| NH4Cl | 0.25 | g | | KCl | 0.34 | g | | MgCl2·6H2O | 4 | g | | MgSO4·7H2O | 3.5 | g | | CaCl2·2H2O | 0.14 | g | | K2HPO4 | 0.14 | g | | NaCl | 6 | g | | Trace elements solution* | 10 | ml | | Vitamin solution** | 10 | ml | | Bacto Yeast Extract (Difco) | 2 | g | | Polypeptone | 2 | g | | Sodium acetate | 1 | g | | Resazurin | 1 | mg | | NaHCO3 | 5 | g | | Cysteine-HCl | 0.5 | g | | Na2S·9H2O | 0.5 | g | | Distilled water | 1 | L | | | | Mix ingredients except for vitamin solution, NaHCO3, cysteine-HCl and Na2S·9H2O, and autoclave under H2/CO2 (80/20) condition. Separately autoclave cysteine-HCl and Na2S·9H2O as a 5% solution under a N2 atmosphere. Aseptically and anaerobically add filter-sterile vitamin and NaHCO3 solutions, and cysteine-HCl and Na2S·9H2O solutions, and pressurize inoculated bottles to 150 kPa with H2/CO2. | | | | | | | | | | *Trace elements solution | | | | Nitrilotriacetic acid (NTA) | 4.1 | g | | | | FeCl3·6H2O | 0.36 | g | | | | MnCl2·4H2O | 0.09 | g | | | | CoCl2·6H2O | 0.15 | g | | | | CaCl2·2H2O | 0.018 | g | | | | ZnCl2 | 0.09 | g | | | | H3BO3 | 0.017 | g | | | | Na2MoO4·2H2O | 0.009 | g | | | | NiCl2·6H2O | 0.018 | g | | | | Distilled water | 1 | L | | | | First dissolve nitrilotriacetic acid and adjust pH to 6.5 with NaOH, and then add minerals. Final pH 7.0. | | | | | | | | | | **Vitamin solution | | | | Biotin | 2 | mg | | | | Folic acid | 2 | mg | | | | Pyridoxine-HCl | 10 | mg | | | | Thiamine-HCl | 5 | mg | | | | Riboflavin | 5 | mg | | | | Nicotinic acid | 5 | mg | | | | Ca-pantothenate | 5 | mg | | | | p-Aminobenzoic acid | 1 | mg | | | | Vitamin B12 | 0.01 | mg | | | | Distilled water | 1 | L |
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