JP2020010725A - 微生物乾燥菌体の製造方法 - Google Patents
微生物乾燥菌体の製造方法 Download PDFInfo
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- JP2020010725A JP2020010725A JP2019195170A JP2019195170A JP2020010725A JP 2020010725 A JP2020010725 A JP 2020010725A JP 2019195170 A JP2019195170 A JP 2019195170A JP 2019195170 A JP2019195170 A JP 2019195170A JP 2020010725 A JP2020010725 A JP 2020010725A
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Classifications
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/04—Preserving or maintaining viable microorganisms
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- A—HUMAN NECESSITIES
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- 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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/125—Casei
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- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
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- Wood Science & Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
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Abstract
Description
ラクトバチルス・カゼイ乾燥菌体を生理食塩水(0.85%NaCl)で段階希釈した。希釈液1mLをBCP加プレートカウント寒天培地で混釈し、37℃、72時間培養した後に、生じたコロニーを計測し、希釈率を乗じ、ラクトバチルス・カゼイ生菌数とした。
ラクトバチルス・カゼイ乾燥菌体の調製(1):
ラクトバチルス・カゼイ(Lactobacillus casei)YIT 9029を、酵母エキス1%、リン酸一カリウム0.1%、リン酸二カリウム0.2%、乳糖2%を含む培地(pH7)で、 37℃、20時間嫌気的に培養した。培養終了後、液温が20℃以下になるまで冷却し、5規定の水酸化ナトリウム溶液でpHを7.0に調整した。この培養液を遠心分離(14000G、4℃、30分間)して得られた菌体を回収して、下記表1に示した組成で全量が1000mLとなるように分散媒を製造し、分散媒に菌体を2.0×1011cfu/mLとなるように懸濁した。菌体懸濁液をトレーに分注し、凍結乾燥法により乾燥菌体を調製した。なお、凍結乾燥は、凍結乾燥機 TAKARA FREEZE-DRYER TF20-80TANNS((株)宝エーテーエム)を用い、棚温−40℃で9時間、その後20℃で80時間の条件で行った。得られた乾燥菌体をミルで粉砕し、通常の大気組成下で脱気を行わずに、0.2gずつカプセル(ヒドロキシプロピルメチルセルロース製)に充填し、脱酸素剤(三菱ガス化学(株)製)と共にアルミパウチに入れ、35℃、4週間保存した後、ラクトバチルス・カゼイの生菌数を測定した。保存開始時の生菌数に対する保存後の生菌数の割合(生残率)を表1に示す。
ラクトバチルス・カゼイ乾燥菌体の調製(2):
分散媒の組成を下記表2に示したものに変更した以外は、実施例1と同様の方法で、ラクトバチルス・カゼイ(Lactobacillus casei)YIT 9029の凍結乾燥菌体を調製し、通常の大気組成下で脱気を行わずに、0.2gずつカプセル(ヒドロキシプロピルメチルセルロース製)に充填し、脱酸素剤(三菱ガス化学(株)製)と共にアルミパウチに入れ、35℃、4週間保存した後、ラクトバチルス・カゼイの生菌数を測定した。保存開始時の生菌数に対する保存後の生菌数の割合(生残率)を表2に示す。
ラクトバチルス・カゼイ乾燥菌体の調製(3):
実施例1の発明方法1と同様の方法でラクトバチルス・カゼイ(Lactobacillus casei)YIT 9029の凍結乾燥菌体を調製し、通常の大気組成下で脱気を行わずに、0.2gずつカプセル(ヒドロキシプロピルメチルセルロース製)に充填し、PTP包装(Press Through Package、アルミシートおよび塩化ビニル製)し、脱酸素剤(三菱ガス化学(株)製)と共にアルミパウチに入れ、表3に示す条件で保存した後、ラクトバチルス・カゼイの生菌数を測定した。保存開始時の生菌数に対する保存後の生菌数の割合(生残率)を表3に示す。なお、2℃1日→35℃2日→30℃6日→22℃6か月の保存条件は、凍結乾燥菌体を30〜35℃で輸送し、その後22℃で保管することを想定したものである。また、保存後の凍結乾燥菌体は、吸湿性が低く、塊が形成されておらず、外観、色調、臭いも保存開始時と変わらず、良好なものであった。
Claims (12)
- 微生物菌体を、保護剤としてのグルタミン酸もしくはその塩および二糖類、抗酸化剤およびキレート剤を含む分散媒中に懸濁した後、乾燥することを特徴とする微生物乾燥菌体の製造方法。
- 分散媒中の含有量が、保護剤が5〜30質量%、抗酸化剤が0.05〜5質量%およびキレート剤が0.5〜5質量%である請求項1に記載の微生物乾燥菌体の製造方法。
- 二糖類がトレハロースである請求項1に記載の微生物乾燥菌体の製造方法。
- 抗酸化剤が、アスコルビン酸またはその塩である請求項1〜3のいずれか1項に記載の微生物乾燥菌体の製造方法。
- キレート剤がクエン酸またはその塩である請求項1〜4のいずれか1項に記載の微生物乾燥菌体の製造方法。
- 微生物菌体がラクトバチルス属に属する微生物の菌体である請求項1〜5のいずれか1項に記載の微生物乾燥菌体の製造方法。
- ラクトバチルス属に属する微生物がラクトバチルス・カゼイ YIT 9029(FERM BP−1366)である請求項6に記載の微生物乾燥菌体の製造方法。
- 乾燥を、凍結乾燥法により行う請求項1〜7のいずれか1項に記載の微生物乾燥菌体の製造方法。
- 乾燥後に、微生物乾燥菌体を脱酸素剤と共にアルミパウチに充填するものである請求項1〜8の何れかに1項に記載の微生物乾燥菌体の製造方法。
- 分散媒の溶媒が水である請求項1〜9の何れか1項に記載の微生物乾燥菌体の製造方法。
- 保護剤としてのグルタミン酸もしくはその塩および二糖類、抗酸化剤およびキレート剤を含む分散媒を用いて微生物菌体を乾燥させた微生物乾燥菌体。
- 保護剤としてのグルタミン酸もしくはその塩および二糖類、抗酸化剤およびキレート剤を含む微生物菌体の乾燥用分散媒。
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