In this study, the water-insoluble cumin seed oil (CSO) loaded gliadin (GN)-nanofibers (NFs) mat was prepared in the presence of ethyl cellulose (EC) and adipic acid (AD) using electrospinning method. The NFs were prepared with mixing 30 % w/v of GN and 15 %w/v of EC at the ratio of 80:20 v/v and 5%w/v of AD were selected as optimum formulation based on electrospinning efficiency and SEM images. In the presence of EC, the spinnability of GN fibers improved. Moreover, in the presence of AD the fibers diameters decreased to 50 % (250 ± 14 nm), and uniformity enhanced in all formulations. AD acted as hydrogen bond cross-linker between mixed biopolymers that led to increase in the viscosity of the polymer solutions. The selected formulation was loaded with the CSO based on 10 %w/v of polymers. Thermal stability of the GN fibers was improved in the presence of EC and AD based on DSC results. The FTIR results confirmed the hydrogen bond crosslinking between polymers and AD. Based on antioxidant activities results, the prepared NFs successfully retarded the release of CSO. CSO loaded NFs showed the strong antibacterial and antifungal activity against Escherichia coli, Staphylococcus aureus, and Aspergillus flavus. NFs had no significant toxicity on the viability of normal human umbilical vein endothelial cell (HUVEC). The results of the current study indicated that the prepared NFs mat may have a great potential for using in active food packaging due to its sustained control release properties for loaded antimicrobial and antioxidant active agent.