Brazilian flora biodiversity opens possibility of studies with natural extracts for bactericidal and biocompatible solutions for use in endodontic therapy. The purpose of this study was to investigate the antimicrobial effect of Punica granatum L. (pomegranate) against pathogens of infections from root canal system. Ethanolic solutions (50% ethanol and 50% water) were prepared in 100 mg/mL of lyophilized extracts derived from the peel, seed and leaf of pomegranate, against the following oral microorganisms: Escherichia coli, Staphylococcus aureus, Streptococcus mutans, Enterococcus faecalis and Candida albicans. The groups were distributed according to the extract tested for root canal irrigation: G1 - 50% ethanol (positive control), G2 - 1% sodium hypochlorite (NaOCl) (negative control), G3 - Solution of the peel, G4 Seed solution, G5 - Solution of the leaf. The antimicrobial capacity of the extracts was evaluated by the agar diffusion test, minimum inhibitory concentration (MIC) and minimum bactericidal / fungicidal concentration (MCB/MFC). The data were analyzed by ANOVA (extracts and microorganisms) and Tukey test (p<0.05). By the diffusion test in agar, it was verified that, independently of the microorganism, the largest inhibition zone was found for the positive control (30.76 ± 4.73 a), followed by peel and leaf, which did not presented significant difference between them (23.32 ± 3.65 and 21.08 ± 2.28 b, respectively). The seed had the lowest of microbial inhibition zone (17.83 ± 6.92 c) (p<0.05). In the isolated comparison for each microorganism, peel and leaf extracts presented antimicrobial activity similar to NaOCl for S. mutans (p = 0.0000), besides having a good antimicrobial effect on the other microorganism. For the E. faecalis and E. coli, the extracts (peel, leaf and seed) had similar inhibition zones (p>0.05), but lower than those obtained by the positive control. Regarding MIC and MBC/MFC, it was found a bactericidal and bacteriostatic effect for all extracts. The P. granatum peel extract had greater inhibition zone against C. albicans (MIC = 6.25 mg/mL and MFC = 12.5 mg/mL), followed by S. aureus (MIC and MBC = 12.5 mg/mL). It may be concluded that the hydroalcoholic extracts of P. granatum L. presented antimicrobial activity against oral pathogens common in endodontic infections, with the highest activity for those obtained from the peel and leaf