Enhancing anti-E. coli O157:H7 activity of composite phage nanofiber film by D-phenylalanine for food packaging

Escherichia coli O157:H7 (E. coli O157) is still one of the main pathogenic strains causing food poisoning. Here in, we established a strategy that can simultaneously remove free E. coli O157 and the biofilms it forms. Composite nanofiber (fP-NF) films were synthesized successfully by co-encapsulating E. coli O157 phages which isolated from domestic sewage and D-phenylalanine (D-Phe) into sodium alginate (SA)/polyethylene oxide (PEO) nanofibers by electrospinning. And dehydrated trehalose was added as a phage protectant. When SA:PEO is 3:9, the spinning solution has the optimum conductivity and viscosity, the average diameter of the obtained fP-NFs is 421.87 nm, and the mechanical strength and transparency of the fP-NF film are the best. The thermal stability of the fP-NF film was good, and the phage activity in the fiber was well preserved. The remaining population of phages in fP-NF membranes was still close to 10⁶ PFU/mL after one week of placement, and the cumulative release efficiencies of phages at both room temperature (25 °C) and refrigerated storage temperature (4 °C) were above 70%. The antibacterial experiment showed that compared with the control group, the addition of D-Phe slightly enhanced the antibacterial activity of the nanofiber films against free E. coli O157, and the bactericidal rate within 8 h was 99.74%. On the other hand, D-Phe significantly enhanced the inhibitory activity of the nanofiber films against E. coli O157 biofilms, with the inhibition rate exceeding 99.99% within 72 h. The application results further confirmed that the addition of D-Phe could significantly enhance the inhibitory activity of nanofiber films on E. coli O157 proliferation and biofilm formation during short-term storage of beef and fruits and vegetables. It provides an important strategy and reference for the development of new active food packaging materials.