JP5896543B2 - フェリチンの単離、使用、および分析の方法 - Google Patents
フェリチンの単離、使用、および分析の方法 Download PDFInfo
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- JP5896543B2 JP5896543B2 JP2014550504A JP2014550504A JP5896543B2 JP 5896543 B2 JP5896543 B2 JP 5896543B2 JP 2014550504 A JP2014550504 A JP 2014550504A JP 2014550504 A JP2014550504 A JP 2014550504A JP 5896543 B2 JP5896543 B2 JP 5896543B2
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- ferritin
- soluble
- iron
- insoluble
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Description
本明細書中で使用される下記の用語およびそれらの語尾変化は、そのような用語が使用される文脈が明白に別の意味を意図していない限り下記に示す意味を有する。
(i)その状態に罹りやすいが、まだそれに罹ったと診断されていない被験者にその状態が起こるのを防ぐこと、
(ii)その状態を抑制すること、例えばその進行を阻みまたは遅らせること、あるいは
(iii)その状態を緩和すること、例えばその状態の退縮を引き起こすこと
が含まれる。
本明細書および特許請求の範囲中の規定量は、本発明の実施と矛盾しない規定量の変形量を包含することを意図している。そのような変形量は、明細書中で概要を述べる手順に従って当業熟練者によって容易に決められ、一般には+/−10〜20%の幅の変動をエンコードする。
フェリチンは、ケージド鉄酸化物ミネラルを有するタンパク質ケージを含有する点で食餌性の鉄の独特の形態である。他の鉄サプリメントおよび食餌性の鉄供給源とは対照的に、フェリチンのタンパク質外被は、使用者の腸を、鉄によって引き起こされるフリーラジカル化学作用(腸を刺激する恐れのある)から保護する。これに加えてフェリチン鉄は、腸から血液中にゆっくり放出され、それもまた鉄によって引き起こされるフリーラジカル化学作用から腸を保護することを可能にする。
植物性材料を出発材料として使用して実質上純粋な形でフェラチンを単離することができる。フェラチンは、植物性材料から、(a)植物性材料を可溶性および不溶性画分に分離するステップと、(b)ステップ(a)の可溶性画分から非フェリチン成分を除去し、それによってフェリチンを単離するステップと、(c)ステップ(b)からの単離されたフェリチンを濃縮するステップとによって単離することができる。
動物ではフェリチンは、肝臓、腎臓、脾臓、および骨髄中に高い量で存在する。本発明は、フェリチンを動物の組織から得ることができると考えている。動物由来の鉄含有フェリチンもまた、食餌において鉄の増加を必要としているヒトおよび他の動物が使用することができる。
植物または動物の何れかの出発材料から本発明の方法によって得られる単離されたフェリチンは、様々な産業用途を有する。この単離されたフェリチンは、具体的には食品、飲料、栄養補給食品、およびサプリメントなどのヒトまたは動物の栄養素用の製品に加えるのに有用である。
本発明の方法は、上記の方法によって単離された植物性または動物性フェリチンを含有する治療用または補給用組成物を、鉄を必要としている生物に任意の既知の手段によって送達することができると考える。その生物は、ヒトを含む哺乳動物などの動物であることができる。あるいはその生物は植物である。好ましい実施形態ではこの組成物は、経口的に送達することができる。
植物抽出物中のフェリチンタンパク質および鉄の濃度の大まかな決定については、ウサギのポリクローナル抗ダイズフェリチン抗血清を用いる定量ウェスタンブロットを使用して以前に示されている(Regland,M.およびTheil,E.C.の論文、Plant Mol.Biol.21:555〜560,1993、Kimata,Y.およびTheil,E.C.の論文、Plant Physiol.104:263〜270,1994、Burton,J.W.等の論文、J.Plant Nutr.21:913〜927,1998)。
植物からフェリチンを単離するために、ほぼ中性に緩衝した中性の塩水溶液を加えた後に、出発植物性材料を可溶性および不溶性画分に分離した。この混合物は、室温で冷やしてもよく、また加熱してもよい。この実施例では、その植物性材料/塩溶液を32から43oFの間(0〜6℃)で一晩、ゆっくり揺り動かす(stirring)か、掻き混ぜて(agitation)撹拌しながら冷やし続けた。免疫分析を妨げない、より極端な温度、pH、および溶媒の条件もまた使用することができる。次に、この植物性材料/塩類溶液抽出物を濾過し、続いて35,000xgを超える連続的または静的遠心分離(static centrifugation)を使用して20分間清澄化させた。
例えば、肝臓、腎臓、脾臓、骨髄、または他のフェリチンを多く含む供給源などの動物性材料からフェリチンを単離することができる。動物性材料からのフェリチンの単離は、(a)動物性材料を可溶性および不溶性画分に分離するステップと、(b)この可溶性画分を熱変性するステップと、(d)この熱変性した可溶性画分から酵素消化によって炭水化物を除去するステップと、(c)この酵素処理され、熱変性された可溶性画分を濃縮するステップとを使用して行うことができる。動物由来の鉄を含有する単離されたフェリチンもまた、食餌において鉄を増やす必要性のあるヒトおよび他の動物によって使用されることができる。
フェリチン含量は、動物または植物性出発材料由来の加工物(すなわち食品、サプリメント、栄養補給食品)を含めた任意のフェリチンを含む物質から求めることができる。この実施例では、豆果由来のフェリチン鉄を含有する食品試料においてフェリチン鉄を定量した。
Claims (15)
- 豆果からフェリチンを単離するための方法において、
(a)前記豆果を可溶性および不溶性豆果の画分に分離するステップと、
(b)前記不溶性豆果の画分に中性の塩類緩衝液を加えて、不溶性豆果の溶液にするステップと、
(c)ステップ(b)の前記不溶性豆果の溶液を清澄化して、可溶性および不溶性豆果の溶液の画分に分けるステップと、
(d)ステップ(c)の前記清澄化された可溶性豆果の溶液を1種類または複数種類のグリコシダーゼ酵素で酵素処理するステップと、
(e)ステップ(d)の前記酵素処理され、清澄化された可溶性豆果の溶液を分別して、非フェリチン成分を除去するステップと、
(f)ステップ(e)の前記分別された可溶性豆果の溶液から単離されたフェリチンを濃縮するステップと
を含むことを特徴とする方法。 - 植物性材料からフェリチンを単離する方法において、
(a)前記植物性材料を可溶性および不溶性画分に分離するステップと、
(b)ステップ(a)の前記可溶性画分から非フェリチン成分を1種類または複数種類のグリコシダーゼ酵素で酵素除去し、それによってフェリチンを単離するステップと、
(c)ステップ(b)からの前記単離されたフェリチンを濃縮するステップと
を含むことを特徴とする方法。 - 請求項2に記載の方法において、前記植物性材料が豆果を含むことを特徴とする方法。
- 請求項2に記載の方法において、前記植物性材料が、豆果の加工由来の廃棄流を含むことを特徴とする方法。
- 請求項2に記載の方法において、前記可溶性画分の分別のステップをさらに含むことを特徴とする方法。
- 請求項2に記載の方法において、溶媒による前記可溶性画分の処理のステップをさらに含むことを特徴とする方法。
- 請求項6に記載の方法において、前記溶媒が有機溶媒であることを特徴とする方法。
- 請求項3に記載の方法において、前記豆果がダイズを含むことを特徴とする方法。
- 請求項2に記載の方法において、前記植物性材料が植物全体を含むことを特徴とする方法。
- 請求項2に記載の方法において、前記植物性材料が、種子、豆、茎、果実、葉、根、および花のうちの1種類または複数種類を含むことを特徴とする方法。
- 請求項2に記載の方法において、可溶性および不溶性の分離の前記ステップが、機械的分離を含むことを特徴とする方法。
- 請求項2に記載の方法において、前記単離されたフェリチンの濃縮が、前記可溶性画分を乾燥することを含むことを特徴とする方法。
- 請求項2に記載の方法において、前記単離されたフェリチンの濃縮が、限外濾過を含むことを特徴とする方法。
- 請求項1に記載の方法において、前記豆果がダイズを含むことを特徴とする方法。
- 豆果加工流からフェリチンを単離するための方法において、
(a)前記豆果加工流を可溶性および不溶性豆果の画分に分離するステップと、
(b)前記不溶性豆果の画分に中性の塩類緩衝液を加えて、不溶性豆果の溶液にするステップと、
(c)ステップ(b)の前記不溶性豆果の溶液を清澄化して、可溶性および不溶性豆果の溶液の画分に分けるステップと、
(d)ステップ(c)の前記清澄化された可溶性豆果の溶液を1種類または複数種類のグリコシダーゼ酵素で酵素処理するステップと、
(e)ステップ(d)の前記酵素処理され、清澄化された可溶性豆果の溶液を分別して、非フェリチン成分を除去するステップと、
(f)ステップ(e)の前記分別された可溶性豆果の溶液から単離されたフェリチンを濃縮するステップと
を含むことを特徴とする方法。
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