JP2007028920A - Fermented milk and method for producing the same - Google Patents
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本発明は、発酵乳およびその製造方法に関し、詳しくは、いわゆる乳難発酵性腸管由来乳酸菌を含有する発酵乳およびその製造方法に関する。 The present invention relates to fermented milk and a method for producing the same, and more particularly to fermented milk containing a so-called milk-fermentable intestinal lactic acid bacterium and a method for producing the same.
発酵乳や乳酸菌飲料は、良好な風味を有し、デザートとして広く消費者に飲食されている。また、これらは、便秘防止、下痢改善などの整腸作用、感染症の予防、免疫賦活作用など、健康上好ましい保健効果も有する健康食品として注目されている。発酵乳や乳酸菌飲料の保健効果は、どのような乳酸菌でも同じような効果を示すものではないと考えられている。 Fermented milk and lactic acid bacteria beverages have a good flavor and are widely eaten and consumed by consumers as desserts. In addition, they are attracting attention as health foods having favorable health effects on health such as bowel control such as prevention of constipation, improvement of diarrhea, prevention of infectious diseases, and immunostimulation. The health effects of fermented milk and lactic acid bacteria beverages are not considered to be similar to any lactic acid bacteria.
近年、胃酸、胆汁酸に抵抗性を示し、人の腸管内で生存可能で、消化管内微生物叢を改善することにより宿主に対して有益な保健効果を示す乳酸菌は、プロバイオティクス（乳酸菌）と呼ばれるようになっている。その多くは、動物の腸管に生息する腸管由来乳酸菌である。代表的な腸管由来乳酸菌としては、Ｌａｃｔｏｂａｃｉｌｌｕｓ ａｃｉｄｏｐｈｉｌｕｓ（以下、ＬａｃｔｏｂａｃｉｌｌｕｓはＬ．と表示する）、Ｌ．ｇａｓｓｅｒｉ、Ｌ．ｊｏｈｎｓｏｎｉｉ、Ｌ．ｃｒｉｓｐａｔｕｓ、Ｌ．ｃａｓｅｉ、Ｌ．ｐａｒａｃａｓｅｉ、Ｌ．ｓａｌｉｖａｒｉｕｓ、Ｌ．ｒｈａｍｎｏｓｕｓ等が知られている。そして、斯かる腸管由来乳酸菌を利用した乳酸菌製剤の製造法（特許文献１）や非加熱食肉製品（特許文献２）が提案されている。 In recent years, lactic acid bacteria that are resistant to gastric acid and bile acids, can survive in the human intestinal tract, and have beneficial health effects on the host by improving the gut microbiota, are probiotics (lactic acid bacteria) and It has come to be called. Many of them are intestinal lactic acid bacteria that inhabit the intestinal tract of animals. Representative intestinal lactic acid bacteria include Lactobacillus acidophilus (hereinafter Lactobacillus is denoted as L.), L. gasseri, L.M. johnsonii, L.M. crispatus, L.M. casei, L .; paracasei, L .; salivarius, L.M. rhamnosus and the like are known. And the manufacturing method (patent document 1) and the non-heated meat product (patent document 2) of the lactic acid bacteria formulation using such intestinal tract-derived lactic acid bacteria are proposed.
上記とは対照的に、これまで、一般的に発酵乳や乳酸菌飲料には、乳中での増殖に適した腸管由来乳酸菌以外の乳酸菌が好まれて使用されている。代表的なものとしては、Ｌ．ｂｕｌｇａｒｉｃｕｓ、Ｓｔｒｅｐｔｏｃｏｃｃｕｓ ｔｈｅｒｍｏｐｈｉｌｕｓ、Ｌ．ｈｅｌｖｅｔｉｃｕｓ、Ｌａｃｔｏｃｏｃｃｕｓ等が知られている。これらの乳酸菌は、胃酸、胆汁酸により死滅し生きた状態で腸管に定着しないことから、前述の整腸作用、下痢予防作用、感染症予防作用、免疫賦活作用などの保健効果は、腸管由来乳酸菌よりも弱いと一般的に考えられている。 In contrast to the above, lactic acid bacteria other than intestinal lactic acid bacteria suitable for growth in milk have been generally used for fermented milk and lactic acid bacteria beverages. Representative examples include L.P. bulgaricus, Streptococcus thermophilus, L. Helveticus, Lactococcus, etc. are known. Since these lactic acid bacteria are killed by gastric acid and bile acids and do not settle in the intestinal tract, the health effects such as the aforementioned intestinal regulating action, diarrhea prevention action, infection prevention action, immunostimulation action, etc. It is generally considered weaker.
本来、動物腸管内に生息する場合が多い腸管由来乳酸菌には、脱脂乳のような乳中での乳酸発酵が極めて悪い菌株が存在する。乳酸発酵が悪いと乳酸菌数が低いのみならず、乳酸酸度も低値となり、発酵乳の製造には適さない。このような乳難発酵性腸管由来乳酸菌は、脱脂乳などの乳中で増殖させて利用することが著しく困難である。また、酵母エキス、大豆エキス、乳タンパク分解物などの増殖促進材を添加することにより増殖する場合もあるが、その場合でも増殖促進剤によって風味が好ましくない状態になることがあり、その添加量は制限される。また、乳難発酵性腸管由来乳酸菌含有発酵乳の製造は、Ｌ．ｂｕｌｇａｒｉｃｕｓやＳｔｒｅｐｔｏｃｏｃｃｕｓ ｔｈｅｒｍｏｐｈｉｌｕｓと混合培養することにより可能であるが、これらは、乳中での発酵に適しているために増殖が早くて競合作用を呈するため、乳難発酵性腸管由来乳酸菌の菌数や腸管由来乳酸菌の総菌数の含有率を高めることは期待できない。 Originally, intestinal lactic acid bacteria often inhabiting in the animal intestinal tract include strains such as skim milk that are extremely poor in lactic acid fermentation. If lactic acid fermentation is poor, not only the number of lactic acid bacteria is low, but also the lactic acid acidity is low, which is not suitable for producing fermented milk. Such a milk fermentable intestinal lactic acid bacterium is extremely difficult to grow and use in milk such as skim milk. In addition, growth may occur by adding growth promoters such as yeast extract, soybean extract, and milk protein degradation product, but even in that case, the flavor may be unfavorable due to the growth promoter, and the amount added. Is limited. In addition, the production of fermented milk containing non-fermentable intestinal lactic acid bacteria is described in L.L. It is possible to mix and culture with bulgaricus and Streptococcus thermophilus, but since these are suitable for fermentation in milk and show rapid growth and a competitive action, the number of lactobacilli-derived lactic acid bacteria derived from milk It cannot be expected to increase the content of the total number of intestinal lactic acid bacteria.
本発明は、上記実情に鑑みなされたものであり、その目的は、基本的には通常の発酵乳の製造方法と何ら変わらない方法で製造することが可能であり、乳難発酵性腸管由来乳酸菌の菌数やプロバイオティクス（腸管由来乳酸菌）の総菌数の含有率が高められた発酵乳およびその製造方法を提供することにある。 The present invention has been made in view of the above circumstances, and its purpose can be basically produced by a method that is not different from the usual method for producing fermented milk. An object of the present invention is to provide fermented milk in which the content of the total number of bacteria and the total number of probiotics (intestinal lactic acid bacteria) is increased and a method for producing the same.
本発明者らは、発酵乳製造において乳難発酵性腸管由来乳酸菌を高濃度に含有し、高い乳酸酸度と腸管由来乳酸菌総数を高濃度に含有させる方法について鋭意検討した結果、乳を主原料とするミックス（脱脂乳も含む）に乳難発酵性腸管由来乳酸菌とともにＬ．ｒｈａｍｎｏｓｕｓを添加することにより通常の発酵乳製造方法と何ら変わらない手順で、乳難発酵性腸管由来乳酸菌を高濃度に含有させ、また高い乳酸酸度を保ち、かつプロバイオティクス（腸管由来乳酸菌）の総数を増大させることを見出し、本発明を完成するに至った。すなわち、本発明は次の通りである。 As a result of intensive studies on a method of containing lactobacilli-derived lactic acid bacteria at a high concentration in fermented milk production and a high concentration of lactic acid bacteria having a high lactate content and a total number of intestinal lactic acid bacteria, the present inventors have used milk as a main raw material. L. to mix (including skimmed milk) together with non-fermentable intestinal lactic acid bacteria. By adding rhamnosus, a milk fermentable gut-derived lactic acid bacterium is contained at a high concentration in a procedure that is not different from a normal fermented milk production method, and a high degree of lactic acidity is maintained, and probiotics (gut-derived lactic acid bacteria) The present inventors have found that the total number can be increased and have completed the present invention. That is, the present invention is as follows.
（１）以下の乳酸菌（Ａ）及び（Ｂ）を含有し、乳酸菌（Ａ）の菌数が少なくとも０．８×１０８ＣＦＵ／ｍｌ、乳酸菌（Ａ）と（Ｂ）との合計菌数が少なくとも４．８×１０８ＣＦＵ／ｍｌであることを特徴とする発酵乳。 (1) Contains the following lactic acid bacteria (A) and (B), the number of lactic acid bacteria (A) is at least 0.8 × 10 8 CFU / ml, and the total number of lactic acid bacteria (A) and (B) is Fermented milk, characterized in that it is at least 4.8 × 10 8 CFU / ml.
（Ａ）１重量％の果糖を添加した脱脂乳に１０７ＣＦＵ／ｍｌになるように乳酸菌を接種し２４時間培養した時の酸度が０．５重量％未満の乳酸酸度である性質を有する乳難発酵性腸管由来乳酸菌 (A) Milk having the property that the acidity when inoculated with lactic acid bacteria to 10 7 CFU / ml in skim milk to which 1% by weight of fructose has been added and cultured for 24 hours is less than 0.5% by weight Non-fermentable intestinal lactic acid bacteria
（Ｂ）ラクトバチルス・ラムノーサス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｒｈａｍｎｏｓｕｓ） (B) Lactobacillus rhamnosus
（２）乳酸菌（Ａ）が、ラクトバチルス・アシドフィルス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ａｃｉｄｏｐｈｉｌｕｓ）ＴＭＣ０３５６菌株（ＦＥＲＭ−１９２３２）である（１）に記載の乳酸菌。 (2) The lactic acid bacterium according to (1), wherein the lactic acid bacterium (A) is Lactobacillus acidophilus TMC0356 strain (FERM-19232).
（３）乳酸菌（Ｂ）が、１重量％の果糖添加した脱脂乳に１０７ＣＦＵ／ｍｌになるように接種し、２４時間培養した時の酸度が１．０重量％以上の乳酸酸度である性質を有するラクトバチルス・ラムノーサス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｒｈａｍｎｏｓｕｓ）である（１）又は（２）に記載の発酵乳。 (3) Lactic acid bacteria (B) are inoculated to skim milk added with 1% by weight of fructose to 10 7 CFU / ml, and when cultured for 24 hours, the acidity is 1.0% by weight or more. The fermented milk according to (1) or (2), which is Lactobacillus rhamnosus having properties.
（４）乳酸菌（Ｂ）の菌数が少なくとも４．０×１０８ＣＦＵ／ｍｌである（１）〜（３）の何れかに記載の発酵乳。 (4) The fermented milk according to any one of (1) to (3), wherein the number of lactic acid bacteria (B) is at least 4.0 × 10 8 CFU / ml.
（５）乳を主原料とする培養基で以下の乳酸菌（Ａ）及び（Ｂ）を混合培養することを特徴とする請求項１に記載の発酵乳の製造方法。 (5) The method for producing fermented milk according to claim 1, wherein the following lactic acid bacteria (A) and (B) are mixed and cultured in a culture medium mainly composed of milk.
（Ａ）１重量％の果糖を添加した脱脂乳に１０７ＣＦＵ／ｍｌになるように乳酸菌を接種し２４時間培養した時の酸度が０．５重量％未満の乳酸酸度である性質を有する乳難発酵性腸管由来乳酸菌 (A) Milk having the property that the acidity when inoculated with lactic acid bacteria to 10 7 CFU / ml in skim milk to which 1% by weight of fructose has been added and cultured for 24 hours is less than 0.5% by weight Non-fermentable intestinal lactic acid bacteria
（Ｂ）ラクトバチルス・ラムノーサス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｒｈａｍｎｏｓｕｓ） (B) Lactobacillus rhamnosus
（６）乳酸菌（Ａ）が、ラクトバチルス・アシドフィルス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ａｃｉｄｏｐｈｉｌｕｓ）ＴＭＣ０３５６菌株（ＦＥＲＭ−１９２３２）である（５）に記載の製造方法。 (6) The production method according to (5), wherein the lactic acid bacterium (A) is Lactobacillus acidophilus TMC0356 strain (FERM-19232).
（７）乳酸菌（Ｂ）が、１重量％の果糖添加した脱脂乳に１０７ＣＦＵ／ｍｌになるように接種し、２４時間培養した時の酸度が１．０重量％以上の乳酸酸度である性質を有するラクトバチルス・ラムノーサス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｒｈａｍｎｏｓｕｓ）である（５）又は（６）に記載の製造方法。 (7) Lactic acid bacteria (B) are inoculated to skim milk added with 1% by weight of fructose at 10 7 CFU / ml, and the acidity when cultured for 24 hours is 1.0% by weight or more. The production method according to (5) or (6), which is Lactobacillus rhamnosus having properties.
（８）発酵乳中の乳酸菌（Ｂ）の菌数が少なくとも４．０×１０８ＣＦＵ／ｍｌである（５）〜（７）の何れかに記載の製造方法。 (8) The production method according to any one of (5) to (7), wherein the number of lactic acid bacteria (B) in the fermented milk is at least 4.0 × 10 8 CFU / ml.
本発明によれば、整腸効果、免疫賦活効果などの保健効果が期待される発酵乳として、基本的には通常の発酵乳の製造方法と何ら変わらない方法で製造することが可能であり、乳難発酵性腸管由来乳酸菌の菌数やプロバイオティクス（腸管由来乳酸菌）の総菌数の含有率が高められた発酵乳を提供することが出来る。 According to the present invention, fermented milk that is expected to have health effects such as intestinal regulation effect, immunostimulatory effect, etc., can be basically produced by a method that is not different from the usual fermented milk production method, Fermented milk in which the number of non-fermentable intestinal lactic acid bacteria and the total content of probiotics (intestinal lactic acid bacteria) are increased can be provided.
先ず、説明の便宜上、本発明に係る発酵乳の製造方法について説明する。本発明の製造方法は、特定の乳酸菌を組合せて使用する点以外は、通常の発酵乳の製造方法と同じである。すなわち、全乳または脱脂乳に脱脂粉乳などを加えて固形分を増やし、更に、安定剤、糖類などを添加し、乳酸菌で発酵させる、発酵乳の一般的な製造方法を採用することが出来る。 First, for convenience of explanation, a method for producing fermented milk according to the present invention will be described. The production method of the present invention is the same as the ordinary method for producing fermented milk except that a specific lactic acid bacterium is used in combination. That is, a general method for producing fermented milk can be employed in which skim milk powder or the like is added to whole milk or skim milk to increase the solid content, and further, stabilizers, sugars and the like are added and fermented with lactic acid bacteria.
本発明において、培養基としては、乳を主原料とする培養基である限り、従来公知の培養基を使用することが出来る。 In the present invention, as a culture medium, a conventionally known culture medium can be used as long as it is a culture medium mainly made of milk.
本発明において使用する乳酸菌（Ａ）は、１重量％の果糖を添加した脱脂乳に１０７ＣＦＵ／ｍｌになるように乳酸菌を接種し２４時間培養した時の酸度が０．５重量％未満の乳酸酸度である性質を有する乳難発酵性腸管由来乳酸菌である。 Lactic acid bacteria (A) used in the present invention have an acidity of less than 0.5% by weight when inoculated with lactobacilli to 10 7 CFU / ml in skim milk to which 1% by weight of fructose has been added and cultured for 24 hours. It is a milk-fermentable intestinal lactic acid bacterium having a property of lactic acidity.
乳酸菌（Ａ）としては、上記の性質を有する限りその種類は特に限定されないが、ラクトバチルス属乳酸菌のうち、特に、本出願により、特開２００４−２７７３８１にて既に提案されたラクトバチルス・アシドフィルス（Ｌ．ａｃｉｄｏｐｈｉｌｕｓ）ＴＭＣ０３５６菌株が好適である。斯かる菌株は、上記の出願の明細書に記載された方法に従い、ラクトバチルス属乳酸菌の中から、以下に示す方法によって選抜することが出来る。 The type of lactic acid bacterium (A) is not particularly limited as long as it has the above-mentioned properties. Among lactic acid bacteria belonging to the genus Lactobacillus, in particular, Lactobacillus acidophilus (Lactobacillus acidophilus already proposed in Japanese Patent Application Laid-Open No. 2004-277381 by the present application). L. acidophilus) TMC0356 strain is preferred. Such a strain can be selected from Lactobacillus lactic acid bacteria by the method described below according to the method described in the specification of the above-mentioned application.
すなわち、ヒトの消化管環境（胃酸による低ｐＨ、小腸での胆汁酸の存在）のモデル試験（耐酸性および耐胆汁性）を経て、生残性を有し、ヒト腸管由来の上皮様細胞Ｃａｃｏ−２株（理研細胞銀行 ＲＣＢ０９８８）と培養した際の培養細胞への付着菌数、炎症性サイトカインＩＬ−６およびＩＬ−８産生量を指標として選抜することが出来る。 That is, after undergoing a model test (acid resistance and bile resistance) of the human digestive tract environment (low pH due to gastric acid, the presence of bile acids in the small intestine), the surviving epithelial cell Caco derived from the human intestine -2 strains (RIKEN Cell Bank RCB0988) can be selected using as an index the number of bacteria attached to the cultured cells and the amount of inflammatory cytokines IL-6 and IL-8 produced when cultured.
選抜したＴＭＣ０３５６菌株は、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寄託さており、その受託番号は「ＦＥＲＭ Ｐ−１９２３２」である。 The selected TMC0356 strain is deposited at the National Institute of Advanced Industrial Science and Technology and the Patent Organism Depositary, and the deposit number is “FERM P-19232”.
本発明において使用する乳酸菌（Ｂ）は、ラクトバチルス・ラムノーサス（Ｌ．ｒｈａｍｎｏｓｕｓ）である。そして、好ましい乳酸菌（Ｂ）は、１重量％の果糖添加した脱脂乳に１０７ＣＦＵ／ｍｌになるように接種し、２４時間培養した時の酸度が１．０重量％以上の乳酸酸度である性質を有するラクトバチルス・ラムノーサス（Ｌ．ｒｈａｍｎｏｓｕｓ）である。 The lactic acid bacteria (B) used in the present invention is L. rhamnosus. A preferred lactic acid bacterium (B) is inoculated to skim milk added with 1% by weight of fructose at 10 7 CFU / ml, and the acidity when cultured for 24 hours is 1.0% by weight or more. Lactobacillus rhamnosus having properties.
乳酸菌（Ａ）と乳酸菌（Ｂ）との使用条件は、得られる発酵乳において、乳酸菌（Ａ）の菌数が少なくとも０．８×１０８ＣＦＵ／ｍｌ、乳酸菌（Ａ）と（Ｂ）との合計菌数が少なくとも４．８×１０８ＣＦＵ／ｍｌであり、好ましくは、発酵乳における乳酸酸度が少なくとも０．６重量％となるように、また、乳酸菌（Ｂ）の菌数が少なくとも４．０×１０８ＣＦＵ／ｍｌとなるように設定される。通常、乳酸菌（Ａ）：乳酸菌（Ｂ）の使用比率は１：１重量比である。 The conditions of use of the lactic acid bacteria (A) and the lactic acid bacteria (B) are as follows: In the obtained fermented milk, the number of lactic acid bacteria (A) is at least 0.8 × 10 8 CFU / ml, and the lactic acid bacteria (A) and (B) The total number of bacteria is at least 4.8 × 10 8 CFU / ml, and preferably the lactic acid acidity in the fermented milk is at least 0.6% by weight, and the number of lactic acid bacteria (B) is at least 4. It is set to be 0 × 10 8 CFU / ml. Usually, the use ratio of lactic acid bacteria (A): lactic acid bacteria (B) is 1: 1 weight ratio.
次に、本発明の発酵乳について説明する。本発明の発酵乳は、例えば、上記のような製造方法で得られ、前述の乳酸菌（Ａ）及び（Ｂ）を含有し、乳酸菌（Ａ）の菌数が少なくとも０．８×１０８ＣＦＵ／ｍｌ、乳酸菌（Ａ）と（Ｂ）との合計菌数が少なくとも４．８×１０８ＣＦＵ／ｍｌであることを特徴とし、好ましい態様においては、発酵乳における乳酸酸度は少なくとも０．６重量％であり、また、乳酸菌（Ｂ）の菌数は少なくとも４．０×１０８ＣＦＵ／ｍｌである。なお、乳酸菌（Ａ）及び（Ｂ）の上限は通常４．０×１０１０ＣＦＵ／ｍｌ、乳酸菌（Ａ）と（Ｂ）との合計菌数は通常８．０×１０１０ＣＦＵ／ｍｌ、乳酸酸度の上限は通常２．２重量％である。 Next, the fermented milk of the present invention will be described. The fermented milk of the present invention is obtained, for example, by the above-described production method, contains the lactic acid bacteria (A) and (B) described above, and the number of lactic acid bacteria (A) is at least 0.8 × 10 8 CFU / ml, the total number of bacteria of lactic acid bacteria (A) and (B) is at least 4.8 × 10 8 CFU / ml. In a preferred embodiment, the lactic acid acidity in the fermented milk is at least 0.6% by weight The number of lactic acid bacteria (B) is at least 4.0 × 10 8 CFU / ml. The upper limit of lactic acid bacteria (A) and (B) is usually 4.0 × 10 10 CFU / ml, and the total number of bacteria of lactic acid bacteria (A) and (B) is usually 8.0 × 10 10 CFU / ml, lactic acid The upper limit of acidity is usually 2.2% by weight.
本発明の発酵乳は、上記のように、乳難発酵性腸管由来乳酸菌の菌数やプロバイオティクス（腸管由来乳酸菌）の総菌数の含有率が高められているため、整腸効果、免疫賦活効果などの保健効果が期待される。 As described above, the fermented milk of the present invention has an increased number of non-fermentable intestinal lactic acid bacteria and a total content of probiotics (intestinal lactic acid bacteria). Health effects such as activation effects are expected.
以下、各種の試験結果に基づいて本発明を更に詳細に説明するが、本発明は、その要旨を超えない限り、以下の実施例に限定されるものではない。 Hereinafter, although this invention is demonstrated further in detail based on various test results, this invention is not limited to a following example, unless the summary is exceeded.
試験例１（各種乳酸菌の培養試験）： Test Example 1 (Culture test of various lactic acid bacteria):
（１）乳酸菌：
Ｌ．ａｃｉｄｏｐｈｉｌｕｓ ＴＭＣ０３５６（ＦＥＲＭ Ｐ−１９２３２）、Ｌ．ｒｈａｍｎｏｓｕｓ ＬＡ−２（ＦＥＲＭ ＢＰ−４４４２）、Ｌ．ｒｈａｍｎｏｓｕｓ ＧＧ（Ａｍｅｒｉｃａｎ ｔｙｐｅ ｃｕｌｔｕｒｅ ｃｏｌｌｅｃｔｉｏｎ ：ＡＴＣＣ５３１０３）、その他乳酸菌はＪＣＭ株（理化学研究所保存菌株）を使用した。
(1) Lactic acid bacteria:
L. acidophilus TMC0356 (FERM P-19232), L. rhamnosus LA-2 (FERM BP-4442), L. Rhamnosus GG (American type culture collection: ATCC 53103) and other lactic acid bacteria were JCM strains (stored in RIKEN).
（２）培養基：
９重量％脱脂粉乳、１重量％果糖、９０重量％水から成る。通常、各成分を溶解し、１２１℃、１５分間オートクレーブ滅菌して調製した発酵乳培養基を使用した。
(2) Culture medium:
It consists of 9% by weight skim milk powder, 1% by weight fructose and 90% by weight water. Usually, fermented milk culture medium prepared by dissolving each component and autoclaving at 121 ° C. for 15 minutes was used.
（３）培養方法および評価：
発酵乳培養基にＭＲＳ（ベクトン・ディッキンソン社製）培地で前培養した乳酸菌液を１重量％接種し１０７ＣＦＵ／ｍｌの乳酸菌数になるように調製した。２つの乳酸菌を混合培養する場合は、各乳酸菌数が１０７ＣＦＵ／ｍｌになるように調製した。３７℃で２４時間培養後、乳酸酸度（中和滴定法）、乳酸菌数（ＢＣＰ平板培養法）を測定した。
(3) Culture method and evaluation:
The fermented milk culture medium was inoculated with 1% by weight of a lactic acid bacterium solution pre-cultured with MRS (manufactured by Becton Dickinson) medium so that the number of lactic acid bacteria was 10 7 CFU / ml. When two lactic acid bacteria were mixed and cultured, the number of each lactic acid bacteria was adjusted to 10 7 CFU / ml. After culturing at 37 ° C. for 24 hours, the lactic acid acidity (neutralization titration method) and the number of lactic acid bacteria (BCP plate culture method) were measured.
（４）培養試験Ａ：
表１は、乳難発酵性腸管由来乳酸菌の代表例であるラクトバチルス アシドフィルス（Lactobacillus acidophilus）ＴＭＣ０３５６菌株の単独培養試験の結果であるが、乳酸酸度が０．３２重量％と低値であり乳成分も凝固せず発酵乳製造は不適であることを示している。
(4) Culture test A:
Table 1 shows the results of a single culture test of Lactobacillus acidophilus TMC0356 strain, which is a representative example of a milk-fermentable intestinal lactic acid bacterium. Does not coagulate, indicating that fermented milk production is unsuitable.
（５）培養試験Ｂ：
表２は、腸管由来乳酸菌の代表例である各乳酸菌を接種した結果を示す。２４時間培養後の乳酸酸度が１．０％の高値を示したのはＬ．ｒｈａｍｎｏｓｕｓのみであった。Ｌ．ｒｈａｍｎｏｓｕｓは他の腸管由来乳酸菌よりも乳を主成分とした培養基で高い乳酸酸度を示す場合が多いことが明らかである。
(5) Culture test B:
Table 2 shows the results of inoculating each lactic acid bacterium, which is a representative example of intestinal tract-derived lactic acid bacteria. L. acidity after culturing for 24 hours showed a high value of 1.0%. Rhamnosus only. L. It is clear that rhamnosus often exhibits higher lactic acidity in a culture medium mainly composed of milk than other intestinal lactic acid bacteria.
（６）培養試験Ｃ：
表３は、乳難発酵性腸管由来乳酸菌の代表例であるＬ．ａｃｉｄｏｐｈｉｌｕｓＴＭＣ０３５６とＬ．ｒｈａｍｎｏｓｕｓを混合培養した際の乳酸酸度を示す。混合培養した際の乳酸酸度は１重量％以上の高い乳酸酸度を示し発酵乳の製造に好適と考えられる。
(6) Culture test C:
Table 3 shows a typical example of lactic acid bacteria derived from difficult-to-fermentable intestinal tract. acidophilus TMC0356 and L. The lactate acidity when rhamnosus is mixed and cultured is shown. The lactic acid acidity at the time of mixed culture shows a high lactic acid acidity of 1% by weight or more, which is considered suitable for the production of fermented milk.
（７）培養試験Ｄ：
表４は、乳難発酵性腸管由来乳酸菌の代表例であるＬ．ａｃｉｄｏｐｈｉｌｕｓ ＴＭＣ０３５６とＬ．ｒｈａｍｎｏｓｕｓを混合培養した際の乳酸菌数を示下す。乳難発酵性腸管由来乳酸菌は１０８ＣＦＵ／ｍｌ以上を示し、かつ、腸管由来乳酸菌総数は１０９ＣＦＵ／ｍｌ以上の高値を示し、プロバイオティクス発酵乳として好適となると考えられる。また、Ｌ．ｒｈａｍｎｏｓｕｓは整腸作用、腸内環境改善、抗変異原性、免疫賦活作用、感染症予防、人に有益な効果がもつことが知られており、その利用が期待される。
(7) Culture test D:
Table 4 shows a typical example of lactic acid bacteria derived from non-fermentable intestinal tract. acidophilus TMC0356 and L. The number of lactic acid bacteria when rhamnosus is mixed and cultured is shown. Lactic acid-fermentable intestinal lactic acid bacteria show 10 8 CFU / ml or more, and the total number of intestinal lactic acid bacteria shows a high value of 10 9 CFU / ml or more, which is considered suitable as probiotic fermented milk. L. Rhamnosus is known to have an intestinal regulating action, intestinal environment improvement, antimutagenicity, immunostimulatory action, infection prevention, and beneficial effects on humans, and its use is expected.
実施例１：
脱脂粉乳（高梨乳業（株）製）２７０ｋｇ、砂糖６０ｋｇ、果糖６０ｋｇを温水２５５０ｋｇに溶解後、均質化し、１１５℃殺菌し、発酵乳培養基を調製した。「Ｌ．ａｃｉｄｏｐｈｉｌｕｓ ＴＭＣ０３５６バルクスターター」３０ｋｇと「Ｌ．ｒｈａｍｎｏｓｕｓ ＧＧ株 バルクスターター」３０ｋｇを接種し、３７℃、２４時間発酵させ、冷却し、発酵乳３０００ｋｇを得た。得られた発酵乳の乳酸酸度は１．０重量％であり、「Ｌ．ａｃｉｄｏｐｈｉｌｕｓ ＴＭＣ０３５６」の菌数は３×１０８ＣＦＵ／ｍｌ、「Ｌ．ｒｈａｍｎｏｓｕｓ ＧＧ株」の菌数は２．７×１０９ＣＦＵ／ｍｌ、腸管由来乳酸菌総数は３×１０９ＣＦＵ／ｍｌであった。
Example 1:
270 kg of skim milk powder (manufactured by Takanashi Milk Industry Co., Ltd.), 60 kg of sugar and 60 kg of fructose were dissolved in 2550 kg of warm water, homogenized, and sterilized at 115 ° C. to prepare a fermented milk culture medium. 30 kg of “L. acidophilus TMC0356 bulk starter” and 30 kg of “L. rhhamnosus GG strain bulk starter” were inoculated, fermented at 37 ° C. for 24 hours, and cooled to obtain 3000 kg of fermented milk. The obtained fermented milk has a lactic acid acidity of 1.0% by weight, the number of bacteria of “L. acidophilus TMC0356” is 3 × 10 8 CFU / ml, and the number of bacteria of “L. rhamnosus GG strain” is 2.7 ×. 10 9 CFU / ml, the total number of intestinal lactic acid bacteria was 3 × 10 9 CFU / ml.
Claims (8)
（Ａ）１重量％の果糖を添加した脱脂乳に１０７ＣＦＵ／ｍｌになるように乳酸菌を接種し２４時間培養した時の酸度が０．５重量％未満の乳酸酸度である性質を有する乳難発酵性腸管由来乳酸菌
（Ｂ）ラクトバチルス・ラムノーサス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｒｈａｍｎｏｓｕｓ） It contains the following lactic acid bacteria (A) and (B), the number of lactic acid bacteria (A) is at least 0.8 × 10 8 CFU / ml, and the total number of lactic acid bacteria (A) and (B) is at least 4. Fermented milk characterized by being 8 × 10 8 CFU / ml.
(A) Milk having the property that the acidity when inoculated with lactic acid bacteria to 10 7 CFU / ml in skim milk to which 1% by weight of fructose has been added and cultured for 24 hours is less than 0.5% by weight Non-fermentable intestinal lactic acid bacteria (B) Lactobacillus rhamnosus
（Ａ）１重量％の果糖を添加した脱脂乳に１０７ＣＦＵ／ｍｌになるように乳酸菌を接種し２４時間培養した時の酸度が０．５重量％未満の乳酸酸度である性質を有する乳難発酵性腸管由来乳酸菌
（Ｂ）ラクトバチルス・ラムノーサス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｒｈａｍｎｏｓｕｓ） The method for producing fermented milk according to claim 1, wherein the following lactic acid bacteria (A) and (B) are mixed and cultured in a culture medium mainly composed of milk.
(A) Milk having the property that the acidity when inoculated with lactic acid bacteria to 10 7 CFU / ml in skim milk to which 1% by weight of fructose has been added and cultured for 24 hours is less than 0.5% by weight Non-fermentable intestinal lactic acid bacteria (B) Lactobacillus rhamnosus
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