Different films were synthesized from starch or polysaccharides extracted from distillers dried grains with soluble (DDGS) in combination with different percentages of linear polyethyleneimine (PEI) hydrochloride polymer to assess the mechanical and antimicrobial properties of the resulting composites. Moreover, a simple method for the extraction of the polysaccharide content from DDGS is reported. The materials obtained were characterized by ATR-FTIR, NMR, and XPS spectroscopy, swelling capacity, and by organic elemental analysis. In particular, the stability of the film prepared with only DDGS in copper ion solutions was improved by the incorporation of PEI. 13C HRMAS NMR studies evidenced the incorporation of the PEI polymer in the new films. Moreover, the release of PEI molecules from the films was studied by 1H NMR experiments in D2O to explain the antimicrobial properties of the PEI-based films against Staphylococcus aureus, with the DDGS-10% PEI films being the most active surface. Furthermore, the incorporation of copper ions into the different films enhanced their antimicrobial activity. Additionally, the starch-10% PEI film exhibited good swelling capacity in deionized water (~1500%), which decreased with the addition of salts (~250%). Instead, the DDGS-10% PEI film showed low swelling capacity in deionized water (~80%), with this capacity increasing with the addition of salts (~250%). The mechanical properties of the films improved considerably when 3% PEI was used.