Medium data

Search for medium no. = [403].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

403   ACETOBACTERIUM SUBMARINUS MEDIUM

Sea salts (Sigma)  30.0     g
Yeast extract (BD-Difco)  0.1     g
Trace element solution (see Medium No. 301)  1.0     ml
Vitamin solution (see below)  1.0     ml
Thiamine solution (see below)  1.0     ml
Vitamin B12 solution (see below)  1.0     ml
Na2WO4·2H2O  30.0     mg
Na2SeO4  0.5     mg
Sodium acetate  1.0     g
Methylamine  2.0     g
Sodium formate  5.0     g
Methanol  5.0     ml
KH2PO4  0.2     g
NH4Cl  0.27    g
Resazurin  0.5     mg
NaHCO3  1.26    g
Na2S·9H2O  0.5     g
Distilled water  1.0     L
Mix ingredients except methanol, NaHCO3 and Na2S·9H2O, bring to a boil and cool down under a stream of N2 gas. Add methanol to the medium and distribute into culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of H2-CO2 (4:1, v/v) gas mixture, seal with butyl rubber stoppers, and autoclave. Separately autoclave 5% Na2S·9H2O solution under a N2 atmosphere. Filer-sterilize 8% NaHCO3 solution and store under a N2-CO2 (4:1, v/v) gas mixture. Before inoculation, add the sterile NaHCO3 and Na2S·9H2O solutions to the medium, and readjust pH to 7.0, if necessary. Pressurize the inoculated bottles to 200 kPa H2-CO2 (4:1, v/v).
Vitamin solution:
p-Aminobenzoic acid  4.0     mg
Biotin  1.0     mg
Nicotinic acid  10.0     mg
DL-Calcium pantothenate  5.0     mg
Pyridoxine·HCl  10.0     mg
Sodium phosphate buffer (10 mM, pH 7.1)  100.0     ml
Thiamine solution:
Thiamine·HCl  10.0     mg
Sodium phosphate buffer (25 mM, pH 3.4)  100.0     ml
Vitamin B12 solution:
Vitamin B12  5.0     mg
Distilled water  100.0     ml