Development of targeted antimicrobial peptides for Escherichia coli: Combining phage display and rational design for food safety application

The growing demand for minimally processed foods has heightened the risk of pathogenic contamination. Balancing antimicrobial efficacy with the preservation of probiotic activity remains a significant challenge. In this study, we employed phage display peptide library screening, combined with next-generation sequencing to identify the HIMPIQA domain, which selectively targets pathogenic Escherichia coli (E. coli) in just one screening round. Using the yXy(PX)₄yXy template, we designed antimicrobial peptides (AMPs) where 'y' represents hydrophobic amino acids, and 'X' represents positively charged residues. The peptide WK, combining HIMPIQA with a Trp/Lys-based antimicrobial domain, demonstrated high specificity and antibacterial activity. Molecular docking revealed WK's binding to FaeG and mechanistic studies showed its ability to disrupt E. coli membranes while maintaining probiotic viability. WK's application as a preservative enhanced the shelf life of yogurt and tomatoes. This innovative approach showcases the potential of phage display in developing targeted AMPs for food preservation and safety.