Partial purification of L.Asparaginase from Cairana moschata

Asparaginase is an enzyme that catalyzes the hydrolysis of asparagine to aspartic acid. Asparaginase is naturally produced by microorganisms. Both the substrate and the product of this enzyme during the reaction play an important role in the metabolism of most organisms. Among the valuable physiological functions of this enzyme are control of expression and proper cellular activity in balancing the body with amino acids. In addition, the enzyme L-asparaginase plays an important role in the treatment of acute lymphoblastic leukemia. The aim of this study was to investigate the kinetic properties of Muscovy duck liver asparaginase enzyme and the effect of different parameters on the activity of this enzyme. In this study, asparaginase enzyme from Muscovy duck liver was partially purified using homogenization methods in Tris buffer pH ۸.۵, centrifugation, ۲۰% and ۶۰% ammonium sulfate precipitation and dialysis at ۳ ° C. After extraction, enzyme activity was estimated according to wriston method with Nessler reagent. In this study, the specific activity of asparaginase enzyme isolated from duck liver was determined to be ۴۰.۳۵ U / mg. The optimum pH of the enzyme was ۷ and its optimum temperature was ۴۰ ° C. The Km and Vmax values of the enzyme were calculated to be ۱۲۷.۲۲۹ mM and ۳.۶۳۹ mM / min, respectively. Due to the side effects of asparaginase prepared from microbial sources, extraction and purification of this enzyme from other sources, including Muscovy duck liver for future studies was suggested.

Keywords:

Partial Purification , Nessler Reagent , Kinetic Parameters