The Biological Activity of the Protease Enzyme Extracted from Basidiomycete Inocutis tammaricis: Antibacterial, antioxidant, and antitumor roles of naturally produced protease enzyme by Inocutis tammaricis

The basidiomycete Inocutis tamaricis is a medicinal fungus belonging to the family Hymenochaetaceae. This study evaluates the biological activities of a protease enzyme extracted from I. tamaricis, focusing on its antimicrobial, antioxidant, and antitumor potential. Antimicrobial activity was assessed using the agar well diffusion method against several pathogens at concentrations of 25, 50, and 100 μg/ml. The results demonstrated that the enzyme has exclusive inhibitory activity against gram-positive bacteria, specifically Enterococcus faecalis and Staphylococcus aureus, but shows no activity against gram-negative bacteria or yeasts. Notably, E. faecalis showed greater sensitivity to the enzyme than S. aureus at all tested concentrations. Antioxidant capacity was evaluated via the DPPH radical scavenging assay. The enzyme showed a dose-dependent response, achieving maximum effectiveness (74.88%) at 200 μg/ml, with activity decreasing proportionally at lower concentrations. Furthermore, antitumor assessments were conducted on the breast cancer cell line (MCF-7) and a normal human dermal fibroblast line (HdFn). The findings revealed that the concentration of 400 μg/ml resulted in the lowest cell viability for both lines (40.5% for MCF-7 and 75.7% for HdFn). The calculated IC50 values were 118.6 μg/ml for the cancerous MCF-7 cells and 781.7 μg/ml for the normal HdFn cells. These results indicate that the protease enzyme from I. tamaricis possesses significant antioxidant properties and selective antitumor activity with low toxicity to normal cells, highlighting its potential for pharmaceutical applications.