# 1053: SULFURIMONAS PARALVINELLA MEDIUM ## Main sol. 1053 20 g NaCl 0.09 g K2HPO4 0.07 g KH2PO4 0.8 g CaCl2 0.25 g NH4Cl 1 g NaNO3 4 g MgSO4 x 7 H2O 3 g MgCl2 x 6 H2O 0.33 g KCl 0.01 g Fe2(SO4)3 x H2O 10 ml **Modified Wolin's mineral solution** 1 g Na2S2O3 x 5 H2O 0.5 ml Sodium resazurin (0.1% w/v) 10 g Sulfur (powdered) 1 g NaHCO3 1 ml **Wolin's vitamin solution (10x)** 1000 ml Distilled water 1. Dissolve ingredients (except sulfur, bicarbonate and vitamins), then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Adjust pH to 6.8 with NaOH and dispense under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials (up to a volume of 20%) that contain already the appropriate amount of sulfur. Autoclave at a temperature of 110°C for 20 min. Add vitamins and bicarbonate to the autoclaved medium from sterile anoxic stock solutions. The solution of vitamins is sterilized by filtration and stored under 100% N2 gas, whereas the solution of bicarbonate is prepared under 80% N2 and 20% CO2 gas mixture and autoclaved. Adjust the pH of the complete medium to 6.5. 2. After inoculation pressurize vessels to 2 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture. ## Modified Wolin's mineral solution 1.5 g Nitrilotriacetic acid 3 g MgSO4 x 7 H2O 0.5 g MnSO4 x H2O 1 g NaCl 0.1 g FeSO4 x 7 H2O 0.18 g CoSO4 x 7 H2O 0.1 g CaCl2 x 2 H2O 0.18 g ZnSO4 x 7 H2O 0.01 g CuSO4 x 5 H2O 0.02 g AlK(SO4)2 x 12 H2O 0.01 g H3BO3 0.01 g Na2MoO4 x 2 H2O 0.03 g NiCl2 x 6 H2O 0.3 mg Na2SeO3 x 5 H2O 0.4 mg Na2WO4 x 2 H2O 1000 ml Distilled water 1. First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH. ## Wolin's vitamin solution (10x) 20 mg Biotin 20 mg Folic acid 100 mg Pyridoxine hydrochloride 50 mg Thiamine HCl 50 mg Riboflavin 50 mg Nicotinic acid 50 mg Calcium D-(+)-pantothenate 1 mg Vitamin B12 50 mg p-Aminobenzoic acid 50 mg (DL)-alpha-Lipoic acid 1000 ml Distilled water