Abstract : The purpose of this study was to identify the chemical compound of Citrus lemon (L.) Osbeck. Essential oil (EO) by gas chromatography–mass spectrometry (GC–MS) and to evaluate their antifungal activity and enzyme inhibition (anticholinesterase, anti-tyrosinase, anti-urease, α-glucosidase, and α-amylase). We identified 24 chemical compounds of EO. The major compounds were D-Limonene (74.66%), followed by α-Fenchene (6.16%), γ-Terpinene (5.10%), and α-Citral (3.92%). The antifungal activity against Fusarium oxysporum indicated that the essential oil exhibited good antifungal activity against the tested fungal strain, notably at 100% concentration, by inhibiting the mycelial growth of F. oxysporum by 27.84%. In the urease and α-glucosidase assays, Citrus lemon essential oil had inhibition concentrations (IC50) values of 148.3±1.10 µg/mL and 192.5±1.20 µg/mL, respectively. For anticholinesterase, anti-tyrosinase, and α-amylase, this oil did not show any efficacy against these enzymes. These results suggest a possible use of Citrus lemon essential oil as a potential source of bioactive molecules with anti-urease, anti-glucosidase, and antifungal properties.