Background: To prevent or inhibit the growth of pathogenic microorganisms and food spoilage factors, many studies have been done by using natural preservatives. The aim of study was to investigate the effect of different concentrations of lysozyme and Nisin on the growth rate and also to determine the minimum inhibitory concentration (MIC) and minimum bactericidal cocentratiin (MBC) of these combinations on the bacteria of Escherichia coli, Staphylococcus aureus, Salmonella typhimorium and Listeria monocytogenes. Methods: In this study, various concentrations of lysozyme and Nisin were set in form of alone concentration and in combination concentrations
(0, 19.53, 39.06, 78.13, 156.25, 312.5, 625, 1250, 2500, 5000) in vitro conditions and 6 pH 5.5, 6, 6.5, 7, 7.5, and 8. Microdilution method at 24°C was done and the combined effect on bacteria growth was read by using ELISA reader. Results: The results showed that lysozyme was less effective on Escherichia coli and Nisin was less effective on Listeria monocytogenes. Moreover, combining lysozyme and Nisin at low pH decreased the MIC. Conclusions: The results of the study showed that the effect of combining lysozyme and Nisin on Staphylococcus aureus is above all other bacteria and at low pH reduces the MIC.