JP2023107155A - Novel lactic acid bacteria and use thereof - Google Patents
Novel lactic acid bacteria and use thereof Download PDFInfo
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- JP2023107155A JP2023107155A JP2022008298A JP2022008298A JP2023107155A JP 2023107155 A JP2023107155 A JP 2023107155A JP 2022008298 A JP2022008298 A JP 2022008298A JP 2022008298 A JP2022008298 A JP 2022008298A JP 2023107155 A JP2023107155 A JP 2023107155A
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Abstract
Description
特許法第３０条第２項適用申請有り （その１） ウェブサイトの掲載日 ２０２１年３月５日 ウェブサイトのアドレス ｈｔｔｐｓ：／／ｗｗｗ．ｊｓｂｂａ．ｏｒ．ｊｐ／２０２１／ ｈｔｔｐｓ：／／ｊｓｂｂａ２．ｂｉｏｗｅｂ．ｎｅ．ｊｐ／ｊｓｂｂａ２０２１／ｓｃｈｅｄｕｌｅ．ｐｈｐ ｈｔｔｐ：／／ｊｓｂｂａ．ｂｉｏｗｅｂ．ｎｅ．ｊｐ／ｊｓｂｂａ２０２１／ｄｏｗｎ－ｌｏａｄ＿ｐｄｆ．ｐｈｐ？ｐ＿ｃｏｄｅ＝２Ｂ０７－１４ （その２） 開催日 ２０２１年３月１８日から２０２１年３月２１日（公開日は２０２１年３月１９日） 集会名、開催場所 日本農芸化学会２０２１年度大会（オンライン開催） （その３） ウェブサイトの掲載日 ２０２１年１０月２２日 ウェブサイトのアドレス ｈｔｔｐｓ：／／ｗｗｗ．ｉｔｃ．ｐｒｅｆ．ｔｏｋｕｓｈｉｍａ．ｊｐ／ ｈｔｔｐｓ：／／ｗｗｗ．ｉｔｃ．ｐｒｅｆ．ｔｏｋｕｓｈｉｍａ．ｊｐ／０２＿ｒｅｓｅａｒｃｈ／１２ａｃｈｉｅｖｅｍｅｎｔ．ｓｈｔｍ ｈｔｔｐｓ：／／ｗｗｗ．ｉｔｃ．ｐｒｅｆ．ｔｏｋｕｓｈｉｍａ．ｊｐ／０６＿ｒｅｓｕｌｔ／ｒｅｓｕｌｔ＿Ｒ０２＿０．ｓｈｔｍ ｈｔｔｐｓ：／／ｗｗｗ．ｉｔｃ．ｐｒｅｆ．ｔｏｋｕｓｈｉｍａ．ｊｐ／０６＿ｒｅｓｕｌｔ／ｒｅｓｕｌｔ＿Ｒ０２／２０２０＿１６＿ｎｉｓｈｉｏｋａ．ｐｄｆ （その４） 掲示日 ２０２１年１０月２２日 掲示場所 徳島県立工業技術センター（徳島県徳島市雑賀町西開１１－２） （その５） 提出日 ２０２１年１０月５日 寄託機関 独立行政法人製品評価技術基盤機構 バイオテクノロジーセンター （その６） ウェブサイトの掲載日 ２０２１年１２月２３日 ウェブサイトのアドレス ｈｔｔｐｓ：／／ｗｗｗ．ｎｉｔｅ．ｇｏ．ｊｐ／ｎｂｒｃ／ ｈｔｔｐｓ：／／ｗｗｗ．ｎｉｔｅ．ｇｏ．ｊｐ／ｎｂｒｃ／ｃａｔａｌｏｇｕｅ／ＮＢＲＣＤｉｓｐＳｅａｒｃｈＳｅｒｖｌｅｔ？ｌａｎｇ＝ｊｐApplied for the application of Article 30,
新規乳酸菌及びその利用に関する。 It relates to a new lactic acid bacterium and its use.
乳酸菌は乳酸を産生する細菌の総称であり、古くから人々の生活に深く関与してきた。例えば、伝統的な発酵食品である漬物、ヨーグルト、味噌、醤油は、乳酸発酵が風味に大きく影響している。乳酸菌が産生する乳酸は、発酵食品の風味形成に影響する他、pHを下げて目的外の有害菌（雑菌）の増殖を抑える効果もある。また、乳酸菌の中には様々な機能性を持つ株が報告されている。例えば、消化液に耐性を有する乳酸菌は、生菌を経口摂取した場合、生きた状態で腸まで届き、産生する乳酸によって、生体に悪影響を及ぼし得る菌（悪玉菌）を抑制して腸内環境を整え、整腸効果を発揮しやすいといわれている。また、腸にとどまる乳酸菌は、より高い整腸作用を示すといわれている。また、血圧低下やリラックス効果を有するγ-アミノ酪酸を産生する株も報告されている。 Lactic acid bacteria are a general term for bacteria that produce lactic acid, and have been deeply involved in people's lives since ancient times. For example, traditional fermented foods such as pickles, yogurt, miso, and soy sauce are greatly influenced by lactic acid fermentation. The lactic acid produced by lactic acid bacteria not only affects the flavor formation of fermented foods, but also has the effect of lowering the pH and suppressing the growth of unintended harmful bacteria (miscellaneous bacteria). In addition, strains with various functions have been reported among lactic acid bacteria. For example, lactic acid bacteria, which are resistant to digestive juices, reach the intestine alive when ingested live, and the lactic acid produced suppresses bacteria (bad bacteria) that can adversely affect the body, thereby improving the intestinal environment. It is said that it is easy to prepare the intestinal function and exert an intestinal regulation effect. In addition, lactic acid bacteria that remain in the intestine are said to exhibit a higher intestinal regulation effect. A strain that produces γ-aminobutyric acid, which has blood pressure-lowering and relaxing effects, has also been reported.
乳酸菌による発酵によって作られる発酵食品として、徳島県では昔から阿波晩茶が生産されている。阿波晩茶は微生物の発酵によって製造される後発酵茶に分類され、後発酵茶は世界中でも一部の地域のみで生産される。阿波晩茶は真夏に茶葉が収穫される。阿波晩茶の製造では、一般的に、収穫された茶葉は釜で茹でて茶葉が持つ酵素を失活させ、揉んでから樽に押し詰めにして、重石を乗せて２～４週間程度、嫌気発酵させる。発酵後、茶葉を天日で1～2日間乾燥させて、阿波晩茶が完成する。阿波晩茶は嫌気発酵により茶葉の成分が変化し、酸味のある独特の風味を形成する。阿波晩茶の嫌気発酵の優占菌は乳酸菌であり、該乳酸菌はカテキン等の抗菌成分が多い環境で生育しており、他とは違った性質を持つ可能性がある。阿波晩茶から分離された乳酸菌として、これまでに、ピロガロール産生乳酸菌（特許文献１）や、感染防御に有用な乳酸菌（特許文献２）が報告されているが、他の機能を有する乳酸菌については知られていない。 As a fermented food made by fermentation with lactic acid bacteria, Awa Bancha has long been produced in Tokushima Prefecture. Awa Bancha is classified as a post-fermented tea produced by fermentation of microorganisms, and post-fermented tea is produced only in some areas of the world. Awa Bancha tea leaves are harvested in the middle of summer. In the production of Awa Bancha, the harvested tea leaves are generally boiled in a kettle to deactivate the enzymes in the tea leaves, then kneaded, pressed into barrels, weighted, and left for 2 to 4 weeks anaerobic. ferment. After fermentation, the tea leaves are dried in the sun for 1-2 days to complete Awa Bancha. Awabancha changes the components of the tea leaves through anaerobic fermentation, forming a unique sour flavor. Lactic acid bacteria are the dominant bacteria in anaerobic fermentation of Awa Bancha, and these lactic acid bacteria grow in an environment rich in antibacterial components such as catechins, and may have properties different from others. As lactic acid bacteria isolated from Awa Bancha, pyrogallol-producing lactic acid bacteria (Patent Document 1) and lactic acid bacteria useful for preventing infection (Patent Document 2) have been reported so far. unknown.
阿波晩茶由来の新たな乳酸菌、これを利用した組成物を提供することを目的とする。 An object of the present invention is to provide a new lactic acid bacterium derived from Awa evening tea and a composition using the same.
本発明者は、徳島県の伝統的な発酵茶に着目し、前記課題に鑑み鋭意検討を行ったところ、阿波晩茶から、消化液耐性、腸管上皮細胞付着性、γ-アミノ酪酸産生性及び茶葉抽出物耐性の少なくともいずれかの作用を有する新規乳酸菌を単離することに成功した。本発明は、該知見に基づき更に検討を重ねた結果完成されたものであり、例えば次の態様を包含する。
項１．ラクチプランチバチルス・ペントーサス（Lactiplantibacillus pentosus）AWA1922株（受託番号NBRC115326）、ラクチプランチバチルス・ペントーサス（Lactiplantibacillus pentosus） AWA1955株（受託番号NBRC115327）、レビラクトバチルス・ブレビス（Levilactobacillus brevis）AWA1978株（受託番号NBRC115323）、レビラクトバチルス・ブレビス（Levilactobacillus brevis）AWA1984株（受託番号NBRC115324）、またはレビラクトバチルス・ブレビス（Levilactobacillus brevis）AWA1985株（受託番号NBRC115325）である乳酸菌。
項２．ラクチプランチバチルス・ペントーサス（Lactiplantibacillus pentosus）AWA1922株（受託番号NBRC115326）、ラクチプランチバチルス・ペントーサス（Lactiplantibacillus pentosus） AWA1955株（受託番号NBRC115327）、レビラクトバチルス・ブレビス（Levilactobacillus brevis）AWA1978株（受託番号NBRC115323）、レビラクトバチルス・ブレビス（Levilactobacillus brevis）AWA1984株（受託番号NBRC115324）及びレビラクトバチルス・ブレビス（Levilactobacillus brevis）AWA1985株（受託番号NBRC115325）からなる群より選択される少なくとも１種の乳酸菌を含有する組成物。
項３．前記組成物が経口組成物である、項２に記載の組成物。
項４．前記組成物が、発酵茶、ヨーグルト、漬物及びサプリメントからなる群より選択される少なくとも１種である、項２または３に記載の組成物。
The inventors of the present invention focused on traditional fermented tea in Tokushima Prefecture and made extensive studies in view of the above problems. We have successfully isolated novel lactic acid bacteria that have at least one of the effects of resistance to tea leaf extract. The present invention was completed as a result of further studies based on the findings, and includes, for example, the following aspects.
Item 3. 3. The composition of
Section 4. Item 4. The composition according to
本開示によれば、新規乳酸菌を提供することができる。特に、本開示によれば、消化液耐性、腸管上皮細胞付着性、γ-アミノ酪酸産生性及び茶葉抽出物耐性の少なくともいずれかの作用を有する新規乳酸菌を提供することができる。また、該乳酸菌は、このように消化液耐性、腸管上皮細胞付着性、γ－アミノ酪酸産生性及び茶葉抽出物耐性の少なくともいずれかの有用作用を有することから、様々な組成物、特に経口組成物に好ましく利用できる。 According to the present disclosure, novel lactic acid bacteria can be provided. In particular, according to the present disclosure, it is possible to provide novel lactic acid bacteria having at least one of digestive juice resistance, intestinal epithelial cell adhesion, γ-aminobutyric acid productivity, and tea leaf extract resistance. In addition, since the lactic acid bacterium has such useful effects as at least one of digestive juice resistance, intestinal epithelial cell adhesion, γ-aminobutyric acid production and tea leaf extract resistance, various compositions, especially oral compositions It can be used favorably for things.
乳酸菌
本開示の乳酸菌は、徳島県で生産されている阿波晩茶から本発明者が単離した、ラクチプランチバチルス・ペントーサス（Lactiplantibacillus pentosus）またはレビラクトバチルス・ブレビス（Levilactobacillus brevis）に属する新規の乳酸菌であり、本発明者が次の通り名付けた、次の５種の菌株である：
ラクチプランチバチルス・ペントーサス AWA1922株、
ラクチプランチバチルス・ペントーサス AWA1955株、
レビラクトバチルス・ブレビス AWA1978株、
レビラクトバチルス・ブレビス AWA1984株、
レビラクトバチルス・ブレビス AWA1985株。
Lactic acid bacterium The lactic acid bacterium of the present disclosure is a novel lactic acid bacterium belonging to Lactiplantibacillus pentosus or Levilactobacillus brevis, isolated by the present inventor from Awa evening tea produced in Tokushima Prefecture. and are the following five strains, named by the inventors as follows:
Lactobacillus pentosus strain AWA1922,
Lactobacillus pentosus strain AWA1955,
Levilactobacillus brevis strain AWA1978,
Revilactobacillus brevis strain AWA1984,
Revilactobacillus brevis strain AWA1985.
前記単離についてより具体的には、阿波晩茶（茶葉）にリン酸緩衝生理食塩水を加えて室温で振盪し、上清を２％寒天含むＭＲＳ寒天平板に塗抹し、嫌気培養し、得られた乳酸菌コロニーについて遺伝子解析を行うことにより、前記菌株を単離、同定した。遺伝子解析は、recA遺伝子に対するマルチプレックスPCR（Polymerase Chain Reaction）または16S rRNA遺伝子の相同性に基づき、前記菌株を同定した。該単離、同定は、より詳細には後述の実施例に従う。 More specifically, for the isolation, phosphate-buffered saline is added to Awa Bancha (tea leaves) and shaken at room temperature, and the supernatant is spread on an MRS agar plate containing 2% agar, anaerobically cultured, and obtained. The strain was isolated and identified by performing genetic analysis on the lactic acid bacteria colonies obtained. Genetic analysis identified the strain based on multiplex PCR (Polymerase Chain Reaction) or 16S rRNA gene homology to the recA gene. The isolation and identification follow the examples below in more detail.
なお、ラクチプランチバチルス・ペントーサス、レビラクトバチルス・ブレビスはいずれも、従来、ラクトバチルス（Lactobacillus）属に分類され、それぞれラクトバチルス・ペントーサス（Lactobacillus pentosus）、ラクトバチルス・ブレビス（Lactobacillus brevis）と称されていたが、近年（２０２０年）、乳酸菌が再分類されており、前記属種名は該再分類に従う。 Both Lactobacillus pentosus and Levilactobacillus brevis have been conventionally classified into the genus Lactobacillus and called Lactobacillus pentosus and Lactobacillus brevis, respectively. However, in recent years (2020), lactic acid bacteria have been reclassified, and the genus and species names follow the reclassification.
本開示の乳酸菌は、独立行政法人製品評価技術基盤機構バイオテクノロジーセンター（千葉県木更津市かずさ鎌足2-5-8）に寄託されている（２０２１年１０月２９日）。具体的には、ラクチプランチバチルス・ペントーサスAWA1922株は受託番号NBRC115326、ラクチプランチバチルス・ペントーサスAWA1955株は受託番号NBRC115327、レビラクトバチルス・ブレビスAWA1978株は受託番号NBRC115323、レビラクトバチルス・ブレビスAWA1984株は受託番号NBRC115324、レビラクトバチルス・ブレビスAWA1985株は受託番号NBRC115325で寄託されている。これらの乳酸菌は、所定の手続によって寄託機関から分譲を受けることができる。これらの乳酸菌は次の特徴を有する。 The lactic acid bacteria of the present disclosure have been deposited with the National Institute of Technology and Evaluation Biotechnology Center (2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture) (October 29, 2021). Specifically, the Lactiprancibacillus pentosus AWA1922 strain has accession number NBRC115326, the Lactiprancibacillus pentosus AWA1955 strain has accession number NBRC115327, the Levilactobacillus brevis AWA1978 strain has accession number NBRC115323, and the Levilactobacillus brevis AWA1984 strain has accession number NBRC115323. No. NBRC115324, Revilactobacillus brevis strain AWA1985 has been deposited under accession number NBRC115325. These lactic acid bacteria can be obtained from a depositary institution according to a prescribed procedure. These lactic acid bacteria have the following characteristics.
コロニーの色：乳白色（ＭＲＳ寒天平板）
コロニーの形状：主に円形～楕円形（ＭＲＳ寒天平板）
菌の形態：桿状
運動性：なし
芽胞形成：なし
生育温度：25～37℃ （良好に発育）
糖の資化性：後述の実施例の表１に示す。
Colony color: milky white (MRS agar plate)
Colony shape: Mainly circular to oval (MRS agar plate)
Fungus morphology: rod-shaped
Motility: None Sporulation: None Growth temperature: 25-37°C (Grows well)
Sugar utilization: Shown in Table 1 in Examples below.
表１に示す菌株は、ラクチプランチバチルス・ペントーサスAWA1922株、AWA1955株、これらと同種に分類される公知の乳酸菌ラクチプランチバチルス・ペントーサス（Lactiplantibacillus pentosus）NBRC106467T株（旧分類に従えばLactobacillus pentosus NBRC106467T）、レビラクトバチルス・ブレビスAWA1978株、AWA1984株、AWA1985株、これらと同種に分類される公知の乳酸菌レビラクトバチルス・ブレビス（Levilactobacillus brevis）NBRC107147T株（旧分類に従えばLactobacillus brevis NBRC107147T）である。 The strains shown in Table 1 are Lactobacillus pentosus AWA1922 strain, AWA1955 strain, and known lactic acid bacteria Lactiplantibacillus pentosus NBRC106467 T strain (according to the old classification, Lactobacillus pentosus NBRC106467 T ), Levilactobacillus brevis AWA1978 strain, AWA1984 strain, AWA1985 strain, known lactic acid bacteria classified into the same species as these, Levilactobacillus brevis NBRC107147 T strain (Lactobacillus brevis NBRC107147 T according to the old classification) be.
また、ラクチプランチバチルス・ペントーサスAWA1922株、AWA1955株は、NBRC106467T株との相違点として、次の特徴も備えている。AWA1922株は、ＭＲＳ液体培地で35℃、24時間培養した際、試験管を振盪すると凝集が生じる。AWA1955株は、ＭＲＳ液体培地で35℃、24時間培養した際、高粘性物質を産生する。これは、これらの２株は菌体外多糖を産生しているためと推察され、AWA1922は細胞外膜に結合した莢膜多糖を、AWA1955は遊離のスライム多糖を産生しているためと考えられる。NBRC106467T株ではこのような凝集は生じず、また、高粘性物質も産生されない。 In addition, the Lactiprancibacillus pentosus AWA1922 and AWA1955 strains also have the following features as differences from the NBRC106467 T strain. When the AWA1922 strain is cultured in an MRS liquid medium at 35°C for 24 hours, aggregation occurs when the test tube is shaken. The AWA1955 strain produces a highly viscous substance when cultured in an MRS liquid medium at 35°C for 24 hours. It is presumed that these two strains produce exopolysaccharide, AWA1922 producing capsular polysaccharide bound to the extracellular membrane, and AWA1955 producing free slime polysaccharide. . The NBRC106467 T strain does not cause such aggregation, nor does it produce a highly viscous substance.
また、ラクチプランチバチルス・ペントーサスAWA1922株、AWA1955株、レビラクトバチルス・ブレビスAWA1978株、AWA1984株、AWA1985株は、次の（１）～（４）からなる群より選択される少なくとも１種の特性を有する。（１）消化液耐性、（２）腸管上皮細胞付着性、（３）γ-アミノ酪酸産生性、（４）茶葉抽出物耐性。 In addition, the Lactiprancibacillus pentosus strain AWA1922, AWA1955 strain, Revilactobacillus brevis strain AWA1978, AWA1984, and AWA1985 exhibit at least one characteristic selected from the group consisting of the following (1) to (4). have. (1) digestive juice resistance, (2) intestinal epithelial cell adhesion, (3) γ-aminobutyric acid production, and (4) tea leaf extract resistance.
前記（１）消化液耐性について、後述の実施例に示す通り、ラクチプランチバチルス・ペントーサスAWA1922株、AWA1955株は、これらと同種に分類される公知の乳酸菌ラクチプランチバチルス・ペントーサスNBRC106467T株（後述の実施例において標準株として使用）と概ね同等以上で、胃液と接触した場合であっても生存し、腸液と接触した場合であっても増殖する。特に、AWA1922株は、NBRC106467T株よりも高い消化液耐性を有する。該消化液耐性の確認、評価は、詳細には後述の実験例に従って行うことができる。 Regarding the (1) digestive juice resistance, as shown in the examples below, the Lactiprancibacillus pentosus AWA1922 strain and the AWA1955 strain are known lactic acid bacteria Lactiprancibacillus pentosus NBRC106467 T strain (described below) that are classified into the same species as these. (used as a standard strain in Examples), survives even when in contact with gastric juice, and proliferates even when in contact with intestinal juice. In particular, the AWA1922 strain has higher digestive juice resistance than the NBRC106467 T strain. Confirmation and evaluation of the digestive juice resistance can be performed in detail according to Experimental Examples described later.
また、レビラクトバチルス・ブレビスAWA1978株、AWA1984株、AWA1985株も、後述の実施例に示す通り、胃液と接触した場合であっても生存し、腸液と接触した場合であっても増殖する。このように、これらの菌株も消化液に対する耐性を有する。また、これらの菌株は、後述の実施例に示す通り、公知の乳酸菌レビラクトバチルス・ブレビスNBRC107147T株（後述の実施例において標準株として使用）よりも消化液に対する耐性が高い。該消化液耐性の確認、評価も、詳細には後述の実験例に従って行うことができる。 In addition, the Revilactobacillus brevis strains AWA1978, AWA1984, and AWA1985 also survive in contact with gastric juice and proliferate in contact with intestinal juice, as shown in Examples below. Thus, these strains are also resistant to digestive fluids. In addition, as shown in Examples below, these strains have higher resistance to digestive fluids than the known lactic acid bacteria Levilactobacillus brevis NBRC107147 T strain (used as a standard strain in Examples below). Confirmation and evaluation of the digestive juice resistance can also be performed in detail according to the experimental examples described later.
また、このようにAWA1922株、AWA1978株、AWA1984株、AWA1985株はいずれも、消化液と接した場合でも生存及び／または増殖し、消化液に対して特に高い耐性がある。これらのことから、これらの乳酸菌は、例えば、生きた状態で胃腸に届きやすいという点で有用であるといえる。 In addition, all of the AWA1922, AWA1978, AWA1984, and AWA1985 strains survive and/or proliferate even in contact with digestive juices, and have particularly high resistance to digestive juices. From these facts, it can be said that these lactic acid bacteria are useful, for example, in that they easily reach the gastrointestinal tract in a living state.
前記（２）腸管上皮細胞付着性について、ラクチプランチバチルス・ペントーサスAWA1922株、AWA1955株は、後述の実施例に示す通り、腸管上皮細胞に対する付着性が高い。このことから、AWA1922株、AWA1955株は、消化管内で一定期間滞留できるといえ、このことから、例えば、腸内環境を整える、アレルギーを改善する、病原菌等の感染を予防する、大腸炎を改善する、脂質代謝を改善する、アレルギーをはじめこれらの悪化を軽減するといった点で有用であるといえる。該付着性の確認、評価は、詳細には後述の実験例に従って行うことができる。 With regard to (2) adhesion to intestinal epithelial cells, Lactipplantibacillus pentosus AWA1922 and AWA1955 strains have high adhesion to intestinal epithelial cells, as shown in Examples below. From this, it can be said that the AWA1922 and AWA1955 strains can be retained in the digestive tract for a certain period of time. It can be said that it is useful in terms of improving lipid metabolism and reducing aggravation of these including allergies. Confirmation and evaluation of the adhesion can be performed in detail according to Experimental Examples described later.
前記（３）γ-アミノ酪酸産生性について、レビラクトバチルス・ブレビスAWA1978株、AWA1984株、AWA1985株は、後述の実施例に示す通り、レビラクトバチルス・ブレビスNBRC107147T株よりもγ-アミノ酪酸を多く産生する。γ-アミノ酪酸は、ＧＡＢＡとも呼ばれ、様々な有用作用を有することが知られている。このことから、AWA1978株、AWA1984株、AWA1985株は、γ-アミノ酪酸をより多く産生するという点で有用である。また、このことから、これらの菌株は、γ-アミノ酪酸に基づく有用作用（例えば、高めの血圧の改善、ストレス緩和、睡眠の質改善、疲労感の軽減、肌弾力の維持、認知機能の改善）を得ることを目的として使用される乳酸菌としても有用である。該産生性の確認、評価は、詳細には後述の実験例に従って行うことができる。 Regarding (3) γ-aminobutyric acid productivity, Levilactobacillus brevis AWA1978 strain, AWA1984 strain, and AWA1985 strain produce γ-aminobutyric acid more than Levilactobacillus brevis NBRC107147 T strain, as shown in Examples below. produce a lot. γ-Aminobutyric acid, also called GABA, is known to have various beneficial effects. Therefore, the AWA1978, AWA1984, and AWA1985 strains are useful in that they produce more γ-aminobutyric acid. In addition, from this fact, these strains have useful effects based on γ-aminobutyric acid (for example, improvement of high blood pressure, stress relief, sleep quality improvement, fatigue reduction, skin elasticity maintenance, cognitive function improvement) ) is also useful as a lactic acid bacterium used for the purpose of obtaining. Confirmation and evaluation of the productivity can be performed in detail according to Experimental Examples described later.
前記（４）茶葉抽出物耐性について、後述の実施例に示す通り、本開示の５菌株はいずれも茶葉抽出物に対して耐性を有する。また、ラクチプランチバチルス・ペントーサスAWA1922株は、後述の実施例に示す通り、NBRC106467T株よりも茶葉抽出物に対する耐性が高い。また、レビラクトバチルス・ブレビスAWA1978株、AWA1984株、AWA1985株は、後述の実施例に示す通り、NBRC107147T株よりも茶葉抽出物に対する耐性が高い。実施例に示す茶葉抽出物には、カテキンをはじめとするポリフェノール等の一般的な緑茶由来の成分（抗菌成分等）が含まれていると考えられ、このように、本開示の５菌株、特にAWA1922株、AWA1978株、AWA1984株、AWA1985株は、茶葉抽出物に対する耐性を有する点で有用である。従って、本開示のこれらの菌株は、例えば茶製品（発酵茶を含む）やこのような抗菌成分等を含有する製品にも好ましく用いることができる。該耐性の確認、評価は、詳細には後述の実験例に従って行うことができる。 With respect to (4) tea leaf extract resistance, all five strains of the present disclosure are resistant to tea leaf extract, as shown in the examples below. In addition, the Lactipplantibacillus pentosus AWA1922 strain has higher resistance to tea leaf extracts than the NBRC106467 T strain, as shown in Examples below. In addition, Revilactobacillus brevis AWA1978, AWA1984, and AWA1985 strains are more resistant to tea leaf extracts than NBRC107147 T strain, as shown in Examples below. The tea leaf extract shown in the examples is thought to contain common green tea-derived components (such as antibacterial components) such as catechins and other polyphenols. AWA1922 strain, AWA1978 strain, AWA1984 strain, and AWA1985 strain are useful in that they have resistance to tea leaf extracts. Therefore, these strains of the present disclosure can also be preferably used in, for example, tea products (including fermented tea) and products containing such antibacterial ingredients and the like. Confirmation and evaluation of the resistance can be performed in detail according to Experimental Examples described later.
このように、本開示は新規の乳酸菌を提供する。該乳酸菌は、例えば、前記（１）～（４）からなる群より選択される少なくとも１種の特性を有する点で少なくとも有用であり、該特性に基づく有用作用を得ることを目的として用いることができる点で有用である。また、本開示の乳酸菌は、様々な組成物に利用することができ、また、例えば、これらの有用作用を備えた組成物を得るために好ましく使用することができる。このように、本開示の乳酸菌は、プロバイオティクス菌としても有用である。また、該組成物として、次の組成物が例示される。 Thus, the present disclosure provides novel lactic acid bacteria. The lactic acid bacterium is, for example, at least useful in that it has at least one property selected from the group consisting of (1) to (4) above, and can be used for the purpose of obtaining useful effects based on the property. useful in that it can In addition, the lactic acid bacteria of the present disclosure can be used in various compositions, and can be preferably used, for example, to obtain compositions with these beneficial effects. Thus, the lactic acid bacteria of the present disclosure are also useful as probiotic bacteria. Moreover, the following composition is illustrated as this composition.
組成物
本開示の組成物は、本開示の乳酸菌、すなわち、AWA1922株、AWA1955株、AWA1978株、AWA1984株及びAWA1985株からなる群より選択される少なくとも１種の乳酸菌と、使用形態、使用態様等に応じた任意の成分とを組み合わせて用いることにより製造することができ、必要に応じて培養（発酵を含む）等を行って製造してもよい。該任意の成分として、可食性成分、薬学的に許容される成分、香粧品学的に許容される成分、培地成分等が例示される。
Composition The composition of the present disclosure contains at least one lactic acid bacterium selected from the group consisting of the lactic acid bacteria of the present disclosure, that is, the AWA1922 strain, the AWA1955 strain, the AWA1978 strain, the AWA1984 strain, and the AWA1985 strain, a usage form, a usage mode, etc. It can be produced by using a combination of optional components according to the requirements, and may be produced by performing culture (including fermentation) and the like as necessary. Examples of the optional ingredients include edible ingredients, pharmaceutically acceptable ingredients, cosmetically acceptable ingredients, medium ingredients, and the like.
該組成物は、経口、非経口の別を問わず、好ましくは経口で使用される。また、該組成物の形態も制限されず、固形状、半固形状、液状のいずれであってもよい。また、例えば本開示の組成物が固形状である場合、これは水等と混合して使用してもよい。 The composition is preferably used orally, whether orally or parenterally. Also, the form of the composition is not limited, and may be solid, semi-solid, or liquid. Also, for example, when the composition of the present disclosure is in solid form, it may be used by mixing with water or the like.
該組成物の使用態様も制限されず、目的に応じて適宜設定すればよく、食品組成物（飲料を含む、保健機能食品（特定保健用食品、栄養機能食品、機能性表示食品等）、サプリメント、病者用食品等を含む）、医薬組成物、医薬部外品組成物、化粧品組成物、飼料組成物、また、食品組成物、医薬組成物、医薬部外品組成物、化粧品組成物、飼料等への添加剤として使用することができ、該組成物には、発酵物製造時に使用可能な発酵スターター等として使用される組成物も包含される。 The mode of use of the composition is not limited, and may be appropriately set according to the purpose. , including foods for sick people), pharmaceutical compositions, quasi-drug compositions, cosmetic compositions, feed compositions, food compositions, pharmaceutical compositions, quasi-drug compositions, cosmetic compositions, It can be used as an additive to feeds and the like, and includes compositions used as fermentation starters and the like that can be used in the production of fermented products.
前記乳酸菌としては、例えば、公知の乳酸菌の培養条件に従い、前記乳酸菌と培地とを混合して乳酸菌を培養し、得られた培養物から遠心分離等の手段によって集菌、分離されたものをそのまま用いてもよく、該培養物（発酵物、上清等を含む）、その粗精製物、精製物、これらの処理物（乾燥物（凍結乾燥物、噴霧乾燥物、加熱乾燥物等）、破砕物等）等を任意に用いてもよい。これらは、乳酸菌培養等における従来公知の手順に従い行うことができる。これらは、１種単独で使用してもよく、２種以上を組み合わせて含まれていてもよい。 As the lactic acid bacterium, for example, the lactic acid bacterium is cultured by mixing the lactic acid bacterium with a medium according to known lactic acid bacterium culture conditions, and the resulting culture is collected by means such as centrifugation and separated. The culture (including fermented products, supernatants, etc.), crudely purified products thereof, purified products thereof, processed products thereof (dried products (freeze-dried products, spray-dried products, heat-dried products, etc.), crushed etc.) may be used arbitrarily. These can be performed according to conventionally known procedures in lactic acid bacteria culture and the like. These may be used individually by 1 type, and may be contained in combination of 2 or more types.
本開示の乳酸菌の培養は、該乳酸菌が生育及び／または増殖可能である限り、いかなる培地を用いて行ってもよく、従来公知の一般的な乳酸菌培養手順に従って、例えば乳酸菌培養用培地を使用したり、該培地を適宜改変して使用することができ、また、この限りにおいて培養方法も制限されない。本開示を制限するものではないが、例えば、乳酸菌培養に一般的に使用されているＭＲＳ培地を用い、本開示の乳酸菌が生育可能な条件で培養する方法が挙げられる。 The lactic acid bacteria of the present disclosure may be cultured using any medium as long as the lactic acid bacteria can grow and/or proliferate. Alternatively, the medium can be modified as appropriate and used, and the culture method is not limited in this respect. Although not intended to limit the present disclosure, for example, a method of culturing under conditions in which the lactic acid bacterium of the present disclosure can grow using an MRS medium commonly used for culturing lactic acid bacteria can be mentioned.
また、本開示を制限するものではながいが、例えば、該培地には、必要に応じて、γ-アミノ酪酸の生成基質であるグルタミン酸やその塩（ナトリウム塩等）等、グルタミン酸やその塩を生成し得る成分、乳酸菌の生育（増殖を含む）を促進できるような成分（例えば、グルコース、澱粉、ショ糖、乳糖、デキストリン、ソルビトール、フラクトース等の炭素源；酵母エキス、ペプトン等の窒素源；ビタミン類、ミネラル類、脂肪酸類、微量金属元素等）等の任意の成分が含まれていてもよい。これらは、１種単独で含まれていてもよく、２種以上を組み合わせて含まれていてもよい。 In addition, although not intended to limit the present disclosure, for example, the medium may optionally contain glutamic acid or a salt thereof, such as glutamic acid or a salt thereof (sodium salt, etc.), which is a substrate for the production of γ-aminobutyric acid. ingredients that can promote the growth (including proliferation) of lactic acid bacteria (e.g., carbon sources such as glucose, starch, sucrose, lactose, dextrin, sorbitol, fructose; nitrogen sources such as yeast extract and peptone; vitamins , minerals, fatty acids, trace metal elements, etc.) may be contained. These may be contained individually by 1 type, and may be contained in combination of 2 or more types.
本開示を制限するものではないが、培養温度は、良好な菌の生育の点から、5～45℃が例示され、好ましくは25～37℃が例示される。培地のｐＨ（培養開始時）も制限されないが、35℃でｐＨ4～8が例示され、好ましはくはｐＨ6～7が例示される。培養時間は、培養条件、培養スケール、目的とする菌量等を考慮して決定すればよく、例えば24～48時間等が例示される。該培養は、好気培養、嫌気培養を問わず、好ましくは嫌気培養が例示される。 Although not intended to limit the present disclosure, the culture temperature is exemplified from 5 to 45°C, preferably from 25 to 37°C, from the viewpoint of good fungal growth. The pH of the medium (at the start of culture) is also not limited, but is exemplified at pH 4-8, preferably pH 6-7 at 35°C. The culture time may be determined in consideration of the culture conditions, culture scale, desired amount of bacteria, and the like, and is exemplified by 24 to 48 hours. The culture may be aerobic culture or anaerobic culture, preferably anaerobic culture.
本開示の乳酸菌の培養には、このように公知の培地等を用いて行ってもよく、該培地の代わりに、または、該培地と共に、例えば食品原料や食品といった前記組成物の原料等（組成物原料）を用いて該乳酸菌を培養してもよい。食品原料や食品としては、本開示を制限するものではないが、茶葉、野菜（白菜、大根、人参、きゅうり、なす、キャベツ等）、乳（ヨーグルト、牛乳、スキムミルク、脱脂粉乳等）、穀類（米、大麦、小麦等）、水産物（鮎、サバ、にしん、フナ、サケ、イワナ等）、畜産物、調味料等が例示される。これらは１種単独で使用してもよく、２種以上を組み合わせて使用してもよい。このような組成物原料を用いる場合も、前述の通り、培養条件は該乳酸菌が生育及び／または増殖可能である限り制限されない。また、組成物原料は、必要に応じて、洗浄、粉砕、圧搾、加熱、混合等の前処理に供されていてもよい。 The culture of the lactic acid bacteria of the present disclosure may be performed using a known medium or the like as described above, and instead of the medium or together with the medium, the raw materials of the composition such as food materials and foods (composition The lactic acid bacteria may be cultured using raw materials). Food raw materials and foods include, but are not limited to, tea leaves, vegetables (Chinese cabbage, radish, carrot, cucumber, eggplant, cabbage, etc.), milk (yogurt, milk, skim milk, powdered skim milk, etc.), cereals ( rice, barley, wheat, etc.), marine products (ayu, mackerel, herring, crucian carp, salmon, char, etc.), livestock products, seasonings, and the like. These may be used individually by 1 type, and may be used in combination of 2 or more type. Even when using such composition raw materials, as described above, the culture conditions are not limited as long as the lactic acid bacteria can grow and/or proliferate. Moreover, the composition raw material may be subjected to pretreatment such as washing, pulverization, pressing, heating, mixing, etc., if necessary.
本開示の組成物は、例えば、このようにして得た培養物（発酵物、粗精製物、精製物、これらの処理物等を含む）そのものであってもよく、該培養物と、前記可食性成分、薬学的に許容される成分、香粧品学的に許容される成分、培地成分等の任意の成分とを更に混合等して製造したものであってもよく、必要に応じて更に培養（発酵）等を行ったものであってもよい。前述の通り、本開示を制限するものではないが、本開示の組成物は、本開示の乳酸菌を用いた発酵を行うための発酵スターターとして使用してもよい。また、本開示を制限するものではないが、前述の通り、レビラクトバチルス・ブレビスAWA1978株、AWA1984株、AWA1985株は、レビラクトバチルス・ブレビスNBRC107147T株よりもγ-アミノ酪酸を多く産生できることから、本開示の組成物は、例えばγ-アミノ酪酸産生増強用の組成物として使用してもよい。 The composition of the present disclosure may be, for example, the culture thus obtained (including fermented products, crudely purified products, purified products, processed products thereof, etc.) itself, and the culture and the above-mentioned possible It may be produced by further mixing, etc., with arbitrary ingredients such as food ingredients, pharmaceutically acceptable ingredients, cosmetically acceptable ingredients, and medium ingredients, and if necessary, further culture (fermentation) etc. may be performed. As previously mentioned, without limiting the present disclosure, the compositions of the present disclosure may be used as fermentation starters for performing fermentations using the lactic acid bacteria of the present disclosure. In addition, although not limiting the present disclosure, as described above, the Levilactobacillus brevis AWA1978, AWA1984, and AWA1985 strains can produce more γ-aminobutyric acid than the Levilactobacillus brevis NBRC107147 T strain. , the composition of the present disclosure may be used, for example, as a composition for enhancing γ-aminobutyric acid production.
本開示の組成物において、前記乳酸菌は生菌の状態であってもよく、死菌の状態であってもよく、目的とする使用形態、使用態様、作用等に応じて適宜決定すればよい。 In the composition of the present disclosure, the lactic acid bacteria may be in the state of viable bacteria or in the state of dead bacteria, which may be appropriately determined according to the intended use form, use mode, action, and the like.
本開示を制限するものではないが、例えば、ヨーグルトをはじめとする乳製品等において、乳酸菌は生菌として摂取されることによって、整腸作用、腸内細菌叢改善作用等による健康維持などに有用であることが一般的に知られている。また、本開示の乳酸菌は、前述の通り、消化液耐性や腸管上皮細胞付着性等の少なくともいずれかの特性を有し、これに基づく整腸作用 、腸内細菌叢改善等の効果が期待される。このような観点からは、本開示の乳酸菌が生菌として該組成物に含有されていることが好ましく例示される。 Although it does not limit the present disclosure, for example, in dairy products such as yogurt, lactic acid bacteria are useful for maintaining health by ingesting live bacteria, such as intestinal regulation and intestinal microflora improving effects. It is generally known that In addition, as described above, the lactic acid bacteria of the present disclosure have at least one of properties such as digestive juice resistance and intestinal epithelial cell adhesion, and based on this, effects such as intestinal regulation and intestinal microflora improvement are expected. be. From such a point of view, it is preferably exemplified that the lactic acid bacteria of the present disclosure are contained in the composition as viable bacteria.
このようにして本開示の組成物を製造することができ、前述の通り、該組成物の形態は固形状、半固形状、液状のいずれであってもよい。例えば、本開示の組成物が食品組成物である場合、一般的に食品として摂取される形態であればよい。また、本開示を制限するものではないが、食品組成物の一実施形態としてサプリメントが例示され、その剤型として錠剤、カプセル剤（マイクロカプセルを含む）、丸剤、粉末状、顆粒状、トローチ状、ペースト状、ゲル状、液状（シロップ、乳剤、懸濁剤を含む）等が例示され、また、糖衣等によりコーティングされていてもよい。 In this manner, the composition of the present disclosure can be produced, and as described above, the composition can be in solid, semi-solid, or liquid form. For example, when the composition of the present disclosure is a food composition, it may be in a form generally ingested as food. In addition, although not intended to limit the present disclosure, supplements are exemplified as one embodiment of the food composition, and their dosage forms are tablets, capsules (including microcapsules), pills, powders, granules, and lozenges. Forms, pastes, gels, liquids (including syrups, emulsions and suspensions) are exemplified, and they may be coated with sugar coating or the like.
また、本開示を制限するものではないが、例えば、本開示の組成物が医薬組成物、医薬部外品組成物または化粧品組成物である場合、これらにおいて許容可能な剤型であれば制限されず、その剤型として錠剤、カプセル剤、丸剤、粉末状、顆粒状、トローチ状、ペースト状、ゲル状、液状（シロップ状、乳状、懸濁状を含む）等の経口組成物、軟膏剤、クリーム剤、ローション剤、ゲル剤、液剤、噴霧剤、スプレー剤、貼付剤（パップ剤、プラスター剤、経皮吸収剤、経粘膜吸収剤）、注射剤（例えば腹腔内注射剤）、座剤等の非経口組成物が挙げられる。また、本開示を制限するものではないが、例えば飼料組成物についても、飼料組成物として許容可能な剤型であれば制限されない。 In addition, although it does not limit the present disclosure, for example, when the composition of the present disclosure is a pharmaceutical composition, a quasi-drug composition, or a cosmetic composition, it is limited as long as it is an acceptable dosage form in these. However, oral compositions such as tablets, capsules, pills, powders, granules, troches, pastes, gels, liquids (including syrups, emulsions, and suspensions), ointments, etc. , creams, lotions, gels, liquids, nebulizers, sprays, patches (plasters, plasters, transdermal absorbers, transmucosal absorbers), injections (e.g., intraperitoneal injections), suppositories and parenteral compositions. In addition, although the present disclosure is not limited, for example, the feed composition is not limited as long as it has an acceptable dosage form as a feed composition.
このことから、前記可食性成分、薬学的に許容される成分、香粧品学的に許容される成分等の任意の成分として、担体（水、エタノール、プロピレングリコール、グリセリン、デキストリン、シクロデキストリン等）、賦形剤、結合剤、滑沢剤、コーティング剤、崩壊剤、崩壊補助剤、希釈剤、安定化剤、保存剤、分散剤、湿潤化剤、溶解剤、溶解補助剤、等張化剤、pH調節剤、着色剤、甘味剤、矯味剤、香料、ゲル化剤、増粘剤、乳化剤、酸味料、アミノ酸類、ビタミン類、ミネラル類をはじめとする各種栄養成分、一般的に食品に使用される成分（前記食品原料、食品を含む）等も包含される。また、培地成分については、前述の通り、本開示の乳酸菌が培養できる限り制限されず、前述の培養の説明が適用される。これらは１種単独で使用してもよく、２種以上を組み合わせて使用してもよく、また、その配合量等も使用形態、使用態様等に応じて適宜決定すればよい。 For this reason, a carrier (water, ethanol, propylene glycol, glycerin, dextrin, cyclodextrin, etc.) can be used as an arbitrary component of the edible component, pharmaceutically acceptable component, cosmetically acceptable component, etc. , excipients, binders, lubricants, coating agents, disintegrants, disintegration aids, diluents, stabilizers, preservatives, dispersants, wetting agents, solubilizers, dissolution aids, tonicity agents , pH adjusters, coloring agents, sweeteners, flavoring agents, flavoring agents, gelling agents, thickeners, emulsifiers, acidulants, amino acids, vitamins, minerals and other nutritional ingredients. Components used (including the above-mentioned food raw materials and foods) are also included. In addition, as described above, the medium components are not limited as long as the lactic acid bacteria of the present disclosure can be cultured, and the above description of culture applies. These may be used singly or in combination of two or more, and the blending amount thereof may be appropriately determined according to the type of use, mode of use, and the like.
本開示を制限するものではないが、経口組成物の具体例として、茶（茶葉、発酵茶葉、茶飲料等を含む）、ヨーグルト、チーズ、乳飲料等の乳製品、キムチ、ぬか漬け、ザワークラウト等の漬物、納豆、チョコレート、キャンディー、ビスケット等の菓子類、パン類、麺類、水産（加工）食品（なれずし等）、畜産（加工）食品（ハム等）、油脂類、調味料、甘酒、清涼飲料、炭酸飲料、美容ドリンク、栄養飲料、果実飲料、野菜飲料等の飲料、飲料の濃縮原液、調整用粉末等の飲料類、サプリメント等の発酵食品、非発酵食品が挙げられる。 Although not intended to limit the present disclosure, specific examples of oral compositions include tea (including tea leaves, fermented tea leaves, tea beverages, etc.), yogurt, cheese, dairy products such as milk beverages, kimchi, rice bran pickles, sauerkraut, etc. pickles, natto, chocolate, candy, biscuits and other sweets, bread, noodles, marine (processed) food (narezushi, etc.), livestock (processed) food (ham, etc.), oils and fats, seasonings, amazake, Beverages such as soft drinks, carbonated drinks, beauty drinks, nutritional drinks, fruit drinks, and vegetable drinks, beverages such as concentrated undiluted solutions of drinks, powders for preparation, fermented foods such as supplements, and non-fermented foods.
本開示を制限するものではないが、例えば、前記乳酸菌が茶葉抽出物（茶葉抽出成分）に対する耐性が高い点からは、本開示によれば、経口組成物として茶が好ましい一例として挙げられる。また、例えば、前記乳酸菌が消化液耐性、腸管上皮細胞付着性及び／またはγ-アミノ酪酸産生性を有する点からは、本開示によれば、整腸作用に一層優れた茶やγ-アミノ酪酸を高含量で含む茶を容易に提供することができる。このような茶は、前述の通り、例えば従来公知の茶（発酵茶を含む）の製造手順に従い、本開示の乳酸菌と該茶の原料（茶葉や茶葉抽出物等）とを混合等して製造することができる。 Although not intended to limit the present disclosure, tea is a preferred oral composition according to the present disclosure, for example, because the lactic acid bacteria are highly resistant to tea leaf extracts (tea leaf extract components). Further, for example, from the point that the lactic acid bacteria have digestive juice resistance, intestinal epithelial cell adhesion and/or γ-aminobutyric acid productivity, according to the present disclosure, tea and γ-aminobutyric acid are more excellent in intestinal regulation. can easily provide a tea containing a high content of As described above, such tea is produced, for example, by mixing the lactic acid bacteria of the present disclosure and the raw materials of the tea (tea leaves, tea leaf extracts, etc.) according to conventionally known tea (including fermented tea) production procedures. can do.
また、本開示を制限するものではないが、本開示の組成物の一実施形態としてヨーグルト、キムチ、ぬか漬け、サプリメント等が例示される。ヨーグルトは、例えば従来公知のヨーグルトの製造手順に従い、乳原料等の原料（食材）と本開示の乳酸菌とを混合等して製造することができる。また、キムチは、例えば従来公知のキムチの製造手順に従い、白菜等の野菜や調味料等の任意の原料と本開示の乳酸菌とを混合等して製造することができる。ぬか漬けは、例えば従来公知のぬか漬けの製造手順に従い大根、白菜、人参、きゅうり、なす等の野菜、米ぬか、塩、酒粕等の任意の原料と本開示の乳酸菌とを混合等して製造することができる。また、本開示を制限するものではないが、例えば従来公知のサプリメント等の製造手順に従い、本開示の乳酸菌の乾燥物と、デンプン、乳糖、セルロース等の成分とを混合等して、タブレット等の形態で本開示の組成物を製造することができる。 Moreover, yogurt, kimchi, rice bran pickles, supplements, etc. are exemplified as an embodiment of the composition of the present disclosure, although the present disclosure is not limited thereto. Yogurt can be produced, for example, by mixing ingredients (ingredients) such as dairy ingredients with the lactic acid bacteria of the present disclosure, according to conventionally known procedures for producing yogurt. In addition, kimchi can be produced, for example, by mixing arbitrary raw materials such as vegetables such as Chinese cabbage and seasonings with the lactic acid bacteria of the present disclosure, according to conventionally known kimchi production procedures. Rice bran pickles are produced, for example, by mixing any raw materials such as vegetables such as radish, Chinese cabbage, carrots, cucumbers, and eggplants, rice bran, salt, sake lees, etc. with the lactic acid bacteria of the present disclosure according to the conventionally known production procedure for bran pickles. be able to. In addition, although it does not limit the present disclosure, for example, according to the manufacturing procedure of conventionally known supplements, etc., the dried product of the lactic acid bacteria of the present disclosure, starch, lactose, cellulose and other ingredients are mixed to produce tablets and the like. The compositions of the present disclosure can be manufactured in the form of:
本開示において、該組成物の対象者（対象動物）も制限されず、ヒト、ヒト以外の哺乳動物等が例示される。ヒト以外の哺乳動物としては、マウス、ラット、モルモット、ウサギ、イヌ、ネコ、サル、ブタ、牛、馬等の動物、好ましくはマウス、ラット、モルモット、ウサギ、イヌ、サル等の動物が例示される。 In the present disclosure, the subject (subject animal) of the composition is also not limited, and is exemplified by humans, mammals other than humans, and the like. Mammals other than humans include animals such as mice, rats, guinea pigs, rabbits, dogs, cats, monkeys, pigs, cows and horses, preferably mice, rats, guinea pigs, rabbits, dogs and monkeys. be.
本開示の組成物の対象者（対象動物）への適用量は特に制限されず、対象者（対象動物）の体格、年齢、症状、適用形態、使用目的等に応じて適宜設定すればよい。例えば本開示の組成物が経口組成物である場合、本開示を制限するものではないが、ヒト（体重６０ｋｇ）に適用される経口組成物１００ｇ中、１０４～１０１１ｃｅｌｌｓが挙げられ、好ましくは１０８～１０１１ｃｅｌｌｓが挙げられる。本開示の組成物が経口組成物以外である場合、組成物中の本開示の乳酸菌量は該範囲を参考にして適宜決定すればよい。 The amount of the composition of the present disclosure to be applied to a subject (subject animal) is not particularly limited, and may be appropriately set according to the physique, age, symptoms, application form, purpose of use, etc. of the subject (subject animal). For example, when the composition of the present disclosure is an oral composition, the present disclosure is not limited, but 10 4 to 10 11 cells are preferably included in 100 g of an oral composition applied to humans (body weight 60 kg). includes 10 8 to 10 11 cells. When the composition of the present disclosure is other than an oral composition, the amount of lactic acid bacteria of the present disclosure in the composition may be appropriately determined with reference to the range.
本開示によれば、前記乳酸菌を配合した組成物を容易に提供できる。また、本開示の乳酸菌は、前記（１）～（４）の少なくともいずれかの特性を備えていることから、該特性に起因する有用作用を得ることを目的とした組成物を容易に提供することができる。 According to the present disclosure, it is possible to easily provide a composition containing the lactic acid bacteria. In addition, since the lactic acid bacterium of the present disclosure has at least one of the properties (1) to (4), it is possible to easily provide a composition aimed at obtaining a useful effect resulting from the property. be able to.
本開示を制限するものではないが、消化液耐性や腸管上皮細胞への付着性に起因する有用作用として、整腸作用、腸内細菌叢改善作用、便通改善作用（おなかの調子を整える等）、アレルギー改善作用、病原菌等の感染予防作用、大腸炎の改善作用、脂質代謝改善作用等が例示される。γ-アミノ酪酸産生能に起因する有用作用として、高めの血圧の改善作用、ストレス緩和作用、睡眠の質改善作用、疲労感の軽減作用、肌弾力の維持作用、認知機能の改善作用といった、γ-アミノ酪酸に基づく作用として従来知られている有用作用が例示される。また、前述の通り、茶葉抽出物耐性を有することにより、本開示の乳酸菌を茶製品にも好ましく適用することが可能であり、特に、前述のような有用作用が付与された発酵茶の製造において好ましく適用することが可能である。このように、本開示によれば、このような健康維持または向上を目的とした組成物を容易に提供することができる。 Although it does not limit the present disclosure, useful effects resulting from resistance to digestive juices and adhesion to intestinal epithelial cells include intestinal regulation action, intestinal microflora improving action, and bowel movement improving action (regulating stomach condition, etc.). , allergy-improving action, infection-preventing action against pathogens, colitis-ameliorating action, lipid-metabolism-improving action, and the like. Useful effects resulting from γ-aminobutyric acid-producing ability include γ Useful actions conventionally known as actions based on -aminobutyric acid are exemplified. In addition, as described above, by having tea leaf extract resistance, the lactic acid bacteria of the present disclosure can be preferably applied to tea products, especially in the production of fermented tea to which the above-mentioned useful effects are imparted. It is possible to apply preferably. Thus, according to the present disclosure, such a composition for maintaining or improving health can be easily provided.
また、前述の通り、本開示の乳酸菌をスターターとして組成物原料と混合等して使用することも可能であり、本開示の乳酸菌を用いて発酵食品を製造することで、発酵食品において、ｐＨが低下することにより目的外の有害菌（雑菌）の増殖を抑制しつつ、該乳酸菌に起因する有用作用を付与することができる。また、本開示の乳酸菌を用いることで、製造過程（発酵過程等を含む）の管理が容易になり、また、最終製品の品質の安定性向上等にもつながる。 In addition, as described above, it is possible to use the lactic acid bacteria of the present disclosure as a starter by mixing it with a composition raw material, etc., and by producing a fermented food using the lactic acid bacteria of the present disclosure, the pH of the fermented food is By decreasing it, it is possible to suppress the growth of unintended harmful bacteria (miscellaneous bacteria) and impart useful effects caused by the lactic acid bacteria. In addition, the use of the lactic acid bacteria of the present disclosure facilitates management of the manufacturing process (including fermentation process and the like), and also leads to improved quality stability of the final product.
以下、例を示して本開示をより詳細に説明するが、本開示はこれらに限定されない。
試験例１
通常の阿波晩茶の製造手順に従い茶葉を発酵して得た阿波晩茶の茶葉（発酵後乾燥前の茶葉（湿重量））1gに10mLのリン酸緩衝生理食塩水（組成：1Lあたり塩化ナトリウム6.5g、リン酸二ナトリウム・12水和物5.15g、リン酸一カリウム0.3g、pH7.2±0.2）を加えて室温（25℃）で1分間振盪し、上清を2%寒天含むMRS寒天平板（Becton, Dickinson and Company社製）に塗抹し、嫌気ジャーを用いて35℃で48時間培養し、得られた複数の乳酸菌コロニーから菌体DNAを抽出し、recA遺伝子に対するマルチプレックスPCRや16SrRNAの相同性の結果に基づき同定を行った。
The present disclosure will be described in more detail below with examples, but the present disclosure is not limited thereto.
Test example 1
1g of Awabancha tea leaves obtained by fermenting tea leaves according to the normal manufacturing procedure of Awabancha (tea leaves before drying after fermentation (wet weight)), 10mL of phosphate buffered saline (composition: sodium chloride per 1L) 6.5 g, disodium phosphate dodecahydrate 5.15 g, monopotassium phosphate 0.3 g, pH 7.2 ± 0.2) and shake for 1 minute at room temperature (25°C). Smear on an agar plate (Becton, Dickinson and Company), culture in an anaerobic jar at 35°C for 48 hours, extract bacterial cell DNA from the resulting multiple lactic acid bacteria colonies, and perform multiplex PCR for the recA gene. Identification was based on the 16S rRNA homology results.
該同定において、ラクチプランチバチルス・プランタルムグループ（L.plantarum、L.pentosus、L.paraplantarum）では、16S rRNA遺伝子の相同性が99.8％程度あり、16S rRNA遺伝子の相同性に基づいて、これらの相違等を同定することが困難であった。このことから、recA遺伝子に対するマルチプレックスPCRを行い、PCR増幅産物について2％アガロースゲル電気泳動を行った。その結果、318bpにラクチプランチバチルス・プランタルム（L.plantarum）、218bpにラクチプランチバチルス・ペントーサス（L.pentosus）、107bpにラクチプランチバチルス・パラプランタルム（L.paraplantarum）のバンドが確認され、これにより、L.plantarumグループを同定すると共に、得られたバンドに基づき、ラクチプランチバチルス・ペントーサスに属する単離株（２株）を、それぞれAWA1922株、AWA1955株と名付けた。一方、該グループ以外の菌種ではバンドが現れなかったため、16S rRNA遺伝子の相同性に基づき、レビラクトバチルス・ブレビス（Levilactobacillus brevis）に属する単離株（３株）を、それぞれAWA1978株、AWA1984株、AWA1985株と名付けた。 In the identification, the homology of the 16S rRNA gene is about 99.8% in the Lactiprantibacillus plantarum group (L.plantarum, L.pentosus, L.paraplantarum), and based on the homology of the 16S rRNA gene, these It was difficult to identify differences, etc. For this reason, multiplex PCR was performed on the recA gene, and 2% agarose gel electrophoresis was performed on the PCR amplification products. As a result, 318bp L.plantarum, 218bp L.pentosus, and 107bp L.paraplantarum bands were confirmed. The L. plantarum group was identified, and based on the bands obtained, the isolates (two strains) belonging to Lactiprantivibacillus pentosus were named AWA1922 strain and AWA1955 strain, respectively. On the other hand, since no band appeared in strains other than this group, based on the homology of the 16S rRNA gene, the isolates (3 strains) belonging to Levilactobacillus brevis were selected as AWA1978 strain and AWA1984 strain, respectively. , named the AWA1985 strain.
これらの菌株を、独立行政法人製品評価技術基盤機構バイオテクノロジーセンター（千葉県木更津市かずさ鎌足2-5-8、２０２１年１０月２９日）に寄託した。AWA1922株は受託番号NBRC115326、AWA1955株は受託番号NBRC115327、AWA1978株は受託番号NBRC115323、AWA1984株は受託番号NBRC115324、AWA1985株は受託番号NBRC115325で寄託されている。 These strains were deposited at the National Institute of Technology and Evaluation Biotechnology Center (2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, October 29, 2021). The AWA1922 strain has been deposited under accession number NBRC115326, the AWA1955 strain under accession number NBRC115327, the AWA1978 strain under accession number NBRC115323, the AWA1984 strain under accession number NBRC115324, and the AWA1985 strain under accession number NBRC115325.
前記乳酸菌５株の糖の資化性を、本分野で公知の通常の評価手順に従い評価した。この際、公知の乳酸菌であるラクチプランチバチルス・ペントーサスNBRC106467T株（Lactobacillus pentosus NBRC106467T）、レビラクトバチルス・ブレビスNBRC107147T株（Lactobacillus brevis NBRC107147T）を、それぞれラクチプランチバチルス・ペントーサス、レビラクトバチルス・ブレビスの標準株として用いた。評価結果を表１に示す。 The saccharide assimilation of the 5 strains of lactic acid bacteria was evaluated according to a normal evaluation procedure known in the art. At this time, known lactic acid bacteria Lactobacillus pentosus NBRC106467 T strain (Lactobacillus pentosus NBRC106467 T ) and Levilactobacillus brevis NBRC107147 T strain (Lactobacillus brevis NBRC107147 T ) were used, respectively. It was used as a standard strain of brevis. Table 1 shows the evaluation results.
表１に示す通り、これらの乳酸菌はいずれも標準株と糖の資化性が異なった。 As shown in Table 1, each of these lactic acid bacteria differed from the standard strain in sugar assimilation.
試験例２
試験例１で得た乳酸菌（５菌株）と前記標準株（２菌株）を用いて、以下の通り、（１）消化液耐性、（２）腸管上皮細胞付着性、（３）γ－アミノ酪酸産生性、及び（４）茶葉抽出物耐性について試験を行った。
Test example 2
Using the lactic acid bacteria (5 strains) obtained in Test Example 1 and the standard strains (2 strains), (1) digestive juice resistance, (2) intestinal epithelial cell adhesion, and (3) γ-aminobutyric acid Productivity and (4) tea leaf extract tolerance were tested.
（１）消化液耐性
（１Ａ）試験手順
塩酸によりpH3.0または2.0に調整し、ペプシンを0.04%含むMRS液体培地（Becton, Dickinson and Company社製）を人工胃液とした。pHは、25℃でｐHメーターLAQUA（型番F-74、堀場製作所製）を用いて測定した。また、0.2%胆汁末、0.01%トリプシン、0.01%パンクレアチンを含むMRS液体培地を人工腸液とした。
(1) Digestive juice resistance
(1A) Test procedure MRS liquid medium (manufactured by Becton, Dickinson and Company) adjusted to pH 3.0 or 2.0 with hydrochloric acid and containing 0.04% pepsin was used as artificial gastric juice. The pH was measured at 25° C. using a pH meter LAQUA (model number F-74, manufactured by Horiba, Ltd.). MRS liquid medium containing 0.2% bile powder, 0.01% trypsin, and 0.01% pancreatin was used as artificial intestinal fluid.
予めMRS液体培地で35℃、24時間培養して得た乳酸菌液100μL（109cfu/mL）をMRS液体培地10mLに接種し、35℃で24時間静置培養して得た乳酸菌培養液を得た。このようにして得た乳酸菌培養液1mLを、pH3.0またはpH2.0の前記人工胃液9mLに加え、35℃で3時間接触させた後、得られた溶液をMRS寒天平板に塗抹し、嫌気ジャーを用いて35℃で24時間培養し、生じたコロニー数をカウントした（人工胃液処理：３時間）。また、人工胃液による処理時間0として、前述のようにして得た乳酸菌培養液と前記人工胃液とを混合後直ちにMRS寒天平板に塗抹した以外は、前述と同様に培養してコロニーをカウントした（人工胃液処理：０）。 100 μL (10 9 cfu/mL) of lactic acid bacteria cultured in advance in MRS liquid medium at 35°C for 24 hours was inoculated into 10 mL of MRS liquid medium, and the lactic acid bacteria culture solution obtained by static culture at 35°C for 24 hours was added. Obtained. 1 mL of the lactic acid bacteria culture solution thus obtained was added to 9 mL of the artificial gastric juice of pH 3.0 or pH 2.0 and brought into contact at 35°C for 3 hours. Using a jar, the cells were cultured at 35°C for 24 hours, and the number of formed colonies was counted (treatment with artificial gastric juice: 3 hours). In addition, the culture was cultured and colonies were counted in the same manner as described above, except that the lactic acid bacteria culture solution obtained as described above and the artificial gastric juice were immediately smeared on an MRS agar plate after mixing the artificial gastric juice with the artificial gastric juice for 0 treatment time ( Artificial gastric juice treatment: 0).
また、前述の通り人工胃液で3時間接触させた乳酸菌液100μLを、前記人工腸液10mLと混合し、35℃で24時間培養し、前述と同様にしてMRS寒天平板を用いて培養し、生じたコロニーをカウントした（人工腸液処理：２４時間）。また、人工腸液による処理時間0として、前述の通り人工胃液で3時間接触させた乳酸菌液と前記人工腸液とを混合後直ちにMRS寒天平板に塗抹し、前述と同様にして培養し、生じたコロニーをカウントした（人工腸液処理：０）。 In addition, 100 μL of the lactic acid bacteria solution that had been brought into contact with the artificial gastric juice for 3 hours as described above was mixed with 10 mL of the artificial intestinal juice, cultured at 35° C. for 24 hours, and cultured using an MRS agar plate in the same manner as described above. Colonies were counted (simulated intestinal fluid treatment: 24 hours). In addition, assuming that the treatment time with the artificial intestinal juice is 0, the artificial intestinal juice was mixed with the lactic acid bacteria solution that had been brought into contact with the artificial gastric juice for 3 hours as described above, and immediately smeared on an MRS agar plate and cultured in the same manner as described above to produce colonies. were counted (simulated intestinal fluid treatment: 0).
（１Ｂ）結果
結果を図１及び２に示す。図１はpH3.0に調整した人工胃液を用いた結果、図２はpH2.0に調整した人工胃液を用いた結果を示す。
(1B) Results Results are shown in FIGS. FIG. 1 shows the results of using artificial gastric juice adjusted to pH 3.0, and FIG. 2 shows the results of using artificial gastric juice adjusted to pH 2.0.
図１及び２に示す通り、ラクチプランチバチルス・ペントーサスAWA1922株、AWA1955株は、その標準株NBRC106467Tと概ね同等以上の消化液に対する耐性が認められ、また、AWA1922株では、AWA1955株よりも、消化液に対して高い耐性が認められた。また、図１及び２に示す通り、レビラクトバチルス・ブレビスAWA1978株、AWA1984株、AWA1985株は、その標準株NBRC107147Tよりも消化液に対して高い耐性が認められた。特に、AWA1922株、AWA1978株、AWA1984株、AWA1985株において消化液に対して顕著に高い耐性が認められた。このことから、これらの菌株は消化液に対して優れた耐性を有し、特に、AWA1922株、AWA1978株、AWA1984株、AWA1985株は消化液に対して非常に高い耐性を有することが分かった。 As shown in Figures 1 and 2, the Lactiprancibacillus pentosus AWA1922 and AWA1955 strains were found to have a resistance to digestive fluids that was roughly equal to or higher than that of the type strain NBRC106467T . High resistance to liquid was recognized. 1 and 2, Revilactobacillus brevis strains AWA1978, AWA1984, and AWA1985 were found to have higher resistance to digestive fluids than the standard strain NBRC107147T . In particular, AWA1922 strain, AWA1978 strain, AWA1984 strain, and AWA1985 strain were found to have remarkably high resistance to digestive fluids. From this, it was found that these strains have excellent resistance to digestive juices, and in particular, AWA1922, AWA1978, AWA1984, and AWA1985 strains have extremely high resistance to digestive juices.
（２）腸管上皮細胞（Caco-2）に対する付着性
（２Ａ）試験手順
MRS液体培地で35℃、24時間培養した乳酸菌培養液を遠心により集菌し、リン酸緩衝生理食塩水に置換した後、バクテリアカウンター（商品名バクテリアカウンター、サンリード硝子社製）を用いて菌数を測定した。このようにリン酸緩衝生理食塩水に置換して得た乳酸菌懸濁液を、20%ウシ胎児血清と非必須アミノ酸（8.9mg/L L-Alanine、15mg/L L-Asparagine・H2O、13.3mg/L L-Aspartic Acid、14.7mg/L L-Glutamic Acid、7.5mg/L Glycine、11.5mg/L L-Proline、10.5mg/L L-Serine）を含む細胞培養用培地（MEM（Minimum Essential Medium）、Thermo Fisher Scientific社製）に1×107個/mLとなるよう添加し、これを乳酸菌懸濁培地とした。別途、20%ウシ胎児血清と非必須アミノ酸（8.9mg/L L-Alanine、15mg/L L-Asparagine・H2O、13.3mg/L L-Aspartic Acid、14.7mg/L L-Glutamic Acid、7.5mg/L Glycine、11.5mg/L L-Proline、10.5mg/L L-Serine）を含むMEM中で培養したヒト結腸癌由来Caco-2細胞を2×105個/mLとなるようの6ウェルプレートに植え5%CO2、37℃で48時間培養した。培養後、培養上清を乳酸菌懸濁培地（2mL）で置換し、35℃で2時間保持した。次いで、乳酸菌懸濁培地を除去し、リン酸緩衝生理食塩水で3回洗浄した。その後、滅菌超純水を添加して懸濁した後、MRS寒天平板に塗抹し、嫌気ジャーを用いて35℃で48時間培養後、コロニー数をカウントした。
(2) Adhesion to intestinal epithelial cells (Caco-2) (2A) test procedure
After culturing the lactic acid bacteria in the MRS liquid medium at 35°C for 24 hours, the bacteria were harvested by centrifugation, replaced with phosphate-buffered saline, and then tested using a bacteria counter (trade name: Bacteria Counter, manufactured by Sunlead Glass Co., Ltd.). measured the number. The lactic acid bacteria suspension thus obtained by substitution with phosphate-buffered saline was mixed with 20% fetal bovine serum and non-essential amino acids (8.9 mg/L L-Alanine, 15 mg/L L-Asparagine·H 2 O, Cell culture medium (MEM (Minimum Essential Medium), manufactured by Thermo Fisher Scientific) was added to 1×10 7 cells/mL, and this was used as a lactic acid bacteria suspension medium. Separately, 20% fetal bovine serum and non-essential amino acids (8.9 mg/L L-Alanine, 15 mg/L L-Asparagine H 2 O, 13.3 mg/L L-Aspartic Acid, 14.7 mg/L L-Glutamic Acid, 7.5 6 wells of human colon cancer-derived Caco-2 cells cultured in MEM containing mg/L Glycine, 11.5mg/L L-Proline, 10.5mg/L L-Serine) at 2×10 5 cells/mL The cells were planted on a plate and cultured at 5% CO 2 and 37° C. for 48 hours. After culturing, the culture supernatant was replaced with a lactic acid bacteria suspension medium (2 mL) and kept at 35°C for 2 hours. Then, the lactic acid bacteria suspension medium was removed and washed three times with phosphate-buffered saline. After that, sterilized ultrapure water was added to suspend the suspension, which was smeared on an MRS agar plate, cultured at 35°C for 48 hours using an anaerobic jar, and the number of colonies was counted.
（２Ｂ）結果
結果を図３に示す。図３に示す通り、ラクチプランチバチルス・ペントーサスAWA1922株、AWA1955株では、その標準株NBRC106467Tよりも腸管上皮細胞Caco-2に対する付着性が高かった。このことから、AWA1922株やAWA1955株は、特に消化管内に一定期間滞留できるといえ、腸内環境を整える点で特に有用であることが分かった。
(2B) Results The results are shown in FIG. As shown in FIG. 3, the Lactiprancibacillus pentosus strain AWA1922 and AWA1955 had higher adhesion to intestinal epithelial cell Caco-2 than the standard strain NBRC106467T . From this, it can be said that the AWA1922 and AWA1955 strains can be retained in the gastrointestinal tract for a certain period of time, and are particularly useful in regulating the intestinal environment.
（３）γ-アミノ酪酸の産生性
（３Ａ）試験手順
5%グルタミン酸ナトリウムを含むMRS液体培地10mLに、MRS液体培地で35℃、24時間培養した乳酸菌培養液を100μL加え、35℃で48時間培養した。得られた培養液を遠心し、上清100μLを10%スルホサリチル酸900μLと混合した後、クエン酸リチウム緩衝液P-21（日本電子株式会社製）で希釈して、全自動アミノ酸分析機JLC500/V2（日本電子株式会社製）を使用してγ-アミノ酪酸の量を測定した。
(3) γ-aminobutyric acid productivity (3A) test procedure
To 10 mL of MRS liquid medium containing 5% sodium glutamate, 100 μL of lactic acid bacteria culture medium cultured in MRS liquid medium at 35° C. for 24 hours was added and cultured at 35° C. for 48 hours. The resulting culture solution was centrifuged, 100 μL of the supernatant was mixed with 900 μL of 10% sulfosalicylic acid, diluted with lithium citrate buffer P-21 (manufactured by JEOL Ltd.), and analyzed using a fully automatic amino acid analyzer JLC500/ V2 (manufactured by JEOL Ltd.) was used to measure the amount of γ-aminobutyric acid.
（３Ｂ）結果
結果を図４に示す。図４に示す通り、レビラクトバチルス・ブレビスAWA1978株、AWA1984株、AWA1985株では、その標準株NBRC107147T株よりも多くのγ-アミノ酪酸を産生することが分かった。なお、ラクチプランチバチルス・ペントーサスAWA1922株、AWA1955株、その標準株NBRC106467Tはいずれも、該試験においてγ-アミノ酪酸の産生は認められなかった。
(3B) Results The results are shown in FIG. As shown in FIG. 4, it was found that the Revilactobacillus brevis AWA1978, AWA1984, and AWA1985 strains produced more γ-aminobutyric acid than the standard strain NBRC107147T strain. In addition, none of Lactipplantibacillus pentosus AWA1922 strain, AWA1955 strain, and its type strain NBRC106467 T produced γ-aminobutyric acid in the test.
（４）茶葉抽出物中での生育性
（４Ａ）試験手順
市販品の一般的な緑茶茶葉に沸騰した超純水を加え茶葉濃度を0.02g/mLとし、グルコースを1%になるよう加え、1分間静置した。このようにして得た溶液の上清を1%グルコースを含む茶葉抽出物とした。得られた茶葉抽出物10mLに、MRS液体培地で35℃、24時間培養した乳酸菌培養液を100μL加え、35℃で24時間培養した後、MRS寒天平板に塗抹してコロニー数を測定した。MRS液体培地で培養した乳酸菌培養液（茶葉抽出液と非接触）をMRS寒天平板に塗抹したときのコロニー数を100%として、茶葉抽出物と接触させた場合の乳酸菌の相対生育率を算出した。
(4) Growth in tea leaf extract (4A) Test procedure Boiled ultrapure water was added to commercially available general green tea leaves to make the tea leaf concentration 0.02 g/mL, glucose was added to 1%, Allow to stand for 1 minute. The supernatant of the solution thus obtained was used as a tea leaf extract containing 1% glucose. To 10 mL of the obtained tea leaf extract, 100 μL of lactic acid bacteria cultured in an MRS liquid medium at 35° C. for 24 hours was added, cultured at 35° C. for 24 hours, and then smeared on an MRS agar plate to measure the number of colonies. The relative growth rate of lactic acid bacteria in contact with the tea leaf extract was calculated assuming that the number of colonies when the lactic acid bacteria culture solution (not in contact with the tea leaf extract) cultured in the MRS liquid medium was smeared on the MRS agar plate was 100%. .
（４Ｂ）結果
結果を図５に示す。茶葉抽出物は抗菌作用を有することが知られているものの、図５に示す通り、いずれの菌株でも緑茶葉抽出成分に対する耐性が認められ、特に、ラクチプランチバチルス・ペントーサスAWA1922株、レビラクトバチルス・ブレビスAWA1978株、AWA1984株、AWA1985株においては各標準株と比較して茶葉抽出物に対する高い耐性が認められた。
(4B) Results The results are shown in FIG. Tea leaf extracts are known to have an antibacterial effect, but as shown in FIG. 5, resistance to green tea leaf extract components was observed in all strains. Brevis strains AWA1978, AWA1984, and AWA1985 were found to be highly resistant to tea leaf extracts compared to each standard strain.
NBRC115323
NBRC115324
NBRC115325
NBRC115326
NBRC115327
NBRC115323
NBRC115324
NBRC115325
NBRC115326
NBRC115327
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