The present study was aimed to evaluate the antimicrobial activities of bioactive compounds synthesized from vermicast isolated actinomycetes species. Specifically, the synthesized bioactive compounds were evaluated for their antimicrobial activity against selected Gram + ve and Gram - ve human pathogenic bacteria including Staphylococcus aureus, Bacillus circulans, Bacillus subtilis, Pseudomonas aeruginosa, and Escherichia coli. Interestingly, a total of five different actinomycetes species were recovered from vermicasts. More interestingly, among these potential actinomycetes species, the bioactive compounds synthesized by isolate AS9 showed a significant antibacterial activity and its mean zone of inhibition was found at 11.3 ± 1.6 mm, 9.5 ± 0.91 mm, and 9.9 ± 1.71mm against S. aureus, B. subtilis, and B. circulans, respectively. Furthermore, according to antibacterial activity and spectrum broadness, three of the actinomycetes strains were selected and characterized by conventional methods. Subsequently, the bioactive compound profiling of these isolated actinomycetes strains performed through GC-MS analysis indicating the presence of the bioactive compounds including 3, octadecene (E), behnic alcohol phenol, 2,4-bis(1,1-dimethyl ethyl) 1-nonadecene, 1-heneicosanol, milbemycin 3-eicosene (E), and 1-docosanol.