DSMZ 193a . PHB/PYRUVATE MEDIUM

Official DSMZ medium: This medium was manually curated by experts from the DSMZ.

Culture medium recipe

Vol.
×
Equipment needed: Autoclave Gassing station Hungate tubes Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 932 ml - -
Solution B 30 ml - -
Solution C 20 ml - -
Solution D 10 ml - -
Solution E 10 ml - -
1 Solution A is sparged with 80% H2 and 20% CO2 gas mixture to reach a pH below 6 (at least 30 min), then distributed under the same gas atmosphere in anoxic Hungate-type tubes or serum vials and autoclaved. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solutions C and D are prepared under 100% N2 gas atmosphere and sterilized by filtration. Solution E is autoclaved separately under 100% N2 gas. To complete the medium appropriate amounts of solutions B to E are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be 6.8 - 7.0. After inoculation, pressurize vessels with 80% H2 and 20% CO2 gas mixture to 1 bar overpressure.
2 Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution, freshly prepared under N2 and filter-sterilized) may stimulate growth at the beginning. For transfers use 5 - 10% inoculum.
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2SO4 3.00 g 3.219 22.662
KH2PO4 0.20 g 0.215 1.577
NH4Cl 0.30 g 0.322 6.018
NaCl 7.00 g 7.511 128.52
MgCl2 x 6 H2O 1.30 g 1.395 6.861
KCl 0.50 g 0.536 7.196
CaCl2 x 2 H2O 0.15 g 0.161 1.095
Selenite-tungstate solution 1.00 ml - -
Trace element solution SL-10 1.00 ml - -
Poly[(R)-3-hydroxybutyric acid]
(SIGMA)
1.00 g 1.073 -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.4e-4 0.002
Distilled water 930.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 1.5 g 50 471.752
Distilled water 30.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na-pyruvate 5 g 250 2271.82
Distilled water 20 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Wolin's vitamin solution 10 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 0.4 g 40 166.54
Distilled water 10.0 ml - -
Solution occurs in 174 other media (first defined in Medium 385).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH 0.5 g 0.5 12.501
Na2SeO3 x 5 H2O 3.0 mg 0.003 0.011
Na2WO4 x 2 H2O 4.0 mg 0.004 0.012
Distilled water 1000.0 ml - -
Solution occurs in 263 other media (first defined in Medium 320).
Compound Amount Unit Conc. [g/L] Conc. [mM]
HCl
(25%)
10.0 ml 2.5 68.568
FeCl2 x 4 H2O 1.5 g 1.5 7.545
ZnCl2 70.0 mg 0.07 0.514
MnCl2 x 4 H2O 100.0 mg 0.1 0.505
H3BO3 6.0 mg 0.006 0.097
CoCl2 x 6 H2O 190.0 mg 0.19 1.463
CuCl2 x 2 H2O 2.0 mg 0.002 0.012
NiCl2 x 6 H2O 24.0 mg 0.024 0.185
Na2MoO4 x 2 H2O 36.0 mg 0.036 0.149
Distilled water 990.0 ml - -
1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.
Solution occurs in 357 other media (first defined in Medium 141).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 2.0 mg 0.002 0.008
Folic acid 2.0 mg 0.002 0.005
Pyridoxine hydrochloride 10.0 mg 0.01 0.049
Thiamine HCl 5.0 mg 0.005 0.015
Riboflavin 5.0 mg 0.005 0.013
Nicotinic acid 5.0 mg 0.005 0.041
Calcium D-(+)-pantothenate 5.0 mg 0.005 -
Vitamin B12 0.1 mg 1.0e-4 7.4e-5
p-Aminobenzoic acid 5.0 mg 0.005 0.036
(DL)-alpha-Lipoic acid 5.0 mg 0.005 0.024
Distilled water 1000.0 ml - -

Metadata

Last modified: 19.08.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Defined medium
Final pH: 6.8 - 7.0
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Gas composition

  • 80 % H2
  • 20 % CO2

Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .

Associated strains

All strains for this medium: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaCl 6.986 119.542
Na-pyruvate 4.99 45.346
Na2SO4 2.994 21.079
Na2CO3 1.497 14.124
MgCl2 x 6 H2O 1.297 6.382
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Cultivation metadata from 1 strains BacDive

oxygen anaerobe 1 strain
temperature mesophilic 1 strain