Step A
Asparaginase activity was found to associate with fraction precipitation at 40-60% saturation. The crude enzyme
extract was brought to 45 % saturation with ammonium sulphate (2.6g) at pH of 8.4 and samples were kept
overnight in cooled temperature. After equilibration is achieved supernatant was subjected to centrifugation at 5000
rpm for 10 min at 40C, after which supernatant was brought to 85% saturation with ammonium sulphate (5.8g) and
again centrifuge at 5000 rpm for 10 min. The precipitate was collected separately and store at 40C until next step
was done.