A novel killer toxin produced by yeast Metschnikowia pulcherrima was purified and added into ready to cook meatballs to enhance their microbial safety and extension of their shelf life. The agent was added into ready to cook meatballs at two different concentrations (1%-K1 and 2%-K2). The results of those two groups were compared to the control group (K0) lacking the killer toxin. Physical, chemical and microbiological analyses were carried out in meat dough and all analyses were repeated at two day intervals during 10 day-storage at +4 ?C. Addition of inhibitor compound in meat dough decreased the numbers of total aerobic mesophillic bacteria, yeast and molds and lactic acid bacteria. Staphylococci/Micrococci, coliform bacteria and total psychrotrophic bacterial counts of the samples were determined as well. Results showed that all indicators of microbial deterioration were found to be higher in K1 group than K2 group, revealing that there was an inverse correlation between the concentration of killer toxin and the number of microorganisms causing spoilage. In addition to 1 log decrease in the number of microorganisms in toxin added groups, the high TBARS values of the control group also showed the effectiveness of the toxin. Toxic effect analysis results showed that the killer toxin had no toxic effect on L929 mouse fibroblast cells after 24h exposure.