DSMZ 991 . CALDISPHAERA MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Filter Water bath Gassing station Serum bottle Hungate tubes Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
(NH4)2SO4 1.30 g 1.275 9.645
KH2PO4 0.28 g 0.275 2.017
MgSO4 x 7 H2O 0.25 g 0.245 0.994
CaCl2 x 2 H2O 0.07 g 0.069 0.467
FeCl3 x 6 H2O 0.02 g 0.02 0.073
Allen's trace element solution 10.00 ml - -
Trisodium citrate x 2 H2O 2.94 g 2.882 9.801
Sodium resazurin
(0.1% w/v)
0.50 ml 4.9e-4 0.002
Sulfur
(powdered)
10.00 g 9.804 305.704
Yeast extract
(OXOID)
0.50 g 0.49 -
Wolin's vitamin solution 10.00 ml - -
Na2S x 9 H2O 0.50 g 0.49 2.041
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except sulfur, yeast extract, vitamins and sulfide), adjust pH to 3.5 with 10 N H2SO4 and 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 (e.g., 20 ml medium in 100 ml bottles) containing already the appropriate amount of sulfur. Sterilize medium by heating cultivation vessels in a water bath to 90 - 100°C for 1 - 2 hours on each of 3 successive days. Add yeast extract, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of complete medium to 4.0 - 4.5, if necessary. After inoculation pressurize vials to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
Solution occurs in 14 other media (first defined in Medium 88).
Compound Amount Unit Conc. [g/L] Conc. [mM]
MnCl2 x 4 H2O 180 mg 0.18 0.91
Na2B4O7 x 10 H2O 450 mg 0.45 -
ZnSO4 x 7 H2O 22 mg 0.022 0.077
CuCl2 x 2 H2O 5 mg 0.005 0.029
Na2MoO4 x 2 H2O 3 mg 0.003 0.012
VOSO4 x 2 H2O 3 mg 0.003 0.015
CoSO4 x 7 H2O 1 mg 1.0e-3 0.006
Distilled water 1000 ml - -
1 Adjust pH of final solution to 2 with 1 N HCl.
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: 11.04.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 4.0 - 4.5
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes, Serum bottle, Water bath
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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]
Sulfur 9.804 305.704
Trisodium citrate x 2 H2O 2.882 9.801
(NH4)2SO4 1.275 9.645
Na2S x 9 H2O 0.49 2.041
Yeast extract 0.49 -
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Cultivation metadata from 1 strains BacDive

pH 2 - 4 1 strain
pH 4 - 6 1 strain
phyla 1 1 strain
nutrition chemoorganotroph 1 strain
oxygen anaerobe 1 strain
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