Satureja calamintha ssp. nepeta (L.) Briq. is a medicinal and aromatic plant used for long time in Algerian traditional medicine. This study was carried out to determine physical characteristics (density and refractive index), total phenolic content (Folin-Ciocalteau assay), antioxidant (DPPH free radical scavenging method) and antimicrobial (disc diffusion assay against Staphylococcus aureus (ATCC 29273), Pseudomonas aeruginosa (ATCC 27853) and Candida albicans (ATCC 10231)) activities of essential oil of Satureja calamintha spp. nepeta (Lamiaceae). The essential oil of aerial parts of S. calamintha was isolated by the hydrodistillation method. Concerning the physical characteristics, essential oil gave a value of density of 0.910 and revealed an refractive index of 1.480. The total phenolic content was in the order of 219±0.06 mg Gallic Acid Equivalents per g of sample. This oil had antioxidant power with a percentage of inhibition of the free radical DPPH (91.81 %) at 500 µg mL-1. Furthermore, the antimicrobial results revealed that this oil exhibit antimicrobial activity with minimum inhibitory concentration that ranged from 0.09 to 1.56 µL mL-1. Overall, the results obtained are promising and open up new perspectives in the field of natural applications which can be used in the food, pharmaceutical and cosmetic industries.