Mineral Medium with Dichloromethane (DSMZ Medium 465c)

[所属分类:培养基配方] [发布时间:2021-6-1] [发布人:网站管理员2] [阅读次数:] [返回]

Mineral Medium with Dichloromethane
(DSMZ Medium 465c)

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

Composition per liter:

Na 2 HPO 4 ·2H 2 O.............................................................................3.5g
KH 2 PO 4 .........................................................................................1.0g
(NH 4 ) 2 SO 4 ....................................................................................0.5g
MgCl 2 ·6H 2 O .................................................................................0.1g
Ca(NO 3 ) 2 ·4H 2 O ..........................................................................0.05g
Bromthymol blue.....................................................................50.0mg
Methanol..................................................................................10.0mL
Trace elements solution SL-4....................................................1.0mL
Dichloromethane..................................................................... variable
pH 7.25 ± 0.2 at 25°C
Trace Elements Solution SL-4:
Composition per liter:
EDTA............................................................................................0.5g
FeSO 4 ·7H 2 O..................................................................................0.2g
Trace elements solution SL-6................................................100.0mL
Trace Elements Solution SL-6:
Composition per liter:
H 3 BO 3 ...........................................................................................0.3g
CoCl 2 ·6H 2 O..................................................................................0.2g
ZnSO 4 ·7H 2 O.................................................................................0.1g
MnCl 2 ·4H 2 O ...............................................................................0.03g
Na 2 MoO 4 ·H 2 O............................................................................0.03g
NiCl 2 ·6H 2 O.................................................................................0.02g
CuCl 2· ·2H 2 O................................................................................0.01g
Preparation of Trace Elements Solution SL-6: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Adjust pH to 3.4.
Preparation of Trace Elements Solution SL-4: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Preparation of Medium:  Add components, except methanol and di-
chloromethane, to 1.0L distilled/deionized water. Adjust pH to 7.25. Au-
toclave for 15 min at 15 psi pressure–121°C. Cool to room temperature.
Inoculate the medium and supply methanol via incubation atmosphere
(up to 10.0mL methanol per liter medium). Subsequently feed small
amounts of dichloromethane (toxic to bacteria at 2 mmol per liter or
less) via incubation atmosphere. Readjust pH of the culture with sterile
1M NaOH as necessary
Use: For the cultivation of Methylophilus leisingeri, Rhizobium
radiobacter, and other dichloromethane-utilizing bacteria.