Herbal medicines are defined as traditionally used natural products. The current study in an attempt try to investigate the antibacterial activity on extended spectrum beta-lactamases (ESBL), methalo beta-lactamases (MBL) producing gram negative bacteria and methicillin resistant Staphylococcus aureus (MRSA) and subsequently, to determine the antioxidant activity of Thymus daenensis. For this reasons, firstly cytotoxicity of T. daenensis was determined by MTT assay. Then, essential oil was subjected for antibacterial and antioxidant activity. Our results demonstrated that 15 mg/ml concentration of T. daenensis inhibited both P. aeruginosa producing MBL and E.coli producing ESBL, while this value was 25 mg/ml concentration for MRSA inhibition. The association between phenolic compound and antioxidant activity was found for the ABTS(•+) method (43.52%) in the lowest level, while, for FRAD and DPPH(•) methods the opposite story occurred (70.5% correlation for DPPH(•) and 50.9% for FRAD). Our findings suggested that T. daenensis has a potential herbal medicine that should be considered as an antibacterial and antioxidant with very low toxicity.