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DSMZ

DSMZ 203 . METHANOTHERMUS MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 203 #

ml
Equipment needed: Autoclave Filter Gassing station Serum bottle
Compound Amount Unit Conc. [g/L] Conc. [mM]
Mineral solution 1 37.5 ml - -
Mineral solution 2 37.5 ml - -
NiCl2 x 6 H2O
(0.1% w/v) 		1.0 ml 9.9e-4 0.008
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4) 		2.0 ml 0.002 0.007
Modified Wolin's mineral solution 10.0 ml - -
Na2SO4 3.4 g 3.37 23.723
Sodium resazurin
(0.1% w/v) 		0.5 ml 5.0e-4 0.002
Na2CO3 1.0 g 0.991 9.351
Yeast extract
(OXOID) 		2.0 g 1.982 -
Trypticase peptone
(BD BBL) 		2.0 g 1.982 -
Wolin's vitamin solution (10x) 1.0 ml - -
Na2S x 9 H2O 0.5 g 0.496 2.063
L-Cysteine HCl x H2O 0.5 g 0.496 2.821
Distilled water 920.0 ml - -
1 Dissolve ingredients except carbonate, yeast extract, Trypticase peptone, vitamins, sulfide and cysteine, then adjust pH to 6.0 with sulfuric acid. 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 serum bottles to 30% of their volume and autoclave. Thereafter, add by injection yeast extract, Trypticase peptone, vitamins (sterilized by filtration), sulfide and cysteine from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of complete medium to 6.5.
2 After inoculation pressurize bottles to 2 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.

Mineral solution 1 #

1000
Solution occurs in 4 other media (first defined in Medium 203).
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 6 g 6 34.448
Distilled water 1000 ml - -

Mineral solution 2 #

1000
Solution occurs in 1 other media (first defined in Medium 203).
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 6.0 g 6 44.09
(NH4)2SO4 6.0 g 6 45.406
NaCl 12.0 g 12 205.339
MgSO4 x 7 H2O 2.4 g 2.4 9.737
CaCl2 x 2 H2O 1.6 g 1.6 10.883
Distilled water 1000.0 ml - -

Modified Wolin's mineral solution #

1000
Solution occurs in 166 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 305 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 - -

Metadata

Last modified: 27.10.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 6.5
Equipment needed: Autoclave, Filter, Gassing station, Serum bottle
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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: 4
Strains with modifications: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Na2SO4 3.37 23.723
Trypticase peptone 1.982 -
Yeast extract 1.982 -
Na2CO3 0.991 9.351
L-Cysteine HCl x H2O 0.496 2.821
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Cultivation metadata from 4 strains BacDive

phyla 1 4 strains
oxygen anaerobe 4 strains
temperature hyperthermophilic 4 strains