An ornithine-rich dodecapeptide with improved proteolytic stability selectively kills gram-negative food-borne pathogens and its action mode on Escherichia coli O157:H7

Ping Zeng, Lanhua Yi, Qipeng Cheng, Jun Liu, Sheng Chen, Kin Fai Chan, Kwok Yin Wong

Research output: Journal article publicationJournal articleAcademic researchpeer-review

13 Citations (Scopus)


Food-borne pathogenic bacteria are dispersed throughout the entire chain of the food industry. However, many food preservatives are limited by poor biocompatibility such as cumulative poisoning. The antimicrobial peptide is increasingly regarded as a promising preservative in food research due to its high bioactivity and low cytotoxicity. In this study, thirteen peptides were designed, synthesized, and screened for application as food preservatives. One of them, termed zp65, whose sequence is GIOAOIIIOIOO-NH2, demonstrated potent bactericidal effect against common Gram-negative strains including enterohemorrhagic Escherichia coli, Salmonella, and Citrobacter freundii. Encouragingly, zp65 showed negligible cytotoxicity to both mammalian cells and Galleria mellonella larvae. Peptide zp65 was prone to form α-helix structure in amphiphilic environments, facilitating its affinity with bacterial membrane. Furthermore, the proteolytic stability of zp65 was much higher than its derivatives consisting of totally natural amino acids. Isothermal titration calorimetry indicated that zp65 has a strong binding affinity to lipopolysaccharide with Kd = 1.3 μM, suggesting its possible action target on the bacterial envelope. Mechanistic studies revealed that this peptide also influenced the membrane potential of E.coli O157:H7 (O157) in a dose-dependent manner. Surprisingly, peptide zp65 did not induce disruption of membrane permeability even at a higher concentration of 4-fold minimal inhibitory concentration. By employing confocal microscopy, peptide zp65 labeled by fluorescein isothiocyanate mainly aggregated on the bacterial membrane. These results suggested that the bactericidal mode of action of zp65 is likely attributed to depolarization of the cell membrane. The minced lean beef experiment indicated that the maximum reduction of O157 reached 1.46 log colony-forming unit (CFU) per gram on day 1 after zp65 treatment at the dosage of 40 μg/g. Compared with the untreated cooked beef sample, the CFU of the zp65-treated group remained at a much lower level after 10-day storage. Subsequently, treatment with zp65 at concentrations above 32 μM also significantly reduced O157 viable counts in fresh tomato juice. And the zp65 treatment could rescue about 40% of Galleria mellonella larvae injected with O157-contaminated tomato juice. The peptide zp65 exhibits great potential and deserves further study as a candidate for food preservative.

Original languageEnglish
Article number109281
JournalInternational Journal of Food Microbiology
Publication statusPublished - 16 Aug 2021


  • Antimicrobial peptides
  • Food preservative
  • Lipopolysaccharide-binding
  • Membrane depolarization
  • Membrane potential

ASJC Scopus subject areas

  • Food Science
  • Microbiology


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