The present study aimed to produce bio-active packaging materials made of whey proteins (WPs) and essential oil (EO) extracted from Thymbra (Satureja capitata, L.), one of the most popular Palestinian wild plants. In this study, two different Thymbra leaves from Nablus and Qabatiya in Palestine were collected and analyzed for EOs by gas chromatography and mass spectrometry. Based on the analysis, two EOs, namely, TEO1 and TEO2, were extracted, and it was found that both samples primarily contain γ-terpinene and carvacrol, whereas p-cymene was detected only in TEO1. The antimicrobial activity of TEO1 and TEO2 was evaluated by microbroth microdilution assays against pathogenic bacteria and yeast. Based on the results, TEO1 exhibited potent antimicrobial activity against the test strains. Besides, TEO1 was chosen to functionalize WP-based films at different concentrations (0.1%, 0.4%, and 0.8% v/v of Film Forming Solutions). Film mechanical property investigation showed a marked reduction in the tensile strength and Young’s modulus at 0.8% TEO1. In contrast, its elongation at break value was significantly (p < 0.05) increased due to the plasticizing effect of the EO. Moreover, the film transparency was found to be significantly (p < 0.05) reduced by increasing TEO1 concentrations. Finally, microbiological investigations indicated that film antimicrobial activity against both gram-positive and gram-negative bacteria increased dose-dependently. The overall results open interesting perspectives for employing these films as preservative materials in food packaging.