This study aimed to develop innovative micro-nano fibers loaded with lemon peel essential oil for food packaging via centrifugal spinning. Lemon peel essential oil (LPO) was extracted from dried lemon peel by hydrodistillation. The major one of the 16 components of the extract detected by GS-MS was limonene (60.4%). Gelatin fibrous mats loaded with three different concentrations of LPO were fabricated with centrifugal spinning and then crosslinked. Micro-nanofibers were characterized in encapsulation efficiency, morphological, chemical, thermal, hydrophobicity and microbiological aspects. The in-vitro evaluation showed that the effect of fibers on grampositive Staphylococcus aureus (S. aureus) ATCC 29213, especially due to the antimicrobial activity of limonene, was greater than that of gram-negative Escherichia coli (E.coli) ATCC 35218. This effect was also consistent with the in-situ evaluation in which micro-nano fibers were applied to contaminated cheeses. Moreover, LPOloaded gelatin centrifugal spun positively affected the shelf life by suppressing the growth of aerobic mesophilic bacteria and yeast molds that cause spoilage in cheese.