1166 .
DESULFONATRONUM THIOAUTOTROPHICUM MEDIUM
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* Medium for Desulfurispira natronophila DSM 22071
Replace formate and pyruvate with 1.60 g/l Na-acetate as substrate and supplement medium with 3.00 g/l sulfur. For sterilization steam sulfur for 3 hours on each of 3 successive days. Add sulfur aseptically to the autoclaved medium while retaining anoxic conditions.
Growth conditions: anaerobic
30 °C
Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final gas composition:
100 % N2
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Final pH:
9.5 - 10.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
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NaCl | 6.00 | g | ||
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K2HPO4 | 1.00 | g | ||
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Na2SO4 | 2.80 | g | ||
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NaHCO3 | 8.00 | g | ||
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Na2CO3 | 22.00 | g | ||
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NH4Cl | 0.20 | g | ||
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MgCl2 x 6 H2O | 0.20 | g | ||
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Trace elements solution (Pfennig & Lippert,1966) | 1.00 | ml | ||
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Selenite-tungstate solution | 1.00 | ml | ||
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Yeast extract | 0.05 | g | ||
| Na-formate* | 3.40 | g | |||
| Na-pyruvate* | 0.55 | g | |||
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Na2S2O3 x 5 H2O | 5.00 | g | ||
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Wolin's vitamin solution (10x) | 1.00 | ml | ||
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Na2S x 9 H2O | 0.24 | g | ||
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Elemental sulfur | 3.00 | g | ||
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Na acetate | 1.60 | g | ||
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Distilled water | 1000.00 | ml | ||
| 1 Dissolve sodium chloride, hydrogenphosphate and sodium sulfate, then sparge solution with 100% N2 gas for 30 - 45 min to make it anoxic. Add and dissolve carbonates, dispense under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add ammonium chloride, magnesium chloride, trace elements, yeast extract, formate, pyruvate, thiosulfate, vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Stock solutions of vitamins and thiosulfate should be sterilized by filtration. Adjust pH of complete medium to 9.5 - 10. | |||||
Trace elements solution (Pfennig & Lippert,1966) #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
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EDTA | 5.00 | g | ||
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FeSO4 x 7 H2O | 2.20 | g | ||
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ZnSO4 x 7 H2O | 0.10 | g | ||
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MnCl2 x 4 H2O | 0.03 | g | ||
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H3BO3 | 0.03 | g | ||
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CoCl2 x 6 H2O | 0.20 | g | ||
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CuCl2 x 2 H2O | 0.03 | g | ||
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NiCl2 x 6 H2O | 0.03 | g | ||
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Na2MoO4 x 2 H2O | 0.03 | g | ||
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Distilled water | 1000.00 | ml | ||
| 1 pH 3.0-4.0 | |||||
Selenite-tungstate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
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NaOH | 0.5 | g |
|
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Na2SeO3 x 5 H2O | 3.0 | mg |
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Na2WO4 x 2 H2O | 4.0 | mg |
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Distilled water | 1000.0 | ml |
Wolin's vitamin solution (10x) #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
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Biotin | 20 | mg |
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Folic acid | 20 | mg |
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Pyridoxine hydrochloride | 100 | mg |
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Thiamine HCl | 50 | mg |
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Riboflavin | 50 | mg |
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Nicotinic acid | 50 | mg |
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Calcium D-(+)-pantothenate | 50 | mg |
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Vitamin B12 | 1 | mg |
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p-Aminobenzoic acid | 50 | mg |
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(DL)-alpha-Lipoic acid | 50 | mg |
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Distilled water | 1000 | ml |