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1011c. DESULFOTHERMUS MJ MEDIUM (H2/CO2)

Culture medium recipe

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* Modified for Sulfurovum aggregans DSM 27205
beta

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: Hungate tubes Autoclave Filter
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 30.00 g 29.412 503.26
2 0.14 g 0.137 0.788
3 0.14 g 0.137 0.934
4 3.40 g 3.333 13.524
5 4.18 g 4.098 20.157
6 0.33 g 0.324 4.34
7 0.25 g 0.245 4.582
8 0.01 g 0.01 0.025
9 Trace element solution 10.00 ml - -
10 1.50 g 1.471 13.875
11 0.50 g 0.49 4.455
12 0.50 g 0.49 4.374
13 0.10 g 0.098 -
14 1.50 g 1.471 5.925
15 Vitamin solution 10.00 ml - -
16 1000.00 ml 978.634 54322.4
17 20.00 mg/l 0.02 -
* 1.00 g/l - -
* 10.00 g/l - -
Instructions:
  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 192 other media (first defined in medium 141 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 1.50 g 1.5 7.848
2 3.00 g 3 12.172
3 0.50 g 0.5 2.958
4 1.00 g 1 17.111
5 0.10 g 0.1 0.36
6 0.18 g 0.18 0.64
7 0.10 g 0.1 0.68
8 0.18 g 0.18 0.626
9 0.01 g 0.01 0.04
10 0.02 g 0.02 0.042
11 0.01 g 0.01 0.162
12 0.01 g 0.01 0.041
13 0.03 g 0.03 0.126
14 0.30 mg 3.0e-4 0.001
15 0.40 mg 4.0e-4 0.001
16 1000.00 ml 998.207 55408.8
17 , required for pH adjustment x - -
Instructions:
  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 366 other media (first defined in medium 141 )
# Compound Amount Unit Conc. [g/L] Conc. [mM]
1 2.0 mg 0.002 0.008
2 2.0 mg 0.002 0.005
3 10.0 mg 0.01 0.049
4 5.0 mg 0.005 0.015
5 5.0 mg 0.005 0.013
6 5.0 mg 0.005 0.041
7 5.0 mg 0.005 0.01
8 0.1 mg 1.0e-4 6.3e-5
9 5.0 mg 0.005 0.036
10 5.0 mg 0.005 0.024
11 1000.0 ml 998.207 55408.8

Metadata

Media status: Official DSMZ medium
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: * 6.0 - 6.5
Equipment needed: Autoclave, Filter, Hungate tubes

Associated strains

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

  Na-pyruvate
  6.0 - 6.5
  1.00 g/l NaNO3
  80% H2, 20% CO2
  Yeast extract
  10.00 g/l sulfur
  Na lactate
  Sodium dithionite

Gas composition

  • 80 % H2
  • 20 % CO2

Light

Intensity Na-pyruvate lux

Molecular composition
* modified beta

Compound Conc. [g/L] Conc. [mM]
Distilled water 998.207 55408.8
NaCl 29.422 503.428
Sulfur 10 311.857
MgCl2 x 6 H2O 4.098 20.157
MgSO4 x 7 H2O 3.363 13.643
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Cultivation metadata from 3 strains BacDive

halophily not 1 strain
halophily slightly 1 strain
pH 4 - 6 1 strain
pH 6 - 8 1 strain
phyla 1 3 strains
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