PRODUCTION AND PARTIAL PURIFICATION OF L-ASPARAGINASE ENZYME FROM BACTERIA

L-asparaginase is a potential anti-leukemic enzyme. It does not affect normal cells in the human body but target only cancerous cells. L- asparaginase had been isolated from many different microbial sources. The demand for the enzyme worldwide especially in health sector had driven researches focusing on enzyme yield per substrate quantity by applying optimization techniques. In the current study soil microbial isolates were screened for potential producers of L-asparaginase using phenol red indicator growth medium. The isolate was characterized by biochemical tests and was found to belong to Bacillus sp. The enzyme was mass produced by submerged fermentation method. Different concentrations of nitrogen and carbon sources, like peptone and lactose were used for optimizing the enzyme production. The enzyme was partially purified by ammonium sulphate precipitation. Dialysis was carried out to remove the excess salt. The dialysis purified enzyme exhibited maximal enzyme activity at a pH of 7 and temperature of 38°C. The activity was found to be 75% greater than the activity of crude enzyme in broth culture. Protein profiling by SDS-PAGE revealed the molecular weight of the protein to be 42 kDa. Keywords: Bacillus sp, Enzyme, L-asparaginase, partial purification.