Keratin-rich byproducts from the poultry, textile, and leather industries pose a significant challenge for sustainable waste management due to their highly recalcitrant nature. While microbial degradation may offer a viable solution, the mechanisms underlying keratin breakdown remain largely unexplored. In this study, we employed a high-resolution proteogenomic approach to characterize the keratinolytic machinery of Onygena corvina, a non-pathogenic saprophytic fungus. Using a membrane agar plate method with insoluble substrates, we obtained secretomes enriched in secreted and substrate-bound proteins during growth on α- and β-keratin-rich substrates, specifically wool and feather meal, as well as on casein (as control) at days 1, 2, and 3.