Monitoring iron uptake by siderophores

Abstract

Iron is an important element for almost all forms of life. In order to get access to this essential nutriment, Pseudomonads produce two major siderophores, pyoverdine PVD and pyochelin (PCH). Uptake of iron in bacterial cells can be monitored accurately using 55 Fe. Bacteria cells are incubated in the presence of either PVD or PCH loaded with 55 Fe. After incubation, extracellular iron ions are separated from those accumulated in the bacteria cells by either centrifugation or filtration on glass microfiber filters, for the PCH and PVD assays, respectively. 55 Fe contained in the harvested cells on the filter or in the cell pellet is counted in scintillation cocktail. The number of moles of 55 Fe transported can be determined using the specific activity of the radionuclide.