Fusarium sp. l-asparaginases: purification, characterization, and potential assessment as an antileukemic chemotherapeutic agent

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 degrees 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.

Automated summary

Asparaginase plays a crucial role in leukemia treatment as part of chemotherapy. It is isolated from Fusarium sp. and characterized as a metalloprotease with specific inhibitory and activating effects on different ions and substrates. The enzyme also exhibits antileukemic activity with a potential as a chemotherapeutic agent.