JP4726129B2 - ビフィズス菌増殖促進ペプチド - Google Patents
ビフィズス菌増殖促進ペプチド Download PDFInfo
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- JP4726129B2 JP4726129B2 JP2006008738A JP2006008738A JP4726129B2 JP 4726129 B2 JP4726129 B2 JP 4726129B2 JP 2006008738 A JP2006008738 A JP 2006008738A JP 2006008738 A JP2006008738 A JP 2006008738A JP 4726129 B2 JP4726129 B2 JP 4726129B2
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- peptide
- bifidobacteria
- growth
- actomyosin
- leu
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Description
また本発明は、〔5〕アクトミオシンあるいはアクトミオシンを含む食肉タンパク質懸濁液をプロテアーゼで処理することにより得られるペプチド含有画分を含有し、ビフィズス菌増殖促進効果を有することが表示されている食品、サプリメント、飼料、医薬品に関するものである。さらに本発明は、〔6〕Glu-Leu-Metの配列で示されるペプチド又はその塩に関するものである。
本発明の有効成分(ペプチド)を経口投与(摂取)する場合、本発明の効果が損なわれない範囲で、上記の各製品に慣用される他の成分を含んでもよい。このような添加剤の例としては、賦形剤(ラクトース、スターチ等)、結合剤(シロップ、ゼラチン等)、香料、崩壊剤等が挙げられる。また、水溶性食物繊維、不溶性食物繊維、オリゴ糖等のビフィズス菌の増殖に有用な成分を添加してもよい。さらに、ビタミンA、ビタミンB1、ビタミンB2、ビタミンB6、ビタミンB12、ビタミンC、ビタミンD、ビタミンE、ナイアシン(ニコチン酸)、パントテン酸、葉酸等のビタミン類、リジン、スレオニン、トリプトファン等の必須アミノ酸類や、カルシウム、マグネシウム、鉄、亜鉛、銅等のミネラル類及び、例えば、α−リノレン酸、EPA、DHA、カゼインホスホペプチド(CPP)、カゼインカルシウムペプチド(CCP)等の人の健康に寄与する物質類、その他の食品や食品添加物として認可されている有用物質の1種又は2種以上が使用できる。
あるいは適当な食品等に添加・加工(混合、加熱等)してもよい。
以下、本発明を実施例で説明する。以下の実施例は、本発明を説明するためにあげた例であり、これにより本発明を限定するものではない。
アクトミオシンは、豚骨格筋から調製した。すなわち、挽き肉にした豚骨格筋(大腿二頭筋)に緩衝液(0.6M KCl/0.04M NaHCO3/0.01m Na2CO3)を加え、混合・遠心分離を繰り返した後、透析による脱塩と凍結乾燥により、アクトミオシン標品を得た。蒸留水10mlにアクトミオシン100mgを入れ、十分に懸濁させた。これに、5種類のプロテアーゼ(パパイン、フィチン、トリプシン、プロテイネースK、プロナーゼE)のうちの1種を10mg添加し、37℃で3時間反応させた。反応終了後、95℃で10分間加熱し、プロテアーゼを失活させた。遠心分離により不溶物を除去した上澄液をアクトミオシン分解物溶液とした。
ビフィズス菌に対するアクトミオシン分解物あるいはペプチドの増殖促進効果は、以下の方法により測定した。まず、110℃で10分間加熱殺菌した10%還元脱脂乳(以下、脱脂乳)5mlに、測定試料(アクトミオシン分解物溶液あるいはペプチド溶液)を50μl添加した。これに、増殖促進効果の指標菌として用いたビフィズス菌の培養液(MRS液体培地で前培養したもの)10μlを接種した。なお、ビフィズス菌にはヒト腸管由来の7菌株(Bifidobacterium bifidum JCM1254、B. breve JCM1192、B. longum JCM1217、B. catenulatum JCM1194、B. angulatum JCM7096、B. pseudocatenulum JCM1200、B. adolescentis JCM7042)を用いて検討したが、脱脂乳中においてペプチドの増殖促進効果が最も顕著にあらわれたB. bifidum JCM1254を最終的な指標菌として用いることとした。ビフィズス菌接種後の脱脂乳は、37℃で48時間培養した後に、pHを測定した。ビフィズス菌は増殖に伴い、乳酸や酢酸を生成し、培地pHを低下させることが知られている。このため、培地pHの低下程度から、ビフィズス菌の増殖程度を判定することができる。
豚骨格筋アクトミオシンのプロテアーゼ分解物5種のビフィズス菌(B. bifidum JCM1254)増殖促進効果を測定した。図1に示した結果のように、アクトミオシンのパパイン分解物に強い活性(脱脂乳のpH低下=ビフィズス菌増殖促進効果)が見られた。
目的ペプチドの精製は、ゲル濾過クロマトグラフィーと逆相高速液体クロマトグラフィー(HPLC)の組み合わせにより実施した。ゲル濾過クロマトグラフィーは、カラムにセファデックスG25Fine(26mm×100mm)を用い、溶出液には蒸留水を使用した。流速は2ml/minとし、溶出液中のペプチドの検出は、215nmの吸光度の測定により行った。アクトミオシン分解物溶液1mlをカラムに負荷し、溶出液を2分間隔(4mlずつ)で分画採取した。各画分は凍結乾燥させた後に、蒸留水に再溶解させ、ビフィズス菌増殖促進効果を判定した。
精製したペプチドのアミノ酸配列は、自動エドマン法(気相法)によるN末端アミノ酸分析により決定した。自動分析装置としてアプライドバイオシステムズ社製プロテインシークエンサーモデル470Aを使用した。また、ペプチドの分子量測定には、島津製作所製質量分析装置QP8000αを使用した。
合成したトリペプチド(Glu-Leu-Met)がビフィズス菌増殖促進効果を示すことを確認すると共に、この活性が単に構成アミノ酸や構成部分ペプチド(ジペプチド)に起因するものではないことを示した。図2は、合成したトリペプチド(Glu-Leu-Met)と部分配列ジペプチド(Glu-Leu, Leu-Met)、そして構成アミノ酸(Glu, Leu, Met, 3種アミノ酸混合)のビフィズス菌増殖促進効果を測定した結果を示したグラフである。それぞれのペプチドとアミノ酸は、脱脂乳中の濃度が0.1mMになるように添加した。Glu-Leu-Metの部分配列であるジペプチド(Glu-Leu, Leu-Met)と構成アミノ酸(Glu, Leu, Met, 3種アミノ酸混合)は、いずれもGlu-Leu-Metより弱い活性しか示さなかった。このことから、Glu-Leu-Metのビフィズス菌増殖促進効果は、構成アミノ酸が単なる窒素源として利用されることにより発現するのではなく、トリペプチド配列に重要な意義があることが確認された。
Claims (6)
- Glu-Leu-Metの配列で示されるペプチド又はその塩を有効成分とするビフィズス菌増殖促進剤。
- 請求項1に記載のビフィズス菌増殖促進剤を含有するビフィズス菌増殖促進効果を有することを特徴とする食品。
- 請求項1に記載のビフィズス菌増殖促進剤を含有するビフィズス菌増殖促進効果を有することを特徴とするサプリメント。
- 請求項1に記載のビフィズス菌増殖促進剤を含有するビフィズス菌増殖促進効果を有することを特徴とする飼料。
- 請求項1に記載のビフィズス菌増殖促進剤を含有するビフィズス菌増殖促進効果を有することを特徴とする医薬品。
- Glu-Leu-Metの配列で示されるペプチド又はその塩。
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Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
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JPH05336954A (ja) * | 1992-06-05 | 1993-12-21 | Eisai Co Ltd | 微生物培養法及び微生物用培地 |
JPH05344865A (ja) * | 1992-06-16 | 1993-12-27 | Yukie Murai | 健康食品 |
JPH08509366A (ja) * | 1993-04-26 | 1996-10-08 | ノボ ノルディスク アクティーゼルスカブ | タンパク質の加水分解方法 |
US5711977A (en) * | 1996-05-06 | 1998-01-27 | Food Industry Research And Development Institute | Bifidobacteria strains with acid, bile salt and oxygen tolerance and their culture method |
JP2000093166A (ja) * | 1998-09-25 | 2000-04-04 | Kyodo Milk Industry Co Ltd | 卵白由来ビフィズス菌増殖促進物質と当該物質を含有する食品 |
JP2003102427A (ja) * | 2001-09-27 | 2003-04-08 | Prima Meat Packers Ltd | ストレス性胃潰瘍予防作用を有する機能性食品素材又は飲食品 |
JP2004057047A (ja) * | 2002-07-26 | 2004-02-26 | Snow Brand Milk Prod Co Ltd | 乳酸菌の生育促進剤及びその製造法 |
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Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH05336954A (ja) * | 1992-06-05 | 1993-12-21 | Eisai Co Ltd | 微生物培養法及び微生物用培地 |
JPH05344865A (ja) * | 1992-06-16 | 1993-12-27 | Yukie Murai | 健康食品 |
JPH08509366A (ja) * | 1993-04-26 | 1996-10-08 | ノボ ノルディスク アクティーゼルスカブ | タンパク質の加水分解方法 |
US5711977A (en) * | 1996-05-06 | 1998-01-27 | Food Industry Research And Development Institute | Bifidobacteria strains with acid, bile salt and oxygen tolerance and their culture method |
JP2000093166A (ja) * | 1998-09-25 | 2000-04-04 | Kyodo Milk Industry Co Ltd | 卵白由来ビフィズス菌増殖促進物質と当該物質を含有する食品 |
JP2003102427A (ja) * | 2001-09-27 | 2003-04-08 | Prima Meat Packers Ltd | ストレス性胃潰瘍予防作用を有する機能性食品素材又は飲食品 |
JP2004057047A (ja) * | 2002-07-26 | 2004-02-26 | Snow Brand Milk Prod Co Ltd | 乳酸菌の生育促進剤及びその製造法 |
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