CYTOTOXIC POTENTIAL OF L-ASPARAGINASE FROM BACILLUS SP. IN VITRO

Abstract

This study reports the cytotoxic potential of L-Asparaginase isolated from Bacillus sp. Bacillus sp. was isolated from local soil/water samples and identified by rapid plate assay and further confirmed by phenotypic characterization. Extracellular L- Asparaginase was isolated from broth culture of Bacillus sp.  and purified by ammonium sulfate precipitation, followed by dialysis and ion exchange and gel filtration chromatography techniques. The purified enzyme was used to study the in vitro cytotoxic potential. Varying concentrations (31.25, 62.5, 125, 250, 500 µg/ml) of purified L-Asparaginase was tested on MCF7, HeLa, HepG2 and 3T3L1cell lines by MTT assay. Curcumin was maintained as a positive control. The results revealed that the enzyme showed a significant cytotoxic activity and a dose dependent effect. Minimum inhibition of (19.44%) was observed at an enzyme concentration of 31.25 µg/ml and maximum inhibition (71.14%) was observed at 500ug/ml against MCF7 cell line. Minimum inhibition of (10.04%) was observed at an enzyme concentration 31.25 µg/ml and maximum inhibition (68.92%) was observed at 500 µg/ml against HeLa cell line. Minimum inhibition of (7.45%) was shown at an enzyme concentration of 31.25 µg/ml and maximum inhibition (68.28 %) was observed at 500 µg/ml against HepG2 cell line. Minimum inhibition of (4.4%) was shown by enzyme concentration 31.25 µg/ml and maximum inhibition (47.4%) was observed at 500 µg/ml against 3T3L1 cell line. Curcumin at 5µM concentration showed an inhibition of 48.13% against MCF-7 cells, 54.42% against HeLa cells, 64.94% against HepG2 cells and 44.5% against 3T3L1 cell lines.