388. THERMODESULFOBIUM ACIDIPHILUM MEDIUM

NH4Cl 0.33 g

KCl 0.33 g

KH2PO4 0.33 g

MgCl2 6 H2O 0.33 g

CaCl2 2 H2O 0.33 g

Na2SO4 2.8 g

Trace mineral solution SL-10 (see below) 1.0 ml

Na-resazurin solution (0.1% w/v) 0.5 ml

Yeast extract 3.0 g

Vitamin solution (see below) 10.0 ml

Na2S 9 H2O 0.45 g

Distilled water 1000.0 ml

Dissolve ingredients except yeast extract, vitamins and sulfide. Adjust pH to 4.5 with H2SO4 and sparge medium with 100% CO2 gas for 10 15 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes  and autoclave. Add yeast extract, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. The sulfide stock solution (3% w/v) should be neutralized before use. The pH of the complete medium should be at 4.5.

Trace element solution SL-10:

HCl (25%; 7.7 M) 10.0 ml

FeCl2 4 H2O 1.5 g

ZnCl2 70.0 mg

MnCl2 4 H2O 100.0 mg

H3BO3 6.0 mg

CoCl2 6 H2O 190.0 mg

CuCl2 2 H2O 2.0 mg

NiCl2 6 H2O 24.0 mg

Na2MoO4 2 H2O 36.0 mg

Distilled water 990.0 ml

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine 0.1 mg

Riboflavin 5.0 mg

Pantotenoic acid 5.0 mg

p-Aminobenzoic acid 5.0 mg

Thiamine-HCl 5.0 mg

Nicotinic acid 5.0 mg

Vitamin B12 0.1 mg

Lipoic acid 5.0 mg

Distilled water 1000.0 ml

pH 7.2-7.4

 

Updated 10.08.2021