Cytotoxic effect of the purified inulinase from locally Isolate Staphylococcus aureus on Hep-2 cancer cell line in vitro

Abstract

Inulinase is an enzyme catalyzing the hydrolysis of inulin, a plant reserve polysaccharide, into fructoses and fructo- oligosaccharides which are widely used in pharmaceutical and food industry. Here we reported inulinase production from Staphylococcus as there is not found any report on inulinase production from Staphylococcus. Eight(32%) isolates of Staphylococcus aureus were isolated from different agricultural rhizosphere soil samples and screened for higher inulinase production, Staphylococcus aureusS₃ gave higher inhibition zone around the colony in comparison with other isolates. Inulinase was  purified to homogeneity with ammonium sulfate at 70% saturation followed by DEAE-Cellulose A-50 ion exchange chromatography and sephadex G-100 gel filtration chromatography with a recovery yield of 21% and 31.63 fold of purification. The results revealed the purified inulinase has a clear cytotoxic activity on cancer cell (Hep-2)with high significances on this tumor cell line during the three incubation periods, suggesting that the cytotoxic effect of this enzyme is a dose and time dependent, The higher level of inhibition(66%) was obtained at a higher concentration 300?g/ml after 72hour of exposure, this result increased the benefit from using the inulinase enzyme as anticancer drug.

Keywords: inulinase, Staphylococcus aureus, Hep-2 cancer cell line