The release of proteolytic enzymes with elastinolytic and collagenolytic activity was investigated in 10 different isolates of Paracoccidioides brasiliensis. The study encompassed both the mycelial and the yeast phase of the fungus. In the mycelial phase, secretion of elastinolytic and collagenolytic enzymes was demonstrated in 70% and 80% of the isolates, respectively. None of the isolates in the yeast phase exhibited elastinolytic activity, but 90% showed collagenolytic activity. These results demonstrate that the dimorphic fungus P. brasiliensis has the ability to release proteolytic enzymes in both growth phases, although with apparently different specificities.