JP7488182B2

JP7488182B2 - Composition for promoting muscle repair

Info

Publication number: JP7488182B2
Application number: JP2020522633A
Authority: JP
Inventors: 忠昭宮崎; 久子中川; 敬弘關
Original assignee: Megmilk Snow Brand Co Ltd
Current assignee: Megmilk Snow Brand Co Ltd
Priority date: 2018-06-01
Filing date: 2019-05-31
Publication date: 2024-05-21
Anticipated expiration: 2039-05-31
Also published as: JPWO2019230957A1; WO2019230957A1

Description

IPOD IPOD FREM BP-10953FREM BP-10953 NPMD NPMD NITE BP-02642NITE BP-02642 NPMD NPMD NITE BP-02643NITE BP-02643 NPMD NPMD NITE BP-02644NITE BP-02644

本発明は筋修復促進用組成物に関する。 The present invention relates to a composition for promoting muscle repair.

骨格筋はヒトの体重の４０％程度を占める最大の器官であり、その機能は全身運動以外にも姿勢の維持やエネルギー代謝、内臓器官の保護と多岐にわたる。Skeletal muscles are the largest organ in humans, accounting for about 40% of body weight, and their functions are diverse, including not only whole-body movement but also maintaining posture, energy metabolism, and protecting the internal organs.

骨格筋は修復機能を有しており、物理的、化学的な損傷を受けると、筋繊維周囲に位置する筋衛星細胞が働くことで損傷部位が修復される。筋衛星細胞は通常静止した状態であるが、骨格筋に損傷が起きると活性化し、筋芽細胞に分化する。筋芽細胞は増殖後、筋管に分化し、損傷部位に融合することで骨格筋が修復される。Skeletal muscles have a repair function, and when they are physically or chemically damaged, satellite cells located around muscle fibers act to repair the damaged area. Satellite cells are normally quiescent, but when skeletal muscles are damaged, they become activated and differentiate into myoblasts. After proliferation, myoblasts differentiate into myotubes and fuse with the damaged area, repairing the skeletal muscle.

骨格筋の修復を促進する化合物として顆粒球コロニー刺激因子（特許文献１）やβ－ヒドロキシ－β－メチルブチレート（特許文献２）、レチノイン酸受容体γ（ＲＡＲγ）アゴニスト（特許文献３）が報告されている。Compounds that have been reported to promote skeletal muscle repair include granulocyte colony-stimulating factor (Patent Document 1), β-hydroxy-β-methylbutyrate (Patent Document 2), and retinoic acid receptor gamma (RARγ) agonists (Patent Document 3).

乳酸菌は代謝の過程で乳酸を産生する細菌であり、古くから様々な発酵食品に利用されている。近年は、プロバイオティクス（生きて腸まで届くことで宿主に有益な効果をもたらす）として、乳酸菌の機能性が注目されている。Lactic acid bacteria are bacteria that produce lactic acid during the metabolic process and have long been used in a variety of fermented foods. In recent years, the functionality of lactic acid bacteria as probiotics (which reach the intestine alive and have a beneficial effect on the host) has been attracting attention.

これまでに、乳酸菌を有効成分とする筋修復促進用組成物はいずれの文献にも開示も示唆もされていない。 To date, no literature has disclosed or suggested a composition for promoting muscle repair that contains lactic acid bacteria as an active ingredient.

国際公開２０１０／１３１３８２号International Publication No. 2010/131382 特表２０１６－５２００５０号Special Publication No. 2016-520050 特表２０１３－５３６８５５号Special Publication No. 2013-536855

本発明は、乳酸菌を有効成分とする新規の筋修復促進用組成物を提供することを課題とする。 The objective of the present invention is to provide a novel composition for promoting muscle repair that contains lactic acid bacteria as an active ingredient.

上記課題を解決するために、本発明者らは鋭意検討した結果、筋修復促進作用を有する乳酸菌を見出し、本発明を完成させた。
すなわち、本発明は乳酸菌を有効成分とする新規の筋修復促進用組成物を提供するものである。また、本発明は産業上利用可能な新規の乳酸菌株を提供するものである。
したがって、本発明は以下の構成を有する。
（１）ラクトバチルス（Ｌａｃｔｏｂａｃｉｌｌｕｓ）に属する菌の菌体及び／又はその培養物を有効成分とする筋修復促進用組成物。
（２）ラクトバチルス（Ｌａｃｔｏｂａｃｉｌｌｕｓ）に属する菌が、ラクトバチルスガセリ（Ｌａｃｔｏｂａｃｉｌｌｕｓｇａｓｓｅｒｉ）、ラクトバチルスデルブルッキーサブスピーシーズデルブルッキー（Ｌａｃｔｏｂａｃｉｌｌｕｓｄｅｌｂｒｕｅｃｋｉｉｓｕｂｓｐ．ｄｅｌｂｒｕｅｃｋｉｉ）、ラクトバチルスロイテリ（Ｌａｃｔｏｂａｃｉｌｌｕｓｒｅｕｔｅｒｉ）、ラクトバチルスヘルベティカス（Ｌａｃｔｏｂａｃｉｌｌｕｓｈｅｌｖｅｔｉｃｕｓ）から選択されるひとつ以上であることを特徴とする（１）に記載の筋修復促進用組成物。
（３）ラクトバチルス（Ｌａｃｔｏｂａｃｉｌｌｕｓ）に属する乳酸菌が、ラクトバチルスガセリＳＢＴ２０５５（ＦＥＲＭＢＰ－１０９５３）、ラクトバチルスデルブルッキーサブスピーシーズデルブルッキーＳＢＴ０４１３（ＮＩＴＥＰ－０２６４２）、ラクトバチルスロイテリＳＢＴ１９２６（ＮＩＴＥＰ－０２６４３）、ラクトバチルスヘルベティカスＳＢＴ１１３８０（ＮＩＴＥＰ－０２６４４）から選択されるひとつ以上であることを特徴とする（１）に記載の筋修復促進用組成物。
（４）前記菌体及び／又はその培養物が、死菌体である、（１）～（３）のいずれか一項に記載の筋修復促進用組成物。
（５）前記死菌体が、菌体破砕物の不溶性画分である、（４）に記載の筋修復促進用組成物。
（６）新規乳酸菌ラクトバチルスデルブルッキーサブスピーシーズデルブルッキーＳＢＴ０４１３。
（７）新規乳酸菌ラクトバチルスロイテリＳＢＴ１９２６。
（８）新規乳酸菌ラクトバチルスヘルベティカスＳＢＴ１１３８０。 In order to solve the above problems, the present inventors conducted extensive research and discovered lactic acid bacteria having the effect of promoting muscle repair, thereby completing the present invention.
That is, the present invention provides a novel composition for promoting muscle repair that contains lactic acid bacteria as an active ingredient, and also provides a novel industrially applicable lactic acid bacteria strain.
Therefore, the present invention has the following configuration.
(1) A composition for promoting muscle repair, comprising as an active ingredient a bacterial cell of a bacterium belonging to Lactobacillus and/or a culture thereof.
(2) The composition for promoting muscle repair according to (1), characterized in that the bacterium belonging to Lactobacillus is one or more selected from Lactobacillus gasseri, Lactobacillus delbrueckii subsp. delbrueckii, Lactobacillus reuteri, and Lactobacillus helveticus.
(3) The composition for promoting muscle repair according to (1), characterized in that the lactic acid bacteria belonging to Lactobacillus are one or more selected from Lactobacillus gasseri SBT2055 (FERM BP-10953), Lactobacillus delbrueckii subsp. delbrueckii SBT0413 (NITE P-02642), Lactobacillus reuteri SBT1926 (NITE P-02643), and Lactobacillus helveticus SBT11380 (NITE P-02644).
(4) A composition for promoting muscle repair described in any one of (1) to (3), wherein the bacterial cells and/or cultures thereof are killed bacterial cells.
(5) The composition for promoting muscle repair described in (4), wherein the killed bacterial cells are an insoluble fraction of disrupted bacterial cells.
(6) A novel lactic acid bacterium, Lactobacillus delbrueckii subsp. delbrueckii SBT0413.
(7) Novel lactic acid bacterium, Lactobacillus reuteri SBT1926.
(8) Novel lactic acid bacterium, Lactobacillus helveticus SBT11380.

本発明によれば、ラクトバチルス（Ｌａｃｔｏｂａｃｉｌｌｕｓ）に属する菌の菌体及び／又はその培養物を有効成分とする筋修復促進用組成物を提供することができる。また、本発明によれば、産業上利用可能な新規の乳酸菌株を提供することができる。According to the present invention, a composition for promoting muscle repair can be provided that contains as an active ingredient a bacterial cell of a bacterium belonging to the genus Lactobacillus and/or a culture thereof. Furthermore, according to the present invention, a novel lactic acid bacteria strain that can be used industrially can be provided.

筋損傷剤反応後（２４時間）のマウス筋芽細胞から筋損傷剤を除去し、様々な乳酸菌加熱菌体を添加して、４８時間培養後に筋芽細胞の増殖率を比較したグラフである。This is a graph comparing the proliferation rates of myoblasts after 48 hours of incubation in which the muscle-damaging agent was removed from mouse myoblasts after reaction with the agent (24 hours), various heat-treated lactic acid bacteria cells were added, and the myoblasts were cultured for 48 hours. 筋損傷剤反応後（２４時間）のマウス筋芽細胞から筋損傷剤を除去し、乳酸菌加熱菌体（ラクトバチルスガセリ）を異なる濃度で添加して、４８時間培養後に筋芽細胞の増殖率を比較したグラフである。This graph shows the results of removing the muscle-damaging agent from mouse myoblasts after reaction with the agent (24 hours), adding heat-treated lactic acid bacteria (Lactobacillus gasseri) at different concentrations, and comparing the proliferation rate of the myoblasts after culturing for 48 hours. 筋損傷剤反応後（２４時間）のマウス筋芽細胞から筋損傷剤を除去し、乳酸菌加熱菌体（ラクトバチルスガセリ）の破砕物の画分を添加して、４８時間培養後に筋芽細胞の増殖率を比較したグラフである。This is a graph comparing the proliferation rate of myoblasts after 48 hours of incubation in which the muscle-damaging agent was removed from mouse myoblasts after reaction with the agent (24 hours), a fraction of disrupted heat-treated lactic acid bacteria (Lactobacillus gasseri) was added, and the cells were cultured for 48 hours.

（ラクトバチルス（Ｌａｃｔｏｂａｃｉｌｌｕｓ）属に属する乳酸菌）
本発明のラクトバチルス（Ｌａｃｔｏｂａｃｉｌｌｕｓ）属に属する乳酸菌は、ラクトバチルス（Ｌａｃｔｏｂａｃｉｌｌｕｓ）属に分類される乳酸菌であればどのようなものでも用いることができる。
具体的には、ラクトバチルスガセリ（Ｌａｃｔｏｂａｃｉｌｌｕｓｇａｓｓｅｒｉ）、ラクトバチルスデルブルッキーサブスピーシーズデルブルッキー（Ｌａｃｔｏｂａｃｉｌｌｕｓｄｅｌｂｒｕｅｃｋｉｉｓｕｂｓｐ．ｄｅｌｂｒｕｅｃｋｉｉ）、ラクトバチルスロイテリ（Ｌａｃｔｏｂａｃｉｌｌｕｓｒｅｕｔｅｒｉ）、ラクトバチルスヘルベティカス（Ｌａｃｔｏｂａｃｉｌｌｕｓｈｅｌｖｅｔｉｃｕｓ）等を例示できるが、これらに限定されるものではない。
本発明のラクトバチルス（Ｌａｃｔｏｂａｃｉｌｌｕｓ）属に属する乳酸菌は、ラクトバチルス（Ｌａｃｔｏｂａｃｉｌｌｕｓ）に属する乳酸菌が、ラクトバチルスガセリＳＢＴ２０５５（ＦＥＲＭＢＰ－１０９５３）、ラクトバチルスデルブルッキーサブスピーシーズデルブルッキーＳＢＴ０４１３（ＮＩＴＥＰ－０２６４２）、ラクトバチルスロイテリＳＢＴ１９２６（ＮＩＴＥＰ－０２６４３）、ラクトバチルスヘルベティカスＳＢＴ１１３８０（ＮＩＴＥＰ－０２６４４）であることが好ましい。
（ラクトバチルス（Ｌａｃｔｏｂａｃｉｌｌｕｓ）属に属する乳酸菌の調製）
ラクトバチルス属に属する乳酸菌は、乳酸菌培養の常法に従って培養し、所望の量を調製すればよい。調製の一態様を以下に示す。ＭＲＳ（ＤＩＦＣＯ）培地を用いてラクトバチルス属に属する乳酸菌を培養し、得られた培養物を遠心分離により集菌することにより菌体を得る。得られた菌体をそのまま用いてもよいし、濃縮、乾燥、凍結乾燥処理に供した菌体を用いることもできる。菌体は加熱乾燥などにより死菌体にしたものを用いることもできる。
（ラクトバチルス属に属する乳酸菌の不溶性画分（沈渣））
本発明のラクトバチルス属に属する乳酸菌の不溶性画分（沈渣）の調製法の一態様を以下に示す。ラクトバチルス属の菌体の凍結乾燥末を緩衝液に懸濁し、８０℃、３０分間程度加熱して加熱菌体を得る。この加熱菌体をフレンチプレスにより破砕する。破砕液を遠心分離して上清を除き、ラクトバチルス属に属する乳酸菌の沈渣を得る。
本発明の筋修復促進用組成物では、有効成分は菌体破砕物の不溶性画分であることができ、詳細には菌体破砕物の緩衝液（例えば、リン酸緩衝液）への不溶性画分であることができる。菌体破砕物の可溶性画分を追加で含んでもよく、含まなくともよい。
（新規乳酸菌株）
本発明は、新規乳酸菌株に関するものである。これらの乳酸菌株とはラクトバチルスに属するラクトバチルスデルブルッキーサブスピーシーズデルブルッキー（Ｌａｃｔｏｂａｃｉｌｌｕｓｄｅｌｂｒｕｅｃｋｉｉｓｕｂｓｐ．ｄｅｌｂｒｕｅｃｋｉｉ）ＳＢＴ０４１３（ＮＩＴＥＰ－０２６４２）、ラクトバチルスロイテリ（Ｌａｃｔｏｂａｃｉｌｌｕｓｒｅｕｔｅｒｉ）ＳＢＴ１９２６（ＮＩＴＥＰ－０２６４３）、ラクトバチルスヘルベティカス（Ｌａｃｔｏｂａｃｉｌｌｕｓｈｅｌｖｅｔｉｃｕｓ）ＳＢＴ１１３８０（ＮＩＴＥＰ－０２６４４）である。以下、同乳酸菌株を「本発明の乳酸菌」、「本発明の乳酸菌株」、又は単にＳＢＴ０４１３、ＳＢＴ１９２６、ＳＢＴ１１３８０株と記載することがある。
これらの乳酸菌株は、２０１８年３月１５日に、独立行政法人製品評価技術基盤機構特許微生物寄託センター（郵便番号２９２－０８１８千葉県木更津市かずさ鎌足２－５－８１２２号室）に、ＳＢＴ０４１３はＮＩＴＥＰ－０２６４２、ＳＢＴ１９２６はＮＩＴＥＰ－０２６４３、ＳＢＴ１１３８０はＮＩＴＥＰ－０２６４４の受託番号で寄託されている。
本発明の乳酸菌は、上記乳酸菌株に制限されず、これらの寄託乳酸菌株と実質的に同等の乳酸菌株であってもよい。実質的に同等の乳酸菌株とは、ラクトバチルスデルブルッキーサブスピーシーズデルブルッキー、ラクトバチルスロイテリ、ラクトバチルスヘルベティカスに属する乳酸菌株であって、寄託乳酸菌株と同程度の高い筋修復促進作用を有する乳酸菌株を言う。
また、実質的に同等の乳酸菌株は、さらに、その１６ＳｒＲＮＡ遺伝子の塩基配列が、上記寄託乳酸菌株の１６ＳｒＲＮＡ遺伝子の塩基配列と９８％以上、好ましくは９９％以上、より好ましくは１００％の相同性を有し、且つ、好ましくは上記寄託乳酸菌株と同一の菌学的性質を有する。さらに、本発明の乳酸菌は、本発明の効果が損なわれない限り、寄託乳酸菌株、又はそれと実質的に同等の乳酸菌株から、変異処理、遺伝子組換え、自然変異株の選択等によって育種された乳酸菌株であってもよい。
（利用方法）
上記の通り、本発明の組成物は濃縮、乾燥、凍結乾燥処理に供した菌体、加熱乾燥などにより得られる死菌体も有効成分とすることができることから、製剤、飲食品、飼料の原料として広く用いることができる。
本発明の組成物の投与対象は特に限定されず、ヒトに対して投与することができるが、投与対象はヒト以外の動物（例えば、イヌ、ネコ、ウマ又はウサギ等）であっても良い。投与対象がヒトである場合は、２０歳未満の未成年、成人、又は６５歳以上の高齢者などに投与することができる。
本発明の組成物の摂取量は、投与対象者の症状、年齢などを考慮してそれぞれ個別に決定されるが、通常成人の場合、０．５－５０００ｍｇであればよく、０．５－５００ｍｇが望ましく、０．５－５０ｍｇが最も望ましい。
（筋修復促進効果の評価方法）
実施例に記載の方法で評価が可能である。すなわち、以下の方法で評価が可能である。
１０ｍｇ/ｍｌラクトバチルスガセリＳＢＴ２０５５加熱菌体をフレンチプレス（Ａｍｉｎｃｏ）で１,２００ｐｓｉＧで３回破砕する。破砕後に、遠心分離（４℃、７０００ｒｐｍ、１５分間）をして上清と沈渣を得る。さらに、上清を０．２２μｍフィルターで濾過した。沈渣は除去した上清と等量のＰＢＳ（－）を添加して、ボルテックス後に遠心分離（４℃、７０００ｒｐｍ、１５分間）し、上清を除去する。これを３回繰り返し、上清と等量のＰＢＳ（－）を添加したものを沈渣として使用する。フレンチプレスに供した１０ｍｇ／ｍｌラクトバチルスガセリＳＢＴ２０５５加熱菌体から得られた破砕物上清と沈渣の濃度を、それぞれ１０ｍｇ／ｍｌ相当量とする。１００μｇ/ｍｌ相当量になるように１０％ＦＢＳ（ＧＩＢＣＯ）、１％ペニシリンーストレプトマイシン（ＳＩＧＭＡ）を含有したＤＭＥＭ（ＳＩＧＭＡ）で希釈し、破砕物上清の懸濁液または沈渣の懸濁液を得る。Ｃ２Ｃ１２マウス筋芽細胞をコラーゲンコートした９６ｗｅｌｌプレートに５，０００ｃｅｌｌｓ/ｗｅｌｌずつ播種して、５％ＣＯ_２インキュベーターにて３７℃、２４時間培養後に、０．５μＭブピバカイン塩酸塩（和光純薬）を２４時間反応させる。その後、ブピバカインを取り除き、未破砕の加熱菌体、破砕物上清、または破砕物沈渣を添加して筋芽細胞を４８時間培養する。培養後、滅菌ＰＢＳ（－）による洗浄を２回行ってから、ｃｅｌｌｃｏｕｎｔｉｎｇｋｉｔ―８（同仁化学）を添加して３７℃、２時間反応させて４５０ｎｍの吸光度を測定する。得られた吸光度の値を（加熱菌体－ブランク）／（加熱菌体非添加－ブランク）×１００の計算式に当てはめて、増殖率を算出する。乳酸菌菌体による筋芽細胞の増殖促進効果は加熱菌体非添加と比べて、増殖率の増加が認められれば、損傷後の筋芽細胞の増殖を促進したと判断することができる。 (Lactic acid bacteria belonging to the genus Lactobacillus)
As the lactic acid bacteria belonging to the genus Lactobacillus of the present invention, any lactic acid bacteria classified into the genus Lactobacillus can be used.
Specific examples include Lactobacillus gasseri, Lactobacillus delbrueckii subsp. delbrueckii, Lactobacillus reuteri, Lactobacillus helveticus, and the like, but are not limited thereto.
The lactic acid bacteria belonging to the genus Lactobacillus of the present invention are preferably Lactobacillus gasseri SBT2055 (FERM BP-10953), Lactobacillus delbrueckii subsp. delbrueckii SBT0413 (NITE P-02642), Lactobacillus reuteri SBT1926 (NITE P-02643), or Lactobacillus helveticus SBT11380 (NITE P-02644).
(Preparation of lactic acid bacteria belonging to the genus Lactobacillus)
The lactic acid bacteria belonging to the genus Lactobacillus may be cultured according to a conventional method for culturing lactic acid bacteria, and a desired amount may be prepared. One embodiment of the preparation is shown below. The lactic acid bacteria belonging to the genus Lactobacillus are cultured using MRS (DIFCO) medium, and the resulting culture is centrifuged to collect the bacteria to obtain the bacteria. The obtained bacteria may be used as it is, or may be subjected to a concentration, drying, or freeze-drying process. The bacteria may be killed by heating and drying, etc., and used.
(Insoluble fraction (sediment) of lactic acid bacteria belonging to the genus Lactobacillus)
The following is an embodiment of the method for preparing the insoluble fraction (sediment) of the lactic acid bacteria belonging to the genus Lactobacillus of the present invention. The freeze-dried powder of the bacteria of the genus Lactobacillus is suspended in a buffer solution and heated at 80°C for about 30 minutes to obtain heated bacteria. The heated bacteria are disrupted by a French press. The disrupted liquid is centrifuged to remove the supernatant, and a sediment of the lactic acid bacteria belonging to the genus Lactobacillus is obtained.
In the composition for promoting muscle repair of the present invention, the active ingredient can be an insoluble fraction of the disrupted bacterial cell, specifically, a fraction of the disrupted bacterial cell that is insoluble in a buffer (e.g., phosphate buffer). A soluble fraction of the disrupted bacterial cell may or may not be additionally contained.
(New lactic acid bacteria strain)
The present invention relates to novel lactic acid bacteria strains, which are Lactobacillus delbrueckii subsp. delbrueckii SBT0413 (NITE P-02642), Lactobacillus reuteri SBT1926 (NITE P-02643), and Lactobacillus helveticus SBT11380 (NITE P-02644), all of which belong to the Lactobacillus genus. Hereinafter, this lactic acid bacteria strain may be referred to as "the lactic acid bacteria of the present invention,""the lactic acid bacteria strain of the present invention," or simply as SBT0413, SBT1926, or SBT11380 strain.
These lactic acid bacteria strains were deposited on March 15, 2018 at the National Institute of Technology and Evaluation, Patent Microorganisms Deposit Center (Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818). SBT0413 was deposited under the accession number NITE P-02642, SBT1926 was deposited under the accession number NITE P-02643, and SBT11380 was deposited under the accession number NITE P-02644.
The lactic acid bacteria of the present invention are not limited to the above-mentioned lactic acid bacteria strains, and may be substantially equivalent to these deposited lactic acid bacteria strains. The substantially equivalent lactic acid bacteria strains refer to lactic acid bacteria strains belonging to Lactobacillus delbrueckii subsp. delbrueckii, Lactobacillus reuteri, and Lactobacillus helveticus, and have the same high muscle repair promoting effect as the deposited lactic acid bacteria strains.
Furthermore, the substantially equivalent lactic acid bacteria strain has a base sequence of its 16S rRNA gene that is 98% or more, preferably 99% or more, more preferably 100% homologous to the base sequence of the 16S rRNA gene of the deposited lactic acid bacteria strain, and preferably has the same bacteriological properties as the deposited lactic acid bacteria strain. Furthermore, the lactic acid bacteria of the present invention may be a lactic acid bacteria strain bred from the deposited lactic acid bacteria strain or a substantially equivalent lactic acid bacteria strain by mutation treatment, genetic recombination, selection of natural mutants, or the like, so long as the effects of the present invention are not impaired.
(How to Use)
As described above, the composition of the present invention can contain as an active ingredient bacterial cells subjected to concentration, drying, or freeze-drying treatment, or killed bacterial cells obtained by heat drying or the like, and therefore can be widely used as a raw material for preparations, foods, beverages, and feeds.
The subject of administration of the composition of the present invention is not particularly limited, and the subject of administration can be humans, but the subject of administration may be animals other than humans (e.g., dogs, cats, horses, rabbits, etc.). When the subject of administration is humans, the composition can be administered to minors under 20 years of age, adults, or elderly people over 65 years of age.
The dosage of the composition of the present invention is determined individually for each subject taking into consideration the symptoms, age, etc., of the subject, but generally, for adults, it is 0.5-5000 mg, preferably 0.5-500 mg, and most preferably 0.5-50 mg.
(Method for evaluating the muscle repair promoting effect)
The evaluation can be performed by the method described in the Examples. That is, the evaluation can be performed by the following method.
10 mg/ml Lactobacillus gasseri SBT2055 heated cells are disrupted three times at 1,200 psiG in a French press (Aminco). After disruption, the cells are centrifuged (4°C, 7000 rpm, 15 minutes) to obtain a supernatant and a sediment. The supernatant is then filtered through a 0.22 μm filter. The sediment is added with an equal amount of PBS(-) to the removed supernatant, vortexed, and centrifuged (4°C, 7000 rpm, 15 minutes) to remove the supernatant. This is repeated three times, and the supernatant is used as the sediment after adding an equal amount of PBS(-). The concentrations of the disrupted supernatant and sediment obtained from 10 mg/ml Lactobacillus gasseri SBT2055 heated cells subjected to French press are equivalent to 10 mg/ml, respectively. The cells were diluted with DMEM (SIGMA) containing 10% FBS (GIBCO) and 1% penicillin-streptomycin (SIGMA) to a concentration equivalent to 100 μg/ml to obtain a suspension of the homogenate supernatant or a suspension of the sediment. C2C12 mouse myoblasts were seeded at 5,000 cells/well on a collagen-coated 96-well plate, and incubated at 37°C for 24 hours in a 5% _CO2 incubator. Then, 0.5 μM bupivacaine hydrochloride (Wako Pure Chemical Industries) was added and the myoblasts were incubated for 48 hours with undisrupted heated cells, homogenate supernatant, or homogenate sediment. After incubation, the cells were washed twice with sterile PBS (-), and cell counting kit-8 (Dojin Chemical Industries) was added and incubated at 37°C for 2 hours to measure the absorbance at 450 nm. The proliferation rate was calculated by applying the obtained absorbance value to the formula (heated cells - blank) / (no heated cells added - blank) x 100. If an increase in proliferation rate is observed in the proliferation-promoting effect of lactic acid bacteria cells on myoblasts compared to the case where heated cells are not added, it can be determined that the proliferation of myoblasts after injury has been promoted.

以下、本発明の実施例をもとにさらに詳細に説明するが、本発明は係る実施例に限定して解釈されるものではない。なお、特に説明のない限り、本明細書において％は重量％を示す。The present invention will be described in more detail below with reference to examples, but the present invention should not be construed as being limited to these examples. In this specification, % means % by weight unless otherwise specified.

＜実施例品１＞乳酸菌加熱菌体
下記（１）の各供試菌をＭＲＳ培地（ＤＩＦＣＯ）に植菌し、３７℃にて１６時間静置培養を行った。培養物を、遠心分離（４℃、７０００ｒｐｍ、１５分間）した後、滅菌水による洗浄と遠心分離を３回繰り返して行い、洗浄菌体を得た。この洗浄菌体を凍結乾燥処理して菌体粉末を得た。菌体粉末を１０ｍｇ/ｍｌになるように滅菌ＰＢＳ（－）で希釈し、８０℃、３０分間加熱して加熱菌体を得た。加熱菌体は１００μｇ/ｍｌになるように１０％ＦＢＳ（ＧＩＢＣＯ）、１％ペニシリン－ストレプトマイシン（ＳＩＧＭＡ）を含有したＤＭＥＭ（ＳＩＧＭＡ）で希釈した。
（１）供試菌
ラクトバチルス属の下記７株を供試菌とした。
ラクトバチルスガセリ（Ｌａｃｔｏｂａｃｉｌｌｕｓｇａｓｓｅｒｉ）ＳＢＴ２０５５（ＦＥＲＭＢＰ―１０９５３）、ラクトバチルスデルブルッキーサブスピーシーズブルガリカス（Ｌａｃｔｏｂａｃｉｌｌｕｓｄｅｌｂｒｕｅｃｋｉｉｓｕｂｓｐ．ｂｕｌｇａｒｉｃｕｓ）ＳＢＴ２１１５、ラクトバチルスデルブルッキーサブスピーシーズデルブルッキー（Ｌａｃｔｏｂａｃｉｌｌｕｓｄｅｌｂｒｕｅｃｋｉｉｓｕｂｓｐ．ｄｅｌｂｒｕｅｃｋｉｉ）ＳＢＴ０４１３（ＮＩＴＥＰ－０２６４２）、ラクトバチルスデルブルッキーサブスピーシーズラクティス（Ｌａｃｔｏｂａｃｉｌｌｕｓｄｅｌｂｒｕｅｃｋｉｉｓｕｂｓｐ．ｌａｃｔｉｓ）ＳＢＴ２０８０、ラクトバチルスロイテリ（Ｌａｃｔｏｂａｃｉｌｌｕｓｒｅｕｔｅｒｉ）ＳＢＴ１９２６（ＮＩＴＥＰ－０２６４３）、ラクトバチルスヘルベティカス（Ｌａｃｔｏｂａｃｉｌｌｕｓｈｅｌｖｅｔｉｃｕｓ）ＳＢＴ１１３８０（ＮＩＴＥＰ－０２６４４）、ラクトバチルスブレビス（Ｌａｃｔｏｂａｃｉｌｌｕｓｂｒｅｖｉｓ）ＳＢＴ２０３５
なお、ＦＥＲＭＢＰ－１０９５３は、独立行政法人産業技術総合研究所特許微生物寄託センターに寄託されている。
＜試験例１＞筋修復を促進する乳酸菌のスクリーニング
実施例品１を以下の試験に供した。
（１）試験方法
Ｃ２Ｃ１２マウス筋芽細胞を１０％ＦＢＳ（ＧＩＢＣＯ）、１％ペニシリン－ストレプトマイシン（ＳＩＧＭＡ）を含有したＤＭＥＭ（ＳＩＧＭＡ）で培養した。Ｃ２Ｃ１２マウス筋芽細胞をコラーゲンコートした９６ｗｅｌｌプレートに５，０００ｃｅｌｌｓ/ｗｅｌｌずつ播種して、５％ＣＯ_２インキュベーターにて３７℃、２４時間培養後に、０．５μＭブピバカイン塩酸塩（和光純薬）を２４時間反応させた。
その後、ブピバカインを取り除き、各加熱菌体を添加して筋芽細胞を４８時間培養した。培養後、滅菌ＰＢＳ（－）による洗浄を２回行ってから、ｃｅｌｌｃｏｕｎｔｉｎｇｋｉｔ―８（同仁化学）を添加して３７℃、２時間反応させて４５０ｎｍの吸光度を測定した。得られた吸光度の値を（加熱菌体添加－ブランク）／（加熱菌体非添加－ブランク）×１００の計算式に当てはめて、増殖率を算出した。乳酸菌菌体による筋芽細胞の増殖促進効果は加熱菌体非添加と比べて、増殖率の増加が認められれば、損傷後の筋芽細胞の増殖を促進したと判断することができる。 Example 1: Heated lactic acid bacteria cells Each test bacterium listed in (1) below was inoculated into MRS medium (DIFCO) and subjected to static culture at 37°C for 16 hours. The culture was centrifuged (4°C, 7000 rpm, 15 minutes), and then washed with sterile water and centrifuged three times to obtain washed cells. These washed cells were freeze-dried to obtain a cell powder. The cell powder was diluted with sterile PBS (-) to 10 mg/ml, and heated at 80°C for 30 minutes to obtain heated cells. The heated cells were diluted with DMEM (SIGMA) containing 10% FBS (GIBCO) and 1% penicillin-streptomycin (SIGMA) to 100 μg/ml.
(1) Test Bacteria The following seven strains of the genus Lactobacillus were used as test bacteria.
Lactobacillus gasseri SBT2055 (FERM BP-10953), Lactobacillus delbrueckii subsp. bulgaricus SBT2115, Lactobacillus delbrueckii subsp. delbrueckii SBT0413 (NITE P-02642), Lactobacillus delbrueckii subsp. lactis subsp. lactis SBT2080, Lactobacillus reuteri SBT1926 (NITE P-02643), Lactobacillus helveticus SBT11380 (NITE P-02644), Lactobacillus brevis SBT2035
FERM BP-10953 has been deposited at the Patent Microorganisms Depositary of the National Institute of Advanced Industrial Science and Technology.
<Test Example 1> Screening of lactic acid bacteria that promote muscle repair Example product 1 was subjected to the following test.
(1) Test method C2C12 mouse myoblast cells were cultured in DMEM (SIGMA) containing 10% FBS (GIBCO) and 1% penicillin-streptomycin (SIGMA). C2C12 mouse myoblast cells were seeded at 5,000 cells/well on a collagen-coated 96-well plate and cultured in a 5% _CO2 incubator at 37°C for 24 hours, after which they were reacted with 0.5 μM bupivacaine hydrochloride (Wako Pure Chemical Industries) for 24 hours.
Thereafter, bupivacaine was removed, each heated bacterial cell was added, and myoblasts were cultured for 48 hours. After the culture, the cells were washed twice with sterile PBS (-), and then cell counting kit-8 (Dojindo Chemical Industries) was added and reacted at 37°C for 2 hours to measure the absorbance at 450 nm. The obtained absorbance value was applied to the formula (heated bacterial cell added-blank)/(heated bacterial cell not added-blank) x 100 to calculate the proliferation rate. If an increase in proliferation rate is observed in the proliferation rate of myoblasts compared to the case where heated bacterial cells are not added, it can be judged that the proliferation of myoblasts after injury has been promoted by the lactic acid bacteria cells.

（２）試験結果
乳酸菌株ごとの筋芽細胞の増殖率を図１に示した。吸光度を測定した結果、７菌株中４菌株の加熱菌体において、筋芽細胞の増殖率の増加が認められた。したがって、特定の株の乳酸菌加熱菌体に損傷後の筋芽細胞の増殖を促進する作用が認められた。 (2) Test results The proliferation rate of myoblasts for each lactic acid bacteria strain is shown in Figure 1. As a result of measuring the absorbance, an increase in the proliferation rate of myoblasts was observed in the heated cells of 4 out of 7 strains. Therefore, it was confirmed that the heated cells of certain strains of lactic acid bacteria have the effect of promoting the proliferation of myoblasts after damage.

＜試験例２＞ラクトバチルスガセリＳＢＴ２０５５加熱菌体の濃度依存的な効果
（乳酸菌菌体の調製）
実施例品１について、菌体の濃度依存的な筋修復促進効果を調べた。
（１）試験方法
１０ｍｇ/ｍｌラクトバチルスガセリＳＢＴ２０５５加熱菌体を１、１０、１００μｇ/ｍｌになるように１０％ＦＢＳ（ＧＩＢＣＯ）、１％ペニシリンーストレプトマイシン（ＳＩＧＭＡ）を含有したＤＭＥＭ（ＳＩＧＭＡ）で希釈した。Ｃ２Ｃ１２マウス筋芽細胞をコラーゲンコートした９６ｗｅｌｌプレートに５，０００ｃｅｌｌｓ/ｗｅｌｌずつ播種して、５％ＣＯ_２インキュベーターに３７℃、２４時間培養後に、０．５μＭブピバカイン塩酸塩（和光純薬）を２４時間反応させた。その後、ブピバカインを取り除き、各濃度加熱菌体を添加して筋芽細胞を４８時間培養した。培養後、滅菌ＰＢＳ（－）による洗浄を２回行ってから、ｃｅｌｌｃｏｕｎｔｉｎｇｋｉｔ―８（同仁化学）を添加して３７℃、２時間反応させて４５０ｎｍの吸光度を測定した。得られた吸光度の値を（加熱菌体－ブランク）／（加熱菌体非添加－ブランク）×１００の計算式に当てはめて、増殖率を算出した。乳酸菌菌体による筋芽細胞の増殖促進効果は加熱菌体非添加と比べて、増殖率の増加が認められれば、損傷後の筋芽細胞の増殖を促進したと判断することができる。 Test Example 2: Concentration-dependent effect of heated Lactobacillus gasseri SBT2055 cells (preparation of lactic acid bacteria cells)
The muscle repair promoting effect of Example Product 1, which depends on the concentration of bacterial cells, was investigated.
(1) Test method 10mg/ml Lactobacillus gasseri SBT2055 heated cells were diluted with DMEM (SIGMA) containing 10% FBS (GIBCO) and 1% penicillin-streptomycin (SIGMA) to 1, 10, and 100μg/ml. C2C12 mouse myoblasts were seeded on a collagen-coated 96-well plate at 5,000 cells/well, and incubated in a 5% _CO2 incubator at 37℃ for 24 hours, after which 0.5μM bupivacaine hydrochloride (Wako Pure Chemical Industries) was reacted for 24 hours. Then, bupivacaine was removed, and heated cells were added at each concentration, and the myoblasts were incubated for 48 hours. After the culture, the cells were washed twice with sterilized PBS(-), and then cell counting kit-8 (Dojindo Laboratories) was added and incubated at 37°C for 2 hours to measure absorbance at 450 nm. The obtained absorbance value was applied to the formula (heated cells-blank)/(no heated cells-blank) x 100 to calculate the proliferation rate. If an increase in proliferation rate is observed in the proliferation-promoting effect of lactic acid bacteria cells on myoblasts compared to the case where heated cells are not added, it can be determined that the proliferation of myoblasts after injury has been promoted.

（２）試験結果
ラクトバチルスガセリＳＢＴ２０５５加熱菌体濃度ごとの筋芽細胞の増殖率を図２に示した。吸光度を測定した結果、ラクトバチルスガセリＳＢＴ２０５５加熱菌体の濃度依存的に、筋芽細胞の増殖率の増加が認められた。したがって、損傷後の筋芽細胞の増殖を促進する作用にはラクトバチルスガセリＳＢＴ２０５５加熱菌体の濃度依存性が認められた。 (2) Test results Figure 2 shows the proliferation rate of myoblasts for each concentration of Lactobacillus gasseri SBT2055 heated cells. As a result of measuring absorbance, it was found that the proliferation rate of myoblasts increases depending on the concentration of Lactobacillus gasseri SBT2055 heated cells. Therefore, it was found that the effect of promoting the proliferation of myoblasts after injury is dependent on the concentration of Lactobacillus gasseri SBT2055 heated cells.

＜試験例３＞ラクトバチルスガセリＳＢＴ２０５５加熱菌体の有効成分の探索
（乳酸菌菌体の調製）
実施例品１について、菌体破砕物の可溶性画分（上清）と不溶性画分（沈渣）の筋修復促進効果を調べた。
（１）試験方法
１０ｍｇ/ｍｌラクトバチルスガセリＳＢＴ２０５５加熱菌体をフレンチプレス（Ａｍｉｎｃｏ）で１,２００ｐｓｉＧで３回破砕した。破砕後に、遠心分離（４℃、７０００ｒｐｍ、１５分間）をして上清と沈渣を得た。さらに、上清は０．２２μｍフィルターで濾過した。沈渣は除去した上清と等量のＰＢＳ（－）を添加して、ボルテックス後に遠心分離（４℃、７０００ｒｐｍ、１５分間）し、上清を除去した。これを３回繰り返し、上清と等量のＰＢＳ（－）を添加したものを沈渣として使用した。フレンチプレスに供した１０ｍｇ／ｍｌラクトバチルスガセリＳＢＴ２０５５加熱菌体から得られた破砕物上清と沈渣の濃度を、それぞれ１０ｍｇ／ｍｌ相当量とした。１００μｇ/ｍｌ相当量になるように１０％ＦＢＳ（ＧＩＢＣＯ）、１％ペニシリンーストレプトマイシン（ＳＩＧＭＡ）を含有したＤＭＥＭ（ＳＩＧＭＡ）で希釈し、破砕物上清の懸濁液または沈渣の懸濁液を得た。Ｃ２Ｃ１２マウス筋芽細胞をコラーゲンコートした９６ｗｅｌｌプレートに５，０００ｃｅｌｌｓ/ｗｅｌｌずつ播種して、５％ＣＯ_２インキュベーターにて３７℃、２４時間培養後に、０．５μＭブピバカイン塩酸塩（和光純薬）を２４時間反応させた。その後、ブピバカインを取り除き、未破砕の加熱菌体、破砕物上清、または破砕物沈渣を添加して筋芽細胞を４８時間培養した。培養後、滅菌ＰＢＳ（－）による洗浄を２回行ってから、ｃｅｌｌｃｏｕｎｔｉｎｇｋｉｔ―８（同仁化学）を添加して３７℃、２時間反応させて４５０ｎｍの吸光度を測定した。得られた吸光度の値を（加熱菌体－ブランク）／（加熱菌体非添加－ブランク）×１００の計算式に当てはめて、増殖率を算出した。乳酸菌菌体による筋芽細胞の増殖促進効果は加熱菌体非添加と比べて、増殖率の増加が認められれば、損傷後の筋芽細胞の増殖を促進したと判断することができる。 <Test Example 3> Search for active ingredients in heated Lactobacillus gasseri SBT2055 cells (preparation of lactic acid bacteria cells)
For Example Product 1, the muscle repair promoting effects of the soluble fraction (supernatant) and insoluble fraction (sediment) of the disrupted bacterial cell were examined.
(1) Test method 10 mg/ml Lactobacillus gasseri SBT2055 heated cells were disrupted three times at 1,200 psiG in a French press (Aminco). After disruption, the cells were centrifuged (4°C, 7000 rpm, 15 minutes) to obtain a supernatant and a sediment. The supernatant was then filtered through a 0.22 μm filter. The sediment was added with an equal amount of PBS(-) to the removed supernatant, vortexed, and centrifuged (4°C, 7000 rpm, 15 minutes), and the supernatant was removed. This was repeated three times, and the supernatant was used as the sediment after adding an equal amount of PBS(-). The concentrations of the disrupted supernatant and sediment obtained from 10 mg/ml Lactobacillus gasseri SBT2055 heated cells subjected to French press were each equivalent to 10 mg/ml. The cells were diluted with DMEM (SIGMA) containing 10% FBS (GIBCO) and 1% penicillin-streptomycin (SIGMA) to a concentration equivalent to 100 μg/ml to obtain a suspension of the homogenate supernatant or a suspension of the sediment. C2C12 mouse myoblasts were seeded at 5,000 cells/well on a collagen-coated 96-well plate, and incubated at 37°C for 24 hours in a 5% _CO2 incubator. Then, 0.5 μM bupivacaine hydrochloride (Wako Pure Chemical Industries) was added and the myoblasts were incubated for 48 hours with undisrupted heated cells, homogenate supernatant, or homogenate sediment. After incubation, the cells were washed twice with sterile PBS (-), and cell counting kit-8 (Dojin Chemical Industries) was added and incubated at 37°C for 2 hours to measure the absorbance at 450 nm. The proliferation rate was calculated by applying the obtained absorbance value to the formula: (heated cells - blank)/(no heated cells added - blank) x 100. If an increase in proliferation rate was observed in the proliferation-promoting effect of lactic acid bacteria cells on myoblasts compared to the case where heated cells were not added, it can be determined that the proliferation of myoblasts after injury was promoted.

（２）試験結果
画分ごとの筋芽細胞の増殖率を図３に示した。吸光度を測定した結果、ラクトバチルスガセリＳＢＴ２０５５加熱菌体の破砕物沈渣が未破砕の菌体と同様に、筋芽細胞の増殖率を増加させた。したがって、ラクトバチルスガセリＳＢＴ２０５５加熱菌体の不溶性画分に、損傷後の筋芽細胞の増殖を促進する効果が認められた。 (2) Test results The proliferation rate of myoblasts in each fraction is shown in Figure 3. The absorbance was measured, and the disrupted sediment of Lactobacillus gasseri SBT2055 heated cells increased the proliferation rate of myoblasts, similar to that of undisrupted cells. Therefore, the insoluble fraction of Lactobacillus gasseri SBT2055 heated cells was found to have the effect of promoting the proliferation of myoblasts after injury.

＜実施例品２＞乳酸菌培養物の調製
ラクトバチルスガセリＳＢＴ２０５５をＭＲＳ液体培地（ＤＩＦＣＯ）にて培養した。対数増殖期にある各培養液を、０．３％の酵母エキスを添加した１０％還元脱脂乳（１１５℃、２０分滅菌）に１％接種し、各々マザーカルチャーを作製した。これに１０％の還元脱脂乳を添加して、１００℃にて１０分間加熱したヨーグルトミックスに２．５％添加して調製した。３７℃で発酵を行い、乳酸酸度０．８５に到達した時点で冷却し、発酵を終了させた。得られた発酵乳を凍結乾燥してラクトバチルスガセリＳＢＴ２０５５菌体培養物の粉末を得た。得られた菌体培養物をリン酸緩衝液に再懸濁し、１×１０^９ｃｅｌｌｓ／ｍｌに調整した。 Example 2: Preparation of lactic acid bacteria culture Lactobacillus gasseri SBT2055 was cultured in MRS liquid medium (DIFCO). Each culture in the logarithmic growth phase was inoculated at 1% into 10% reduced skim milk (sterilized at 115°C for 20 minutes) containing 0.3% yeast extract, and each mother culture was prepared. 10% reduced skim milk was added to this, and 2.5% was added to a yogurt mix heated at 100°C for 10 minutes to prepare it. Fermentation was carried out at 37°C, and when the lactic acid acidity reached 0.85, it was cooled to terminate the fermentation. The obtained fermented milk was freeze-dried to obtain a powder of Lactobacillus gasseri SBT2055 bacterial culture. The obtained bacterial culture was resuspended in phosphate buffer and adjusted to 1 x ¹⁰⁹ cells/ml.

＜実施例品３＞乳酸菌菌体の調製
ラクトバチルスガセリＳＢＴ２０５５をＭＲＳ液体培地（ＤＩＦＣＯ）に植菌し、３７℃にて１６時間静置培養を行った。培養物を、４℃、７０００ｒｐｍで１５分間遠心分離した後、滅菌水による洗浄と遠心分離を３回繰り返して行い、洗浄菌体を得た。この洗浄菌体を凍結乾燥処理して菌体粉末を得た。 Example 3: Preparation of lactic acid bacteria Lactobacillus gasseri SBT2055 was inoculated into MRS liquid medium (DIFCO) and subjected to static culture at 37° C. for 16 hours. The culture was centrifuged at 4° C. and 7000 rpm for 15 minutes, and then washed with sterilized water and centrifuged three times to obtain washed bacteria. The washed bacteria was freeze-dried to obtain a bacterial powder.

＜実施例品４＞錠剤の製造
実施例品３にて調製した菌体粉末１部に脱脂粉乳４部を混合し、この混合粉末を打錠機により１ｇずつ常法により打錠して、本発明のラクトバチルスガセリＳＢＴ２０５５の菌体２００ｍｇを含む錠剤をそれぞれ調製した。 Example 4: Preparation of tablets One part of the bacterial powder prepared in Example 3 was mixed with 4 parts of skim milk powder, and this mixed powder was compressed into tablets of 1 g each using a tablet press in the usual manner to prepare tablets each containing 200 mg of Lactobacillus gasseri SBT2055 bacterial cells of the present invention.

＜実施例品５＞散剤の製造
ラクトバチルスガセリＳＢＴ２０５５をＭＲＳ液体培地（ＤＩＦＣＯ）５Ｌに摂取後、３７℃、１８時間静置培養を行った。培養終了後、７０００ｒｐｍで１５分間遠心分離を行い、培養液の１／５０量の濃縮菌体を得た。次いで、この濃縮菌体を、脱脂粉乳１０重量％、グルタミン酸ソーダ１重量％を含む分散媒と同量混合し、ｐＨ７に調整後、凍結乾燥を行った。得られた凍結乾燥物を６０メッシュのふるいで整粒化し、凍結乾燥菌末を製造した。第１３改正日本薬局方解説書製剤総則「散剤」の規定に準拠し、この凍結乾燥菌末１ｇにラクトース（日局）４００ｇ、バレイショデンプン（日局）６００ｇを加えて均一に混合し、散剤を得た。 Example 5: Preparation of powder Lactobacillus gasseri SBT2055 was taken into 5L of MRS liquid medium (DIFCO), and then statically cultured at 37°C for 18 hours. After the culture was completed, centrifugation was performed at 7000 rpm for 15 minutes to obtain concentrated cells in an amount of 1/50 of the culture liquid. Then, this concentrated cell was mixed with an equal amount of a dispersion medium containing 10% by weight of skimmed milk powder and 1% by weight of sodium glutamate, and the pH was adjusted to 7, followed by freeze-drying. The obtained freeze-dried product was granulated with a 60-mesh sieve to produce freeze-dried powder. In accordance with the provisions of the 13th Revised Japanese Pharmacopoeia Commentary on Preparations General Provisions "Powder", 400g of lactose (JP) and 600g of potato starch (JP) were added to 1g of this freeze-dried powder and mixed uniformly to obtain a powder.

＜実施例品６＞カプセル剤の製造
表１に示した配合により原料を混合し、造粒により顆粒状とした後、空カプセルに１０ｍｇずつ充填して、カプセル剤を製造した。 Example 6: Production of Capsules The raw materials were mixed according to the ratio shown in Table 1, granulated into granules, and then 10 mg of the mixture was filled into empty capsules to produce capsules.

＜実施例品７＞スティック状健康食品の製造
実施例品１の粉末３０ｇに、ビタミンＣとクエン酸の等量混合物４０ｇ、グラニュー糖１００ｇ、コーンスターチと乳糖の等量混合物６０ｇを加えて混合した。混合物をスティック状袋に詰め、スティック健康食品を製造した。 Example Product 7: Manufacturing of stick-shaped health food 40 g of an equal mixture of vitamin C and citric acid, 100 g of granulated sugar, and 60 g of an equal mixture of cornstarch and lactose were added to 30 g of the powder of Example Product 1 and mixed. The mixture was packed into a stick-shaped bag to manufacture a stick health food.

＜実施例品８＞飲料の製造
表２に示した配合により原料を混合し、容器に充填した後、加熱殺菌して、果汁飲料を製造した。

Example 8: Production of beverage The ingredients were mixed according to the composition shown in Table 2, filled into containers, and then heat sterilized to produce a fruit juice beverage.

本発明によれば、ラクトバチルスに属する乳酸菌菌体及び／又は乳酸菌培養物を有効成分とする筋修復促進剤を提供することができる。According to the present invention, a muscle repair promoter can be provided that contains lactic acid bacteria cells and/or lactic acid bacteria cultures belonging to Lactobacillus as active ingredients.

＜寄託生物材料への言及＞
（１）ラクトバチルスガセリＳＢＴ２０５５
イ当該生物材料を寄託した寄託機関の名称及び住所
独立行政法人製品評価技術基盤機構特許生物寄託センター
日本国千葉県木更津市かずさ鎌足２－５－８１２０号室（郵便番号２９２－０８１８）
ロイの寄託機関に生物材料を寄託した日付
１９９６年３月２７日
２００８年２月２６日（原寄託によりブダペスト条約に基づく寄託への移管日）
ハイの寄託機関が寄託について付した受託番号
ＦＥＲＭＢＰ－１０９５３
（２）ラクトバチルスデルブルッキーサブスピーシーズデルブルッキーＳＢＴ０４１３
イ当該生物材料を寄託した寄託機関の名称及び住所
独立行政法人製品評価技術基盤機構特許微生物寄託センター
日本国千葉県木更津市かずさ鎌足２－５－８１２２号室（郵便番号２９２－０８１８）
ロイの寄託機関に生物材料を寄託した日付
２０１８年２月２１日
ハイの寄託機関が寄託について付した受託番号
ＮＩＴＥＰ－０２６４２（なお、原寄託日後、原寄託に基づくブダペスト条約に基づく寄託への移管申請を２０１９年５月２７日に行い、生存確認試験の完了後に、受領番号を通知する書面を受領した。受領番号は、ＮＩＴＥＡＢＰ－０２６４２である。）
（３）ラクトバチルスロイテリＳＢＴ１９２６
イ当該生物材料を寄託した寄託機関の名称及び住所
独立行政法人製品評価技術基盤機構特許微生物寄託センター
日本国千葉県木更津市かずさ鎌足２－５－８１２２号室（郵便番号２９２－０８１８）
ロイの寄託機関に生物材料を寄託した日付
２０１８年２月２１日
ハイの寄託機関が寄託について付した受託番号
ＮＩＴＥＰ－０２６４３（なお、原寄託日後、原寄託に基づくブダペスト条約に基づく寄託への移管申請を２０１９年５月２７日に行い、生存確認試験の完了後に、受領番号を通知する書面を受領した。受領番号は、ＮＩＴＥＡＢＰ－０２６４３である。）
（４）ラクトバチルスヘルベティカスＳＢＴ１１３８０
イ当該生物材料を寄託した寄託機関の名称及び住所
独立行政法人製品評価技術基盤機構特許微生物寄託センター
日本国千葉県木更津市かずさ鎌足２－５－８１２２号室（郵便番号２９２－０８１８）
ロイの寄託機関に生物材料を寄託した日付
２０１８年２月２１日
ハイの寄託機関が寄託について付した受託番号
ＮＩＴＥＰ－０２６４４（なお、原寄託日後、原寄託に基づくブダペスト条約に基づく寄託への移管申請を２０１９年５月２７日に行い、生存確認試験の完了後に、受領番号を通知する書面を受領した。受領番号は、ＮＩＴＥＡＢＰ－０２６４４である。） <Reference to deposited biological material>
(1) Lactobacillus gasseri SBT2055
Name and address of the depository institution to which the biological material was deposited: National Institute of Technology and Evaluation Patent Organism Depositary Center, Room 120, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan (postal code: 292-0818)
Date of deposit of biological material in the depositary institution in the Republic of Korea March 27, 1996 February 26, 2008 (date of transfer to deposit under the Budapest Treaty by the original deposit)
The deposit number given by the depository institution in Hai is FERM BP-10953.
(2) Lactobacillus delbrueckii subsp. delbrueckii SBT0413
Name and address of the depository institution to which the biological material was deposited: National Institute of Technology and Evaluation Patent Microorganisms Depository Center, Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan (postal code: 292-0818)
The date of deposit of the biological material at the depository in Loi is February 21, 2018. The depository in Hai assigned the deposit accession number NITE P-02642. (After the original deposit date, an application for transfer to a deposit under the Budapest Treaty based on the original deposit was submitted on May 27, 2019, and a written notice notifying the accession number was received after the completion of the viability confirmation test. The accession number is NITE ABP-02642.)
(3) Lactobacillus reuteri SBT1926
Name and address of the depository institution to which the biological material was deposited: National Institute of Technology and Evaluation Patent Microorganisms Depository Center, Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan (postal code: 292-0818)
The date of deposit of the biological material at the depositary institution in Loi is February 21, 2018. The depositary institution in Hai assigned the deposit the accession number NITE P-02643. (After the original deposit date, an application for transfer to a deposit under the Budapest Treaty based on the original deposit was submitted on May 27, 2019, and a letter notifying the accession number was received after the completion of the viability confirmation test. The accession number is NITE ABP-02643.)
(4) Lactobacillus helveticus SBT11380
Name and address of the depository institution to which the biological material was deposited: National Institute of Technology and Evaluation Patent Microorganisms Depository Center, Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan (postal code: 292-0818)
The date of deposit of the biological material at the depositary institution in Loi is February 21, 2018. The accession number given by the depositary institution in Hai to the deposit is NITE P-02644. (After the original deposit date, an application for transfer to a deposit under the Budapest Treaty based on the original deposit was made on May 27, 2019, and a written notice notifying the accession number was received after the completion of the viability confirmation test. The accession number is NITE ABP-02644.)

Claims

A composition for promoting muscle repair, comprising as an active ingredient a bacterial cell of a bacterium belonging to Lactobacillus and/or a culture thereof,
The composition for promoting muscle repair is characterized in that the bacterium belonging to Lactobacillus is one or more selected from Lactobacillus gasseri, Lactobacillus delbrueckii subsp. delbrueckii, Lactobacillus reuteri, and Lactobacillus helveticus.

The lactic acid bacteria belonging to Lactobacillus are selected from the group consisting of Lactobacillus gasseri SBT2055 (FERM BP-10953), Lactobacillus delbrueckii subsp. delbrueckii SBT0413 (NITE B P-02642), Lactobacillus reuteri SBT1926 (NITE B P-02643), and Lactobacillus helveticus SBT11380 (NITE B P-02644). The composition for promoting muscle repair according to claim 1, characterized in that the lactic acid bacteria belonging to Lactobacillus are selected from the group consisting of Lactobacillus gasseri SBT2055 (FERM BP-10953), Lactobacillus delbrueckii subsp. delbrueckii SBT0413 (NITE B P-02642), Lactobacillus reuteri SBT1926 (NITE B P-02643), and Lactobacillus helveticus SBT11380 (NITE B P-02644).

The composition for promoting muscle repair according to claim 1 or 2, wherein the bacterial cells and/or cultures thereof are killed bacterial cells.

The composition for promoting muscle repair according to claim 3, wherein the killed bacteria is an insoluble fraction of disrupted bacteria.

Novel lactic acid bacterium, Lactobacillus delbrueckii subsp. delbrueckii SBT0413 (NITE BP-02642) .

Novel lactic acid bacterium, Lactobacillus reuteri SBT1926 (NITE BP-02643) .

Novel lactic acid bacterium, Lactobacillus helveticus SBT11380 (NITE BP-02644) .