452. BIOMAIBACTER MEDIUM (BbM)

Solution 1:

NaCl 0.3 g

MgCl₂ × 6 H₂O 0.1 g

CaCl₂ × 2 H₂O 0.11 g

NH₄Cl 0.3 g

KH₂PO₄ 0.41 g

Na₂HPO₄ 0.53 g

NaHCO₃ 4.0 g

D-Glucose 1.0 g

Yeast extract 0.5 g

Vitamin solution (see below) 1.0 ml

Trace element solution (see below) 1.0 ml

Distilled water 940.0 ml

Solution 2:

Na₂S × 9 H₂O 0.48 g

Distilled water 10.0 ml

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 DL-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

Trace element solution:

FeCl₂ × 4 H₂O 1.5 g

ZnCl₂ 68.0 mg

MnCl₂ × 4 H₂O 100.0 mg

H₃BO₃ 62.0 mg

CoCl₂ × 6 H₂O 120.0 mg

Na₂MoO₄ × 2 H₂O 24.0 mg

HCl (0.05 M) 1000.0 ml

pH 7.0-7.2

Solution 1 is boiled before sterilization for a few minutes being flushed with gas mixture of 80% N₂ and 20% CO₂ with sodium bicarbonate added until an equilibrium pH of 6.9 - 7.1. Solution 1 is autoclaved under this gas mixture at 121°C for 15 min. Solution 2 is autoclaved under 100% N₂ at 121°C for 15 min. Vitamin solution is filter sterilized.