Purification of L-glutaminase Produced from Pseudomonas aeruginosa P48 and Evaluate Its Anticancer Activity

Abstract

     The global need for the treatment of cancer diseases has been directed towards the use of enzymes, as these diseases have worsened in recent years for several reasons, including chemical and physical pollutants. L-glutaminase is one of the enzymes used in the treatment of tumors, as it converts the amino acid glutamine into glutamic acid, which helps inhibit the growth of cancer cells. Therefore, this study aimed to produce and purify L-glutaminase from a local isolate of P. aeruginosa P48 and evaluate its anticancer activity. The result of PCR with the 16SrRNA gene for identification of P. aeruginosa P48 showed that the 16SrRNA gene (956bp) was presented as a single band on agarose gel as an indicator that this species was P. aeruginosa isolate. The L-glutaminase from P. aeruginosa P48 was purified utilizing ion exchange chromatography after precipitation via ammonium sulfate (0-75%), with a yield of 108.8%, a purification fold 2.189 and the specific activity was 9.5453U/mg protein. The enzyme has a molecular weight of 39 KDa. The results of the cytotoxic impact utilizing the MTT experiment indicated that partially purified L-glutaminase caused a reduction in cell viability (p ≤ 0.05) at a dose-dependent manner on HL-60 cell lines, with a calculating IC₅₀ of 143.9 µg/ml, compare with normal cell line (HdFn Cell Line) at IC₅₀ of 187.3 µg/ml. After conducting an investigation of its anticancer activity, the findings were demonstrated that L-glutaminase has potential therapeutic uses in the treatment of cancer.