| NH4Cl | 0.33 | g |
| KH2PO4 | 0.33 | g |
| KCl | 0.33 | g |
| CaCl2·2H2O | 0.33 | g |
| MgCl2·6H2O | 0.33 | g |
| MgSO4·7H2O | 4 | g |
| NaCl | 250 | g |
| Coenzyme M | 0.142 | g |
| Bacto Yeast Extract (Difco) | 0.5 | g |
| Vitamin solution* | 10 | ml |
| Trace elements solution** | 1 | ml |
| Trimethylamine-HCl | 5 | g |
| Resazurin | 1 | mg |
| NaHCO3 | 5 | g |
| Na2S·9H2O | 0.5 | g |
| Distilled water | 1 | L |
| | Mix ingredients except for vitamin solution, trimethylamine-HCl, NaHCO3 and Na2S·9H2O, and autoclave under a N2/CO2 atmosphere (80/20). Separately autoclave Na2S·9H2O solution as 5% solution under a N2 atmosphere. Prior to inoculation, add the filter-sterile vitamin, 50% trimethylamine-HCl and 5% NaHCO3 solutions, and Na2S·9H2O. Adjust pH of the medium to 7.4 with filter-sterile 5% Na2CO3 solution (approximately final concentration of Na2CO3 is 0.5 g/L). |
| | | | | | |
| | *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 |
| | | | | | |
| | **Trace elements solution |
| | | Nitrilotriacetic acid (NTA) | 12.8 | g |
| | | FeCl3·6H2O | 1.35 | g |
| | | MnCl2·4H2O | 0.1 | g |
| | | CoCl2·6H2O | 0.024 | g |
| | | CaCl2·2H2O | 0.1 | g |
| | | ZnCl2 | 0.1 | g |
| | | CuCl2·2H2O | 0.025 | g |
| | | H3BO3 | 0.01 | g |
| | | Na2MoO4·2H2O | 0.024 | g |
| | | NaCl | 1 | g |
| | | NiCl2·6H2O | 0.12 | 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. |