Purification and comparison of intracellular proteinase A in Candida spp. isolates from Malaysian and Iranian patients and infected mice

摘要

Objective. - To compare the specific intracellular proteinase A activity in clinical isolates of Candida species isolated from Iranian and Malaysian patients, the blood and kidneys of mice infected by Candida cells isolated from these human patients.Patients and methods. - The intracellular proteinase A was extracted using glass beads and ultracentrifugation from yeasts cells and purified by ion-exchange chromatography (DEAE-cellulose), followed by affinity chromatography (ConA-agarose). Purity of proteinase A was controlled by SDS-PAGE and its identification was realized by western blot. Enzyme activity was performed using azocasein as substrate.Results. - Intracellular proteinase A enzyme activity was higher in Candida albicans (C. albicans) than in non-albicans Candida isolates from Malaysian and Iranian patients, mice blood and mice kidneys (P < 0.05). In clinical Candida spp. from human patients, proteinase A activity was significantly higher in Malaysian samples than in their Iranian counterparts (P < 0.05). However, Candida spp. isolates obtained from blood and kidneys of mice infected by human clinical strains of Candida spp. showed no significant differences in proteinase A activity between Malaysian and Iranian samples (P > 0.05). On the other hand, in both clinical and control yeast isolates obtained from Malaysian and Iranian patients, significant differences were found between systemic and non-systemic samples (P < 0.05) but this difference was not observed in mice blood and kidneys.