Fonsecaea pedrosoi is the major etiologic agent of chromoblastomycosis. The virulence of F. pedrosoi is a meagerly explored phenomenon. The ability to interact with host cells and the production of hydrolytic enzymes are thought to be important virulence mechanisms of fungal pathogens. Here, we measured the production of two distinct lipolytic enzymes, phospholipase and esterase, by three clinical strains of F. pedrosoi isolated from chromoblastomycosis lesions, as well as their capabilities to interact with epithelial cells. All the strains were excellent esterase producers, generating elevated hydrolytic halos after 5 days of growth. Conversely, phospholipase activity was detected only after 10 days, except for the most recent strain of F. pedrosoi (Magé) in which measurable phospholipase activity was detected on day 5. The ability to interact with epithelial cells was also investigated. Regarding the adhesion capability, an indirect connection was observed in relation to the adaptation time of each strain in axenic culture, in which Magé strain showed the best adhesion ability followed by LDI 11428 and 5VPL strains. Both 5VPL and Magé strains were also detected inside the epithelial cells, while the LDI 11428 strain was rarely detected in cytoplasmatic vacuolar compartments. Moreover, these F. pedrosoi strains were able to cause injury in epithelial cells.