Abstract

The emergence, persistence and spread of antibiotic-resistant human pathogenic bacteria heralds a growing global health crisis. Drug-resistant strains of gram-negative bacteria, such as Pseudomonas aeruginosa, are especially dangerous and the medical and economic burden they impose underscore the critical need for finding new antimicrobials. Recent studies have demonstrated that plant-expressed bacteriocins of the colicins family can be efficient antibacterials against all major enteropathogenic strains of E. coli. We extended our studies of colicin-like bacteriocins to pyocins, which are produced by strains of P. aeruginosa for ecological advantage against other strains of the same species. Using a plant-based transient expression system, we expressed six different pyocins, namely S5, PaeM, L1, L2, L3 and one new pyocin, PaeM4, and purified them to homogeneity. Among these pyocins, PaeM4 demonstrated the broadest spectrum of activity by controlling 53 of 100 tested clinical isolates of P. aeruginosa. The activity of plant-made pyocins was confirmed in the agar drop, liquid culture susceptibility and biofilm assays, and in the Galleria mellonella animal infection model.

Highlights

  • Pseudomonas aeruginosa is a ubiquitous Gram-negative, rod-shaped, asporogenous, monoflagellated bacterium belonging to the class Gammaproteobacteria, and is characterized in part by its high nutritional versatility

  • Because of the unmet need for new natural non-antibiotic antibacterials for controlling pathogenic Pseudomonas aeruginosa, we conducted a similar investigation of the colicin analogues pyocins, which are produced by members of the genus Pseudomonas

  • We examined the activity of pyocins in young, one day old P. aeruginosa biofilms grown under iron-limited conditions

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Summary

Introduction

Pseudomonas aeruginosa is a ubiquitous Gram-negative, rod-shaped, asporogenous, monoflagellated bacterium belonging to the class Gammaproteobacteria, and is characterized in part by its high nutritional versatility. P. aeruginosa is an opportunistic pathogen and can cause diseases in plants and animals, including humans. In the latter, it can cause life-threatening chronic infections (pneumonia, ventilator-associated pneumonia, catheter-related infections, burn wound infections, and sepsis) especially in patients with a weakened immune system. It can cause life-threatening chronic infections (pneumonia, ventilator-associated pneumonia, catheter-related infections, burn wound infections, and sepsis) especially in patients with a weakened immune system This bacterium is one of the six pathogens causing hospital ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter) infections, which readily develop resistance to antibiotics [1].

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