The medical importance of honey has been extensively demonstrated. Although high osmolarity, acidity and hydrogen peroxide (H2O2) proved to be the most prevalent factors in honey’s activity, the underlying antimicrobial mechanism remains obscure. Our aim is to provide insight into the physiological changes and genetic responses in honey-treated bacteria, thus improving our understanding of this natural product as a potential novel antimicrobial. A model honey composed of sugars, gluconic acid, and H2O2as they are accumulated in honey after enzymatic reaction happens, was used the investigation of honey’s activity. The bactericidal action of the model was tested on E. coli K-12 strain MG1655. Flow cytometry (FC) and Atomic Force Microscopy (AFM) identified physiological changes such as membrane potential, blebbing, and cell lysis. Reactive Oxygen Species (ROS) accumulation was observed in individual cells by FC. Transposon Directed Insertion Sequencing (TraDIS) identified mutants’ fitness over a time course of E. coli treatment by model honey. The loss of selenocysteine (selAB) and formate dehydrogenase (fdhDE) mutants, proved the redox- balancing activity as essential for the repression of ROS in stressed cells. High susceptibility of energy metabolism (atpABD) and peptidoglycan synthesis (prc) mutants, indicated the strain unable to maintain the reductive cell environment necessary for cellular activities, post honey exposure. Our findings identified some of the honey’s targets when acting as an antimicrobial. The synergies observed support the use of honey as an antimicrobial; however, the identification of mutations that led to enhanced resistance to honey is an important finding that needs further study.

  • This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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