The chemical composition, antimicrobial, and antibiofilm activities of the essential oil from the fruits of Turkish endemic Prangos hulusii were studied. Essential oil was analyzed by GC and GC/MS. Thirty-nine compounds were identified, which represent 98.6% of the total composition of the essential oil. Monoterpene hydrocarbons and sesquiterpene hydrocarbons were the most represented classes of volatiles with 49.8% and 21.0%, respectively. The main constituents of the essential oil were (E)-methyl cinnamate (22.9%), Î±-phellandrene (13.8%), and (E)-Î²-ocimene (12.9%). (E)-methyl cinnamate was detected in Prangos spp. for the first time. Antimicrobial activities against three Gram-positive bacteria, three Gram-negative bacteria, and three yeast strains were evaluated using disk diffusion test and microdilution method. Antibiofilm experiments were done by spectrophotometric microplate method and the results of Staphylococcus aureus, Escherichia coli, and Candida albicans were visualized via scanning electron microscopy. The essential oil had an antimicrobial effect and showed an antibiofilm effect on these strains.