DSMZ 843 . TREPONEMA AZOTONUTRICIUM MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 1.00 g 1 17.112
KCl 0.50 g 0.5 6.707
MgCl2 x 6 H2O 0.40 g 0.4 1.968
CaCl2 x 2 H2O 0.10 g 0.1 0.68
NH4Cl 0.30 g 0.3 5.609
KH2PO4 0.20 g 0.2 1.47
Na2SO4 0.15 g 0.15 1.056
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Trypticase peptone
(BD BBL)
1.00 g 1 -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 -
Na2CO3 1.50 g 1.5 14.153
Sugar mix 30.00 ml - -
Yeast autolysate 50.00 ml - -
Seven vitamins solution 1.00 ml - -
Co-factors solution 20.00 ml - -
DL-Dithiothreitol
(DTT)
0.40 g 0.4 2.592
Distilled water 900.00 ml - -
1 Dissolve ingredients (except carbonate, sugar mix, yeast autolysate, vitamins, co-factors, and DTT), sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Distribute under same gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume (e.g., 4.5 ml per Hungate-type tube) and autoclave. After sterilization add carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere and sugar mix, yeast autolysate, vitamins, co-factors and DTT from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and sterilized by filtration.
2 Check pH of complete medium and adjust to 7.2 - 7.4, if necessary.
3 Note: Basal medium and yeast autolysate have to be prepared freshly and cannot be stored for longer periods.
Solution occurs in 1 other media (first defined in Medium 843).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Cellobiose 1.70 g 56.667 165.547
Sucrose 1.70 g 56.667 165.547
Maltose x H2O 1.80 g 60 175.287
Trehalose 1.90 g 63.333 185.023
D-Xylose 0.75 g 25 166.522
Distilled water 30.00 ml - -
Equipment needed: Centrifuge
Compound Amount Unit Conc. [g/L] Conc. [mM]
Yeast extract
(dry baker's yeast, SIGMA YSC-1)
14 g 280 -
Distilled water 50 ml - -
1 Suspend 14.0 g of dry baker's yeast (SIGMA YSC-1) in 50 ml distilled water and allow cells to autolyse for 48 h at 55°C. Thereafter, cell debris are removed by centrifugation (3.700 x g for 30 min) and the amber-colored supernatant is stored in aliquots at -20°C.
Equipment needed: Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Thiamine pyrophosphate 2.50 g 2.5 5.426
Pyridoxal hydrochloride 0.25 g 0.25 1.228
Pyridoxal phosphate 0.25 g 0.25 1.012
Ca-folinate 0.05 g 0.05 0.098
ß-NAD 0.05 g 0.05 -
Coenzyme A 0.05 g 0.05 0.065
FAD 0.05 g 0.05 0.064
Haemin
(dissolved separately in 10 mM NaOH)
65.00 mg 0.065 0.1
Nicotinamide 25.00 mg 0.025 0.205
Folic acid 2.50 mg 0.003 0.006
Riboflavin 0.50 mg 5.0e-4 0.001
Distilled water 1000.00 ml - -
1 Adjust pH to 7.0 with 1 N NaOH against indicator paper, filter sterilize and store under 100% N2 gas atmosphere.
Solution occurs in 262 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 173 other media (first defined in Medium 385).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH 0.5 g 0.5 12.501
Na2SeO3 x 5 H2O 3.0 mg 0.003 0.011
Na2WO4 x 2 H2O 4.0 mg 0.004 0.012
Distilled water 1000.0 ml - -
Solution occurs in 78 other media (first defined in Medium 503).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Vitamin B12 100 mg 0.1 0.074
p-Aminobenzoic acid 80 mg 0.08 0.583
D-(+)-biotin 20 mg 0.02 0.082
Nicotinic acid 200 mg 0.2 1.625
Calcium pantothenate 100 mg 0.1 0.21
Pyridoxine hydrochloride 300 mg 0.3 1.459
Thiamine-HCl x 2 H2O 200 mg 0.2 0.536
Distilled water 1000 ml - -

Metadata

Last modified: 07.04.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.2 - 7.4
Equipment needed: Autoclave, Centrifuge, Filter, Gassing station, Hungate tubes
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Gas composition

  • 80 % N2
  • 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: 2

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Yeast extract 14 -
Trehalose 1.9 5.551
Maltose x H2O 1.8 5.259
Cellobiose 1.7 4.966
Sucrose 1.7 4.966
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Cultivation metadata from 2 strains BacDive

pH 6 - 8 1 strain
phyla 1 1 strain
oxygen anaerobe 1 strain
temperature mesophilic 1 strain