DSMZ 119a . METHANOBREVIBACTER MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Autoclave Centrifuge Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.50 g 0.499 3.663
MgSO4 x 7 H2O 0.40 g 0.399 1.618
NaCl 0.40 g 0.399 6.824
NH4Cl 0.40 g 0.399 7.456
CaCl2 x 2 H2O 0.05 g 0.05 0.339
Trace element solution SL-10 1.00 ml - -
FeSO4 x 7 H2O solution (0.1% w/v) 2.00 ml - -
Yeast extract
(OXOID)
1.00 g 0.997 -
Na-acetate 1.00 g 0.997 12.154
Na-formate 2.00 g 1.994 29.321
Clarified rumen fluid 200.00 ml - -
Fatty acid mixture 20.00 ml - -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 0.002
NaHCO3 4.00 g 3.988 47.473
L-Cysteine HCl x H2O 0.50 g 0.499 2.838
Na2S x 9 H2O 0.50 g 0.499 2.076
Distilled water 780.00 ml - -
1 Dissolve ingredients except bicarbonate, cysteine and sulfide. Sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve bicarbonate, then dispense medium under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes to 30% of their volume and autoclave. Add cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Prior to use check pH of complete medium and adjust to 6.8 - 7.0, if necessary.
2 After inoculation add sterile 80% H2 and 20% CO2 gas mixture to 1 bar overpressure.
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 27 other media (first defined in Medium 1310).
1 Rumen fluid from cow or sheep (obtained from fistulated animals or abattoir refuse) is filtered through muslin, autoclaved at 121°C for 15 min and then centrifuged at 27,000 g for 20 min. The supernatant is made anoxic by sparging with 100% N2 gas for 15 min, dispensed under same gas atmosphere into anoxic serum vials to 30% of volume and then stored frozen at -20°C.
Solution occurs in 7 other media (first defined in Medium 119).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Isobutyric acid 23 ml - -
DL-2-Methylbutyric acid 27 ml - -
Valeric acid 27 ml - -
Isovaleric acid 27 ml - -
Distilled water 896 ml - -
1 Adjust pH to 7.5 with concentrated NaOH.
Solution occurs in 7 other media (first defined in Medium 119).
Compound Amount Unit Conc. [g/L] Conc. [mM]
FeSO4 x 7 H2O 1 g 1 3.597
H2SO4
(0.1 N)
1000 ml - -
1 The ferrous sulfate solution is not stable and should be freshly prepared.

Metadata

Last modified: 27.10.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 6.8 - 7.0
Equipment needed: Autoclave, Centrifuge, 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]
NaHCO3 3.988 47.473
Na-formate 1.994 29.321
Na-acetate 0.997 12.154
Yeast extract 0.997 -
KH2PO4 0.499 3.663
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature mesophilic 1 strain