PRODUCTION, CHARACTERIZATION AND ANTICANCEROUS ACTIVITY OF L-ASPARAGINASE FROM BACILLUS SP

L-asparaginase, an enzyme that catalyzes the hydrolysis of L-asparagine to L-aspartate and ammonia, is widely used as an antineoplastic agent in the treatment of Acute Lymphoblastic Leukemia (ALL) and in the food industry. Microorganisms producing the enzyme were isolated from soil and cultured in Luria-Bertani (LB) media. The optimal conditions for enzyme production were determined to be 40°C, pH 6, and 48 hours of incubation. The addition of ammonium sulfate, sucrose, and cobalt chloride enhanced enzyme production. The enzyme was purified through salt dialysis, ion-exchange chromatography, and gel filtration. Gram staining and biochemical tests identified the producer as a Bacillus species. Characterization of the purified enzyme showed an activity of 0.27 U/mL at 30 minutes of incubation, 2.93 U/mL at 50 mM substrate concentration, 0.36 U/mL at pH 6, and 0.44 U/mL at room temperature. SDS-PAGE analysis revealed a molecular weight of 45 kDa. MTT assay using the 3T3 cell line demonstrated 62.37% inhibition, indicating significant anticancer potential. These findings highlight L-asparaginase as a promising candidate for therapeutic applications.