JP6435462B2 - 微生物の培養方法 - Google Patents
微生物の培養方法 Download PDFInfo
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- JP6435462B2 JP6435462B2 JP2014069424A JP2014069424A JP6435462B2 JP 6435462 B2 JP6435462 B2 JP 6435462B2 JP 2014069424 A JP2014069424 A JP 2014069424A JP 2014069424 A JP2014069424 A JP 2014069424A JP 6435462 B2 JP6435462 B2 JP 6435462B2
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- microorganism
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- antibiotic
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- 229960003250 telithromycin Drugs 0.000 description 1
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Description
<1> 微生物群から特定の微生物を分離して培養するための方法であって、
前記微生物群が抗生物質非存在下にて対数増殖する期間の一部又は全部において、前記微生物群を抗生物質存在下にて液体培養する工程と、
前記工程にて液体培養した微生物群を、前記抗生物質非存在下にて分離培養する工程とを含み、
かつ、前記抗生物質が、核酸合成阻害剤、タンパク質合成阻害剤、細胞壁合成阻害剤及び葉酸合成阻害剤からなる群から選択される少なくとも2種の抗生物質の組み合わせである方法。
<2> 前記抗生物質が、核酸合成阻害剤、タンパク質合成阻害剤、細胞壁合成阻害剤及び葉酸合成阻害剤の組み合わせである、<1>に記載の方法。
<3> 前記抗生物質が、ノルフロキサシン、ホスホマイシン、テトラサイクリン及びトリメトプリムの組み合わせである、<1>に記載の方法。
<4> 前記分離培養が塗抹培養である、<1>〜<3>のうちのいずれか1に記載の方法。
<5> 前記微生物群が腸内常在菌群である、<1>〜<4>のうちのいずれか1に記載の方法。
<6> 特定の微生物を生産するための方法であって、
<1>〜<5>のうちのいずれか1に記載の方法によって、微生物群から前記特定の微生物を分離して培養する工程と、
前記工程にて分離培養した前記特定の微生物を回収する工程と
を含む方法。
<7> 微生物群から新規な微生物をスクリーニングするための方法であって、
<1>〜<5>のうちのいずれか1に記載の方法によって、微生物群から前記特定の微生物を分離して培養する工程と、
前記工程にて分離培養した前記特定の微生物の特性を分析する工程と、
前記工程にて得られた特性と公知の微生物の特性とを比較し、一致しない場合には、前記特定の微生物は新規な微生物であると判定する工程と
を含む方法。
本発明の微生物の培養方法は、微生物群から特定の微生物を分離して培養するための方法であって、
前記微生物群が抗生物質非存在下にて対数増殖する期間の一部又は全部において、前記微生物群を抗生物質存在下にて液体培養する工程と、
前記工程にて液体培養した微生物群を、前記抗生物質非存在下にて分離培養する工程とを含み、
かつ、前記抗生物質が、核酸合成阻害剤、タンパク質合成阻害剤、細胞壁合成阻害剤及び葉酸合成阻害剤からなる群から選択される少なくとも2種の抗生物質の組み合わせである方法である。
DNAジャイレース阻害作用等の核酸合成を阻害する作用を有する抗生物質であり、例えば、ノルフロキサシン、オフロキサシン、エノキサシン、塩酸シプロフロキサシン、塩酸ロメフロキサシン、レボフロキサシン、ガレノキサシン、フレロキサシン、シタフロキサシン、トスフロキサシントシル酸塩、スパルフロキサシン、ガチフロキサシン、モキシフロキサシン等のキノロン系抗生物質、リファンピシン等のリファンピシン系抗生物質が挙げられる。
タンパク質の合成を阻害する作用を有する抗生物質であり、例えば、テトラサイクリン、ドキシサイクリン、ミノサイクリン、オキシテトラサイクリン、デメクロサイクリン、クロルテトラサイクリン等のテトラサイクリン系抗生物質、ストレプトマイシン、カナマイシン、ゲンタマイシン、トブラマイシン、アミカシン、ジベカシン、アルべカシン等のアミノグリコシド系抗生物質、エリスロマイシン、クラリスロマイシン、ロキシスロマイシン、アジスロマイシン、ジョサマイシン、ロキタマイシン、キタサマイシン、アセチルスピラマイシン、テリスロマイシン、リンコマイシン、クリンダマイシン、
タクロリムス等のマクロライド・ケトライド系抗生物質、クロラムフェニコール等のクロラムフェニコール系抗生物質が挙げられる。
細胞壁の合成を阻害する作用を有する抗生物質であり、例えば、ホスホマイシン等のホスホマイシン系抗生物質、ペニシリン、ベンジルペニシリン、フェノキシメチルペニシリン、ベンジルペニシリンベンザチン、メチシリン
、オキサシリン、クロキサシリン、ジクロキサシリン、アンピシリン、アモキシシリン、バカンピシリン、タランピシリン、カルベニシリン、チカルシリン、メズロシリン、テモシリン、アパラシリン、ピペラシリン、スルタミシリン、アゾシリン、ピブメシリナム等のペニシリン系抗生物質、セファゾリン、セファロチン、セファピリン、セファレキシン、セファラジン、セファドロキシル、セフマンドール、セフロキシム、セフォニシド、セフォラニド、セファクロル、セフプロジル、セフポドキシム、ロラカルベフ、セフトリアキソン、セフォタキシム、セフチゾキシム、セフタジジム、セフォペラゾン、セフスロジン、セフチブテン、セフィキシム、セフェタメット、セフジトレン
ピボキシル、セフェピム、セフピロム、セフォキシチン、セフォテタン、セフメタゾール、セフブペラゾン、セフミノクス、ラタモキセフ、フロモキセフ等のセフェム・オキサセフェム系抗生物質、ファロペネム、イミペネム、メロペネム等のペネム・カルバペネム系抗生物質、アズトレオナム、カルモナム等のモノバクタム系抗生物質が挙げられる。
葉酸の合成(代謝)を阻害する作用を有する抗生物質であり、例えば、トリメトプリム等のトリメトプリム系抗生物質、プロントジル、スルファモノメトキシン、スルファジアジン、スルファジメトキシン、スルファセタミド、スルファドキシン、スルファニルアミド、スルフィソミジン、スルフィソキサゾール、スルファメトキサゾール、スルファジミジン、スルファメラジン、スルファキノキサリン等のスルホンアミド系抗生物質が挙げられる。
第一の寒天培地と第二の寒天培地を区画する工程であって、両寒天培地で培養される各微生物を通過させないが微生物が産出する物質は通過させるフィルターで区画する工程と、
前述の液体培養した微生物群を第一の寒天培地で培養する工程とを含む、
特定の微生物を共生微生物と共培養するための方法。
前述の通り、公知の分離培養(例えば、特定の抗生物質を添加した寒天培地を用いた塗抹培養)においては培養することのできない非優占的な微生物を増殖させることができる。従って、本発明は、非優占的な微生物(特定の微生物)を生産するための下記方法を提供することもできる。
前記工程にて分離培養した前記特定の微生物を回収する工程と
を含む方法。
前述の通り、公知の分離培養においては培養することのできない非優占的な微生物を培養することができる。さらに後述の実施例において示す通り、このようして得られた非優占的な微生物の中から、新規な微生物を同定することもできる。従って、本発明は、微生物群から新規な微生物をスクリーニングするための下記方法を提供することもできる。
前記工程にて分離培養した前記特定の微生物の特性を分析する工程と、
前記工程にて得られた特性と公知の微生物の特性とを比較し、一致しない場合には、前記特定の微生物は新規な微生物であると判定する工程と
を含む方法。
糞便は、各被験者(ヒト)から採取した後、速やかに冷蔵にて保管したものを使用した。そして、各新鮮糞便を希釈液Aにて102倍希釈することにより、腸内常在菌混合懸濁液として調製した。希釈液Aの組成を表1に示す。
腸内常在菌混合懸濁液を液体培養するために用いたNT液体培地、及び、塗抹培養(分離培養)等するために用いたNT寒天培地の組成を表2に示す。また、NT液体培地の成分である「5×ソルト」の組成を表3に示す。
前記NT液体培地には、表4に記載の抗生物質を、単独又は組み合わせて添加して用いた。
Claims (6)
- 微生物群から特定の微生物を分離して培養するための方法であって、
前記微生物群が抗生物質非存在下にて対数増殖する期間の一部又は全部において、前記微生物群を抗生物質存在下にて液体培養する工程と、
前記工程にて液体培養した微生物群を、前記抗生物質非存在下にて分離培養する工程とを含み、
かつ、前記抗生物質が、核酸合成阻害剤、タンパク質合成阻害剤、細胞壁合成阻害剤及び葉酸合成阻害剤の組み合わせである方法。 - 前記抗生物質が、ノルフロキサシン、ホスホマイシン、テトラサイクリン及びトリメトプリムの組み合わせである、請求項1に記載の方法。
- 前記分離培養が塗抹培養である、請求項1又は2に記載の方法。
- 前記微生物群が腸内常在菌群である、請求項1〜3のうちのいずれか1項に記載の方法。
- 特定の微生物を生産するための方法であって、
請求項1〜4のうちのいずれか1項に記載の方法によって、微生物群から前記特定の微生物を分離して培養する工程と、
前記工程にて分離培養した前記特定の微生物を回収する工程と
を含む方法。 - 微生物群から新規な微生物をスクリーニングするための方法であって、
請求項1〜4のうちのいずれか1項に記載の方法によって、微生物群から前記特定の微生物を分離して培養する工程と、
前記工程にて分離培養した前記特定の微生物の特性を分析する工程と、
前記工程にて得られた特性と公知の微生物の特性とを比較し、一致しない場合には、前記特定の微生物は新規な微生物であると判定する工程と
を含む方法。
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