Enterococcus faecalis is a facultative anerobic bacterium which can causesecondary periapical infection and is very resistant to numerous antimicrobial substances normally used during the root canal treatment. Earthworm (Lumbricusrubellus) possess antimicrobial peptide, known as Lumbricin-1 which is known tohinder the growth of Gram positive and Gram negative bacteria as well as fungi, but rarely caused resistance. This research was an experimental laboratory study conducted to observe the antibacterial activity of Lumbricin-1 contained in earthworm powder towards the growth of E. faecalis in vitro. Enterococcusfaecalis was cultured on CHROMagar VRE media and incubated anaerobically for24-48 hours in the temperature of 37⁰C. The bacterium was identified by observing the colour of the colony of the bacterium growing on the CHROMagarVRE medium and Gram staining, while antibacterial activity test was performedusing disk diffusion method. Statistical analysis using one way ANOVA and Duncan test showed that there was a significant difference (p< 0, 05) between test and control group. The result of the study showed that earthworm powder possessed strong antibacterial activity towards the growth of Enterococcusfaecalis.