Analysis of Iron Requirements and Siderophore Production

  • Denise M. Burnside
  • Nicholas P. CianciottoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1921)


This chapter describes the methods for inducing, detecting, and purifying the Legionella pneumophila siderophore. The first protocol details the methods by which L. pneumophila is cultured to facilitate production of the siderophore, rhizoferrin. This chapter then describes how to purify siderophore from culture supernatants through sequential reversed-phase/weak-anion exchange chromatography and high-performance liquid chromatography. The next section describes assays which allow the detection of the iron-binding capability and the biological activity of the purified siderophore. Lastly, this chapter describes the growth of L. pneumophila in chemically defined liquid medium (CDM) containing various iron sources as a method to assess the iron requirements of L. pneumophila.

Key words

Legionella pneumophila CDM BYE BCYE Iron Siderophore HPLC Rhizoferrin Bioassay CAS assay Iron salts 



This work was supported by NIH grant AI034937 awarded to NPC.


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Microbiology and ImmunologyNorthwestern University Medical SchoolChicagoUSA

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