JP6046636B2 - プロバイオティクス果実飲料 - Google Patents
プロバイオティクス果実飲料 Download PDFInfo
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- JP6046636B2 JP6046636B2 JP2013549838A JP2013549838A JP6046636B2 JP 6046636 B2 JP6046636 B2 JP 6046636B2 JP 2013549838 A JP2013549838 A JP 2013549838A JP 2013549838 A JP2013549838 A JP 2013549838A JP 6046636 B2 JP6046636 B2 JP 6046636B2
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- probiotic
- acid
- fruit
- casei
- bacterial strain
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Classifications
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- A—HUMAN NECESSITIES
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/02—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation containing fruit or vegetable juices
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
- A23L2/84—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter using microorganisms or biological material, e.g. enzymes
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Description
(a)少なくとも1種のプロバイオティクス細菌株を増殖中に酸適応化させ;
(b)工程(a)で得られた少なくとも1種の酸適応プロバイオティクス細菌株を、果実飲料中に接種し;
(c)任意により、プロバイオティクス果実飲料として包装する;
ことを含む、製造方法。
(i) 少なくとも1種の酸適応プロバイオティクス細菌株を、pH安定剤の非存在下、25℃から43℃の範囲で、好ましくは25℃から40℃の範囲で、初期pH値が約6.0〜約7.0である好適な培地中で増殖させ、ここで当該培地は、当該培地のpH値を約5.0〜約4.0まで到達させることが可能な組成を有し;
(ii) 少なくとも1種の酸適応プロバイオティクス細菌株を回収し;
(iii)任意により、少なくとも1種の酸適応プロバイオティクス細菌株を濃縮し;
及び、
(iv)任意により、少なくとも1種の酸適応プロバイオティクス細菌株を凍結又は凍結乾燥する。
(a) 少なくとも1種の酸適応プロバイオティクス細菌株を、pH安定剤の非存在下、25℃から43℃の範囲で、好ましくは25℃から40℃の範囲で、初期pH値が約6.0〜約7.0である好適な培地中で増殖させ、ここで当該培地は、当該培地のpH値を約5.0〜約4.0まで到達させることが可能な組成を有し;
(b) 少なくとも1種の酸適応プロバイオティクス細菌株を回収し;
(c)任意により、少なくとも1種の酸適応プロバイオティクス細菌株を濃縮し;
及び、
(d)任意により、少なくとも1種の酸適応プロバイオティクス細菌株を凍結又は凍結乾燥する。
本文中の「果実ジュース」なる用語は、新鮮な果実を、熱や溶媒を加えることなく、機械的に絞る又は潰すことにより自然に得られる液体のことをいう。「果実ジュース」は1種類の果物からなっていてもよく、又は1以上の果物の混合物からなっていてもよい。
MRS培地(ディフコ(Difco)、ref.288110)に冷凍保存されたL.カゼイ431登録商標(L. casei 431)株を1%(V/V)となるように接種した。37℃、終夜、pHが3.9以下となるまで培養し、使用時まで5℃で保存した。終夜培養した培養物を、MRS培地(ディフコ(Difco)) 4リットル(1%V/V)に接種し、37℃、終夜、pHが3.9以下となるまでインキュベートし、使用時まで5℃で保存した。当該予備接種工程は、発酵に十分な接種量が確保できるまで行った。400lの適切な培地が入ったバイオリアクターに、予備接種工程で得られた全量を、10℃で接種した。温度を35℃まで上昇させて発酵を開始し、発酵に続いて炭水化物の代謝による乳酸産生が起こるとともにpHが低下するのを監視した。当該pHの低下に伴い細胞は徐々に低pHへ適応する。酸適応細胞をpH5.0、4.5及び4.0の各点において遠心分離により回収した。発酵時間は回収の時点に応じ10〜17時間であった(図1)。
濃縮物の細胞全量は直ちに液体窒素(-196℃)で凍結し、使用時まで-55℃で保存した。
最大の生存細胞数を求めるため、以下のように最適な回収基準を決定した。バイオリアクターを予備接種により接種し、実施例1で述べたように発酵を開始した。予備接種(pH4.5相当)の11時間後から、30分毎にpHが4.0を下回るまで発酵物のサンプルを取出し、コロニー形成単位(CFU)数を決定することにより生存数を評価した(図2)。CFUはMRS寒天培地で約10倍希釈した後、37℃、3日間嫌気性条件でインキュベートすることにより決定した。
実施例1で述べた手順により生産されたL.カゼイ431登録商標(L. casei 431)株を、以下の組成による、異なる果実による飲料に対する接種に使用した。
31% イチゴピューレ
26% バナナピューレ
ブドウジュース、オレンジジュース、マンゴピューレ、モモピューレ及びブルーベリーピューレを加え合計100%とする。
32% パイナップルジュース
32% オレンジジュース
26% バナナピューレ
7% マンゴピューレ
3% ライムピューレ
16%の異なる赤色果実(ラズベリーピューレ、ブラックカラントジュース、ブルーベリーピューレ)
グレープジュース、イチゴピューレ及びバナナピューレを加え合計100%とする。
実施例1で説明された手順により生産されたL.カゼイ431登録商標(L. casei 431)菌を、100%オレンジジュース(pH3.8)に接種した。プロバイオティクスF-DVS培養物をペプトン生理食塩水で希釈し、オレンジジュースの包装単位に対し1% v/vとなるよう接種し、初期細胞数1.5×108CFU/mlを得た。当該ジュースに、pH4.0、4.5、5.0適応の、及び対照として非適応のL.カゼイ431登録商標(L. casei 431)を接種した。ジュースは遮光条件下8℃で42日間保存され、細胞数は通常どおり、サンプルをMRS寒天培地で約10倍希釈した後、37℃、3日間嫌気性条件でインキュベートすることにより決定した。
実施例1に従って産生されたL.カゼイ431登録商標(L. casei 431)菌を-50℃の冷凍ペレットとして保存し、通常の生存率試験を行った。細胞数は、解凍培養物をMRS寒天培地で5プレートを約10倍希釈後、37℃、3日間嫌気性条件でインキュベートすることにより決定した。図7に示すように、-50℃、6か月間保存した場合、非適応、及びpH4.0、4.5、5.0適応のL.カゼイ431登録商標(L. casei 431)のいずれを用いた場合も、細胞数は非常に安定していた。
-50℃、6か月間、冷凍ペレットとして保存後、100%オレンジジュース中、pH4.0適応L.カゼイ431登録商標(L. casei 431)の25バッチの生存率を決定した。プロバイオティクスF-DVS培養物をペプトン生理食塩水で希釈し、オレンジジュースのパック単位に対し1% v/vとなるよう接種し、初期細胞数1.5×108CFU/mlを得た。対照として、他の包装単位の100%オレンジジュースに対し非適応L.カゼイ431登録商標(L. casei 431)を加え、-50℃で6か月間保存した。
ジュースは遮光条件下8℃で42日間保存され、一定間隔でのL.カゼイ431登録商標(L. casei 431)サンプルの細胞数は、プレートをMRS寒天培地で約10倍希釈した後、37℃、3日間嫌気性条件でインキュベートすることにより決定した。
実施例1の方法により産生されたL.カゼイ431登録商標(L. casei 431)を用いて100%オレンジジュース(pH3.8)に接種を行った。プロバイオティクスF-DVS培養物をペプトン生理食塩水で希釈し、おおよその初期細胞数を確定した。果実ジュースの包装単位に対し1% v/vとなるよう接種したところ初期細胞数は1e08 CFU/mlであった。当該ジュースに、pH4.0に適応の、及び対照として非適応の「対照」L.カゼイ431登録商標(L. casei 431)を接種した。接種済のジュースは遮光条件下8℃で70日間保存し、一定間隔でサンプルを採取し、採取サンプルの細胞数は、プレートをMRS寒天培地で約10倍希釈した後、37℃、3日間嫌気性条件でインキュベートすることにより決定した。
欧州特許0113055号
米国特許出願2010/0086646号
PCT特許出願2010/132017号
Claims (7)
- プロバイオティクス果実飲料の製造方法であって、以下の工程:
(a)少なくとも1種のプロバイオティクス細菌株を増殖中に酸適応化させ、ここで前記工程(a)が以下の工程:
(i) ラクトバシラス(Lactobacillus)及びビフィドバクテリウム(Bifidobacterium)から成る群から選ばれる、前記少なくとも1種のプロバイオティクス細菌株を、pH安定剤の非存在下、25℃から43℃の範囲で、初期pH値が6.0〜7.0であり、そして少なくとも2%の糖及び0.4%の酵母抽出物を含む好適な培地中で増殖させ、ここで前記培地は、前記培地のpH値を5.0〜4.0まで到達させることが可能な組成を有し;
(ii) 前記少なくとも1種の酸適応プロバイオティクス細菌株を回収し;そして
(iii) 前記少なくとも1種の酸適応プロバイオティクス細菌株を濃縮する
ことを含み;
(b)前記工程(a)で得られた少なくとも1種の酸適応プロバイオティクス細菌株を、果実飲料中に接種し;そして
(c)任意により、プロバイオティクス果実飲料として包装する
ことを含む、製造方法。 - 前記工程(a)における酸適応化が、前記少なくとも1種のプロバイオティクス細菌株を、pH安定剤の非存在下、25℃から40℃の範囲で増殖させることを含む、請求項1の方法。
- さらに前記少なくとも1種の酸適応プロバイオティクス細菌株を凍結又は凍結乾燥することを含む、請求項1又は2の方法。
- 前記工程(b)において、前記少なくとも1種の酸適応プロバイオティクス細菌株の含有量(果実飲料1ml当たりの初期CFU値)が、果実飲料1ml当たり1×104〜1×1010CFUとなるように接種される、請求項1〜3の何れか1項の方法。
- 最低30日間、8℃で保存後の前記少なくとも1種の酸適応プロバイオティクス細菌株の生存率が、果実飲料1ml当たりの初期CFU値に対し少なくとも50%である、請求項1〜4の何れか1項の方法。
- 前記プロバイオティクス果実飲料のpH値が3.2〜4.2である、請求項1〜5の何れか1項の方法。
- 前記プロバイオティック果実飲料が、20重量%〜99.99重量%の果実ジュース及び/又は果実ピューレを含む、請求項1〜6の何れか1項の製造方法。
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