This study focuses on the extraction of phenolic compounds from the fermentation of Phanerochaete chrysosporium and Gloeophyllum trabeum. The main goal was to synthesize phenol/chitosan microspheres and PVA films and characterized using FTIR, TGA, DSC, SEM, and mechanical tests to evaluate their physical, chemical, and mechanical properties for antimicrobial packaging applications. Homogeneous chitosan microspheres loaded with lignin-derived phenols were obtained, showing controlled release of antimicrobial compounds. The incorporation of phenolic microspheres into PVA/chitosan films resulted in significant improvements in mechanical properties: the films exhibited an elastic modulus of 36.14 ± 3.73 MPa, tensile strength of 12.01 ± 1.14 MPa, and elongation at break of 65.19 ± 5.96%. Thermal tests revealed that chitosan-containing films had enhanced thermal stability, with decomposition temperatures (T10) reaching 116.77 °C, compared to 89.28 °C for pure PVA. In terms of antimicrobial activity, PVA/chitosan/phenol films effectively reduced Lactobacillus growth and milk acidity, maintaining quality for up to 96 h at room temperature, outperforming controls with acetic acid and H2O2. The films also inhibit yeast growth for one week. In conclusion, phenols can be effective antimicrobial agents in dairy, but their use should be monitored. Additionally, PVA/chitosan-phenol films offer biodegradability, antimicrobial properties, and sustainability for diverse applications.