442. DESULFOTHERMOBACTER MEDIUM (DstbM)

NH4Cl 0.33 g

KCl 0.33 g

MgCl2 × 6 H2O 0.33 g

CaCl2 × 2 H2O 0.33 g

KH2PO4 0.33 g

Na2SO4 2.80 g

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

Selenite-tungstate solution (see below) 1.0 ml

Wolin's vitamin solution (see below) 1.0 ml

Na2S × 9 H2O 0.50 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 3.0 mg

Na2WO4 × 2 H2O 4.0 mg

Distilled water 1000.0 ml

Wolin's vitamin solution

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine-HCl 10.0 mg

Thiamine-HCl 5.0 mg

Riboflavin 5.0 mg

Nicotinic acid 5.0 mg

Ca-D-pantothenate 5.0 mg

Vitamin B12 0.10 mg

p-Aminobenzoic acid 5.0 mg

(±)-α-Lipoic acid 5.0 mg

Distilled water 1000.0 ml

Dissolve all ingredients except vitamins and sulfide, then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 – 45 min to make it anoxic. Adjust pH to 4.5 with HCl, dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Vitamins should be sterilized by filtration. Adjust pH of complete medium to 4.5 - 4.8, if necessary.

 

Updated 26.05.2022