Medium with Polyhydroxybutyric Acid as Carbon Source (DSMZ Medium 474)

[所属分类:培养基配方] [发布时间:2021-5-20] [发布人:网站管理员2] [阅读次数:] [返回]

Medium with Polyhydroxybutyric Acid
as Carbon Source
(DSMZ Medium 474)

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

Composition per liter:

Agar............................................................................................16.0g
Na 2 HPO 4 .....................................................................................2.44g
KH 2 PO 4 .......................................................................................1.52g
(NH 4 ) 2 SO 4 ....................................................................................0.5g
MgSO 4 ·7H 2 O................................................................................0.2g
CaCl 2 ·2H 2 O ................................................................................0.05g
PHB solution............................................................................66.0mL
Trace elements solution SL-4..................................................10.0mL
pH 6.9 ± 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.

PHB Solution:
Composition per 100.0mL:
Poly-ß-hydroxybutyric acid (PHB)...............................................3.0g
Preparation of PHB Solution: Add poly-ß-hydroxybutyric acid
(PHB) to 100.0mL distilled/deionized water. Stir overnight. Sonicate
until a white homogenous suspension is obtained. Autoclave for 5 min
at 15 psi pressure–121°C. Cool to room temperature.
Preparation of Medium: Add components, except PHB solution, to
1.0L distilled/deionized water. Adjust pH to 6.9. Autoclave for 15 min at
15 psi pressure–121°C. Cool to 50°C. Use 500.0mL to prepare bottom
layer of a double agar plate by aseptically pouring 10.0mL amounts
into sterile Petri dishes. Allow to solidify. Warm the PHB solution to
50°C. Aseptically add 33 mL of sterile PHB solution to the remaining
500.0mL of the medium. Mix thoroughly. Pour the PHB containing
agar as a top layer over the solidified base agar.
Use: For the cultivation of Comamonas testosteroni.