453. SULFURIMONAS MEDIUM (SmM)

KH₂PO₄ 0.33 g

NH₄Cl 0.33 g

KCl 0.33 g

MgCl₂ × 6H₂O 0.33 g

CaCl₂ × 6H₂O, 0.16 g

NaCl 10.0 g

NaHCO₃ 2.0 g

Trace element solution (see below) 1.0 ml

Vitamin solution (see below) 1.0 ml

Elemental sulfur 5.0 g

KNO₃ 10 mM

Trace element solution:

(NH₄)₂Fe(SO₄)₂ × 6 H₂O (Mohr's salt) 784.0 mg

HCl (concentrated) 5.0 ml

CoCl₂ × H₂O 238.0 mg

(NH₄)₂Ni(SO₄)₂ × 6 H₂O 395.0 mg

Na₂MoO₄ × 2 H₂O 24.0 mg

Na₂WO₄ × 2 H₂O 33.0 mg

ZnSO₄ × 7 H₂O 144.0 mg

CuCl₂ × 2 H₂O 2.0 mg

Na₂SeO₄ 94.0 mg

H₃BO₃ 6.0 mg

MnCl₂ × 4 H₂O 99.0 mg

Distilled water 995.0 ml

Mohr's salt is dissolved firstly in concentrated HCl, then is mixed with distilled water and other salts are dissolved in the sequence indicated.

Vitamin solution:

Biotin 20.0 mg

Folic acid 20.0 mg

Pyridoxine 0.1 mg

Riboflavin 50.0 mg

Pantotenoic acid 50.0 mg

p-Aminobenzoic acid 50.0 mg

Thiamine-HCl 50.0 mg

Nicotinic acid 50.0 mg

Vitamin B₁₂ 1.0 mg

Lipoic acid 50.0 mg

Distilled water 1000.0 ml

The anoxic medium is prepared by boiling and cooling it under N₂ flow. No reducing agents were added. The isolation medium pH of 7.5–8.0 (measured at 25 °C). Elemental sulfur as electron donor is added to the Hungate’s tubes prior sterilization; potassium nitrate from anoxic sterile stock solution is added as electron acceptor just before inoculation.