Abstract : The bacteriocin activity test using proteolytic enzymes aims to confirm the test, namely to ensure that the antibacterial substance produced by Lactobacillus plantarum is bacteriocin. As an antibacterial, bacteriocins have properties that can be degraded by proteolytic enzymes. The parameters measured in this study were the inhibition zone test on the indicator pathogenic bacteria Escherichia coli, Salmonella typhi. and Staphylococcus aureus. The analysis of the research data was the analysis of Completely Randomized Design (CRD). The results showed that increasing storage time had a significant effect (P<0.05) on bacteriocin activity (inhibition zone diameter) against pathogenic bacteria Escherichia coli, Salmonella typhi. and Staphylococcus aureus. Optimum fermented chicken egg bacteriocin activity at room temperature storage for 96 hours can be seen from the formation of clear zones on the indicator medium. The data obtained showed that Escherichia coli and Salmonella typhi were sensitive and had larger inhibition zones than Staphylococcus aureus. Based on the inhibition zone test, inhibition was greater for Gram-negative bacteria (Escherichia coli and Salmonella typhi) when compared to Gram-positive bacteria (Staphylococcus aureus).