276. PROTEINIVORAX MEDIUM

KH₂PO₄ 0.30 g

MgSO₄ × 7 H₂O 0.12 g

CaCl₂ × 2 H₂O 0.03 g

NaCl 20.0 g

Tryptone 3.0 g

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

Na₂CO₃ 60.0 g

NaHCO₃ 50.0 g

Na-Thioglycolate 1.0 g

Distilled water 1000.0 ml

Dissolve ingredients (except carbonates and thioglycolate), boil medium for 1 min., then cool to room temperature under N₂ gas atmosphere. Add carbonates and adjust pH to 8.5 – 9.0. Dispense under same gas atmosphere in culture vessels and autoclave. After autoclaving at 121°C for 15 min add thio-glycolate from a sterile anoxic stock solution sterilized by filtration.

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

The trace element solution preparation: FeCl₂ × 4 H₂O is dissolved firstly in HCl, and then is mixed with water and other salts are dissolved in the sequence indicated.