Intracellular L-Asparaginase from Bacillus sp. PG02: Purification, Biochemical Characterization and Evaluation of Optimum pH and Temperature

Corresponding Author: Mahsa Rahimzadeh Molecular Medicine Research Center, Department of Biochemistry, Faculty of Medicine, Hormozgan University of Medical Sciences, Emam Hossein Boulevard, Bandar Abbas, Iran P.O. Box: 7919693116 Tel; +98-761-6688024 Fax: +98-761-6668478 Email: [email protected] [email protected] Abstract: Bacterial L-asparaginases are amidohydrolases that act on Lasparagine and produce L-aspartate and ammonia. These enzymes have been used in treatment of lymphoblastic leukemia. In the present study, a novel strain, Bacillus sp. PG02 was explored for the production of intracellular L-asparaginase enzyme. The nitrogen source for L-asparaginase production was L-asparagine. New intracellular L-asparaginase was purified using ion exchange chromatography and the purity was assessed using SDS-PAGE. Kinetic parameters km and Vmax and thermal properties were studied using L-asparagine as the substrate. SDS-PAGE analysis showed apparent molecular weight of approximately 38 kDa. The enzyme was active in a wide pH ranges (5-10) and it was maximally active at pH 7.5. Bacillus PG02 L-asparaginase was optimally active at 40°C. Thermal inactivation studies exhibited t1/2 of 32.5 min in 37°C. Also T50 and ∆G of inactivation were measured. The results revealed that the enzyme had appropriate characteristics and thus could be a potential candidate for medical and basic investigations.