Objective: The medicinal value of plants lies in some chemical substrates that produce a definitive physiological action on the human body. Ankaferd comprises a standardized mixture of the plants Thymus vulgaris, Glycyrrhiza glabra, Vitis vinifera, Alpinia officinarum and Urtica dioica. The aim of this study was to assess the in vitro antimicrobial activities of Ankaferd medicinal plant extract. Material and Methods: The antimicrobial activity assay was performed by agar well diffusion to assess the antagonistic activity of Ankaferd against 26 indicator strains including human pathogens and food spoiling gram-negative and gram-positive bacteria. Results: Ankaferd was active against all bacteria tested while nisin, the only commercial bacteriocin for food preservation, was inactive against gram-negative indicator strains. Besides a high inhibitory activity against gram-positive and gram-negative bacteria, including human pathogens and food spoiling bacteria, Ankaferd was more stable than nisin in different heat and enzyme treatments. Conclusion: Antibacterial activity of Ankaferd can be extended to extreme environmental conditions such as the potential use of the preparation for the treatment of infectious diseases and preservation of different type foods from food-born pathogens or food spoiling bacteria. The demonstration of the anti-infective properties of Ankaferd adds a new value to its haemostatic effect in the healing of infected hemorrhagic wounds, as well as opening new avenues for its potential use for anti-infective actions and food preservation.