Yellow Wasp Polistes flavus venom toxins were isolated and purified on a Sepharose CL-6B 200 column. Purified proteins were investigated for its antibacterial and antifungal activity against 13 infectious microbial pathogens. Paper disc diffusion and serial micro-dilution assays were performed for the determination of inhibition zone (DIZ) diameters and minimal inhibitory concentration, respectively. Triton X-100 (0.1%) proved to be a good solubilizing agent for toxin/proteins. Higher protein solubilization was observed in the supernatant than in the residue, except TCA (tri-chloroacetic acid). The elution pattern of purified and homogenized sting glands exhibited two major peaks at 280 nm in fraction No. 41-61 and 81-101. The total yield of protein was 69.21% and specific activity was determined in each fraction. Molecular weights in protein fractions were ranging from 6-70 kD. MIC (Minimum Inhibitory Concentration) values were 12.3 μg/ml against K. pneumonia 12.3 μg/ml against E. coli and L. acidophilus, 24.6 μg/ml against B. cereus; 49.24 μg/ml against S. aureus and M. luteus. By agar disc diffusion method, the diameter of inhibition zones in mm in presence of yellow wasp toxins is at a concentration range of 98.56-6.9 μg/ml E. coli 18.36 ± 0.14, Bacillus cereus 14.566 ± 0.21, L. acidophilus 18.10 ± 0.21, Micrococcus luteus 18.76 ± 0.19, S. aeurus 17.36 ± 0.43, Klebsiella pneuminiae 19.56 ± 0.21, Salmonella typhi 19.96 ± 0.31, Vibrio cholera 21.83 ± 0.22, Pseudomonas aeruginosa 22.90 ± 0.09, Aspergillus niger 21.66 ± 0.16, Candida albicans 23.33 ± 0.26, Rhizopus stolonifer 24.96 ± 0.16 respectively. The antibacterial and antifungal activity of venom toxin may be due to action on cell membrane, its destruction and cell lysis. The wasp toxins may be used as strong biological agents to control microbes.