JP5357042B2 - 骨強化用食品素材 - Google Patents
骨強化用食品素材 Download PDFInfo
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- JP5357042B2 JP5357042B2 JP2009538919A JP2009538919A JP5357042B2 JP 5357042 B2 JP5357042 B2 JP 5357042B2 JP 2009538919 A JP2009538919 A JP 2009538919A JP 2009538919 A JP2009538919 A JP 2009538919A JP 5357042 B2 JP5357042 B2 JP 5357042B2
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- protein fraction
- milk protein
- milk
- bone
- degradation product
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- Fodder In General (AREA)
Description
本発明の乳タンパク質画分又は乳タンパク質画分分解物は、骨強化作用を有するので、骨疾患の予防又は治療、骨強化等を目的とした骨強化剤として、また、骨疾患の予防又は治療用もしくは骨強化用医薬品、飲食品及び飼料の有効成分として有用である。
(1) 乳由来であること。
(2) ソディウムドデシルサルフェート−ポリアクリルアミドゲル(SDS-PAGE)電気泳動で,分子量6,000〜150,000ダルトンの範囲のタンパク質含有する画分であること。
(3) 構成アミノ酸組成中に塩基性ア本発明は、下記の構成からなる発明である。
ミノ酸を12〜14重量%含有し、かつ、塩基性アミノ酸/酸性アミノ酸比が0.5〜0.7の範囲であること。
(4) 骨芽細胞における石灰化促進作用を有すること。
(B) 前記乳タンパク質画分をタンパク質分解酵素で分解して得られる乳タンパク質画分分解物。
(C)(A)又は(B)に記載の乳タンパク質画分又は乳タンパク質画分分解物を含有する骨強化剤。
(D)(A)又は(B)に記載の乳タンパク質画分又は乳タンパク質画分分解物を配合した骨強化用医薬品。
(E)(A) 又は(B)に記載の乳タンパク質画分又は乳タンパク質画分分解物を配合した骨強化用飲食品。
(F)(A) 又は(B)に記載の乳タンパク質画分又は乳タンパク質画分分解物を配合した骨強化用飼料。
従って、本発明の骨強化作用を有する乳タンパク質画分又は乳タンパク質画分分解物を有効成分とする骨強化剤、及び、骨強化作用を有する乳タンパク質画分又は乳タンパク質画分分解物を配合した骨強化用医薬品、飲食品及び飼料は、ヒトや動物の生体内で骨芽細胞の石灰化を促進することから、骨強化作用を有し、また骨粗鬆症等の骨量減少の抑制に有効である。
下記の方法(特許第3112637号参照)により、骨強化が認められる市販品の乳タンパク質画分を調製した。
陽イオン交換樹脂のスルホン化キトパール(富士紡績製) 0.5リットルを充填した直径10cmのカラムを脱イオン水で充分洗浄した。このカラムに未殺菌脱脂乳 50リットルを流速100 ml /minで通液した後、このカラムを脱イオン水で充分洗浄し、0.95M塩化ナトリウムを含む 0.05Mリン酸緩衝液(pH 7.0) 2.5リットルを通液して樹脂に吸着したタンパク質を溶出した。そして、この溶出液を逆浸透(RO)膜処理により、脱塩し、濃縮した後、凍結乾燥して粉末状の乳タンパク質画分を得た。この操作を2回繰り返して104gのタンパク質画分を得た。このタンパク質画分の等電点は 7.0〜8.5 であって、このタンパク質画分に含まれる塩基性アミノ酸は17.8%であった。
参考例1及び実施例1で得られた各タンパク質画分について、Danjoらの方法 [Danjo.A. et al., Biochem. Biophys. Res.Commun., vol.360, pp.199-204, 2007]にしたがって調べた。すなわち、マウス頭蓋冠由来骨芽細胞様細胞MC3T3-E1を5x103 cell/cm2の細胞密度となるように、αMEM+10% FBS培地に懸濁し、24 ウエルの培養皿 へ播種した。12時間静置後、石灰化誘導培地(αMEM+10 % FBS 50μg/ml+5 mMβ-glycerophosphoacid)に交換するとともに、参考例1で得られた乳タンパク質及び実施例1で得られた本発明品をそれぞれ10-10〜10-5Mの濃度で添加した。培地を3日毎に交換し、28日間培養を続けた。
培養14、21、28 日目に、4%パラフォルムアルデヒド含有0.1M リン酸緩衝液(pH 7.4)で10分間、室温で固定した。0.01M PBS で洗浄後、2% アリザリンレッドS水溶液で15分、室温で反応させ、カルシウム染色を行った。骨の無機成分の一つであるリン酸を染色するために、固定後、蒸留水で洗浄し、5% 硝酸銀水溶液で1時間、室温で反応させてvon Kossa 染色を行った。石灰化小結節内のカルシウムイオン量の定量にはカルシウムCキット(和光純薬工業社製)を用いた。
実施例1で得られた本発明品は、用量依存的に骨芽細胞の石灰化を促進する特性が認められた.参考例1で得られた乳タンパク質にも同様の作用が認められるものの、本発明品と比較するとその作用は弱いことが判明した。
参考例1及び実施例1で得られた各タンパク質画分について、動物実験により骨強化作用を調べた。動物実験には4週齢のWistar系雌ラットを用いた。1週間の予備飼育後、卵巣摘出手術を施し、その後、カルシウム欠乏食で5週間飼育して動物実験に供した。なお、卵巣を摘出し、カルシウム欠乏食で5週間飼育したラットは、明らかに骨粗鬆症状態にあった。この骨粗鬆症状態を惹起したラットを1群6匹ずつ、乳タンパク質画分無添加の対照群(A群)、参考例1で得られた乳タンパク質画分0.5重量%投与群(B群)、実施例1で得られた乳タンパク質画分0.1重量%投与群(C群)、実施例1で得られた乳タンパク質画分0.5重量%投与群(D群)、実施例1で得られた乳タンパク質画分1.0重量%投与群(E群)の5試験群に分け、それぞれ表1に示す試験飼料で4ヶ月飼育した。なお、各試験飼料の窒素含量(17.06%)が同様となるようカゼインで調整した。また、各試験飼料については、100g当たり、カルシウム 300mg、リン 230mg及びマグネシウム 50mgを配合した。
なお、実験結果は示さなかったが、実施例3及び実施例4で得られた乳タンパク質画分分解物を用いた場合も同様の効果が認められた。
Claims (6)
- 次の(1)〜(4)の特性を有する乳タンパク質画分。
(1) 乳由来であること。
(2) ソディウムドデシルサルフェート−ポリアクリルアミドゲル(SDS-PAGE)電気泳動で、分子量6,000〜150,000ダルトンの範囲のタンパク質を含有する画分であること。
(3) 構成アミノ酸組成中に塩基性アミノ酸を12〜14重量%含有し、かつ、塩基性アミノ酸/酸性アミノ酸比が0.5〜0.7の範囲であること。
(4) 骨芽細胞における石灰化促進作用を有すること。 - 請求項1に記載の乳タンパク質画分をタンパク質分解酵素で分解して得られる骨芽細胞における石灰化促進作用を有するタンパク質分解物。
- 請求項1又は2に記載の乳タンパク質画分又は乳タンパク質画分分解物を含有する骨強化剤。
- 請求項1又は2に記載の乳タンパク質画分又は乳タンパク質画分分解物を配合した骨強化用医薬品。
- 請求項1又は2に記載の乳タンパク質画分又は乳タンパク質画分分解物を配合した骨強化用飲食品。
- 請求項1又は2に記載の乳タンパク質画分又は乳タンパク質画分分解物を配合した骨強化用飼料。
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CA2823505A1 (en) | 2011-01-26 | 2012-08-02 | Megmilk Snow Brand Co., Ltd. | Agent for improving peripheral sensitivity of skin |
JP6140164B2 (ja) * | 2012-07-31 | 2017-05-31 | 雪印メグミルク株式会社 | 骨強化用チーズ組成物及びその製造方法 |
MX366535B (es) * | 2012-07-31 | 2019-07-12 | Megmilk Snow Brand Co Ltd | Producto novedoso de leche en polvo y metodo para preparar el mismo. |
KR102555277B1 (ko) * | 2022-11-15 | 2023-07-14 | 주식회사 에이치피오 | 뼈 형성 촉진 또는 골 손실 예방용 조성물 |
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ES2547402T3 (es) | 2015-10-06 |
CN101842026A (zh) | 2010-09-22 |
EP2210507B1 (en) | 2015-09-09 |
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AU2008320270A1 (en) | 2009-05-07 |
MY160058A (en) | 2017-02-15 |
US8420599B2 (en) | 2013-04-16 |
KR20100100830A (ko) | 2010-09-15 |
WO2009057280A1 (ja) | 2009-05-07 |
CN101842026B (zh) | 2014-02-19 |
KR101705905B1 (ko) | 2017-02-10 |
NZ584888A (en) | 2012-08-31 |
CA2704221C (en) | 2014-12-02 |
EP2210507A1 (en) | 2010-07-28 |
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JPWO2009057280A1 (ja) | 2011-03-10 |
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