368. METHANOBACTERIUM VETERUM MEDIUM

Solution 1:

Na-acetate 3 H2O 0.5 g

(NH4)2SO4 0.45 g

K2HPO4 0.29 g

KH2PO4 0.18 g

MgSO4 7 H2O 0.12g

CaCl2 2 H2O 0.06 g

NaCl 0.05

Resazurin 0.01 g

Na2CO3 5.0 g

Trace element solution (see below) 10.0 ml

Vitamin solution (see below) 10.0 ml

Distilled water 965.0 ml

Solution 2 (reducing agents):

L-Cysteine-HCl H2O 0.5 g

Na2S 9 H2O 0. 5 g

Distilled water 10.0 ml

Trace element solution:

Nitrilotriacetic acid 1.5 g

MgSO4 7 H2O 3.0 g

MnSO4 7 H2O 0.5 g

NaCl 1.0 g

FeSO4 7 H2O 0.1 g

CoSO4 0.18 g

CaCl2 2 H2O 0.1 g

ZnSO4 0.18 g

CuSO4 0.01 g

KAl(SO4)2 12 H2O 0.02 g

H3BO3 0.01 g

Na2MoO4 2 H2O 0.01 g

NiCl2 0.025 g

Na2SeO3 5 H2O 0.3 mg

Distilled water 1000.0 ml

pH 7.0-7.4

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 (thioctic) acid 5.0 mg

Distilled water 1000.0 ml

pH 7.2-7.4

Prepare medium in anaerobic conditions, blowing through with N2 without O2 up to sterilization. Solutions of reducing agents (10.0 ml) and others solutions add to base medium after separate sterilization at 121C for 15 min. Sterilize vitamin solution by filtration.

Cultivate in a gas mixture of 80% H2 and 20% CO2.

 

Updated 10.08.2021