This paper examines the synthesis of gamma-butyrolactone compounds 2, 3, 4, 5, and 9, and their potential antimicrobial nature and ability. The structures of all the compounds mentioned above were determined and confirmed with elemental analyses, and IR, H-1-NMR, and C-13-NMR spectroscopy. The in vitro antimicrobial activity of compounds 2, 3, 4, 5, and 9 was examined and evaluated against the following pathogens: Enterobacter aerogenes, Enterobacter cloacae, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus agalactiae, Aspergillus niger, and Candida albicans. Compound 5 (20 mg/mL) demonstrated high antimicrobial activity against S. epidermidis, which was closely comparable to the antimicrobial activity of streptomycin.