JP4720975B2 - エグジゴバクテリウム(Exiguobacterium)属の細菌株、培養方法および使用 - Google Patents
エグジゴバクテリウム(Exiguobacterium)属の細菌株、培養方法および使用 Download PDFInfo
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- JP4720975B2 JP4720975B2 JP2004560550A JP2004560550A JP4720975B2 JP 4720975 B2 JP4720975 B2 JP 4720975B2 JP 2004560550 A JP2004560550 A JP 2004560550A JP 2004560550 A JP2004560550 A JP 2004560550A JP 4720975 B2 JP4720975 B2 JP 4720975B2
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- PLXBWHJQWKZRKG-UHFFFAOYSA-N Resazurin Chemical compound C1=CC(=O)C=C2OC3=CC(O)=CC=C3[N+]([O-])=C21 PLXBWHJQWKZRKG-UHFFFAOYSA-N 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-N Sulfurous acid Chemical compound OS(O)=O LSNNMFCWUKXFEE-UHFFFAOYSA-N 0.000 description 1
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Classifications
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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
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
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- 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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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/56—Lactic acid
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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Description
しかしながら、この目録において与えられた種は、本発明の株とは全く異なっている。
非特許文献2において出版された論文には、Farrowらが、種々の種間の比較研究を記載する。エグジゴバクテリウム(Exiguobacterium)属が言及されているが、種に関しては特定化されていない。
非特許文献3におけるFruhlingらによる論文は、海水(pond water)から単離されたExiguobacterium undae sp.nov.およびExiguobacterium antarcticum sp.nov.の株を報告する。
与えられたExiguobacterium属のこれらの株および他の種は、本発明の株とははっきり区別される系統発生論的な位置を有する。
EMBLデータベースドキュメントのアクセション番号D55730号は、Exiguobacterium acetylicumクローンの16S rRNAを与え、これもまた、本発明ものとは異なる種に対応する。
これらの文書はそれぞれExiguobacterium auriantiacum(NDCDO)およびExiguobacterium undaeの16S rRNAの配列を報告する。データベースドキュメントに関連して上記に与えられた説明も利用する。
本発明者らによる得られたサンプルの研究によって、種々の工業分野において非常に興味深い性質を示すエグジゴバクテリウムの新種を単離するに至った。
−その成長および所望の代謝物の生成に適した条件下に、上記規定の細菌株を培養する工程と、
−生成された代謝物を回収し、次いで、所望の代謝物を単離し、それを精製する工程と
を包含する。
単離は深海水熱系のサンプルを用いて行われた。
1gのNH4Cl、0.3gのKH2PO4、0.3gのK2HPO4、25gのNaCl、0.2gのCaCl2、0.1gのKCl、3gのMgCl2、0.5gのCH3COONa、0.5gのシステイン−HCl、0.1gの酵母菌抽出物(Difco Laboratories)、10mlのBalchミネラル溶液(1)および1mgのレザズリンを(蒸留水1リットルあたりに)含む基本培地(basic medium)が用いられる。pHは10M KOHを用いて7.3に調整され、培地は窒素流下に煮沸され、周囲温度に冷却される。
KH2PO4 6 g
(NH4)2SO4 6 g
NaCl 12 g
MgSO4・7H2O 2.6 g
CaCl2・2H2O 0.16 g
蒸留H2O十分量 1000 ml
Balch微量元素溶液
ニトリロ酢酸 1.5 g
MnSO4・2H2O 0.5 g
MgSo4・7H2O 3 g
NaCl 1 g
FeSO4・7H2O 0.1 g
CoCl2・6H2O 0.1 g
CaCl2・2H2O 0.1 g
ZnCl2 0.1 g
CuSO4・5H2O 0.01 g
AlK(SO4)2 0.01 g
H3BO3 0.01 g
Na2MoO4 0.01 g
蒸留H2O十分量 1000ml
培地のpHは10M KOHを用いて7.3に調整される。
株10Cは、非胞子形成性、通性嫌気性、グラム陽性の棹形態の細菌であり、45℃、pH7および0〜2%のNaClで至適に生育する。
糖の発酵は、L(+)ラクテートを必然的に生成する(発酵されたグルコースのモルあたりラクテートの約2モル)。適切な生育条件下に、ホルメート(formate)、アセテートおよびエタノールの生成物が観察される。
株10CはDNA中の(グアニン+シトシン)含有率が50.4mol%であることを特徴とする。
(1.糖の発酵によるバイオマスの生成)
方法は、非再生培地(non-renewed medium)で行われる。
酵母菌抽出物/プロテインヒドロリアーゼ(protein hydrolyzate)
計算値
NH4Cl 1g/l
NaCl 0.5g/l
KH2PO4 0.3g/l
K2HPO4 0.3g/l
MgCl2・6H2O 0.2g/l
KCl 0.1g/l
CaCl2・2H2O 0.1g/l
糖および酵母菌抽出物の濃度は得ることが望まれる細胞の濃度に依存する。
複数のバッチを一緒に連結することによって連続して形成されたバッチ方式で研究が行われた。これは、「供給−採取(feed-harvest)」または「繰り返しバッチ(repeat batch)」システムであり、これは、発酵槽を新しい培地で充填する工程、次いで、細菌をバッチ培養する工程、次いで、発酵マスト(must)を空にして、次のバッチのインキュベーションのためにタンクを開始時の状態にし、再度充填等をする工程の3工程を一緒に連続的に連結することによって概略的に表され得る。
他の実験では、上記のデンプン基質および緩衝された培地(ただし、リットルあたり10gのデンプンを有する)を用いて、95%超のL(+)ラクテートおよび痕跡量のホルメートを与えるデンプンの転換が得られる。
Claims (5)
- エグジゴバクテリウム属であり、かつ、DNA配列の少なくとも一部が、2002年12月5日に、第I−2962号のもとで、Collection Nationale de Cultures de Microorganismes(C.N.C.M)(英語名:French national collection of microorganism cultures)に寄託された株のゲノムDNAまたはプラスミドDNAとハイブリダイズするDNA配列を有し、これらの株は、40〜50℃程度の温度、5.4〜9.15のpHで成育する熱抵抗性、糖分解性およびペプチド分解性であり、かつL(+)ラクテートを生成することが可能であり、40〜50℃程度の温度、5.4〜9.15のpHの生育特性を有し、45℃の至適成育温度を有し、DNAにおけるグアニンおよびシトシンを合わせた含有率が約50mol%であることを特徴とし、さらに、16S rRNAについてのDNAの配列番号1:
- 2002年12月5日に番号I−2962のもとでC.N.C.Mに寄託された、請求項1に記載の細菌株。
- 請求項1または2に記載の細菌株を培養する方法であって、
該方法は、通性嫌気性条件下に、37℃で5.4〜9.15のpHで、この株によってエネルギー源として用いられる糖を含む基本培地において行われることを特徴とする、方法。 - 食物発酵処理における請求項1または2に記載の細菌株の使用。
- L(+)ラクテート等の代謝物を生成する方法であって、
請求項1または2に記載の細菌株を、その成長および所望の代謝物の生成に適した条件下に培養する工程と、
生成された代謝物を回収し、所望の代謝物を単離し、そして、それを精製する工程と
を包含することを特徴とする方法。
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FR0215865A FR2848564B1 (fr) | 2002-12-13 | 2002-12-13 | Souches bacteriennes du genre exiguobacterium, procede de culture et applications |
FR0215865 | 2002-12-13 | ||
PCT/FR2003/003665 WO2004055173A1 (fr) | 2002-12-13 | 2003-12-10 | Souches bacteriennes du genre exiguobacterium, procede de culture et applications |
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AU (1) | AU2003296812A1 (ja) |
CA (1) | CA2509637A1 (ja) |
DE (1) | DE60318484T2 (ja) |
FR (1) | FR2848564B1 (ja) |
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CN112195121B (zh) * | 2019-09-30 | 2021-10-12 | 中国农业科学院生物技术研究所 | 一种耐高温微小杆菌及其应用 |
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WO2002006452A2 (en) * | 2000-07-18 | 2002-01-24 | National Research Council Of Canada | Cloning, sequencing and expression of a comamonas cyclopentanone 1,2-monooxygenase-encoding gene in escherichia coli |
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HK1084972A1 (en) | 2006-08-11 |
DE60318484D1 (de) | 2008-02-14 |
AU2003296812A1 (en) | 2004-07-09 |
CA2509637A1 (fr) | 2004-07-01 |
ATE382679T1 (de) | 2008-01-15 |
FR2848564B1 (fr) | 2006-12-01 |
EP1570050A1 (fr) | 2005-09-07 |
FR2848564A1 (fr) | 2004-06-18 |
DE60318484T2 (de) | 2008-12-24 |
WO2004055173A1 (fr) | 2004-07-01 |
JP2006509522A (ja) | 2006-03-23 |
US20070009553A1 (en) | 2007-01-11 |
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