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DSMZ

DSMZ 507 . CARBOXYDOTHERMUS MEDIUM

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

Culture medium recipe

Vol.
×
Medium for Thermogladius calderae DSM 22663

* Modification: Supplement medium with 10.00 g/l sulfur and increase amount of yeast extract to 1.00 g/l. Sulfur is sterilized separately by steaming for 3 h on each of three successive days.

Growth conditions: 3-7 days anaerobic 85 °C

Main sol. 507 #

ml
Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KCl 0.33 g 0.323 4.331
MgCl2 x 6 H2O 0.52 g 0.509 2.503
CaCl2 x 2 H2O 0.29 g 0.284 1.93
NH4Cl 0.33 g 0.323 6.037
KH2PO4 0.33 g 0.323 2.373
Trace element solution SL-11 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
NaHCO3 1.00 g 0.978 11.648
Yeast extract* 1.00 g - -
Wolin's vitamin solution (10x) 1.00 ml - -
Neutralized sulfide solution 3% (w/v) 20.00 ml - -
Sulfur* 10.00 g - -
Distilled water 1000.00 ml - -
1 Dissolve ingredients except bicarbonate, yeast extract, vitamins and sulfide, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add yeast extract, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Neutralize the sodium sulfide with sterile H2SO4 (see medium 29). After completing the medium adjust pH to 6.8-7.0.
2 Inoculated vessels are pressurized with sterile carbon monoxide gas to 2 bar overpressure.

Trace element solution SL-11 #

1000
Solution occurs in 22 other media (first defined in Medium 722).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2-EDTA x 2 H2O 5.2 g 5.2 15.467
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 1000.0 ml - -
1 Dissolve EDTA in 800 ml distilled water, adjust pH to 7 using 2 N NaOH and add ferrous chloride. After ferrous chloride has dissolved add other compounds. Finally adjust pH to 6.0 and bring volume to 1000 ml.

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 - -

Neutralized sulfide solution 3% (w/v) #

100
Solution occurs in 19 other media (first defined in Medium 28).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 3 g 30 124.905
Distilled water 100 ml - -
1 The sulfide solution is prepared in a 250 ml screw-capped bottle with a butyl rubber septum and a magnetic stirrer. The solution is bubbled with nitrogen gas, closed and autoclaved for 15 min. at 121°C. After cooling to room temperature the pH is adjusted to about 7.0 by adding of sterile 2 M H2SO4 drop-wise with a syringe without opening the bottle.

Metadata

Last modified: 27.10.22
Source: DSMZ
Taxonomic range:  Bacteria
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 6.8 - 7.0
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Gas composition

  • 66 % CO
  • 33 % N2

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

Associated strains

All strains for this medium: 9
Strains with modifications: 5
Thermogladius calderae DSM 22663

  Yeast extract
  1 g Yeast extract
  10 g Sulfur

Molecular composition
* modified beta

Compound Conc. [g/L] Conc. [mM]
Sulfur 9.785 305.106
NaHCO3 0.978 11.648
Yeast extract 0.978 -
Na2S x 9 H2O 0.587 2.444
MgCl2 x 6 H2O 0.509 2.503
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Cultivation metadata from 9 strains BacDive

pH 6 - 8 3 strains
pH 8 - 10 2 strains
pH 4 - 6 1 strain
phyla 2 9 strains
nutrition organoheterotroph 1 strain
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