Pathogenic microorganisms posed perniciousness for postharvest fruits and vegetables, as well as brought potential risks for human health. In this work, pullulan/polyvinyl alcohol (PUL/PVA) nanofibers incorporated with thymol-loaded porphyrin metal-organic framework nanoparticles (THY@PCN-224 NPs) were developed for antibacterial food packaging. PCN-224 MOFs not only act as thymol loading carriers but also highly produce singlet oxygen (1O2) with bactericidal activity. PUL/PVA nanofiber was a promising sustainable substrate because of its good flexibility, biocompatibility and biodegradability. The loading capacity of PCN-224 for thymol was about 20%. The THY@PCN/PUL/PVA nanofibers exhibited synergistic antibacterial activities against E. coli (~99%) and S. aureus (~98%) under light irradiation. The cell viability assays and fruit preservation study demonstrated good biosafety of the polymeric film. The results suggested that this novel nanofiber has potential application prospects for food packaging. Display omitted •Pullulan/polyvinyl alcohol (PUL/PVA) biopolymer nanofibers were fabricated.•PUL/PVA nanofibers showed good availability, biodegradability and biosafety.•PCN-224 has photosensitive bactericidal activity under light irradiation.•Thymol loaded into PCN-224 nanoparticles achieved synergistic antibacterial activity.•Polymer-based THY@PCN@PUL/PVA nanofibers have potential in active packaging.