Development and application of an assay for uranyl complexation by fungal metabolites, including siderophores

Joanna C. Renshaw, Verity Halliday, Geoffrey D. Robson, Anthony P J Trinci, Marilyn G. Wiebe, Francis R. Livens, David Collison, Robin J. Taylor

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Abstract

An assay to detect UO2 2+ complexation was developed based on the chrome azurol S (CAS) assay for siderophores (B. Schwyn and J. B. Neilands, Anal. Biochem. 160:47-56, 1987) and was used to investigate the ability of fungal metabolites to complex actinides. In this assay the discoloration of two dyed agars (one containing a CAS-Fe3+ dye and the other containing a CAS-UO2 2+ dye) caused by ligands was quantified. The assay was tested by using the siderophore desferrioxamine B (DFO), and the results showed that there was a regular, reproducible relationship between discoloration and the amount of siderophore added. The ratio of the discoloration on the CAS-UO2 2+ agar to the discoloration on the CAS-Fe3+ agar was independent of the amount of siderophore added. A total of 113 fungi and yeasts were isolated from three soil samples taken from the Peak District National Park. The fungi were screened for the production of UO2 2+ chelators by using the CAS-based assay and were also tested specifically for hydroxamate siderophore production by using the hydroxamate siderophore auxotroph Aureobacterium flavescens JG-9. This organism is highly sensitive to the presence of hydroxamate siderophores. However, the CAS-based assay was found to be less sensitive than the A. flavescens JG-9 assay. No significant difference between the results for each site for the two tests was found. Three isolates were selected for further study and were identified as two Pencillium species and a Mucor species. Our results show that the new assay can be effectively used to screen fungi for the production of UO2 2+ chelating ligands. We suggest that hydroxamate siderophores can be produced by mucoraceous fungi.

Original languageEnglish
Pages (from-to)3600-3606
Number of pages7
JournalApplied and Environmental Microbiology
Volume69
Issue number6
Publication statusPublished - 1 Jun 2003

Keywords

  • development and application
  • assay
  • uranyl complexation
  • fungal metabolites

Cite this

Renshaw, J. C., Halliday, V., Robson, G. D., Trinci, A. P. J., Wiebe, M. G., Livens, F. R., Collison, D., & Taylor, R. J. (2003). Development and application of an assay for uranyl complexation by fungal metabolites, including siderophores. Applied and Environmental Microbiology, 69(6), 3600-3606.