Vibrioferrin production by the food spoilage bacterium Pseudomonas fragi

Abstract

Pseudomonas fragi is a meat and milk spoilage bacterium with high iron requirements, however mechanisms of iron acquisition remain largely unknown. The aim of this work was to investigate siderophore production as an iron acquisition system for P. fragi. A vibrioferrin siderophore gene cluster was identified in 13 P. fragi and experiments were conducted with a representative strain of this group (F1801). Chromeazurol S assays showed P. fragi F1801 produced siderophores under iron starvation at optimum growth and refrigeration temperature. Conversely, supplementation of low iron media with 50 μM FeCl₃ repressed transcription of the vibrioferrin genes and siderophore production. Disruption of the siderophore receptor (pvuA) caused polar effects on downstream vibrioferrin genes, resulting in impaired siderophore production of the ΔpvuA mutant. Growth of this mutant was compared to growth of a control strain (Δlip) with wild-type vibrioferrin genes in low iron media supplemented with iron chelators 2,2'-bipyridyl or apo-transferrin. While 25 μM 2,2'-bipyridyl caused impaired growth of ΔpvuA, growth of the mutant was completely inhibited by 2.5 μM apo-transferrin, but could be restored by FeCl_3-addition. In summary, this work identifies a vibrioferrin-mediated iron acquisition system of P. fragi, which is required for growth of this bacterium under iron-starvation.