Ureolytic bacteria culture

Sporosarcina pasteurii, a basophilic ureolytic bacterium, was used in the present work. It was activated in a sterile liquid medium, which consists of 20 g/L yeast extract, 10 g/L NH₄Cl, 20 g/L urea, 10 mg/L MnSO₄⋅H₂O, 24 mg/L NiCl₂⋅6H₂O. The surrounding pH for the sterile liquid medium was adjusted to 8.8 using 1 M solution of NaOH. The activated ureolytic bacteria were mixed with glycerol using a ratio of 7:3 and stored at –20 °C. They were subjected to shaking culture at 30 °C and 180 rpm for 30 h. Further, the chemicals of urea, MnSO₄⋅H₂O, NiCl₂⋅6H₂O, NaOH, and Cu(NO₃)₂⋅3H₂O were diluted to given concentrations, respectively, and applied to the subsequent test tube experiments.