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DSMZ

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

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

Culture medium recipe

Vol.
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Medium for Sulfurovum aggregans DSM 27205

* Modification: 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.

Growth conditions: 2-3 days microaerobic 32 °C

Main sol. 1011c #

ml
Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 30.00 g 29.644 507.26
K2HPO4 0.14 g 0.138 0.794
CaCl2 x 2 H2O 0.14 g 0.138 0.941
MgSO4 x 7 H2O 3.40 g 3.36 13.631
MgCl2 x 6 H2O 4.18 g 4.13 20.317
KCl 0.33 g 0.326 4.374
NH4Cl 0.25 g 0.247 4.618
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 1.482 13.985
Na-pyruvate* 0.50 g 0.494 4.49
Na-lactate* 0.50 g 0.494 4.409
Yeast extract* 0.10 g 0.099 -
Na2S2O3 x 5 H2O 1.50 g 1.482 5.972
Wolin's vitamin solution (10x) 1.00 ml - -
Na-dithionite solution (5% w/v)* 1.00 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, vitamins and dithionite), 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 and dithionite from sterile anoxic stock solutions prepared under 80% N2 and 20% CO2 gas mixture. Solutions of vitamins, thiosulfate and dithionite are sterilized by filtration. Adjust pH of the complete medium to 6.7.
2 After inoculation pressurize vessels to 2 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.

Modified Wolin's mineral solution #

1000
Solution occurs in 167 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.

Wolin's vitamin solution (10x) #

1000
Solution occurs in 306 other media (first defined in Medium 120).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 20 mg 0.02 0.082
Folic acid 20 mg 0.02 0.045
Pyridoxine hydrochloride 100 mg 0.1 0.486
Thiamine HCl 50 mg 0.05 0.148
Riboflavin 50 mg 0.05 0.133
Nicotinic acid 50 mg 0.05 0.406
Calcium D-(+)-pantothenate 50 mg 0.05 0.21
Vitamin B12 1 mg 1.0e-3 7.4e-4
p-Aminobenzoic acid 50 mg 0.05 0.365
(DL)-alpha-Lipoic acid 50 mg 0.05 0.242
Distilled water 1000 ml - -

Na-dithionite solution (5% w/v) #

10
Solution occurs in 7 other media (first defined in Medium 829).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH
(1 N) 		1.0 ml - -
Na2S2O4 0.5 g 50 287.175
Distilled water 9.0 ml - -
1 Add 1 N NaOH to distilled water and make the solution anoxic by sparging with 100% N2 gas. Then dissolve the Na-dithionite and filter sterilize the solution into anoxic Hungate tubes. Store the prepared solution in a dark and cool place. Prepare only small amounts of stock solution, as Na-dithionite decomposes rapidly.

Metadata

Last modified: 17.10.22
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

  • 73 % H2
  • 18 % CO2
  • 7 % N2
  • 2 % O2

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

Associated strains

All strains for this medium: 5
Strains with modifications: 4
Sulfurovum aggregans DSM 27205

  10 g Sulfur powder
  1 g NaNO3
-  Na-dithionite solution (5% w/v)
  Yeast extract
  Na-pyruvate
  Na-lactate
  6.0 - 6.5
  73% H2, 18% CO2, 7% N2, 2% O2

Molecular composition
* modified beta

Compound Conc. [g/L] Conc. [mM]
Sulfur powder 9.881 38.524
MgCl2 x 6 H2O 4.13 20.317
Na2CO3 1.482 13.985
Na2S2O3 x 5 H2O 1.482 5.972
NaNO3 0.988 11.626
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Cultivation metadata from 5 strains BacDive

pH 4 - 6 1 strain
pH 6 - 8 1 strain
phyla 1 5 strains
oxygen anaerobe 2 strains
oxygen microaerophile 1 strain
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