In this study, chemical composition and in vitro antibacterial and antifungal effects of essential oils extracted from fruit peel of Citrus aurantium L. grown in West Anatolian ecological conditions, were investigated. Antibacterial and antifungal activities were determined with the use of Minimum Inhibitory Concentration (MIC) method. Essential oils were obtained in a clevenger apparatus and their compositions were analysed by Gas Chromatography-Mass Spectrometry (GC-MS). According to the GC-MS results, dl-limonene (72.51%) and hexasiloxane (13.28%) were the major components in the essential oil obtained from C. aurantium Central population. Limonene (77.27%) and hexasiloxane (6.19%) dominated in the samples originated from Germencik region. Limonene (79.77%) was found to be in the highest concentration in the essential oil obtained from C. aurantium Koçarlı population. Limonene (95.70%) followed by β‑myrcene (0.76%) were the major components of the essential oil obtained from Citrus aurantium Nazilli population. MIC values ranged from 2.5 to 5 µL/mL for gram (−) bacteria and from 0.3125 to 5 µL/mL for gram (+) bacteria. Essential oil from Germencik region generally showed the best effect against fungi. Essential oil obtained from Citrus aurantium fruit peel could be an alternative to synthetic antimicrobial agents.