JP4426506B2 - New lactic acid bacteria and beverages using new lactic acid bacteria - Google Patents
New lactic acid bacteria and beverages using new lactic acid bacteria Download PDFInfo
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- JP4426506B2 JP4426506B2 JP2005191352A JP2005191352A JP4426506B2 JP 4426506 B2 JP4426506 B2 JP 4426506B2 JP 2005191352 A JP2005191352 A JP 2005191352A JP 2005191352 A JP2005191352 A JP 2005191352A JP 4426506 B2 JP4426506 B2 JP 4426506B2
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Description
本発明は、乳酸菌及びその乳酸菌を利用した飲料に関し、更に詳しくは、免疫賦活作用の高い乳酸菌及びその乳酸菌を利用した飲料に関する。 The present invention relates to a lactic acid bacterium and a beverage using the lactic acid bacterium, and more particularly to a lactic acid bacterium having a high immunostimulatory effect and a beverage using the lactic acid bacterium.
インターフェロン（ＩＦＮ）は、抗ウイルス活性を有する分子量約２０，０００の蛋白質の総称である。ＩＦＮにはα、β、γの３種の型が存在し、生物活性として、抗ウイルス作用、抗腫瘍作用、免疫応答調節作用等が知られている。なかでもＩＦＮ−γはマクロファージ、ナチュラルキラー細胞、細胞障害性Ｔ細胞などの免疫担当細胞の分化誘導・活性化に重要な役割を果たす。 Interferon (IFN) is a general term for proteins having antiviral activity and having a molecular weight of about 20,000. There are three types of IFNs, α, β, and γ, and their biological activities are known to include antiviral action, antitumor action, and immune response regulating action. Among these, IFN-γ plays an important role in inducing and activating differentiation of immunocompetent cells such as macrophages, natural killer cells, and cytotoxic T cells.
現在、遺伝子組み換え型ＩＦＮや、精製ＩＦＮがウイルス感染、ウイルス性肝炎、癌に対する治療薬等として使用されているが、ＩＦＮを生体内へ直接投与するような治療法では多様な副作用が生じる。 Currently, recombinant IFN and purified IFN are used as therapeutic agents for viral infections, viral hepatitis, cancer, etc., but treatment methods such as direct administration of IFN into a living body have various side effects.
そのため、生体内でのＩＦＮ−γ産生を増強させることによって、ウイルス感染の防御や、癌等の改善・治療を図るための多くの物質や組成物が開発されている。 Therefore, many substances and compositions have been developed for the purpose of protecting against viral infection and improving / treating cancer and the like by enhancing IFN-γ production in vivo.
下記の特許文献１には、エンテロコッカス属に属する微生物の菌体が、インターフェロン（ＩＦＮ）の産生を増強させる作用を有することが開示されている。この特許文献１に記載されている菌種は、健常者の腸内およびサイレージから分離された乳酸菌の一種であるため、副作用の無い安全な菌種である。
しかしながら、上記の特許文献１の菌体は、ＩＦＮ−γの産生を増強させる作用を有することが知られているが、未だ効果が不充分である。このため、更に産生を増強させる作用を有する菌体の開発が望まれている。 However, although the above-mentioned bacterial cells of Patent Document 1 are known to have an action of enhancing the production of IFN-γ, the effect is still insufficient. For this reason, development of the microbial cell which has the effect | action which further enhances production is desired.
本発明は、上記のような課題に鑑みてなされたものであり、生体のＩＦＮ−γ産生増強作用を有する新規の乳酸菌及び新規乳酸菌を利用した飲料を提供することを目的とする。 This invention is made | formed in view of the above subjects, and it aims at providing the drink using the novel lactic acid bacteria which have an IFN-gamma production enhancement effect of a biological body, and novel lactic acid bacteria.
本発明者らは、上記課題を解決するために鋭意研究を重ねた結果、ロイコノストック（ｌｅｕｃｏｎｏｓｔｏｃ）属、特に、メセントロイデス（ｍｅｓｅｎｔｅｒｏｉｄｅｓ）に分類される菌体又はその処理物がＩＦＮ−γの産生増強作用を有することを見出し、本発明を完成するに至った。より具体的には、本発明は以下のようなものを提供する。 As a result of intensive studies to solve the above-mentioned problems, the present inventors have found that microbial cells classified into the genus Leuconostoc, particularly mesenteroides, or processed products thereof are IFN-γ. The present invention has been completed. More specifically, the present invention provides the following.
（１） ロイコノストック属に分類される乳酸菌を含有する飲料。 (1) Beverages containing lactic acid bacteria classified as genus Leuconostoc.
本発明の飲料によれば、ロイコノストック属に分類される乳酸菌が、生体内のＩＦＮ−γ産生を増強する効果を有するため、これを含有する飲料についても、ＩＦＮ−γを増強する効果がある。また、使用される乳酸菌は、漬け物から分離された菌種であるため、副作用の無い安全な菌種である。 According to the beverage of the present invention, since the lactic acid bacteria classified into the genus Leuconostoc have an effect of enhancing IFN-γ production in the living body, the effect of enhancing IFN-γ also for beverages containing the same. is there. Moreover, since the lactic acid bacteria used are the bacterial species isolated from the pickles, they are safe bacterial species with no side effects.
（２） 前記乳酸菌が発酵して製造される（１）記載の飲料。 (2) The beverage according to (1), wherein the lactic acid bacteria are fermented and produced.
（３） 前記乳酸菌が、ロイコノストック属メセントロイデス種に分類される乳酸菌である（１）又は（２）記載の飲料。 (3) The beverage according to (1) or (2), wherein the lactic acid bacterium is a lactic acid bacterium classified as Leuconostoc genus Mecentroides.
（４） 前記乳酸菌が、ＦＥＲＭ（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、以下同じ） Ｐ−２０５００（受託番号）である（１）から（３）いずれか記載の飲料。 (4) The beverage according to any one of (1) to (3), wherein the lactic acid bacterium is FERM (National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary, hereinafter the same) P-20500 (accession number).
（５） 前記乳酸菌が、ＦＥＲＭ Ｐ−２０５０１（受託番号）である（１）から（３）いずれか記載の飲料。 (5) The beverage according to any one of (1) to (3), wherein the lactic acid bacterium is FERM P-20501 (accession number).
（６） 受託番号がＦＥＲＭ Ｐ−２０５００である乳酸菌株。 (6) Lactic acid strain whose accession number is FERM P-20500.
（７） 受託番号がＦＥＲＭ Ｐ−２０５０１である乳酸菌株。 (7) Lactic acid strain whose accession number is FERM P-20501.
なお、本発明における「飲料」には、乳酸菌を含有する飲料（以下、「乳酸菌含有飲料」という）、および乳酸菌により発酵させた飲料（以下、「乳酸菌発酵飲料」という）が含まれる。 The “beverage” in the present invention includes beverages containing lactic acid bacteria (hereinafter referred to as “lactic acid bacteria-containing beverages”) and beverages fermented with lactic acid bacteria (hereinafter referred to as “lactic acid bacteria fermented beverages”).
本発明によれば、生体内のＩＦＮ−γ産生増強作用を有する新規の乳酸菌を提供し、更に、感染防御、アレルギー改善作用を有する飲料を提供することができる。 ADVANTAGE OF THE INVENTION According to this invention, the novel lactic acid bacteria which have the IFN-gamma production enhancement effect in the living body can be provided, and also the drink which has an infection defense and allergy improvement effect can be provided.
以下、本発明について詳しく説明する。 The present invention will be described in detail below.
＜乳酸菌含有飲料の製造＞
本発明に係る乳酸菌含有飲料は、果実、果汁、糖類、酸味料、水等の原料を所定量調合した調合液に、ロイコノストック属に分類される乳酸菌を含有させ、製造することができる。
<Manufacture of lactic acid bacteria-containing beverages>
The lactic acid bacteria-containing beverage according to the present invention can be produced by containing a lactic acid bacterium classified as genus Leuconostoc in a preparation solution prepared by mixing a predetermined amount of raw materials such as fruits, fruit juices, sugars, acidulants and water.
＜乳酸菌発酵飲料の製造＞
また、乳酸菌発酵飲料は、果実、果汁、乳製品、豆乳などを単独、または２種以上を調合して製造することができ、乳酸菌による発酵は、原料を調合した後、行うことができ、また、調合する前に行うこともできる。また、原料の一部を調合し、発酵させた後、残りの原料を調合することもできる。更に、乳酸菌発酵飲料を製造後、新たに乳酸菌を添加し製造することもできる。
<Manufacture of lactic acid bacteria fermented beverages>
In addition, lactic acid bacteria fermented beverages can be produced by mixing fruits, fruit juices, dairy products, soy milk, etc., alone or in combination of two or more, and fermentation with lactic acid bacteria can be performed after preparing the raw materials, It can also be done before compounding. Moreover, after mixing a part of raw material and making it ferment, the remaining raw material can also be prepared. Furthermore, after manufacturing a lactic acid bacteria fermented beverage, a lactic acid bacterium can be newly added and manufactured.
発酵は、発酵温度が２０℃以上４０℃以下、好ましくは２５℃以上３７℃以下であり、発酵時間は、５時間以上１２０時間以下、好ましくは１２時間以上６０時間以下で行う。 Fermentation is performed at a fermentation temperature of 20 ° C. or higher and 40 ° C. or lower, preferably 25 ° C. or higher and 37 ° C. or lower, and a fermentation time of 5 hours or longer and 120 hours or shorter, preferably 12 hours or longer and 60 hours or shorter.
本発明においてはロイコノストック属に分類される種々の乳酸菌を用いることができるが、特に、高いＩＦＮ−γ産生増強作用を有するため、メセントロイデス種を用いることが好ましく、更に、受託番号がＦＥＲＭ Ｐ−２０５００である乳酸菌株、受託番号がＦＥＲＭ Ｐ−２０５０１である乳酸菌株が好ましく用いられる。乳酸菌は、調合液１ｍｌあたり１０４個以上１０８個以下添加するのが好ましい。 In the present invention, various lactic acid bacteria classified into the genus Leuconostoc can be used. In particular, since it has a high IFN-γ production enhancing action, it is preferable to use a Mecentroides species, and the accession number is A lactic acid strain having FERM P-20500 and a lactic acid strain having a deposit number of FERM P-20501 are preferably used. It is preferable to add 10 4 or more and 10 8 or less lactic acid bacteria per 1 ml of the preparation solution.
以下に、受託番号がＦＥＲＭ Ｐ−２０５００である乳酸菌株、受託番号がＦＥＲＭ Ｐ−２０５０１である乳酸菌株の分離手段及び菌学的、生理学的性質を示す。 The separation means and mycological and physiological properties of the lactic acid strain whose accession number is FERM P-20500 and the lactic acid strain whose accession number is FERM P-20501 are shown below.
＜受託番号がＦＥＲＭ Ｐ−２０５００の分離手段＞
漬け物１０ｇに生理食塩水９０ｍｌを加え混合し、更に生理食塩水で段階的に希釈して乳酸菌選択培地（５％馬脱繊維素血液添加ＢＬ寒天培地、１．５％寒天含有ＭＲＳ培地、ＰＥＳ培地、Ｍ−Ｅ培地、Ｍｍａ−ＬＢＳ培地）に塗末し培養した。培養条件は嫌気条件下にて２０℃〜３７℃とした。菌の集落が形成されたら、別の同種の平板培地に画線塗布し同様に培養した。更に同じ操作を繰り返し、単一集落を形成する乳酸菌を分離した。
<Separation means with accession number FERM P-20500>
90 g of physiological saline was added to 10 g of pickled vegetables, mixed, and further diluted stepwise with physiological saline to select lactic acid bacteria selection medium (BL agar medium supplemented with 5% horse defibrinated blood, MRS medium containing 1.5% agar, PES medium) , M-E medium, Mma-LBS medium). The culture conditions were 20 ° C. to 37 ° C. under anaerobic conditions. When fungal colonies were formed, streaks were applied to another plate medium of the same kind and cultured in the same manner. Further, the same operation was repeated to isolate lactic acid bacteria forming a single settlement.
＜受託番号がＦＥＲＭ Ｐ−２０５０１の分離手段＞
上記ＦＥＲＭ Ｐ−２０５０１と同様の方法により分離を行った。
<Separation means with accession number FERM P-20501>
Separation was performed in the same manner as in FERM P-20501.
＜菌学的性質及び生理学的性質＞
本発明の乳酸菌である受託番号がＦＥＲＭ Ｐ−２０５００である乳酸菌株の１６ＳリボソームＲＮＡ遺伝子の塩基配列を配列表の配列番号１に示す。受託番号がＦＥＲＭ Ｐ−２０５０１である乳酸菌株の１６ＳリボソームＲＮＡ遺伝子の塩基配列を配列表の配列番号２に示す。既知微生物の塩基配列のデータベース上で、これらの１６ＳリボソームＲＮＡ遺伝子の塩基配列について相同性検索を行ったところ、受託番号がＦＥＲＭ Ｐ−２０５００である乳酸菌株、及び受託番号がＦＥＲＭ Ｐ−２０５０１の乳酸菌株はいずれも、既知のロイコノストック・メセントロイデスに属する微生物の塩基配列と９９％の相同性を有していた。従って、本発明の乳酸菌はいずれもロイコノストック・メセントロイデスに属する微生物と同定された。
<Mycological and physiological properties>
The base sequence of the 16S ribosomal RNA gene of the lactic acid strain whose accession number is FERM P-20500, which is the lactic acid bacterium of the present invention, is shown in SEQ ID NO: 1 of the Sequence Listing. The base sequence of the 16S ribosomal RNA gene of the lactic acid strain whose accession number is FERM P-20501 is shown in SEQ ID NO: 2 of the Sequence Listing. When a homology search was performed on the base sequences of these 16S ribosomal RNA genes on the base sequence database of known microorganisms, a lactic acid strain having an accession number of FERM P-20500 and a lactic acid bacterium having an accession number of FERM P-20501 All the strains had 99% homology with the base sequences of known microorganisms belonging to Leuconostoc mescentroides. Therefore, all the lactic acid bacteria of the present invention were identified as microorganisms belonging to Leuconostoc mescentroides.
また、相同性が９９％であり、完全に一致している既知微生物が検索されなかったことから、新規な乳酸菌であることが確認された。 Moreover, since the homologue was 99% and a known microorganism that was completely matched was not searched for, it was confirmed to be a novel lactic acid bacterium.
本発明で使用する野菜果汁は、特に限定されないが、人参、カボチャ、キャベツ、セロリ、ブロッコリー、ケール、トマト、ピーマン、アスパラガス、ホウレンソウ、カリフラワーなどを挙げることができる。これらは、１種類の野菜果汁であってもよいが、２種類以上の野菜果汁の混合液であってもよい。特に、官能的に相性がよいため、人参、カボチャ、キャベツ、ケール、トマトが、好ましく用いられる。また、これらは、１種類の野菜果汁であってもよいが、２種類以上の野菜果汁の混合液であってもよい。 The vegetable juice used in the present invention is not particularly limited, and examples thereof include carrot, pumpkin, cabbage, celery, broccoli, kale, tomato, pepper, asparagus, spinach and cauliflower. These may be one kind of vegetable juice, but may be a mixed liquid of two or more kinds of vegetable juice. In particular, ginseng, pumpkin, cabbage, kale, and tomato are preferably used because they are functionally compatible. These may be one kind of vegetable juice, but may be a mixture of two or more kinds of vegetable juice.
また、果実としては、特に限定されないが、オレンジ、みかん、グレープフルーツ、レモン、ポンカン、デコポン、リンゴ、ぶどう、メロン、キウイ、イチゴ、ブルーベリー、パイナップル、バナナ、マンゴー、洋ナシ、桃などを挙げることができ、特にオレンジ、みかん、グレープフルーツ、レモン、リンゴ、洋ナシ、桃が好ましく用いられる。また、これらは、１種類の果実であってもよいが、２種類以上の果実の混合液であってもよい。また、発酵を促進するため、発酵前にｐＨ調整剤を添加することもある。 In addition, the fruit is not particularly limited, and examples include orange, orange, grapefruit, lemon, ponkan, decapon, apple, grape, melon, kiwi, strawberry, blueberry, pineapple, banana, mango, pear and peach. In particular, orange, tangerine, grapefruit, lemon, apple, pear and peach are preferably used. These may be one kind of fruit, but may be a mixed liquid of two or more kinds of fruit. Moreover, in order to accelerate | stimulate fermentation, a pH adjuster may be added before fermentation.
また、乳製品としては、特に限定されないが、牛乳、低脂肪乳、脱脂乳、濃縮乳、加工乳、脱脂粉乳、全粉乳、生クリーム、バター、乳蛋白濃縮物、ホエー、植物性クリームなどを挙げることができ、特に牛乳、低脂肪乳、脱脂乳、濃縮乳、加工乳、脱脂粉乳、全粉乳、生クリームが好ましく用いられる。また、これらは、１種類の乳製品であってもよいが、２種類以上の乳製品の混合液であってもよい。 The dairy product is not particularly limited, but includes milk, low fat milk, skim milk, concentrated milk, processed milk, skim milk powder, whole milk powder, fresh cream, butter, milk protein concentrate, whey, vegetable cream, etc. In particular, cow's milk, low fat milk, skim milk, concentrated milk, processed milk, skim milk powder, whole milk powder and fresh cream are preferably used. These may be one kind of dairy product, but may be a mixture of two or more kinds of dairy products.
これらは、野菜果汁、果実、乳製品、豆乳を単独で用いることもでき、２種以上を併用することもできる。また、本発明にかかる飲料は、粉砕物（果肉等）や砂のう等の組織や繊維素等の不溶性の固形分を含んでいてもよい。 These can use vegetable fruit juice, a fruit, dairy products, and soy milk independently, and can also use 2 or more types together. Moreover, the drink concerning this invention may contain insoluble solid content, such as structure | tissues, such as a ground material (fruit pulp etc.) and a sandbag, and a fiber.
また、本発明にかかる飲料には、上記の乳酸菌が、充分なＩＦＮ−γ産生を増強させる効果を奏するために、酸化防止剤、香料、各種エステル類、有機酸類、有機酸塩類、無機酸類、無機酸塩類、無機塩類、色素類、乳化剤、保存料、調味料、甘味料、酸味料、果汁エキス類、野菜エキス類、花蜜エキス類、ｐＨ調整剤、品質安定剤、ビタミン類などの添加剤を単独、あるいは併用して配合してもよい。 In addition, in the beverage according to the present invention, the lactic acid bacterium described above has an effect of enhancing sufficient IFN-γ production, so that antioxidants, fragrances, various esters, organic acids, organic acid salts, inorganic acids, Additives such as inorganic acid salts, inorganic salts, pigments, emulsifiers, preservatives, seasonings, sweeteners, acidulants, fruit juice extracts, vegetable extracts, nectar extracts, pH adjusters, quality stabilizers, vitamins, etc. May be used alone or in combination.
例えば甘味料としては、砂糖、ぶどう糖、果糖、異性化液糖、グリチルリチン、ステビア、アスパルテーム、フラクトオリゴ糖、ガラクトオリゴ糖、スクラロース、アセスルファムカリウム、水あめ等が挙げられる。酸味料としては、天然成分から抽出した果汁類のほか、クエン酸、酒石酸、リンゴ酸、乳酸、フマル酸、リン酸が挙げられる。クエン酸もしくはリンゴ酸を飲料中に０．１〜５ｇ／Ｌ、好ましくは０．５〜２ｇ／Ｌ含有するのがよい。酸化防止剤としては、Ｌ−アスコルビン酸、Ｌ−アスコルビン酸ナトリウム、エリソルビン酸、エリソルビン酸ナトリウム、が挙げられる。飲料中に、０．００５〜０．５質量％、好ましくは０．０１〜０．１質量％含有するのがよい。 Examples of sweeteners include sugar, glucose, fructose, isomerized liquid sugar, glycyrrhizin, stevia, aspartame, fructooligosaccharide, galactooligosaccharide, sucralose, acesulfame potassium, syrup and the like. Examples of acidulants include fruit juices extracted from natural ingredients, citric acid, tartaric acid, malic acid, lactic acid, fumaric acid, and phosphoric acid. Citric acid or malic acid is contained in the beverage in an amount of 0.1 to 5 g / L, preferably 0.5 to 2 g / L. Examples of the antioxidant include L-ascorbic acid, sodium L-ascorbate, erythorbic acid, and sodium erythorbate. It is good to contain 0.005-0.5 mass% in a drink, Preferably it is 0.01-0.1 mass%.
以下、実施例を用いて本発明を更に詳細に説明するが、本発明は以下の実施例に限定されるものではない。 EXAMPLES Hereinafter, although this invention is demonstrated further in detail using an Example, this invention is not limited to a following example.
＜試験例１：乳酸菌の属・種間でのＩＦＮ−γ産生増強作用の比較＞
［乳酸菌死菌体の調製］
表１に示す各菌種を代表的培地としてＭＲＳ培地に接種し、２０〜３７℃で一夜培養（前培養）した。この前培養した溶液を波長６６０ｎｍでの吸光度（ＯＤ６６０）が１．０となるように調製し、ＭＲＳ培地に１／１００（Ｖ／Ｖ）（菌数：１０６個／ｍＬ）の割合で接種した。２０〜３７℃で１０時間以上培養（本培養）し、生菌数約１０７〜１０９個／ｍＬの培養液を得た。得られた培養液を８，０００ｒｐｍ（約１０，０００×ｇ）で２０分間の遠心分離をして集菌し、これを蒸留水で２回洗浄して菌体を得た。この菌体を、用いた液体培地量の１／１００量の蒸留水で懸濁し、１１０℃で１０分間加熱して死菌体懸濁液を得た。次に、凍結乾燥により乾燥処理し、死菌体菌末を得た。
<Test Example 1: Comparison of IFN-γ production enhancing action between genus / species of lactic acid bacteria>
[Preparation of dead lactic acid bacteria]
Each bacterial species shown in Table 1 was inoculated into MRS medium as a representative medium and cultured overnight (preculture) at 20 to 37 ° C. The pre-culture solution absorbance at a wavelength 660 nm (OD660) is prepared so that the 1.0, MRS medium 1/100 (V / V): inoculated at a ratio of (number of cells 10 6 cells / mL) did. Culture (main culture) was performed at 20 to 37 ° C. for 10 hours or more to obtain a culture solution having about 10 7 to 10 9 viable bacteria / mL. The obtained culture solution was centrifuged at 8,000 rpm (about 10,000 × g) for 20 minutes to collect cells, which were washed twice with distilled water to obtain bacterial cells. This microbial cell was suspended in 1/100 of distilled water of the amount of the liquid medium used and heated at 110 ° C. for 10 minutes to obtain a dead cell suspension. Next, it was dried by lyophilization to obtain dead bacterial cell powder.
［試験方法］
脾臓を１２週齢の雄・ＢＡＬＢ／ｃマウスから摘出し、培養液によって洗浄・濾過した後、細胞浮遊液を調製（１×１０７細胞数／ｍＬ）した。この細胞浮遊液を９６穴平底プレートに２００μＬ／ウェルまき、上記で得た乳酸菌死菌体（最終濃度１００μｇ／ｍＬ）の存在下で３７℃、４８時間培養した。培養終了後、培養上清中のＩＦＮ−γ量を酵素結合免疫測定（ＥＬＩＳＡ）法により測定した。結果を図１に示す。
[Test method]
The spleen was removed from 12-week-old male BALB / c mice, washed and filtered with a culture solution, and a cell suspension was prepared (1 × 10 7 cells / mL). This cell suspension was seeded on a 96-well flat bottom plate at 200 μL / well, and cultured at 37 ° C. for 48 hours in the presence of the killed lactic acid bacteria (final concentration 100 μg / mL) obtained above. After completion of the culture, the amount of IFN-γ in the culture supernatant was measured by enzyme linked immunoassay (ELISA). The results are shown in FIG.
乳酸菌無添加区のＩＦＮ−γ産生量を１として、各乳酸菌属、種間での比較を行った。図中の相対ＩＦＮ−γ産生量は、下記式（１）により計算した。 Comparison was made between each lactic acid bacteria genus and species, assuming that the amount of IFN-γ produced in the lactic acid bacteria-free group was 1. The relative IFN-γ production amount in the figure was calculated by the following formula (1).
相対ＩＦＮ−γ産生量＝（各乳酸菌株無添加区のＩＦＮ−γ濃度）／（無添加区のＩＦＮ−γ濃度）・・・（１） Relative IFN-γ production amount = (IFN-γ concentration in each lactic acid bacterial strain-free group) / (IFN-γ concentration in the non-added group) (1)
属間の比較では、ラクトバチルス属（Ｌａｃｔｏｂａｃｉｌｌｕｓ属）、ビフィドバクテリウム属（Ｂｉｆｉｄｂａｃｔｅｒｉｕｍ属）、ペディオコッカス属（Ｐｅｄｉｏｃｏｃｃｕｓ属）、ストレプトコッカス属（Ｓｔｒｅｐｔｏｃｏｃｃｕｓ属）、エンテロコッカス属（Ｅｎｔｅｒｏｃｏｃｕｓ属）は相対ＩＦＮ−γ産生量の属内平均値が１〜３の値を示したのに対して、ロイコノストック属は約９と最も高いＩＦＮ−γ産生増強能を示した。さらにロイコノストック属の中でも、メセントロイデス種に分類される受託番号がＦＥＲＭ Ｐ−２０５００である乳酸菌株、受託番号がＦＥＲＭ Ｐ−２０５０１である乳酸菌株には、特に高いＩＦＮ−γ産生増強作用が認められた。 In comparison between genera, the genus Lactobacillus (genus Lactobacillus), the genus Bifidobacterium (genus Bifidbacterium), the genus Pediococcus (genus Pediococcus), the genus Streptococcus (genus Streptococcus), the relative genus Enterococcus (IFococcus) While the genus average value of γ production showed a value of 1 to 3, the genus Leuconostoc showed the highest ability to enhance IFN-γ production of about 9. Furthermore, among the genus Leuconostoc, the lactic acid strain whose accession number is FERM P-20500 and the lactic acid strain whose accession number is FERM P-20501 is classified into the Mecentroides species, and the IFN-γ production enhancing action is particularly high. Was recognized.
＜実施例１、２＞
［野菜発酵汁の調製］
人参混濁濃縮汁（Ｂｘ４２、濃縮率７倍）を、水を用いてＢｘ２０になるように調製した。この調整液を、１０５℃で３０分間処理した後、ＦＥＲＭ Ｐ−２０５００（実施例１）、又は、ＦＥＲＭ Ｐ−２０５０１（実施例２）により発酵させた。この発酵は、乳酸菌培養用培地で培養したものをスターターとし、１０６〜１０８ｃｅｌｌｓ／ｍｌとなるように接種した後、静置培養をすることにより行った（発酵温度は３０℃、発酵時間は４８時間）。そして、実施例１、２の野菜発酵汁を得た。
<Examples 1 and 2>
[Preparation of vegetable fermented juice]
Ginseng turbid concentrated juice (Bx42, concentration rate 7 times) was prepared using water to become Bx20. This adjustment solution was treated at 105 ° C. for 30 minutes and then fermented with FERM P-20500 (Example 1) or FERM P-20501 (Example 2). This fermentation was carried out by inoculating the cells cultured in a lactic acid bacteria culture medium as a starter and inoculating them at 10 6 to 10 8 cells / ml, followed by stationary culture (fermentation temperature is 30 ° C., fermentation time) 48 hours). And the vegetable fermented juice of Examples 1 and 2 was obtained.
＜試験例２：乳酸菌野菜発酵汁の免疫賦活作用の検討＞
６週齢の雄・Ｂａｌｂ／ｃマウスを用いて検討した。実施例１、２の野菜発酵汁をそれぞれマウスに１０ｍＬ／ｋｇの用量で経口投与した。野菜発酵汁の投与期間は２回／日の頻度で１週間行った。投与終了後、免疫抑制処理として１２時間の拘束ストレスをマウスに負荷した後、脾細胞のＩＦＮ−γ産生量をＥＬＩＳＡ法により測定した。結果を図２に示す。
<Test Example 2: Examination of immunostimulatory action of lactic acid bacteria vegetable fermented juice>
The study was conducted using 6-week-old male Balb / c mice. Each of the fermented vegetable juices of Examples 1 and 2 was orally administered to mice at a dose of 10 mL / kg. The administration period of the fermented vegetable juice was performed twice a day for one week. After completion of administration, mice were subjected to 12 hours of restraint stress as an immunosuppressive treatment, and the amount of IFN-γ produced by splenocytes was measured by ELISA. The results are shown in FIG.
ニンジンエキスのみを投与した対照群１００％として、実施例１、実施例２の比較を行った。対照群と比較し、実施例１、実施例２の野菜発酵汁を投与したマウスは、ＩＦＮ−γ産生能が有意に増強されていることが確認できる。 Example 1 and Example 2 were compared as a control group 100% administered with only carrot extract. As compared with the control group, it can be confirmed that the mice administered with the vegetable fermented juice of Example 1 and Example 2 have significantly enhanced IFN-γ production ability.
Claims (4)
Lactic acid strain (accession number FERM P-20501) classified into Leuconostoc genus Mecentroides.
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| JP4810483B2 (en) * | 2007-03-28 | 2011-11-09 | 株式会社喜源バイオジェニックス研究所 | Fermented food containing plant-based lactic acid bacteria |
| FI20075539L (en) * | 2007-07-12 | 2009-01-13 | Valio Oy | Lactic acid bacteria with proinflammatory properties |
| JP2014000026A (en) * | 2012-06-18 | 2014-01-09 | Kanazawa Univ | Lactobacillus composition for controlling gastrointestinal immunity and lactic acid-fermented food product for controlling gastrointestinal immunity |
| CN105637084B (en) * | 2013-10-17 | 2019-08-06 | 株式公司染色体创药研究所 | Novel lactic acid bacteria, the innate immunity activator using novel lactic acid bacteria as effective component and the diet product containing novel lactic acid bacteria |
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| CN104585826B (en) * | 2015-01-19 | 2016-06-08 | 中国食品发酵工业研究院 | A kind of fermented ginseng goods and preparation method thereof |
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