Methanogenium olentangyi Medium (DSMZ Medium 287)

[所属分类:培养基配方] [发布时间:2021-5-25] [发布人:网站管理员2] [阅读次数:] [返回]

Methanogenium olentangyi Medium
(DSMZ Medium 287)

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per liter:

NaHCO 3 ........................................................................................5.0g
NH 4 Cl ...........................................................................................2.7g
Na-acetate.....................................................................................2.5g
NaCl............................................................................................0.61g
K 2 HPO 4 .........................................................................................0.3g
KH 2 PO 4 .........................................................................................0.3g
(NH 4 ) 2 SO 4 ....................................................................................0.3g
CaCl 2 ·H 2 O ..................................................................................0.14g
MgSO 4 ·7H 2 O..............................................................................0.13g
Resazurin ...................................................................................1.0mg
Trace elements solution...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
L -Cysteine-HCl·H 2 O solution..................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
pH 6.9 ± 0.2 at 25°C
Trace Elements Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O................................................................................0.5g
CoSO 4 ·7H 2 O...............................................................................0.18g
ZnSO 4 ·7H 2 O...............................................................................0.18g
FeSO 4 ·7H 2 O..................................................................................0.1g
CaCl 2 ·2H 2 O...................................................................................0.1g
NiCl 2 ·6H 2 O...............................................................................0.025g
KAl(SO 4 ) 2 ·12H 2 O.......................................................................0.02g
CuSO 4 ·5H 2 O...............................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·4H 2 O ..........................................................................0.01g
Na 2 SeO 3 ·5H 2 O...........................................................................0.3mg
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with
KOH. Add remaining components. Add distilled/deionized water to
1.0L. Adjust pH to 7.0 with KOH.
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl........................................................................10.0mg
Thiamine-HCl·2H 2 O..................................................................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. Filter ster-
ilize.
L -Cysteine-HCl·H 2 O Solution:
Composition per 10.0mL:
L -Cysteine-HCl·H 2 O.....................................................................0.3g
Preparation of  L -Cysteine-HCl·H 2 O Solution:  Add  L -cyste-
ine·HCl·H 2 O to distilled/deionized water and bring volume to 10.0mL.
Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi
pressure–121°C. Cool to room temperature.
Na 2 S·9H 2 O Solution :
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.3g
Preparation of Na 2 S·9H 2 O Solution: Add Na 2 S·9H 2 O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
Preparation of Medium: Prepare and dispense medium under 80%
H 2 + 20% CO 2 gas atmosphere. Add components, except  L -cysteine-
HCl·H 2 O solution, Na 2 S·9H 2 O solution, and vitamin solution, to dis-
tilled/deionized water and bring volume to 970.0mL. Mix thoroughly.
Adjust pH to 6.9. Sparge with 80% H 2 + 20% CO 2 . Autoclave for 15 min
at 15 psi pressure–121°C. Aseptically and anaerobically add 10.0mL  L -
cysteine-HCl·H 2 O solution, 10.0mL Na 2 S·9H 2 O solution, and 10.0mL vi-
tamin solution. Mix thoroughly. Aseptically and anaerobically distribute
into sterile tubes or bottles. After inoculation, flush and repressurize the
gas head space of culture bottles with sterile 80% H 2 + 20% CO 2 to 1 bar
overpressure.
Use:  For the cultivation of Methanoculleus olentangyi=Methanoge-
nium olentangyi.