Fusarium sp. L-asparaginases: purification, characterization, and potential assessment as an antileukemic...

Asparaginases important role in the treatment of leukemia. It is part of chemotherapy in the treatment of leukemia in the last three decades. L-Asparaginase is isolated from Fusarium sp. isolated from soil and purified using ammonium sulfate precipitation and Sephadex G 100. Characterization of the crude enzyme revealed it is a metalloprotease inhibited by EDTA. Hg²⁺, Cd²⁺, and Pb²⁺ also inhibited the enzyme. Mg²⁺, Zn²⁺, and Ca²⁺ activated L-asparaginase. Furthermore, kinetic studies of purified enzyme were carried out. V_max and K_m were 0.031 M and 454 U/mL, respectively. The optimum temperature was 30 °C and the optimum pH was 7. Concerning substrate specificity, gelatin and casein in addition to L-asparagine were tested. The enzyme was found to be nonspecific that could hydrolyze all tested substrates at different rates. The maximum enzyme activity was recorded in the case of L-asparagine, followed by casein and gelatin, respectively. The molecular weight of L-asparaginase was 22.5 kDa. The antileukemic cytotoxicity assay of the enzyme against RAW2674 leukemic cell lines by MTT viability test was estimated. The enzyme exhibited antileukemic activity with IC₅₀ of 50.1 UmL^-1. The current work presents additional information regarding the purification and characterization of the enzyme produced by Fusarium sp. and its evaluation as a potential antileukemic chemotherapeutic agent.