In this study, the chemical composition and biological activity ofAllium scorodoprasumsubsp.jajlae(Vved.) Stearn were investigated for the first time, focusing on its antimicrobial, antioxidant, and antibiofilm properties. A GC-MS analysis was employed to evaluate the composition of its secondary metabolites, identifying linoleic acid, palmitic acid, and octadecanoic acid 2,3-dihydroxypropyl ester as the major compounds in ethanol extract. The antimicrobial activity ofA. scorodoprasumsubsp.jajlaewas assessed against 26 strains, including standard, food isolate, clinical isolate, and multidrug-resistant ones, as well as threeCandidaspecies using the disc diffusion method and the determination of the minimum inhibitory concentration (MIC). The extract showed strong antimicrobial activity againstStaphylococcus aureusstrains, including methicillin-resistant and multidrug-resistant strains, as well asCandida tropicalisandCandida glabrata. Its antioxidant capacity was evaluated using the DPPH method, revealing a high level of antioxidant activity in the plant. Additionally, the antibiofilm activity ofA. scorodoprasumsubsp.jajlaewas determined, demonstrating a reduction in biofilm formation for theEscherichia coliATCC 25922 strain and an increase in biofilm formation for the other tested strains. The findings suggest potential applications ofA. scorodoprasumsubsp.jajlaein the development of novel antimicrobial, antioxidant, and antibiofilm agents.