Antimicrobial resistance (AMR) is a major global health concern. Virgin coconut oil (VCO) shows potential as a natural antimicrobial due to its medium-chain fatty acids, including lauric, caprylic, and capric acids. This study optimized bromelain-based VCO production by adding Ca2+ and Mg2+ ions (as CaCl2 and MgCl2) and evaluated its antimicrobial activity. The optimal bromelain concentration (8% b/v) produced VCO with a 32.05% yield and good-quality parameters, including moisture content, free fatty acids (FFA), peroxide value, iodine value, and lauric acid content. The addition of 2.0 mM Ca2+ increased the VCO yield to 39.89%, while 1.5 mM Mg2+ yielded 36.58%; each effector also improved VCO quality during bromelain-assisted extraction. These results indicate that the addition of CaCl2 and MgCl2 effectors effectively enhanced bromelain activity, resulting in high-quality VCO. The enzymatic VCO exposed inhibition zones of 6.98, 4.58, and 3.37 mm to Staphylococcus aureus, Escherichia coli and Candida albicans, respectively, while VCO from heating extraction appeared to have inhibition zones of 5.19, 3.61, and 3.01 mm. Meanwhile for antibiotic control using chloramphenicol for S. aureus and E. coli showed inhibition zones of 14.01 and 17.98 mm, respectively, then fluconazole antibiotic for Candida albicans gave an inhibition zone of 5.18 mm. Although less potent than antibiotics, enzymatic VCO demonstrated enhanced antimicrobial potential and quality.