Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
Trisodium citrate x 2 H2O | 2.94 | g | ||
|
(NH4)2SO4 | 1.30 | g | ||
|
KH2PO4 | 0.28 | g | ||
|
MgSO4 x 7 H2O | 0.25 | g | ||
|
CaCl2 x 2 H2O | 0.07 | g | ||
|
FeCl3 x 6 H2O | 0.02 | g | ||
|
Allen's trace element solution | 10.00 | ml | ||
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
Sulfur
(powdered) |
10.00 | g | ||
|
Yeast extract
(OXOID) |
0.50 | g | ||
|
Wolin's vitamin solution (10x) | 1.00 | ml | ||
|
Na2S x 9 H2O | 0.50 | g | ||
|
Distilled water | 1000.00 | ml | ||
1 Dissolve ingredients except sulfur, yeast extract, vitamins and sulfide, then adjust the pH to 3.5 with 4 N H2SO4. Sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, then dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials which contain already the appropriate amount of sulfur, only to 30% of their volume to allow for a large headspace. Autoclave at 105°C for 20 min. Prior to inoculation add yeast extract, vitamins (sterilized by filtration) and sulfide from anoxic stock solutions prepared under 100% N2 gas atmosphere. The final pH of the complete medium should be around 4.0. | |||||
2 After inoculation, pressurize the cultivation vessels to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture. |
Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
MnCl2 x 4 H2O | 180 | mg | ||
|
Na2B4O7 x 10 H2O | 450 | mg | ||
|
ZnSO4 x 7 H2O | 22 | mg | ||
|
CuCl2 x 2 H2O | 5 | mg | ||
|
Na2MoO4 x 2 H2O | 3 | mg | ||
|
VOSO4 x 2 H2O | 3 | mg | ||
|
CoSO4 x 7 H2O | 1 | mg | ||
|
Distilled water | 1000 | ml | ||
1 Adjust pH of final solution to 2 with 1 N HCl. |
Compound | Amount | Unit | |
---|---|---|---|
|
Biotin | 20 | mg |
|
Folic acid | 20 | mg |
|
Pyridoxine hydrochloride | 100 | mg |
|
Thiamine HCl | 50 | mg |
|
Riboflavin | 50 | mg |
|
Nicotinic acid | 50 | mg |
|
Calcium D-(+)-pantothenate | 50 | mg |
|
Vitamin B12 | 1 | mg |
|
p-Aminobenzoic acid | 50 | mg |
|
(DL)-alpha-Lipoic acid | 50 | mg |
|
Distilled water | 1000 | ml |