JP5753990B2 - Natto bacteria having anti-rotavirus activity and fermented products thereof, and foods, animal feeds and pharmaceuticals containing them - Google Patents
Natto bacteria having anti-rotavirus activity and fermented products thereof, and foods, animal feeds and pharmaceuticals containing them Download PDFInfo
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Description
本発明は、納豆菌（Bacillus subtilis natto）並びに納豆菌で発酵させた発酵物に関し、特に抗ロタウィルス活性を有し、ロタウィルス（Rotavirus）による下痢発症を予防する納豆菌及びその発酵物に関する。また、これらが含まれた食品、動物飼料及び医薬品に関する。 The present invention relates to a Bacillus subtilis natto and a fermented product fermented with Bacillus natto, and more particularly to a Bacillus natto having anti-rotavirus activity and preventing the onset of diarrhea due to Rotavirus and the fermented product thereof. The present invention also relates to foods, animal feeds and pharmaceuticals containing these.
一般にヒトの９０％程度が乳幼児期にロタウィルスに感染し、そのうち１０％程度が嘔吐、発熱、下痢による脱水症状のために入院、治療を行っている。そのため地域差があるが、全世界で年間７０万人程度が亡くなっていると推定されている。また、ロタウィルスには家畜、ペットも感染しやすく、その場合には死亡率も高く経済的損失が大きい。 In general, about 90% of humans are infected with rotavirus in early childhood, and about 10% are hospitalized and treated for dehydration due to vomiting, fever, and diarrhea. Therefore, although there are regional differences, it is estimated that about 700,000 people die every year worldwide. In addition, livestock and pets are easily infected with rotavirus, in which case mortality is high and economic loss is great.
予防方法としては米国で２００６年にワクチンが開発・承認されたが、日本国内では未承認であり、治療方法に関しては未だ対症療法が中心となっている。 As a preventive method, a vaccine was developed and approved in the United States in 2006, but it has not been approved in Japan, and symptomatic treatment is still the main method of treatment.
他方、プロバイオティクスでの腸管免疫増強作用による予防方法としては、ラクトバチルス・ラムノサス（Lactobacillus rhamnosus）の免疫増強作用（特許文献１）、ラクトバチルス・ロイテリ（Lactobacillus reuteri）の免疫増強作用（特許文献２）、ラクトバチルス属・ロイコノストック属・ペディオコッカス属（特許文献３）の免疫増強作用と乳酸菌を利用した予防方法などが提案されている。 On the other hand, as a preventive method by intestinal immunity enhancing action by probiotics, Lactobacillus rhamnosus immune enhancing action (Patent Document 1), Lactobacillus reuteri immune enhancing action (Patent Document) 2) The immunopotentiating action of Lactobacillus genus, Leuconostoc genus, Pediococcus genus (Patent Document 3) and the prevention method using lactic acid bacteria have been proposed.
しかしながら乳酸菌による発酵物の代表である乳製品ヨーグルト、チーズ等には乳アレルギーや乳糖不耐性の問題があり、プロバイオティクスとして利用できない場合がある。 However, dairy yogurts, cheeses and the like, which are representative of fermented products by lactic acid bacteria, have problems of milk allergy and lactose intolerance and may not be used as probiotics.
そこで本発明は、乳アレルギーや乳糖不耐性の問題が生ずるおそれがなく、ロタウィルスによる下痢発症の予防に効果的な菌体、これが含まれた食品、動物飼料及び医薬品、等を提供することを目的にしている。 Accordingly, the present invention provides a fungus body effective in preventing the development of diarrhea due to rotavirus, a food, animal feed, a pharmaceutical, and the like that are free from problems of milk allergy and lactose intolerance, and the like. It is aimed.
以上の目的を達成するために本発明者らは鋭意研究を重ねた結果、乳アレルギーや乳糖不耐性の問題が生ずるおそれのない納豆菌の中で、納豆菌（Bacillus subtilis natto）ＡＳ３９（寄託番号：ＦＥＲＭ Ｐ−２１８９５）が抗ロタウィルス活性を有し、ロタウィルスによる下痢発症を予防することを見出した。また、この納豆菌（Bacillus subtilis natto）ＡＳ３９（寄託番号：ＦＥＲＭ Ｐ−２１８９５）の抗ロタウィルス活性が、乳酸菌に比べて十分なものであることを見出した。 In order to achieve the above-mentioned object, the present inventors have conducted intensive research. As a result, among the Bacillus natto bacteria that are unlikely to cause problems of milk allergy and lactose intolerance, Bacillus subtilis natto AS39 (deposit number) : FERM P-21895) has anti-rotavirus activity and has been found to prevent the development of diarrhea due to rotavirus. Moreover, it discovered that the anti-rotavirus activity of this Bacillus subtilis natto AS39 (deposit number: FERM P-21895) was sufficient compared with lactic acid bacteria.
請求項１記載の発明は、
抗ロタウィルス活性を有する納豆菌（Bacillus subtilis natto）であって、前記納豆菌が、寄託番号：ＦＥＲＭ Ｐ−２１８９５号として寄託された納豆菌ＡＳ３９株であることを特徴とする納豆菌である。
The invention described in claim 1
What natto (Bacillus subtilis natto) der having anti-rotavirus activity, the Bacillus natto, accession number: is natto, which is a FERM P-21895 No. natto AS39 strain deposited as .
請求項２記載の発明は、
請求項１記載の納豆菌で発酵させた発酵物からなる発酵食品である。
The invention according to claim 2
A fermented food comprising a fermented product fermented with Bacillus natto according to claim 1 .
請求項３記載の発明は、
請求項１記載の納豆菌で発酵させた発酵物を含む食品である。
The invention described in claim 3
A food comprising a fermented product fermented with Bacillus natto according to claim 1 .
請求項４記載の発明は、
請求項１記載の納豆菌で発酵させた発酵物を含む動物用飼料である。
The invention according to claim 4
An animal feed comprising a fermented product fermented with Bacillus natto according to claim 1 .
請求項５記載の発明は、
請求項１記載の納豆菌で発酵させた発酵物を含む医薬品である。
The invention according to claim 5
A pharmaceutical product comprising a fermented product fermented with Bacillus natto according to claim 1 .
本発明によれば、乳アレルギーや乳糖不耐性の問題が生ずるおそれがなく、ロタウィルスによる下痢発症の予防に効果的な菌体、これが含まれた食品、動物飼料及び医薬品、等を提供することができる。 According to the present invention, there is no possibility of causing milk allergy or lactose intolerance, and a fungus body effective in preventing the onset of diarrhea due to rotavirus, foods containing the same, animal feed and pharmaceuticals, and the like are provided. Can do.
この発明の納豆菌ＡＳ３９（寄託番号：ＦＥＲＭ Ｐ−２１８９５）は、標準的な乳酸菌と比較したときにパイエル板細胞のＩｇＡ抗体産生促進作用が高いことが確認できた。すなわち、この発明の抗ロタウィルス活性を有し、ロタウィルスによる下痢発症を予防する納豆菌ＡＳ３９（寄託番号：ＦＥＲＭ Ｐ−２１８９５）は、乳酸菌と比べて、パイエル板細胞のＩｇＡ抗体産生促進作用が高く、このことから、乳酸菌と比べて高い腸管免疫増強作用を発揮できるものと考えられる。 It was confirmed that the Bacillus natto AS39 (deposit number: FERM P-21895) of the present invention has a high IgA antibody production promoting effect on Peyer's patch cells when compared with standard lactic acid bacteria. That is, the Bacillus natto AS39 (deposit number: FERM P-21895) which has the anti-rotavirus activity of the present invention and prevents the onset of diarrhea due to rotavirus has an effect of promoting IgA antibody production of Peyer's patch cells as compared with lactic acid bacteria. Therefore, it is considered that the intestinal immunity enhancing action is higher than that of lactic acid bacteria.
そして、この発明によれば、乳アレルギーや乳糖不耐性の問題が生ずるおそれがなく、乳酸菌と比べて、腸管免疫増強作用やパイエル板細胞のＩｇＡ抗体産生促進作用が十分な納豆菌ＡＳ３９（寄託番号：ＦＥＲＭ Ｐ−２１８９５）を有効成分とした、抗ロタウィルス活性を有しロタウィルスによる下痢発症を予防する食品、動物飼料、医薬品を提供することが可能になる。 And according to this invention, there is no possibility that the problem of milk allergy or lactose intolerance will occur, and compared with lactic acid bacteria, the intestinal immunity enhancing action and the Peyer's plate cell IgA antibody production promoting action are sufficient. : FERM P-21895) as an active ingredient, it is possible to provide foods, animal feeds, and pharmaceuticals that have anti-rotavirus activity and prevent the onset of diarrhea due to rotavirus.
本発明の抗ロタウィルス活性を有し、ロタウィルスによる下痢発症を予防する納豆菌ＡＳ３９（寄託番号：ＦＥＲＭ Ｐ−２１８９５）は、日本各地の田の稲わらから選抜したものである。 The Bacillus natto AS39 (deposit number: FERM P-21895), which has anti-rotavirus activity and prevents the development of diarrhea due to rotavirus, is selected from rice straws in various parts of Japan.
納豆菌としての同定については、納豆菌から製造した納豆の糸引き、味、香り等の官能検査に特に異常が認められなかったことと、１６ＳｒＲＮＡをコードするＰＣＲ産物のシーケンスを行い、その塩基配列が市販納豆菌と一致したことにより確認した。 Regarding the identification as natto bacteria, no abnormalities were found in the sensory tests such as stringing, taste, fragrance etc. of natto produced from natto bacteria, and the sequence of the PCR product encoding 16S rRNA was performed, and its base sequence Was confirmed to be consistent with commercial natto bacteria.
納豆菌は蒸煮した大豆上で３５〜４５℃の温度で良好に生育することが出来、これを１５〜２４時間発酵させたものは納豆として知られている。また納豆菌は豆腐、豆乳、醤油、大豆タンパク質等の大豆加工食品や、おから、大豆ミール、大豆煮汁等の食物残渣も発酵させることが出来る。従ってこれらの発酵物は加工食品、飼料、医薬品としても利用可能である。 Natto bacteria can grow well on steamed soybeans at a temperature of 35 to 45 ° C., and fermented for 15 to 24 hours is known as natto. Natto can also ferment soybean processed foods such as tofu, soy milk, soy sauce, and soy protein, and food residues such as okara, soybean meal, and soybean soup. Therefore, these fermented products can be used as processed foods, feeds and pharmaceuticals.
そこで、本発明の納豆菌ＡＳ３９（寄託番号：ＦＥＲＭ Ｐ−２１８９５）で発酵させた発酵物からなる発酵食品、や当該発酵物を含む食品、動物用飼料、医薬品を提供することができる。 Therefore, fermented foods made of fermented products fermented with Bacillus natto AS39 (deposit number: FERM P-21895) of the present invention, foods containing the fermented products, animal feeds, and pharmaceuticals can be provided.
発酵食品としては典型的に納豆があり、この他に、前述したように、豆腐、豆乳、醤油、大豆タンパク質等の大豆加工食品や、おから、大豆ミール、大豆煮汁等の食物残渣を、本発明の納豆菌ＡＳ３９（寄託番号：ＦＥＲＭ Ｐ−２１８９５）で発酵させた発酵物などが含まれる。 Fermented foods typically include natto. In addition, as described above, processed foods such as tofu, soy milk, soy sauce, and soy protein, and food residues such as okara, soy meal, and soy broth The fermented material fermented with Bacillus natto AS39 (deposit number: FERM P-21895) of the invention is included.
また、発酵物を含む食品としては前述した納豆などの発酵物を利用した加工食品や、納豆菌ＡＳ３９（寄託番号：ＦＥＲＭ Ｐ−２１８９５）で発酵させた前記の発酵物からなる加工食品、納豆菌ＡＳ３９（寄託番号：ＦＥＲＭ Ｐ−２１８９５）で発酵させた前記の発酵物を含む加工食品などがある。これらは、スナック菓子などの菓子類として提供する形態や、顆粒状、バー（棒）形態、粉末、カプセル状、錠剤形態、等、種々の態様、形態で提供することができる。 In addition, as foods containing fermented products, processed foods using fermented products such as natto mentioned above, processed foods made of the fermented products fermented with natto bacteria AS39 (deposit number: FERM P-21895), natto bacteria There is a processed food containing the fermented product fermented with AS39 (deposit number: FERM P-21895). These can be provided in various forms and forms such as forms provided as confectionery such as snack confectionery, granules, bars (bars), powders, capsules, tablets, and the like.
動物飼料としては、納豆菌ＡＳ３９（寄託番号：ＦＥＲＭ Ｐ−２１８９５）で発酵させた前記の発酵物からなる動物飼料や、納豆菌ＡＳ３９（寄託番号：ＦＥＲＭ Ｐ−２１８９５）で発酵させた前記の発酵物を含む動物飼料を、家畜飼料やペットフードなどとして提供することができる。 As animal feed, animal feed comprising the fermented product fermented with Bacillus natto AS39 (deposit number: FERM P-21895) or fermented with Bacillus natto AS39 (deposit number: FERM P-21895). Animal feed including food can be provided as livestock feed or pet food.
医薬品としては、納豆菌ＡＳ３９（寄託番号：ＦＥＲＭ Ｐ−２１８９５）で発酵させた前記の発酵物からなる医薬品や、納豆菌ＡＳ３９（寄託番号：ＦＥＲＭ Ｐ−２１８９５）で発酵させた前記の発酵物を含む医薬品を、顆粒状、バー（棒）形態、粉末、カプセル状、錠剤形態、等、種々の態様、形態で提供することができる。 As a medicine, the medicine comprising the fermented product fermented with Bacillus natto AS39 (deposit number: FERM P-21895) or the fermented product fermented with Bacillus natto AS39 (deposit number: FERM P-21895). The pharmaceuticals to be included can be provided in various forms and forms such as granular, bar (bar) form, powder, capsule form, tablet form and the like.
以下、実施例により、本発明の納豆菌ＡＳ３９（寄託番号：ＦＥＲＭ Ｐ−２１８９５）の抗ロタウィルス活性ならびに下痢発症予防の特徴を説明する。 Hereinafter, examples illustrate the anti-rotavirus activity and characteristics of preventing diarrhea onset of Bacillus natto AS39 (deposit number: FERM P-21895) of the present invention.
７週齢のＢＡＬＢ／ｃマウスを解剖し、パイエル板細胞を単離・調整した。次に５×１０６ｃｅｌｌ／ｍｌの前記パイエル板細胞に納豆菌を１００μｌ（凍結乾燥物２００μｇ／ｍｌ）添加し、ＲＰＭＩ培地で３７℃、５％ＣＯ２の条件下で培養した。 Seven week old BALB / c mice were dissected and Peyer's patch cells were isolated and prepared. Next, 100 μl (freeze-dried product 200 μg / ml) of Bacillus natto was added to 5 × 10 6 cells / ml of the Peyer's plate cells, and cultured in RPMI medium under conditions of 37 ° C. and 5% CO 2 .
培養７日目に培養上清をサンプリングし、ＩｇＡ抗体の産生量をＥＬＩＳＡ法により測定した。比較例として無添加区（パイエル板細胞のみの培養区）、標準的な乳酸菌であるビフィドバクテリウム・インファンティス１２２２Ｔ添加区、及び標準的な納豆菌である三浦株とし、稲わらから分離した納豆菌ＡＳ１４株、ＡＳ３８株、ＡＳ３９株、ＡＳ１１５株、ＡＳ１７３株のＩｇＡ抗体産生量を表１に示す。
表１に示されるように納豆菌ＡＳ１４株、ＡＳ３８株、ＡＳ３９株、ＡＳ１１５株、ＡＳ１７３株はＩｇＡ抗体の産生を促進した。特にＡＳ３９株は顕著な抗体産生量を示した。 As shown in Table 1, the Bacillus natto strain AS14, AS38, AS39, AS115, and AS173 promoted IgA antibody production. In particular, the AS39 strain showed a significant amount of antibody production.
ＡＳ３９株は標準的な乳酸菌であるビフィドバクテリウム・インファンティス１２２２Ｔに比較してパイエル板細胞のＩｇＡ抗体産生促進作用が高いことを確認できた。 It was confirmed that the AS39 strain had a higher IgA antibody production promoting effect on Peyer's patch cells compared to Bifidobacterium infantis 1222T, which is a standard lactic acid bacterium.
７週齢のＢＡＬＢ／ｃ雌マウスを３群に分け、菌無添加の標準飼料、標準的な納豆菌である納豆菌三浦株を０．１％添加した標準飼料、納豆菌ＡＳ３９株を０．１％添加した標準飼料を自由に摂取させて飼育した。４週間後に雄マウスと交配させ、その後ロタウィルスを経口免疫した。出産した５日目の仔マウスにロタウィルスを経口感染させて、その後の下痢発症の有無を観察した。各群における下痢発症率を表２に示す。
納豆菌ＡＳ３９株を添加した標準飼料で飼育した母マウスから産まれた仔マウスの下痢発症率は有意に低下した。特にＡＳ３９群における仔マウスの下痢発症率は、他の２群に比べ特に低い数値を示している。これはパイエル板細胞からのＩｇＡ抗体産生の高い納豆菌ＡＳ３９株を摂取した母マウスの母乳中のロタウィルス抗体が増強され、その母乳を飲んだ仔マウスの下痢発症がロタウィルス抗体により抑えられたものと推察される。 The incidence of diarrhea in pups born from mother mice bred with a standard feed supplemented with Bacillus natto strain AS39 was significantly reduced. In particular, the incidence of diarrhea in pup mice in the AS39 group is particularly low compared to the other two groups. This is because rotavirus antibody in the breast milk of mother mice that ingested the Bacillus natto strain AS39, which produces high IgA antibodies from Peyer's patch cells, was enhanced, and the development of diarrhea in pups that had taken the breast milk was suppressed by rotavirus antibodies. Inferred.
以上より、ＩｇＡ抗体産生促進作用を有する納豆菌、特にＡＳ３９株を食品、動物飼料、医薬に利用することで、ロタウィルスによる下痢発症を効果的に予防又は治療することができると考えられる。 From the above, it is considered that diarrhea caused by rotavirus can be effectively prevented or treated by using natto bacteria having an action of promoting IgA antibody production, particularly AS39 strain, for foods, animal feeds and medicines.
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