T. capitatus is widely used in traditional medicine in Tunisia. The main goal of this study was to evaluate the phytochemical composition, the fatty acids profile, the antioxidant, antibacterial, and antifungal activities as well as the cytotoxic potential of the essential oil (EO) of this plant. The identification and the quantification of the different constituents of the tested EO was determined by gas chromatography-mass spectrometry (GC-MS). Antioxidant activities were evaluated by spectrophotometric methods and chemical tests. HCT 116 cells were used to evaluate the cytotoxic effect of the EO. The microdilution method was conducted to evaluate the antibacterial activity. Poisoned food method was used to test the antifungal activities against fungi species such Aspergillus niger and Aspergillus flavus. The EO presented several components, mainly monoterpenes. Results revealed that T. capitatus EO is not cytotoxic and showed excellent antioxidant activity with a dose dependent manner. Regarding antimicrobial activity, T. capitatus EO was efficient against all tested bacteria and fungi.