Research Article Open Access

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

Fatemeh Izadpanah Qeshmi1, Mahsa Rahimzadeh1, Sedigheh Javadpour1 and Manijeh Poodat1
  • 1 Hormozgan University of Medical Sciences, Iran

Abstract

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 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.

American Journal of Biochemistry and Biotechnology
Volume 12 No. 1, 2016, 12-19

DOI: https://doi.org/10.3844/ajbbsp.2016.12.19

Submitted On: 26 August 2015 Published On: 26 November 2015

How to Cite: Qeshmi, F. I., Rahimzadeh, M., Javadpour, S. & Poodat, M. (2016). Intracellular L-Asparaginase from Bacillus sp. PG02: Purification, Biochemical Characterization and Evaluation of Optimum pH and Temperature. American Journal of Biochemistry and Biotechnology, 12(1), 12-19. https://doi.org/10.3844/ajbbsp.2016.12.19

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Keywords

  • L-Asparaginase
  • Bacillus PG02
  • Thermal Stability
  • Kinetics