The current study describes the phytochemical profile, antimicrobial, mutagenic, and antimutagenic activity of Perovskia atriplicifolia Benth. essential oil, collected in Pakistan. The sample of essential oil was obtained from aerial parts of the plant by hydrodistillation and analyzed by gas chromatography mass spectrometry. From the 18 compounds identified, the major compounds were camphor (28.91%), limonene (16.72%), alpha-globulol (10.21%), trans-caryophyliene (9.30%), and alpha-humulene (9.25%). Antimicrobial activity of the oil was evaluated using agar diffusion method and agar dilution method. The antimicrobial test results showed that the oil had a significant potential antimicrobial activity against 10 bacteria and 5 fungal strains. Furthermore, the mutagenic and antimutagenic activity of the oil was investigated through the Salmonella/microsome test system, with and without S9 metabolic fraction in Salmonella typhimurium TA98 and TA100. None of the tested concentrations of oil was found mutagenic. However, all tested concentrations did show an increase in antimutagenic activity with or without S9 fraction against 2-aminofluorene and daunomicina, but not sodium azide. Results presented here suggest that the essential oil of P. atriplicifolia possesses antimicrobial properties and is therefore a potential source of antimicrobial ingredients for the food and pharmaceutical industry. In addition, that it also has antimutagenic activity raises the importance of this essential oil in this area.