Clostridium thermoaceticum Medium (TYE-CO) (DSMZ Medium 316)

[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 Medium
(TYE-CO)
(DSMZ Medium 316)

培养基配方
Composition per liter:
Trypticase™................................................................10.0g
Yeast extract...............................................................3.0g
Na2HPO4 ·12H2O.............................................................2.8g
NH4Cl ......................................................................1.0g
KH2PO4 .....................................................................0.3g
MgCl2 ·6H2O................................................................0.2g
FeSO4 ·7H2O...............................................................1.0mg
Resazurin .................................................................1.0mg
Trace elements solution ..................................................10.0mL
Na2S·9H2O solution.......................................................10.0mL
Vitamin solution...........................................................5.0mL
pH 7.0 ± 0.2 at 25°C
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl............................................................10.0mg
Thiamine-HCl·2H2O.........................................................5.0mg
Riboflavin ................................................................5.0mg
Nicotinic acid.............................................................5.0mg
D -Ca-pantothenate.........................................................5.0mg
p-Aminobenzoic acid........................................................5.0mg
Lipoic acid ...............................................................5.0mg
Biotin ....................................................................2.0mg
Folic acid.................................................................2.0mg
Vitamin B 12 ..............................................................0.1mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Sparge
with 80% H2 + 20% CO2 . Filter sterilize.
Trace Elements Solution:
Composition per liter:
MgSO4 ·7H2O.................................................................3.0g
Nitrilotriacetic acid........................................................1.5g
NaCl.........................................................................1.0g
MnSO4 ·2H2O.................................................................0.5g
CoSO4 ·7H2O................................................................0.18g
ZnSO4 ·7H2O................................................................0.18g
CaCl2 ·2H2O.................................................................0.1g
FeSO4 ·7H2O.................................................................0.1g
NiCl2 ·6H2O...............................................................0.025g
KAl(SO4)2 ·12H2O...........................................................0.02g
H3BO3 ......................................................................0.01g
Na2MoO4 ·4H2O..............................................................0.01g
CuSO4 ·5H2O................................................................0.01g
Na2SeO3 ·5H2O..............................................................0.3mg
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L. Mix thoroughly.
Na2S·9H2O Solution :
Composition per 10.0mL:
Na2S·9H2O......................................................................0.6g
Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
Preparation of Medium: Add components, except vitamin solu-
tion, and Na2S·9H2O solution, to distilled/deionized water and bring
volume to 985.0mL. Mix thoroughly. Sparge with 100% CO2 . Auto-
clave for 15 min at 15 psi pressure–121°C. Cool to 25°C while sparg-
ing with 100% CO2 . Aseptically and anaerobically add 10.0mL
vitamin solution, and 10.0mL of sterile Na2S·9H2O solution. Mix thor-
oughly. Aseptically and anaerobically distribute into sterile tubes or
flasks.
Use:  For the cultivation of Moorella thermoacetica=Clostridium ther-
moaceticum.