PARTIAL PURIFICATION AND CYTOTOXIC ACTIVITY OF L-ASPARAGINASE ISOLATED FROM Escherichia coli

Abstract

L-Asparaginase (E.C.3.5.1.1) is an important natural product that possesses a broad spectrum of antitumor activity. In the present study, L-asparaginase partially purified from local isolate of Escherichia coli that were grown aerobically for four hours and anaerobically for 18hrs on M9 medium contain L-asparagine as sole nitrogen source. Extraction of the enzyme was done by sonication. Qualitative and quantitative assays for L-Asparaginase production were determined using colorimetric and nesslerization methods respectively. The purification steps involve dialysis of crude extract and DEAE-Cellulose Ion exchange chromatography. The enzyme was purified 3.39-folds and showed a final specific activity of 0.18 U/mg with a 41% yield. The crude extract and DEAE-Cellulose fractions showed slight growth inhibition against RD cell line (human rhambdomyo sarcoma).