In this study, electrospinning technique was employed to fabricate eugenol loaded gelatin nanofibers (ELGNs) by using gelatin as an active material to encapsulate eugenol. The results indicated that eugenol could be successfully encapsulated within nanofibers, as verified by SEM, FTIR, and zeta potential analyses. The average zeta potential value of ELGNs was determined to be − 15.08 mV. The average diameter of gelatin nanofibers was enlarged with eugenol encapsulation, increasing to diameter of 125 nm. ELGNs had a smooth and uniform fiber morphology observed by SEM. The ELGNs demonstrated a potent antibacterial activity by significant (P < 0.05) reduction in the number of total mesophilic aerobic bacteria counts by 1.62 log and psychrophilic aerobic bacteria counts by 1.33 log in fresh beef samples treated with ELGNs. Moreover, the bioactive ELGNs maintained textural properties (hardness–4.75 N, gumminess–1.75 N, and chewiness–3.87 N × mm at 9th day of storage) and sensory properties, exhibiting advantages as antimicrobial packaging materials that maintained the quality and freshness of the raw meat, along with the extension of its shelf life to 9 days. Significant (P < 0.05; 0.01) negative and positive correlations were found among the microbiological, textural, and sensory parameters, which explained and confirmed the reason for the effectiveness of the nanofiber treatment. In conclusion, our results suggested that the electrospinning approach in this study could be an effective technique to encapsulate eugenol within gelatin nanofibers and the electrospun ELGN mats could be a novel bio-degradable active packaging material with good antibacterial effects.