The chemical composition and the biological activities of Mentha pulegium leaves extracts were evaluated. The main components of M. pulegium EO were menthone (41.7%), cis‐isopulegone (31.71%) and isomenthone (15.03%). The methanol extract of M. pulegium was rich in phenols and flavonoids. The EO showed much better antioxidant activity than the methanol extract against the DPPH and ABTS radicals scavenging. The EO showed strong antibacterial and antifungal activities against 10 tested microorganisms (Escherichia coli, Salmonella typhimurium, Staphylococcus aureus, Pseudomonas aeruginosa, Aeromonas hydrophila, Listeria monocytogenes, Bacillus cereus, Aspergillus niger, Aspergillus flavus, Candida albicans) tested whereas the methanol extract almost remained inactive. The IZ diameters and minimal inhibitrice concentration values for tested microorganisms were in the range of 15–30 mm and 0.05–0.8% (v/v), respectively. Future studies should focus on to determine the usage ability of M. pulegium EO in foods as natural preservative in effective dose. Practical Applications The replacement of synthetic additives by natural active compounds fosters research on the screening of medicinal plants as sources of new antioxidants. Moreover, such compounds can be used for extension of product shelf life by inhibiting the growth of foodborne spoilage bacteria. This study focused on the evaluation of biological activities of M. pulegium EO because of its application in food industries as a reasonably safe agent. Thus, this species might be a good candidate for further investigation in developing new antioxidants, as well as preservatives in food against the well‐known causal agents of foodborne diseases and food spoilage, particularly E. coli, S. typhimurium, L. monocytogenes, P. aeruginosa and S. aureus.