<p>In the past few years, the effective application of waterborne polyurethane (WPU) in the automotive industry, paint, packaging, and many other industries has drawn the attention of researchers to the synthesis of new WPU nanocomposite through structural modification and additive incorporation. In this research, layered double hydroxide (LDH) compounds were intercalated with glycine (Gly) to obtain the multifunctional nanoplates. The nanoplates were incorporated into the WPU matrix to create versatile antibacterial and oxygen barrier films while maintaining the essential characteristics of polyurethane films. The structure, mechanical performance, antibacterial activity, morphology, and water absorption behavior of the resulting nanocomposite films were thoroughly analyzed to assess their suitability for food packaging applications. The results show that LDH and LDH-Gly nanoplates improve the mechanical features of waterborne polyurethane-based films. Additionally, the modified films exhibited superior antibacterial activity compared to pure WPU films, showing increased resistance against both Gram-positive and Gram-negative bacteria. Furthermore, the incorporation of LDH-Gly nanoplates significantly reduced the water uptake and oxygen transmission rate (OTR), indicating improved barrier performance.</p>