DSMZ 1011c . DESULFOTHERMUS MJ MEDIUM (H2/CO2)

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

Culture medium recipe

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* Modified for Sulfurovum aggregans DSM 27205

Supplement medium with 1.00 g/l NaNO3 and omit pyruvate, lactate, yeast extract and dithionite. Upon autoclaving add 10.00 g/l sterile sulfur powder (sterilized by steaming for 3 hours on each of 3 successive days) and adjust pH to 6.0 - 6.5. After inoculation do not pressurize with 80% H2 and 20% CO2 gas mixture and add sterile air in an amount that is equivalent to a volume of 10% of the headspace.

Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 30.00 g 29.412 503.281
K2HPO4 0.14 g 0.137 0.788
CaCl2 x 2 H2O 0.14 g 0.137 0.934
MgSO4 x 7 H2O 3.40 g 3.333 13.524
MgCl2 x 6 H2O 4.18 g 4.098 20.157
KCl 0.33 g 0.324 4.34
NH4Cl 0.25 g 0.245 4.582
Fe(NH4)2(SO4)2 x 6 H2O 0.01 g 0.01 -
Modified Wolin's mineral solution 10.00 ml - -
Na2CO3 1.50 g 1.471 13.875
Na-pyruvate* 0.50 g 0.49 4.455
Na-lactate* 0.50 g 0.49 4.374
Yeast extract* 0.10 g 0.098 -
Na2S2O3 x 5 H2O 1.50 g 1.471 5.925
Wolin's vitamin solution 10.00 ml - -
Sulfur powder* 10.00 g - -
NaNO3* 1.00 g - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, pyruvate, lactate, yeast extract, thiosulfate and vitamins), then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials up to a volume of 20% and autoclave. Add pyruvate, lactate, yeast extract, thiosulfate and vitamins to the autoclaved medium from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Solutions of vitamins and thiosulfate are sterilized by filtration. Adjust pH of the complete medium to 6.7. Prior to inoculation reduce medium with 10 - 20 mg/l sodium dithionite, added from a 5% (w/v) solution freshly prepared under N2 and filter sterilized. After inoculation pressurize vessels to 2 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
Solution occurs in 170 other media (first defined in Medium 141).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Nitrilotriacetic acid 1.50 g 1.5 7.848
MgSO4 x 7 H2O 3.00 g 3 12.172
MnSO4 x H2O 0.50 g 0.5 2.958
NaCl 1.00 g 1 17.112
FeSO4 x 7 H2O 0.10 g 0.1 0.36
CoSO4 x 7 H2O 0.18 g 0.18 1.161
CaCl2 x 2 H2O 0.10 g 0.1 0.68
ZnSO4 x 7 H2O 0.18 g 0.18 0.626
CuSO4 x 5 H2O 0.01 g 0.01 0.04
AlK(SO4)2 x 12 H2O 0.02 g 0.02 0.042
H3BO3 0.01 g 0.01 0.162
Na2MoO4 x 2 H2O 0.01 g 0.01 0.041
NiCl2 x 6 H2O 0.03 g 0.03 0.231
Na2SeO3 x 5 H2O 0.30 mg 3.0e-4 0.001
Na2WO4 x 2 H2O 0.40 mg 4.0e-4 0.001
Distilled water 1000.00 ml - -
1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH.
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: 12.04.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: * 6.0 - 6.5
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Gas composition

  • 72 % H2
  • 18 % CO2
  • 9 % Air

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

Associated strains

All strains for this medium: 3
Strains with modifications: 2
Sulfurovum aggregans DSM 27205

  10 g Sulfur powder
  1 g NaNO3
  Dithionite
  Yeast extract
  Na-pyruvate
  Na-lactate
  6.0 - 6.5
  72% H2, 18% CO2, 9% Air

Molecular composition
* modified beta

Compound Conc. [g/L] Conc. [mM]
Sulfur powder 9.804 38.222
MgCl2 x 6 H2O 4.098 20.157
Na2CO3 1.471 13.875
Na2S2O3 x 5 H2O 1.471 5.925
NaNO3 0.98 11.535
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Cultivation metadata from 3 strains BacDive

pH 4 - 6 1 strain
pH 6 - 8 1 strain
phyla 1 3 strains
oxygen anaerobe 2 strains
oxygen microaerophile 1 strain
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