DSMZ 474 . MEDIUM WITH POLYHYDROXYBUTYRIC ACID AS CARBON SOURCE

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Culture medium recipe

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Medium for Comamonas testosteroni DSM 6781

This strain uses the standard medium without modifications.


Growth conditions: 30 °C
Equipment needed: Autoclave
1 Prepare a suspension of 3.0 g poly-ß-hydroxybutyric acid (PHB) in 100 ml distilled water, stir over night and sonicate until a white homogenous suspension is obtained. Autoclave for 5 minutes at 121°C. Prepare double layer agar plates. First layer: medium 457 with 16.0 g agar per l.
Compound Amount Unit Conc. [g/L] Conc. [mM]
Medium 457 1000.0 ml - -
Agar 16.0 g/l 16 47.572
2 Second layer: medium 457 with agar, supplemented with 6.6 ml sterile PHB-suspension per 100 ml.
Poly-ß-hydroxybutyric acid solution 3%(w/v) 6.6 ml - -
Medium 457 100.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
poly-ß-hydroxybutyric acid 3 g 30 -
Distilled water 100 ml - -
Solution occurs in 6 other media (first defined in Medium 457).
Equipment needed: Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2HPO4 2.44 g 2.44 17.188
KH2PO4 1.52 g 1.52 11.17
(NH4)2SO4 0.50 g 0.5 3.784
MgSO4 x 7 H2O 0.20 g 0.2 0.811
CaCl2 x 2 H2O 0.05 g 0.05 0.34
Trace element solution SL-4 10.00 ml - -
Distilled water 1000.00 ml - -
1 Adjust pH to 6.9.
2 Prepare a separate solution of the phosphates and autoclave separately. Combine the two solutions after cooling.
3 Rehydrate and cultivate lyophilized cells in complex medium (e.g. medium 1, 220 or 535). After this reactivation, cultivate in mineral medium 457 with the appropriate carbon source.
Solution occurs in 21 other media (first defined in Medium J340).
Compound Amount Unit Conc. [g/L] Conc. [mM]
EDTA 0.5 g 0.5 1.711
FeSO4 x 7 H2O 0.2 g 0.2 0.719
Trace element solution SL-6 100.0 ml - -
Distilled water 900.0 ml - -
Solution occurs in 55 other media (first defined in Medium 27).
Compound Amount Unit Conc. [g/L] Conc. [mM]
ZnSO4 x 7 H2O 0.10 g 0.1 0.348
MnCl2 x 4 H2O 0.03 g 0.03 0.152
H3BO3 0.30 g 0.3 4.851
CoCl2 x 6 H2O 0.20 g 0.2 1.54
CuCl2 x 2 H2O 0.01 g 0.01 0.059
NiCl2 x 6 H2O 0.02 g 0.02 0.154
Na2MoO4 x 2 H2O 0.03 g 0.03 0.124
Distilled water 1000.00 ml - -
Solution occurs in 20 other media (first defined in Medium 1).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Peptone 5 g 5 -
Meat extract 3 g 3 -
Agar , for solid medium 15 g 15 -
Distilled water 1000 ml - -
1 Adjust pH to 7.0.
2 For Bacillus strains the addition of 10.0 mg MnSO4 x H2O is recommended for sporulation.
Solution occurs in 22 other media (first defined in Medium 220).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Casein peptone 15 g - -
Soy peptone 5 g - -
NaCl 5 g - -
Agar 15 g - -
Distilled water 1000 ml - -
1 Adjust pH to 7.3. Medium is identical with Tryptone Soya Agar (Oxoid CM 131).
Solution occurs in 18 other media (first defined in Medium 535).
Equipment needed: Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Trypticase soy broth
(BBL 11768, Oxoid CM129 or Merck 5459)
30 g - -
Agar 20 g - -
Distilled water 1000 ml - -
1 pH 7.3
2 Autoclave at 121°C for 15 min.

Metadata

Last modified: 24.02.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Defined medium
Final pH: n.d.
Equipment needed: Autoclave
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Associated strains

All strains for this medium: 1
Comamonas testosteroni DSM 6781

No modifications necessary for this strain.

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Agar 16 47.572
Na2HPO4 0.244 1.719
poly-ß-hydroxybutyric acid 0.198 -
KH2PO4 0.152 1.117
(NH4)2SO4 0.05 0.378
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
temperature mesophilic 1 strain