JP2010512755A - 生きたプロバイオティクスを含む乾燥食物製品 - Google Patents
生きたプロバイオティクスを含む乾燥食物製品 Download PDFInfo
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- JP2010512755A JP2010512755A JP2009541634A JP2009541634A JP2010512755A JP 2010512755 A JP2010512755 A JP 2010512755A JP 2009541634 A JP2009541634 A JP 2009541634A JP 2009541634 A JP2009541634 A JP 2009541634A JP 2010512755 A JP2010512755 A JP 2010512755A
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- probiotic
- protein
- dry
- food
- sugar
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Abstract
Description
本明細書で使用する場合、以下の各用語は、このセクションでそれと関連する意味を有する。
本明細書で述べるようにプロバイオティック物質を調製するため、単独またはいくつかの微生物の混合物を選択することができる。プロバイオティック微生物として、任意の微生物を選択することができる。好ましくは、ヒトまたは動物の健康および福祉に有益な作用を及ぼす微生物を使用する。
ここで、以下の実施例を参照して本開示の対象を説明する。これらの実施例は、例示目的のためだけに提供され、対象はこれらの実施例に限定されることなく、本明細書で提供する教示の結果として明白な全ての変形を包含する。
乾燥した安定なプロバイオティック物質の調製
基本処方
300gのトレハロース(Cargill Minneapolis, MN)を1000mlの水に加えて完全に溶解させた。標準的な家庭用ブレンダーを用いて激しく混合しながら大豆タンパク質単離物(100g、Fearn Natural Foods, Mequon, WI)および大豆加水分解物(20g、Sigma-Aldrich, St. Louis, MO)を加えた。次に、該スラリーにアルギン酸ナトリウム(20g)を混合し、室温に冷却した。次いで、滑らかで均一な濃厚ゲルが得られるまで、激しく混合中のスラリーにラクトバシラス・パラカゼイ(Lactobacillus paracasei)(発酵収穫物から直接得た100gの凍結濃縮物)を添加した。このヒドロゲルの組成を表1に示す。
5gのリン酸水素カルシウムを基本配合物(formula)に混合した後、5gのグルコノラクトンを混合し、次の4時間かけて室温でスラリーを硬化させた(固体ヒドロゲル)。この堅いゲルをチーズグラインダーで薄くて長いリーフ(leaf)にスライスした。薄いリーフをトレイ(13×10インチ(33×25cm))上に載せて凍結乾燥機(Virtis Advantage, Virtis, Gardiner, NY)内に置いた。凝縮器を−70℃に設定し、棚温度を+40℃に設定した。次に真空を開始し、外部の真空コントローラー(Thyr-Cont, Electronic, GmbH)を用いて約5000mTOR(約700Pa)で制御した。湿潤製品温度が下がり、約−5℃〜0℃で安定化した。製品温度が暖かくなり始めたとき(湿潤物質に接続された1対の温度センサーで測定)、100mTOR(13Pa)のフル減圧が確立されるまでチャンバーの大気圧を徐々に下げた。真空を高めるこの時間の始めから終わりまで、製品温度を慎重に−5℃〜+5℃に維持した。フル減圧を確立した24時間後に、乾燥製品を凍結乾燥機から取り出した。この乾燥プロトコル後のプロバイオティック物質の水分活性(Aw)は0.05だった(HygroPalm Awl, Rotonic Huntington, NYで測定した)。
小さいハートまたは星形の形を有するマフィンプレート中に上記基本処方スラリーを注ぎ、次の4時間かけて室温で硬化させた(固体ヒドロゲル)。この成形ヒドロゲルを含むプレートを凍結乾燥機内に置いて上述したように乾燥させた。
5gのCaCl2と300gのトレハロースを含む1000mlの浴(0〜5℃で維持)内に18G針を備えた注射器を用いて上記基本配合物を押し出し、または滴下した。スラリーを注入しながら、CaCl2浴を穏やかに撹拌した。マトリックスのストリングまたは液滴を30分間架橋させてから収集し、ペーパータオル上に吸い取らせた。ストリングまたは液滴をまず該物質の水分活性が0.5に下がるまで、従来型オーブン内で35℃にて乾燥させてから、約24時間の最終乾燥のため凍結乾燥機に物質を移した。次に、標準的なコーヒーグラインダーを用いて乾燥液滴またはストリング(Aw=0.06)を微細粉末に粉砕し、50〜250ミクロンのふるいに通してふるい分けした。
ラクトバシラス・アシドフィラス(発酵収穫物から直接得た20%の固体濃縮物)の濃縮ペースト1200mlに600gのトレハロース(Cargill Minneapolis, MN)を加えて完全に溶解させた。標準的な家庭用ミキサーを用いて激しく撹拌しながら、大豆タンパク質単離物(200g、Fearn Natural Foods, Mequon, WI)と大豆加水分解物(20g、Sigma-Aldrich, St. Louis, MO)を加えた。次に、アルギン酸ナトリウム(40g)をスラリーに混ぜ合わせて室温に冷ました。滑らかな濃厚かつ均一なゲルを得た後、5gの炭酸カルシウムを加えた。直後に、半湿潤フリー流動粉末が得られるまで激しく混合しながら、1000gの卵白(Sigma-Aldrich, St. Louis, MO)をゆっくり添加した。この顆粒化プロバイオティック粉末を従来型オーブン内で40℃にて2時間乾燥させた後、24時間真空乾燥させて乾燥(Aw<0.06)顆粒化マトリックス粒子を得た。50〜500ミクロンの一連のふるいに通して顆粒化粒子を特定の大きさの範囲にふるい分けすることができる。このプロバイオティック顆粒化マトリックス粒子の組成を表2に示す。
ラクトバシラス・ラムノサスマイクロマトリックス粒子を上述した通りに製造した。発酵槽から収穫して遠心分離した後の濃縮ラクトバシラス・ラムノサスのCFU数は5×1010/gペーストであり、固形分は24%だった。このプロバイオティクスを全ての保護成分と混合し、ヒドロゲルを形成し、ヒドロゲルを小糸にチョップし、凍結乾燥機内で24時間乾燥させ、粉砕し、50〜250ミクロンの粒径にふるい分けした後、CFU数は9.4×109/g乾燥マイクロマトリックスだった。これは、製造プロセス中のプロバイオティック活性の0.28対数損失に相当する(図1に示すように)。
上述した通りにラクトバシラス・ラムノサスプロバイオティックマイクロマトリックス粒子を調製して乾燥させた。乾燥マイクロマトリックス粒子を40℃および33%の相対湿度に設定した温度および湿度制御インキュベーター内に4週間置いた。10%のトレハロース溶液中でのみ、または本明細書で述べる保護混合物中で乾燥させた細菌の生存能力を1週間間隔で測定した。図2は、保護混合物がトレハロース単独で乾燥させた細菌の生存能力に有意な保護を与えたことを示す。
上述した通りにラクトバシラス・ラムノサス(LGG)、ラクトバシラス・アシドフィラス(LA5)またはラクトバシラス・パラカゼイ(LPC)を含有するマイクロマトリックス粒子(50〜250ミクロン)を調製して乾燥させた。次に、マイクロマトリックス粒子を人工胃液(充満した胃−12%の無脂肪スキムミルク、2%のグルコース、1%の酵母エキスおよび0.05%のシステイン;pH2;または空の胃−0.32%のペプシン、0.2%の塩化ナトリウム、pH1.2)に2時間曝露した。人工胃液への曝露前後に細菌の生存能力を記録した。図3は、人工胃液条件内のマイクロマトリックス物質中のプロバイオティック細菌の有意な保護を実証する。
ドラムタンブラー内で40℃の温度にて脂肪ベース湿分バリア(20%のホホバ油、70%のココアバターおよび10%の蜜ろうの混合物)でプロバイオティック物質をコーティングした。乾燥物質をドラムタンブラー内で撹拌しながら、噴霧ノズルを用いて湿分バリアを噴霧して均一なコーティングを保証した。コーティングの総量は、プロバイオティックフレークおよびトリートでは約20%(未コーティングプロバイオティック物質の)であり、プロバイオティックマイクロマトリックス粒子では40〜50%だった。
プロバイオティックペットフードの調製
市販のイヌ用ペットフードをまず従来型オーブン内で0.1の水分活性まで乾燥させてからドラムタンブラー内でプロバイオティックマイクロマトリックス粒子でコーティングした。ペレットをまず約5%の脂肪ベース湿分バリア(40%の鶏脂、40%のココアバターおよび20%の蜜ろうの混合物)を噴霧してからマイクロマトリックス粒子(通常、108CFU/gの用量を与える全ペットフードの0.1〜0.5%)と混合し、最後に脂肪ベース湿分バリアのさらなる被覆物を噴霧した。コーティングの総量は約15%(ペットフードの)だった。コーティング時間は約30分だった。
いくつかのプロバイオティック微生物を用いた魚飼料の調製
いくつかのプロバイオティクスの混合物を用いて魚用ペレット飼料を調製した。実施例1で述べた通りに、ラクトバシラス・ラムノサス、ラクトバシラス・アシドフィラスおよびビフィドバクテリウム・ラクティス(DSN20215)の混合物を含有するプロバイオティックマイクロマトリックス粒子を調製した。市販の魚飼料をまず従来型オーブン内で0.1の水分活性まで乾燥させてからドラムタンブラー内にてプロバイオティックマイクロマトリックス粒子でコーティングした。ペレットにまず約5%の脂肪ベース湿分バリア(40%の魚油、40%のココアバターおよび20%の蜜ろうの混合物)を噴霧してからマイクロマトリックス粒子(通常、107cfu/gの用量を与える全魚飼料の0.1〜0.5%)と混合し、最後に脂肪ベース湿分バリアのさらなる被覆物を噴霧した。コーティングの総量は(魚飼料の)約10%だった。
プロバイオティック「チップス」の調製(図4に示す通り)
実施例1で述べた通りに、ラクトバシラス・ラムノサスとラクトバシラス・アシドフィラスの混合物でプロバイオティックチップスを調製した。標準的な10〜20%の装填の代わりに約1〜2%のプロバイオティックペーストと共に基本配合物を加えた。5gのリン酸水素カルシウムを基本配合物に混合した後、5gのグルコノラクトンを混ぜ合わせ、このスラリーを1インチ(2.5cm)径の長いプラスチックチューブに流し込み、次の4時間かけて室温で硬化させた(固体ヒドロゲル)。堅い「ホットドッグ」形ゲルを薄円板にスライスし、トレイ(13×10インチ(33×25cm))上に載せて凍結乾燥機内に置き、実施例1で述べた通りに乾燥手順を施した。乾燥プロバイオティックチップス製品を凍結乾燥機から取り出し、1食当たり108CFUの有効用量を含むおいしいパリパリしたトリートを提供する小さいアルミニウム箔パックに窒素下で詰めた。図4は、本明細書で述べるラムノサスの1食当たり108CFUを含むプロバイオティックチップスの例を示す。
プロバイオティックシリアルの調製
市販の朝食用シリアルをまず従来型オーブン内で0.1の水分活性まで乾燥させた。標準的な10〜20%装填のプロバイオティクスを含むプロバイオティックチップスを製造し、さらに実施例1および4で述べた通りに湿分バリアでコーティングした。次に、シリアルを0.1〜0.5%のプロバイオティックチップスと混合した(108cfu/gの用量を与えるため)。
飲料の容器またはビンの蓋内のプロバイオティックマトリックス物質の代替送達形態
一般に、実施例1または実施例4の通りに製造したプロバイオティック物質の全ての形態が同様によく働き、この物質の利用と適用の高い汎用性を確証している。食用食品と共に小さいバッグまたは紙「スティック」包装で、あるいは任意の他の様式で、本明細書で述べた通りに製造したプロバイオティック物質を提供することができる。消費用液体食品と混合すべく、ヨーグルト容器または飲料のビンの蓋内で乾燥マイクロマトリックス粒子またはチップスを提供することもできる。
以下の参考文献を本明細書で引用する。
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Claims (23)
- 少なくとも1種の糖と、少なくとも1種のタンパク質と、少なくとも1種の多糖との乾燥マトリックス中に保存された生存可能なプロバイオティック細菌を含むプロバイオティック物質。
- 前記乾燥マトリックス中の前記糖が二糖である、請求項1に記載のプロバイオティック物質。
- 前記二糖が、スクロースおよびトレハロースから成る群より選択される、請求項2に記載のプロバイオティック物質。
- 前記乾燥マトリックス中の前記タンパク質が、卵白、大豆タンパク質単離物、大豆加水分解物またはそれらの組合せから成る群より選択される、請求項1に記載のプロバイオティック物質。
- 前記多糖が、架橋性ゲルであり、最も好ましくは異なる粘度のアルギン酸塩の組合せ、アガロース、ペクチン、およびキトサンから成る群より選択される、請求項1に記載のプロバイオティック物質。
- 前記乾燥マトリックスが、20%w/v〜60%w/vのトレハロース、2%w/v〜20%w/vの卵白または大豆タンパク質単離物および0.5%w/v〜10%w/vのアルギン酸塩を含む、請求項1に記載のプロバイオティック物質。
- 前記プロバイオティック微生物が、乾燥物質1グラム当たり105〜1014個の生存可能な微生物を含む、請求項1に記載のプロバイオティック物質。
- 追加の湿分バリアでコーティングされている、請求項1に記載のプロバイオティック物質。
- 撥水または防水特性を有する任意の食品グレード物質を含む、請求項8に記載の湿分バリア。
- 油ベース物質の混合物を含む、請求項8に記載の湿分バリア。
- 前記プロバイオティック微生物が、製造後、高温かつ高湿度条件への長期曝露後、および胃内曝露後にその最初の生物活性の大部分を保持している、請求項1に記載のプロバイオティック物質。
- 少なくとも1種の糖と、少なくとも1種のタンパク質と、少なくとも1種の多糖とを含む乾燥マトリックス中に保存された生存可能なプロバイオティック細菌を含む1種または数種のプロバイオティック物質を含む、ヒト用食物製品。
- 湿潤、半湿潤、または半乾燥形態である、請求項12に記載のヒト用食物製品。
- 粉末、粒状、ペレット、タブレット、カプセル、コロイド懸濁液または液体形態である、請求項12に記載のヒト用食物製品。
- バー、液体配合物、またはシリアルもしくは別の食物製品に添加される、請求項12に記載のヒト用食物製品。
- トリート、栄養補助食品添加物または医薬食品添加物の形態である、請求項12に記載のヒト用食物製品。
- 少なくとも1種の糖と、少なくとも1種のタンパク質と、少なくとも1種の多糖とを含む乾燥マトリックス中に保存された生存可能なプロバイオティック細菌を含む1種または数種のプロバイオティック物質を含む、動物飼料製品。
- 湿潤、半湿潤、または半乾燥形態である、請求項17に記載の動物飼料製品。
- 粉末、粒状、ペレット、タブレット、カプセル、コロイド懸濁液または液体形態である、請求項17に記載の動物飼料製品。
- バー、液体配合物、またはシリアルもしくは別の飼料製品に添加される、請求項17に記載の動物飼料製品。
- トリート、栄養補助食品添加物または医薬飼料添加物の形態である、請求項17に記載の動物飼料製品。
- 前記動物が、陸生または水生動物から選択される、請求項17に記載の動物飼料製品。
- プロバイオティック食物または飼料物質の製造方法であって、
a)プロバイオティック微生物の調製物を少なくとも1種の糖、少なくとも1種のタンパク質および少なくとも1種の多糖と混合して、所望の形状及び形態の滑らかで均一なゲルを得る工程;および
b)前記予め形成した固体ゲルを0.1未満の水分活性まで真空乾燥させる工程
を含む方法。
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Also Published As
Publication number | Publication date |
---|---|
JP5285617B2 (ja) | 2013-09-11 |
US9072310B2 (en) | 2015-07-07 |
CA2673120A1 (en) | 2008-06-26 |
US20140377409A1 (en) | 2014-12-25 |
US20100074994A1 (en) | 2010-03-25 |
US9480276B2 (en) | 2016-11-01 |
EP2117354A4 (en) | 2012-02-22 |
EP2117354B1 (en) | 2018-08-08 |
US8460726B2 (en) | 2013-06-11 |
US20130287896A1 (en) | 2013-10-31 |
WO2008076975A1 (en) | 2008-06-26 |
EP2117354A1 (en) | 2009-11-18 |
CA2673120C (en) | 2012-08-07 |
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