<p>This study successfully prepared polylactic acid (PLA) composite films containing <i>Litsea cubeba</i> oil (LCO, 9 wt%) and different concentrations of nano-TiO₂ (0 wt%, 1 wt%, 2 wt%, and 3 wt%), and evaluated their efficacy in preserving fresh-cut jackfruit. The results showed that LCO and nano-TiO₂ could be successfully incorporated into the PLA matrix, which improved the composite films’ structure, flexibility, water and UV resistance, and antioxidant and antimicrobial activities. Notably, the composite film with 2 wt% nano-TiO₂ content exhibited the best overall performance among all the packaging materials. Experiments on fresh-cut jackfruit preservation revealed that the composite films suppressed the proliferation of microorganisms like total bacterial count, mold and yeast, and to varying degrees, inhibited their growth and slowed down the quality deterioration of fresh-cut jackfruit. In addition, composite film with 2 wt% nano-TiO₂ content was the most effective in inhibiting microorganisms, respiratory rate, enzyme activity, and malondialdehyde (MDA) content as well as in maintaining weight loss, color, firmness, flavor, and vitamin C (VC) content, ultimately extended the shelf-life of fresh-cut jackfruit from 6 to 15 days at 10&#xa0;°C. This work demonstrates the feasibility of bioactive PLA films as safe antimicrobial packaging for tropical fruits.</p>