This study is designed to evaluate the antibacterial potential of various extracts of mature green fruits of Artocarpus chama with the identification of bioactive compounds. The antibacterial efficacy was tested against eight pathogenic bacteria using standard antibiotics as the positive control. Antibacterial bioassay was performed by measuring zone of inhibition, Minimum Inhibitory Concentrations (MIC), and Minimum Bactericidal Concentrations (MBC). The most effective extract was further subjected to preliminary phytochemical tests, Thin Layer Chromatography (TLC), and Fourier Transform Infrared analysis. Ethyl acetate extract (EAAC) was found to be the most effective extract producing an inhibition zone of 45.50 ± 0.50 mm against Pseudomonas aeruginosa MTCC 2453. Effective spots (ES) (Rf=0.37) of EAAC separated by TLC analysis were subjected to indirect contact bio-autography for evaluating their efficacies against the tested bacterial strains. Values of MBC/MIC for the extract ranged from 1.20 to 1.80 (<2) indicating their bactericidal property. Gas Chromatography-Mass Spectroscopy analysis of ES was performed for the identification of bioactive antibacterial compounds. Alkaloids, saponins, steroids, terpenoids, and an array of functional groups in EAAC were detected. MICs of ES ranged from 6-10 µg/ml. Four bioactive compounds in ES were detected in GC-MS analysis. Thus, this study revealed that ethyl acetate extract of A. chama fruits contained several bioactive compounds with promising bactericidal properties against tested strains of both Gram-positive and Gram-negative pathogenic bacteria. © 2022 DSR Publishers/The University of Jordan. All Rights Reserved.