Abstract: In the present study fungal colonies were isolated from different protein sources by enrichment culture technique and the fungal isolates were screened for protease activity. Isolate G, the most potent producers of protease were identified as Aspergillus funiculosus G. Smith. The isolate G was found to show highest protease production (activity) in an optimized medium (Yeast extract 0.5%, lactose 0.1% and KNO3 1%) at pH 7.0, temperature 37°C and 5 days of incubation period. The crude enzyme of the isolate G was found to show its highest activity at pH 7.5 and temperature 35°C in enzyme substrate reaction. The molecular weight of enzyme of the isolate Aspergillus funiculosus (G) was found 51 kDa, which was determined by SDS-PAGE technique.