Methanomicrobium mobile Medium (DSMZ Medium 161)

[所属分类:培养基配方] [发布时间:2021-5-26] [发布人:网站管理员2] [阅读次数:] [返回]

Methanomicrobium mobile Medium
(DSMZ Medium 161)

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 ........................................................................................2.0g
Yeast extract..................................................................................1.0g
Trypticase™..................................................................................1.0g
NaCl..............................................................................................0.6g
Cysteine-HCl·H 2 O........................................................................0.5g
Na 2 S·9H 2 O....................................................................................0.5g
K 2 HPO 4 .........................................................................................0.3g
KH 2 PO 4 .........................................................................................0.3g
(NH 4 ) 2 SO 4 .....................................................................................0.3g
MgSO 4 ·7H 2 O..............................................................................0.13g
CaCl 2 ·2H 2 O...............................................................................0.008g
FeSO 4 ·7H 2 O..............................................................................0.002g
Resazurin ..................................................................................0.001g
Rumen fluid, clarified............................................................300.0mL
Fatty acid mixture....................................................................20.0mL
Trace elements solution ...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
pH 6.5 ± 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
CaCl 2 ·2H 2 O...................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
NiCl 2 ·6H 2 O...............................................................................0.025g
KAl(SO 4 ) 2 ·12H 2 O.......................................................................0.02g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·4H 2 O ..........................................................................0.01g
CuSO 4 ·5H 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. Dissolve by adjusting pH
to 6.5 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L. Mix thoroughly.
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. Sparge
with 80% H 2 + 20% CO 2 . Filter sterilize.
Fatty Acid Mixture:
Composition per 20.0mL:
Valeric acid ...................................................................................0.5g
Isovaleric acid...............................................................................0.5g
α-Methylbutyric acid....................................................................0.5g
Isobutyric acid...............................................................................0.5g
Distilled water..........................................................................20.0mL
Preparation of Fatty Acid Mixture:  Add components to 20.0mL
distilled/deionized water. Mix thoroughly.
Preparation of Medium: Prepare and dispense medium under an
oxygen-free 80% H 2 + 20% CO 2  gas mixture. Add components, except
vitamin solution, to 990.0mL distilled/deionized water. Mix thorough-
ly. Sparge with 80% H 2 + 20% CO 2 . Adjust pH to 6.5 with concentrat-
ed NaOH. Autoclave for 15 min at 15 psi pressure–121°C. Cool to
25°C while sparging with 80% H 2 + 20% CO 2 . Aseptically and anaer-
obically add 10.0mL sterile vitamin solution. Mix thoroughly. Asepti-
cally and anaerobically distribute into sterile tubes or flasks.
Alternately the medium can be distributed to tubes under anaerobic
conditions and autoclaved in tubes prior to addition of vitamin solu-
tion. Appropriate amounts of the vitamin solution can then be added to
each tube by syringes. After inoculation, pressurize culture vessels to
2 bar 80% H 2 + 20% CO 2 overpressure.
Use:  For the cultivation of Methanomicrobium mobile.