320. THERMOSULFURIMONAS MEDIUM

NaCl 18.0 g

MgCl2 6 H2O 4.0 g

KCl 0.33 g

CaCl2 2 H2O 0.33 g

(NH4)2SO4 0.5 g

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

Selenite-tungstate solution (see below) 1.0 ml

NaHCO3 2.5 g

KH2PO4 0.33 g

Vitamin solution (see below) 1.0 ml

Na-acetate 3 H2O 1.5 g

Na2S2O3 5 H2O 3.5 g

Yeast extract 0.2 g

Distilled water 1000.0 ml

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

Selenite-tungstate solution:

NaOH 0.5 g

Na2SeO3 5 H2O 3 mg

Na2WO4 2 H2O 4 mg

Distilled water 1000.0 ml

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine-HCl 10.0 mg

Thiamine-HCl 2 H2O 5.0 mg

Riboflavin 5.0 mg

Nicotinic acid 5.0 mg

D-Ca-pantothenate 5.0 mg

Vitamin B12 0.1 mg

p-Aminobenzoic acid 5.0 mg

Lipoic acid 5.0 mg

Distilled water 1000.0 ml

Dissolve ingredients except bicarbonate, phosphate, vitamins, acetate, thiosulfate and yeast extract, then sparge medium with 80% H2 and 20% CO2 gas mixture for 3045 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add phosphate, vitamins, acetate, thiosulfate and yeast extract from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Stock solutions of thiosulfate and vitamins should be sterilized by filtration. The pH of the complete medium should be at 6.56.8.

 

Updated 05.07.2021