Antibacterial activity and multi-target mechanism of harmane against Escherichia coli O157:H7 and its application on ready-to-eat leafy greens

Escherichia coli O157:H7 has caused many foodborne disease outbreaks and resulted in unimaginable economic losses. With the evolution of food consumption, people prefer natural Preservatives. In this study, the natural agent harmane exhibited potential activity against E. coli O157:H7 (MIC = 64 μg/mL). It exhibited bacteriostatic mode at 1 X and 2 X MIC treatment, and bactericidal mode at 4 X MIC treatment. Moreover, it showed good in vitro stability in sheep plasma, low in vitro hemolysis and no in vivo acute toxicity with LD₅₀ > 50 mg/kg. Moreover, harmane significantly decreased the colony number of E. coli O157:H7 in fresh-cut lettuce samples after 5 days of storage without affecting appearance. The mechanism study elucidated that harmane significantly decomposed the mature biofilm by reducing exopolysaccharide contents, and killed the viable Bacterial cells in biofilm. The cell wall was damaged by harmane via interacting with peptidoglycan. Fluorescent staining and intracellular macromolecular leakage assays showed that irreversible destruction to membrane permeability and integrity. When entering the cell, harmane could defeat the redox balance, suppress metabolic activity and target to ribosome. These findings not only revealed the application potential of harmane as new natural preservative, but also preliminarily elucidated the multi-target mechanism, providing a new strategy for controlling E. coli O157:H7 in the food industry.

Biofilm; Cell membrane; Cell wall; Escherichia coli O157:H7; Harmane; Transcriptomics.