid bacterium whose viable count decreases (becomes bacteriostatic or dies) at 10 ° C. or lower, for example, Lactobacillus casei. Can be mentioned.
本発明における乳酸菌を含有する飲料において、乳酸菌の生菌数は、前述のとおり１．０×１０６／ｍｌ以上であることを要するが、上限を１．０×１０９／ｍｌ未満とすることが好ましい。乳酸菌の生菌数が１．０×１０９／ｍｌ以上であると、乳酸菌による乳酸生成が過剰となり、一定期間以上保存した際の酸度が過度に大きくなってしまう場合がある。 In the beverage containing lactic acid bacteria in the present invention, the number of viable bacteria of lactic acid bacteria is required to be 1.0 × 10 6 / ml or more as described above, but the upper limit should be less than 1.0 × 10 9 / ml. Is preferred. When the number of viable bacteria of lactic acid bacteria is 1.0 × 10 9 / ml or more, lactic acid production by lactic acid bacteria becomes excessive, and the acidity when stored for a certain period or more may become excessively large.
また、本発明の乳酸菌を含有する飲料は、ここまでに掲げた成分のほかに、味質の調整や安定性の付与のため、乳酸菌を含有する飲料に一般的に用いられる各種食品素材や食品添加物を添加することができる。例えば、野菜汁、野菜ペースト、果汁、果実ペースト、穀物抽出物、茶抽出物、コーヒー抽出物、香辛料抽出物等の食品素材、甘味料、増粘剤、乳化剤、酸味料、ビタミン類、香料等の食品添加物が挙げられる。 In addition to the ingredients listed so far, the beverage containing the lactic acid bacteria of the present invention includes various food materials and foods commonly used for beverages containing lactic acid bacteria in order to adjust the taste and impart stability. Additives can be added. For example, vegetable juice, vegetable paste, fruit juice, fruit paste, grain extract, tea extract, coffee extract, spice extract and other food materials, sweeteners, thickeners, emulsifiers, acidulants, vitamins, fragrances, etc. Food additives.
本発明の乳酸菌を含有する飲料は、これら飲料で通常用いられる方法により製造することができ、一例として、殺菌した乳成分含有培地に乳酸菌スターターを接種・培養し、これに、乳酸カルシウムや各種食品素材及び食品添加物を含有し殺菌したシロップとを混合した後、均質化処理することで得られる。 The beverage containing the lactic acid bacteria of the present invention can be produced by a method usually used in these beverages. As an example, a lactic acid bacteria starter is inoculated and cultured in a sterilized milk component-containing medium. It is obtained by mixing sterilized syrup containing raw materials and food additives and then homogenizing.
本発明の乳酸菌を含有する飲料は、ｐＨを酸性としても乳酸菌生菌数の減少が抑えられるため、酸味料や果汁、野菜汁を含む飲料であっても、冷蔵条件で通常よりも長期間保存可能な生きた乳酸菌を含有する飲料を製造することができる。 Since the beverage containing the lactic acid bacteria of the present invention can suppress the decrease in the number of viable lactic acid bacteria even if the pH is acidic, even beverages containing acidulants, fruit juices, and vegetable juices are stored for a longer period of time than usual under refrigerated conditions. Beverages containing possible live lactic acid bacteria can be produced.
以上のようにして得られた本発明の乳酸菌を含有する飲料は、個別の包装容器に充填することで最終製品が得られ、冷蔵条件で例えば２週間以上の長期間保存したとしても、味質等に大きく影響を及ぼすことなく乳酸菌生菌数の減少が抑えられた飲料を得ることができる。 Even if the beverage containing the lactic acid bacteria of the present invention obtained as described above is filled into individual packaging containers to obtain the final product and stored for a long period of time, such as 2 weeks or longer under refrigerated conditions, Thus, it is possible to obtain a beverage in which a decrease in the number of viable lactic acid bacteria is suppressed without greatly affecting the above.
以下、本発明の内容を実施例、比較例等を用いて具体的に説明するが、本発明はこれらに何ら限定されるものではない。なお、特に記載のない限り「％」とは「質量％」を、「部」は「質量部」をそれぞれ意味するものとする。 Hereinafter, the content of the present invention will be specifically described with reference to Examples and Comparative Examples, but the present invention is not limited to these. Unless otherwise specified, “%” means “mass%”, and “part” means “part by mass”.
実験例１：乳酸カルシウムの効果
（実施例１）
脱脂粉乳を１３．５％含む乳培地を調製し加熱殺菌後、ラクトバチルス・カゼイ及びストレプトコッカス・サーモフィルスのスターター液を該培地に接種し、３０℃で４８時間培養し、発酵液を得た。一方、ぶどう糖果糖液糖、酸味料、安定剤を含むシロップ液ベースに、乳酸カルシウムを最終濃度０．３％となるように添加し、加熱殺菌及び冷却を行った。その後、前記工程を経た乳酸カルシウム含有シロップ液ベース９６．５部、及び、前記で調製した発酵液３．５部を混合し、均質化処理することにより、実施例１の乳酸菌生菌を３．０×１０７／ｍｌ含有する飲料を得た。
Experimental Example 1: Effect of calcium lactate (Example 1)
A milk medium containing 13.5% nonfat dry milk was prepared and sterilized by heating. Then, a starter solution of Lactobacillus casei and Streptococcus thermophilus was inoculated into the medium and cultured at 30 ° C. for 48 hours to obtain a fermentation broth. On the other hand, calcium lactate was added to a syrup base containing glucose fructose liquid sugar, acidulant, and stabilizer to a final concentration of 0.3%, followed by heat sterilization and cooling. Thereafter, 96.5 parts of the calcium lactate-containing syrup solution base having undergone the above steps and 3.5 parts of the fermentation solution prepared above were mixed and homogenized to obtain 3. A beverage containing 0 × 10 7 / ml was obtained.
（比較例１）
実施例１と同じｐHとなるようクエン酸ナトリウムを添加したこと以外、実施例１と同様の原料及び製法により、比較例１の乳酸菌生菌を３．１×１０７／ｍｌ含有する飲料を得た。
(Comparative Example 1)
A beverage containing 3.1 × 10 7 / ml of live lactic acid bacteria of Comparative Example 1 was obtained by the same raw materials and production method as in Example 1 except that sodium citrate was added to the same pH as in Example 1. It was.
実施例１及び比較例１の乳酸菌を含有する飲料について、製造直後のｐＨ及び乳酸菌生菌数を測定し、１０℃で１、４、７、１４、２１及び２８日間保存した後のそれぞれの乳酸菌生菌数を測定し、２１日間保存後のｐＨを測定した。その結果を表１（ｐＨ）及び図１（乳酸菌生菌数）に示す。 About the drink containing the lactic acid bacteria of Example 1 and Comparative Example 1, the pH immediately after production and the number of living lactic acid bacteria were measured, and each lactic acid bacterium after being stored at 10 ° C. for 1, 4, 7, 14, 21, and 28 days The number of viable bacteria was measured, and the pH after storage for 21 days was measured. The results are shown in Table 1 (pH) and FIG. 1 (live lactic acid bacteria count).
実験例１の結果、乳酸カルシウムの代わりにクエン酸ナトリウムでｐＨを調整した比較例１の飲料では、１０℃で４日間保存した時点で乳酸菌生菌数が急減し、その後斬減し続けたが、乳酸カルシウムを添加した実施例１の飲料は、１０℃で２８日間保存しても乳酸菌生菌数を一定以上保っていた。また、比較例１の飲料では、１０℃で２８日間保存した時点で、生菌数が、乳等省令において無脂乳固形分３．０％未満の乳酸菌飲料として成分規格上必要とされる１．０×１０６／ｍｌを下回った。
なお、実験例１のいずれの飲料においても、ここで用いた添加物が味質、風味及び物性に大きな影響を与えることがなかった。
以上より、実験例１において、実施例１の飲料は、乳酸菌の減少を効果的に抑制できることが示された。
As a result of Experimental Example 1, in the beverage of Comparative Example 1 in which the pH was adjusted with sodium citrate instead of calcium lactate, the number of viable lactic acid bacteria rapidly decreased when stored at 10 ° C. for 4 days, and then continued to decrease. The beverage of Example 1 to which calcium lactate had been added maintained the number of viable lactic acid bacteria above a certain level even when stored at 10 ° C. for 28 days. Moreover, in the drink of Comparative Example 1, when stored at 10 ° C. for 28 days, the number of viable bacteria is required in terms of ingredient specifications as a lactic acid bacteria drink having a nonfat milk solid content of less than 3.0% according to a ministerial ordinance such as milk. Less than 0.0 × 10 6 / ml.
In any beverage of Experimental Example 1, the additive used here did not greatly affect the taste, flavor and physical properties.
From the above, in Experimental Example 1, it was shown that the beverage of Example 1 can effectively suppress the reduction of lactic acid bacteria.
実験例２：乳酸カルシウムの添加量の検討
（実施例２Ａ〜２Ｄ）
脱脂粉乳を１０％含む乳培地を調製し加熱殺菌後、ラクトバチルス・カゼイのスターター液を該培地に接種し、３０℃で４８時間培養し、発酵液を得た。一方、ぶどう糖果糖液糖、酸味料、安定剤を含むシロップ液ベースに、乳酸カルシウムの最終濃度を下記表２に掲げる濃度となるように添加し、加熱殺菌及び冷却を行った。その後、前記工程を経た乳酸カルシウム含有シロップ液ベース９５部、及び、前記で調製した発酵液５部を混合後、ｐＨを４．３に調整することで、実施例２Ａ〜２Ｄの乳酸菌生菌を１．２×１０８／ｍｌ含有する飲料を得た。
Experimental Example 2: Examination of addition amount of calcium lactate (Examples 2A to 2D)
A milk medium containing 10% nonfat dry milk was prepared, heat-sterilized, inoculated with a starter solution of Lactobacillus casei, and cultured at 30 ° C. for 48 hours to obtain a fermentation broth. On the other hand, the final concentration of calcium lactate was added to a syrup liquid base containing glucose fructose liquid sugar, acidulant, and stabilizer so as to have the concentrations listed in Table 2 below, and heat sterilization and cooling were performed. Then, after mixing 95 parts of the calcium lactate-containing syrup solution base and 5 parts of the fermented solution prepared as described above, the pH was adjusted to 4.3, whereby the living lactic acid bacteria of Examples 2A to 2D were obtained. A beverage containing 1.2 × 10 8 / ml was obtained.
（比較例２）
乳酸カルシウムを添加しないこと以外、実施例２と同様の原料及び製法により、比較例２の乳酸菌生菌を１．２×１０８／ｍｌ含有する飲料を得た。
(Comparative Example 2)
A beverage containing 1.2 × 10 8 / ml of live lactic acid bacteria of Comparative Example 2 was obtained by the same raw materials and production method as in Example 2 except that calcium lactate was not added.
実施例２Ａ〜２Ｄ及び比較例２の乳酸菌を含有する飲料について、１０℃で７、１４及び２１日間保存した後のそれぞれの乳酸菌生菌数を測定した。その結果を図２に示す。 About the drink containing the lactic acid bacteria of Example 2A-2D and the comparative example 2, each lactic acid bacteria viable count after storing for 7 to 14 days at 10 degreeC was measured. The result is shown in FIG.
実験例２の結果から、実施例２Ａ〜２Ｄのいずれの飲料についても、乳酸カルシウムを添加していない比較例２の飲料に比べて、乳酸菌生菌数の減少を抑制していることが示された。特に、乳酸カルシウム濃度が０．３％以上である実施例２Ｂ〜２Ｄの各飲料は、２１日間冷蔵保存をしても、乳酸菌生菌数が保存開始時とほぼ同程度又はそれ以上であることが示された。
From the results of Experimental Example 2, it is shown that any of the beverages of Examples 2A to 2D suppresses the decrease in the number of living lactic acid bacteria compared to the beverage of Comparative Example 2 in which calcium lactate is not added. It was. In particular, the beverages of Examples 2B to 2D having a calcium lactate concentration of 0.3% or more have approximately the same or higher viable number of lactic acid bacteria than that at the start of storage even after refrigerated storage for 21 days. It has been shown.
実験例３：菌株別の乳酸カルシウムの効果
（実施例３Ａ〜３Ｄ）
脱脂粉乳を１０％含む乳培地を調製し加熱殺菌後、下記表３に掲げる乳酸菌のスターター液を該培地に接種し、３０℃で４８時間培養し、発酵液を得た。一方、ぶどう糖果糖液糖、酸味料、安定剤及び乳酸カルシウムを含むシロップ液ベース９５部を調製し、加熱殺菌及び冷却を行った。その後、前記工程を経たシロップ液ベース９５部、及び、前記で調製した発酵液５部を混合し、クエン酸ナトリウムによりｐＨを４．３付近に調整し、均質化処理することにより、実施例３Ａ〜３Ｄの乳酸菌生菌を含有する飲料を得た。なお、当該飲料における乳酸カルシウム濃度は０．９％である。
Experimental Example 3: Effect of calcium lactate by strain (Examples 3A to 3D)
A milk medium containing 10% skim milk powder was prepared and heat-sterilized, and then the starter solution of lactic acid bacteria listed in Table 3 below was inoculated into the medium and cultured at 30 ° C. for 48 hours to obtain a fermentation broth. On the other hand, 95 parts of a syrup solution base containing glucose fructose liquid sugar, acidulant, stabilizer and calcium lactate was prepared, and heat sterilized and cooled. Thereafter, 95 parts of the syrup base obtained through the above step and 5 parts of the fermented liquid prepared above were mixed, adjusted to a pH of about 4.3 with sodium citrate, and homogenized to give Example 3A. A beverage containing ~ 3D live lactic acid bacteria was obtained. The calcium lactate concentration in the beverage is 0.9%.
（比較例３Ａ〜３Ｄ）
乳酸カルシウムを添加しないこと以外、実施例３Ａ〜３Ｄと同様の原料及び製法により、比較例３Ａ〜３Ｄの乳酸菌生菌を含有する飲料を得た。
(Comparative Examples 3A to 3D)
Except not adding calcium lactate, the drink containing the lactic-acid-bacteria bacterium of Comparative Examples 3A-3D was obtained by the same raw material and manufacturing method as Example 3A-3D.
実施例３Ａ〜３Ｄ及び比較例３Ａ〜３Ｄの乳酸菌を含有する飲料について、製造直後の乳酸菌生菌数を測定した後、１０℃で７、１４及び２１日間保存した後のそれぞれの乳酸菌生菌数を測定した。その結果を図３〜６に示す。
なお、ラクトバチルス・カゼイNYC13株は特許出願人が保有している菌株であり、他のJCM株は全て独立行政法人理化学研究所より入手した。
For beverages containing the lactic acid bacteria of Examples 3A to 3D and Comparative Examples 3A to 3D, the number of living lactic acid bacteria immediately after production was measured, and then the number of living lactic acid bacteria after storage at 10 ° C. for 7, 14 and 21 days Was measured. The results are shown in FIGS.
The Lactobacillus casei NYC13 strain is a strain owned by the patent applicant, and all other JCM strains were obtained from RIKEN.
実験例３の結果から、実施例３Ａ〜３Ｄのラクトバチルス属のいずれの菌株に対しても、乳酸カルシウム添加による乳酸菌生菌数の減少抑制効果が認められた。 From the results of Experimental Example 3, the effect of suppressing the decrease in the number of living lactic acid bacteria by adding calcium lactate was observed for any strain of the genus Lactobacillus of Examples 3A to 3D.
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