444. ISACHENKONIA MEDIUM

NH4Cl 0.5 g

NaCl 20.0 g

CaCl2 × 6 H2O 0.033 g

KH2PO4 0.2 g

MgCl2 × 6 H2O 0.1 g

Na2CO3 50.0 g

NaHCO3 60.0 g

Glucose 3.0 g

Yeast extract 0.05 g

Trace element solution (see below) 1 ml

Vitamin solution (see below) 10 ml

Na2S × 9 H2O 3.0 g

pH 9.2

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 it 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 100 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 medium is prepared anaerobically. NaHCO3, Na2CO3 and vitamins, Na2S × 9 H2O are added after boiling and cooling of the medium under 100% CO2 in gas phase. Adjust the pH between 9.0 and 9.2 with sterile NaOH after autoclaving. Yeast extract must be added to the final solution before inoculation.

 

Updated 26.05.2022