Aspergilli are the main causal agent of invasive apergillosis chest disease. A total of 80 isolates of Aspergillus flavus (20), A. fumigatus (15), A. niger (30) and A. terreus (15) isolates, previously recovered from aspergillosis suspected patients at Assiut university hospitals, were assayed for their enzymes and toxins profile. The results revealed that, all of the tested isolates had the ability to utilize calf lung tissue in addition, to produce catalase, peroxidase. Meanwhile, 82.5-90% isolates produced protease, lipase, urease, phospholipase whereas, 70% of isolates exhibited hemolytic activities. TLC of the cleaned extracts of the tested isolates exhibited the ability of all A. flavus isolates assayed to produce aflatoxins B1 & G1, 53%, moreover, A. fumigatus isolates produced fumagillin and gliotoxin. On the other side, 43.3 % & 23.3% of A. niger isolates produced ochratoxins & gliotoxin respectively. Virulence assay of 10 μl of gliotoxin standard and cleaned extracts of A. fumigatus toxic isolates showed necrotic area on Guinea Pigs lungs compared to the control. Thus, opportunistic fungi isolated from aspergillosis patients possess high enzymatic and toxic profile that might play an important role in their mycopathy.