We investigated the antioxidant and antimicrobial potential of sequentially extracted onion polysaccharide fractions namely HBSS, CHSS, DASS and CASS. The different antioxidant assays indicated that ACLPs exhibited potentially appreciable antioxidant activity in a dose-dependent manner. Among all the fractions, CHSS rendered the highest antioxidant action towards ABTS radical cations (97.52%), Fe2+ chelating (98.94%) and superoxide anion radical scavenging (76.27%). Whereas, HBSS possessed the highest potential for DPPH radicals (93.68%), hydroxyl radicals (65.12%) as well as for reducing power (0.559). CASS exhibited the highest lipid peroxidation inhibition (86.43%), while, DASS showed the best β-carotene bleaching inhibition (92.26%). Furthermore, regardless of the bacterial strain, DASS represented the strongest antibacterial activity on the basis of largest inhibition zone, the lowest minimal inhibitory concentration and maximum inhibition of bacterial growth in liquid medium. Overall results indicated that ACLPs hold a promise as potential natural antioxidant additives and antimicrobial agents for formulating the functional foods with potential applications in the medical and food industries.