Engineering phage endolysins and receptor-binding proteins for foodborne pathogen control and detection: A review and...

Antimicrobial resistance exacerbates food safety risks, highlighting the need for novel, matrix-tolerant control and detection tools. Phage-derived endolysins and receptor-binding proteins (RBPs) offer high specificity, yet their performance is often reduced in complex food environments compared with buffer assays. In this review, we summarize engineering principles and food-system evidence that govern efficacy, including matrix-driven attenuation (e.g., viscosity, protein/fat adsorption, ionic strength, and proteolysis) and developability constraints linked to manufacturing and formulation. We also summarize practical endpoints for food applications spanning sanitation of food-contact surfaces, biocontrol on raw commodities, and culture-free pathogen detection. On this basis, we propose ISELI-CL (In Silico-Experiment-Learning-Iterative Closed Loop), a structured framework that combines computational design and active learning with high-throughput validation to balance activity, manufacturability, and food matrix compatibility. By treating buffer-to-food translation as a quantifiable domain shift, ISELI-CL is intended to guide candidate prioritization and matrix-aware validation of mechanism-informed phage proteins in food microbiology.