Abstract: In recent years, tendency to use of natural antimicrobial agents in food industry has increased. Pomegranate peels containing phenolic compounds and anti-microbial agents, are counted as valuable source for extraction of these compounds. In this study, the extraction of pomegranate peel extract was carried out at different ethanol/water ratios (40:60, 60:40, and 80:20), temperatures (25, 40, and 55 ˚C), and time durations (20, 24, and 28 h). The extraction yield, phenolic compounds, flavonoids, and anthocyanins were measured. Antimicrobial activity of pomegranate peel extracts were determined against some food-borne microorganisms such as Salmonella enteritidis, Escherichia coli, Listeria monocytogenes, Staphylococcus aureus, Aspergillus niger, and Saccharomyces cerevisiae by agar diffusion and MIC methods. Results showed that at ethanol/water ratio 60:40, 25 ˚C and 24 h maximum amount of phenolic compounds (349.518 mg gallic acid/g dried extract), flavonoids (250.124 mg rutin/g dried extract), anthocyanins (252.047 mg cyanidin3glucoside/100 g dried extract), and the strongest antimicrobial activity were obtained. All extracts’ antimicrobial activities were demonstrated against every tested microorganisms. Staphylococcus aureus showed the highest sensitivity among the tested microorganisms.