Intracellular L-Asparaginase from Bacillus sp. PG02: Purification, Biochemical Characterization and Evaluation of Optimum pH and Temperature
Fatemeh Izadpanah Qeshmi, Mahsa Rahimzadeh, Sedigheh Javadpour and Manijeh Poodat
DOI : 10.3844/ajbbsp.2016.12.19
American Journal of Biochemistry and Biotechnology
Volume 12, Issue 1
Bacterial L-asparaginases are amidohydrolases that act on L-asparagine 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 intra-cellular 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 40Â°C. Thermal inactivation studies exhibited t1/2 of 32.5 min in 37Â°C. Also T50 and âG 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.
© 2016 Fatemeh Izadpanah Qeshmi, Mahsa Rahimzadeh, Sedigheh Javadpour and Manijeh Poodat. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.