Biodegradable films based on polymers from renewable resources have become a feasible technology to preserve the quality (texture, color, flavor) and safety of food. The addition of antimicrobial agents to films can prevent the growth of pathogenic microorganisms that affect meat and poultry products. In this study, a biodegradable film with sodium alginate (SA), taro starch (MS), and nisin (Nis) was optimized to have high tensile strength (TS), breaking force (BF), and a low water vapor permeability (WVP) using a Box-Behnken response surface design, and its antimicrobial effect was evaluated in relation to its use as a packaging material for chicken meat. The OB was characterized via analysis of its mechanical, physical, and chemical properties; in addition, the total migration of Nis was also analyzed, along with its retention ability, the kinetics of the release of Nis into food simulants, and its antimicrobial activity against Listeria monocytogenes in vitro and on inoculated chicken meat. The resulting optimal OB was produced with 1.9% MS, 1% glycerol (G), and 2,369 IU/mL of Nis, and displayed adequate TS and WVP. The OB significantly reduced the microbial load and helped extend the shelf life of the chicken meat under refrigeration by up to 15 d. Total migration and the kinetics of the release of Nis showed that the OB can be used on hydrophilic and acidic foods, making it a natural alternative for use in food packaging.