The aerial parts of Satureja metastasiantha were hydrodistilled for 3 h using a Clevenger-type apparatus. The essential oils were analyzed by gas chromatography/flame ionization detector and gas chromatography/mass spectrometry, simultaneously, the main compounds of which were characterized as p-cymene (22.3%), thymol (21.0%), carvacrol (18.45%), and gamma-terpinene (12.1%). Antioxidant capacity, acetylcholinesterase and butyryl-cholinesterase inhibition effects, and antimicrobial and antifungal properties of the species were evaluated. The anticholinesterase activity of the essential oil of S. metastasiantha was observed with 30% inhibition at 200 mu g/mL. The essential oil of the species showed activity against Staphylococcus aureus with 128 mu g/mL minimum inhibitory concentration value.