Clostridium thermoaceticum II Medium (DSMZ Medium 527)

[所属分类:培养基配方] [发布时间:2020-5-15] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Clostridium thermoaceticum II Medium
(DSMZ Medium 527)

培养基配方
Composition per 1010mL:
Solution B..................................................................600.0mL
Solution C..................................................................300.0mL
Solution A..................................................................100.0mL
Solution D...................................................................10.0mL
pH 6.9 ± 0.2 at 25°C
Solution A:
Composition per 100.0mL:
Glucose.......................................................................18.0g
Preparation of Solution A:  Add glucose to distilled/deionized wa-
ter and bring volume to 100.0mL. Mix thoroughly. Sparge with 100%
N2 gas mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool
to 25°C.
Solution B:
Composition per 600.0mL:
Yeast extract................................................................5.0g
Tryptone.....................................................................5.0g
Pyruvic acid.................................................................1.8g
(NH4)2SO4 ...................................................................1.0g
MgSO4 ·7H2O................................................................0.25g
Fe(NH4)2 (SO4)2 ·6H2O......................................................0.04g
Co(NO3)2 ·6H2O.............................................................0.03g
Na2WO4 ·2H2O...............................................................3.3mg
Na2MoO4 ·4H2O..............................................................2.4mg
ZnCl2 ......................................................................1.4mg
Resazurin ..................................................................1.0mg
Na2SeO3 ·5H2O..............................................................0.3mg
NiCl2 ·6H2O................................................................0.2mg
Preparation of Solution B:  Add components to distilled/deionized
water and bring volume to 600.0mL. Mix thoroughly. Sparge with
100% CO2 gas mixture. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 25°C.
Solution C:
Composition per 300.0mL:
NaHCO3 .....................................................................16.8g
K2HPO4 ......................................................................7.0g
KH2PO4 ......................................................................5.5g
Preparation of Solution C:  Add components to distilled/deionized
water and bring volume to 300.0mL. Mix thoroughly. Sparge with
100% CO2 gas mixture. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 25°C.
Solution D:
Composition per 10.0mL:
Cysteine solution ...........................................................5.0mL
Na2S·9H2O solution..........................................................5.0mL
Preparation of Solution D:  Combine 5.0mL cysteine soluiton and
5.0mL Na2S·9H2O solution. Mix thoroughly. Sparge with 100% N2 gas
mixture. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O..................................................................0.5g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Sparge with N2 .
Cysteine Solution:
Composition per 100.0mL:
L -Cysteine·HCl·H2O.......................................................5.0g
Preparation of Cysteine Solution:  Add  L -cysteine·HCl·H2O to
distilled/deionized water and bring volume to 100.0mL. Mix thorough-
ly. Sparge with 100% N2 .
Preparation of Medium: Aseptically and anaerobically combine
100.0mL solution A, 600.0mL solution B, 300.0mL solution C, and
10.0mL solution D. Aseptically and anaerobically distribute into sterile
tubes or flasks.
Use: For the cultivation of Moorella thermoacetica=Clostridium ther-
moaceticum.