This study focuses on the isolation and characterization of bioactive compounds from marine microbial species, specifically targeting their potential as antibacterial agents. A marine sample was collected from Ain Sokhna, leading to the identification of six bacterial isolates (J8-J13) evaluated for their antagonistic activity against seven bacterial pathogens. Among these, Bacillus strain J13 exhibited the highest antibacterial activity, with a zone of inhibition measuring 30 ± 0.1 mm against Bacillus cereus, and notable inhibition against Pseudomonas aeruginosa (28 ± 0.1 mm), Bacillus subtilis (28 ± 0.2 mm), Salmonella typhimurium (27 ± 0.2 mm), Escherichia coli (25 ± 0.2 mm), and Klebsiella pneumoniae (23 ± 0.2 mm). The chemical analysis of J13's metabolites was conducted using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and gas chromatography-mass spectrometry (GC-MS), revealing several bioactive compounds, including bis(2-ethylhexyl) phthalate. The Minimum Inhibitory Concentration (MIC) values ranged from 15.62 to 31.25 µg/ml, indicating the efficacy of these compounds at low concentrations. Advanced biotechnological techniques such as genomic sequencing and metabolomic profiling could further elucidate the mechanisms underlying the antibacterial properties of these compounds, maximizing their potential for therapeutic applications. By integrating these approaches, this research not only contributes valuable insights into combating antibiotic resistance but also paves the way for developing innovative treatments that could significantly impact human health and well-being.