Cytotoxicity of Collagenases and Elastases Purified from Candida Species on Some Carcinoma Cell Lines

The purpose of this work was to assay for the cytotoxicity of collagenases and elastases purified from Candida parapsilosis and Candida krusei on some carcinoma cell lines. Four Candida species and Saccharomyces cerevisiae were tested for their collagenolytic and elastinolytic enzyme activities. C. parapsilosis and C. krusei were proved to be high producers for the two enzymes in the culture filtrates. The four enzymes (collagenases and elastases from C. parapsilosis and C. krusei) were purified to full homogeneity using (NH4)2 SO4 precipitation, anion exchange column chromatography by DEAE- cellulose and gel filtration using Sephadex G100. The molecular masses of the four enzymes were determined by SDS-PAGE in parallel with all purification steps. However, the molecular masses of the single band of the purified enzymes were apparently determined to be, 97.2 and 53.4 KDa for collagenase and elastase in C. parapsilosis, 66.4 KDa and 23.5 KDa for collagenase and elastase in C. krusei. The four enzymes were characterized by studying the effect of temperature, pH, protease inhibitors substrate specificity. Cytotoxicity assay of the four enzymes either in crude or in purified state was done singly or in combination mixtures using three carcinoma cell lines. It was found that larynx carcinoma cell line (HEP2) was the only sensitive cancer cells to proteases treatments, while colon carcinoma cell line (HCT116) and breast carcinoma cell line (MCF7) were not affected by enzyme treatments. Combination mixture of the four enzymes recorded the most potent cytotoxic effect on larynx carcinoma cell line (HEP2) with minimum value of IC50 (3.1 Uml-1)