Isolation, purification, and photostimulation of protease enzyme from selected fungal isolates

Protease enzymes play a vital role in biotechnology and industry due to their ability to hydrolyze peptide bonds in proteins. Microbial proteases, particularly those derived from fungi, are favored for their ease of cultivation, high yield, and stability under extreme conditions. Despite the importance of fungal proteases, there is limited data on the comparative production capacities and environmental responsiveness of different fungal strains, especially under photostimulation and varying nutrient conditions. Understanding how these factors influence protease yield is essential for optimizing large-scale enzyme production. This study aimed to isolate, purify, and photostimulate the protease enzyme from selected fungal isolates. It also intended to assess their protease activity, pH and temperature stability, and the influence of various carbon sources and light wavelengths on enzyme productivity. A total of 35 soil samples were collected from five regions in Egypt. Fungal species were isolated, identified, and screened for protease activity. Enzyme activity was quantified under different incubation periods, pH levels, temperatures, carbon sources, and light exposures. Enzyme units (U/ml) were used to evaluate productivity. Five fungal species were identified, with Trichoderma viridi demonstrating the highest enzyme activity and stability across most conditions. Dextrose and red light significantly enhanced enzyme production. Optimal temperature and pH varied by species, with enzyme synthesis declining above 90°C. The study provides comprehensive insights into environmental and nutritional optimization for fungal protease production, highlighting T. viridi as a promising candidate for industrial enzyme synthesis under photostimulated conditions.