Phospholipase A2 (PLA2) represents a major venom component of snakes and bees and exhibits a broad range of biological effects including myotoxicity, neurotoxicity, hemolysis, cardiotoxicity, anticoagulant and antiplatelet activities. Melittin, is the main component and the major pain producing substance of honeybee venom. The aim of the present study was to differentiate between snake and bee venoms using Aqueous Olive Leaf Extract (AOLE) employing fluorescence techniques. Tryptophan, which is contained in both snake and bee venoms is fluorescent at UV wavelength and hence widely used as a tool to monitor conformational changes in proteins and to draw inferences regarding local structure and dynamics. Fluorescence spectroscopy and molecular modeling have been used to analyze enzyme activity in the absence and presence of AOLE and to verify potential binding of AOLE components to the enzyme. Changes in the fluorescence intensities with blue and red shifts were obtained with bee and snake venoms, respectively. Binding of AOLE constituents near the active site of the enzyme could be evidenced and possible modes of interaction are discussed. The fluorescence method used was rapid and sensitive and was able to differentiate between snake and bee venoms utilizing AOLE.