237. Desulfurococcus fermentans medium

NH₄Cl 0.33 g

KH₂PO₄ 0.33 g

KCl 0.33 g

CaCl₂ × 2 H₂O 0.44 g

MgCl₂ × 6 H₂O 0.7 g

NaCl 0.5 g

Trace elements SL-10 (see below) 1.0 ml

Vitamin solution (see below) 10.0 ml

Yeast extract 0.2 g

Starch 5.0 g

Resazurin 1.0 mg

NaHCO₃ 0.8 g

Na₂S × 9 H₂O 0.5 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

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine 10.0 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

Dissolve ingredients (except vitamins, bicarbonate and sulfide), boil medium for 1 min, then cool to room temperature under 80% N₂ and 20% CO₂ gas atmosphere. Adjust pH to 6.2-6.4 and autoclave at 121°C 15 min. After autoclaving add vitamins from an anoxic stock solution sterilized by filtration and bicarbonate from a sterile stock solution prepared under 80% N₂ and 20% CO₂ gas mixture (autoclave at 121°C 15 min). Prior to inoculation reduce medium by adding sulfide from a sterile, anoxic stock solution prepared under N₂ (autoclave at 121°C 15 min).

 

Updated 1.02.2024