Cytotoxic Effect of Free and Immobilized Elastase from Klebsiella Pneumoniae In Vitro And In Vivo

Abstract

Forty-eight of Klebsiella pneumoniae isolates were collected from different sources in hospitals of Wasit Governorate. The isolate represents a potential source of elastase enzyme. Elastase was purified by three steps of ammonium sulfate precipitation, ion-exchange chromatography, and gel-filtration chromatography. The optimal temperature and pH for elastase were 37 C and 7 respectively, and its stable on the same conditions. The activity of this elastase was found to decrease when the temperature was higher than 40 C, and also inhibited by ZnCl2, FeCl2, and MnCl2 ions. Immobilized elastase on TiO2 NPs, showed low cytotoxicity to normal cell line in vitro, but it had anti-cancer activity against A375 cancer cell line and showed high significant against it within IC50 of (92.48, 78.16) for free and immobilize elastase respectively. High Content Screening provides multiparametric study of chemical toxicity at the cancer cells. In addition, immobilized elastase was low cytotoxicity when used it in vivo.