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

Renshaw, Joanna C. and Halliday, Verity and Robson, Geoffrey D. and Trinci, Anthony P J and Wiebe, Marilyn G. and Livens, Francis R. and Collison, David and Taylor, Robin J. (2003) Development and application of an assay for uranyl complexation by fungal metabolites, including siderophores. Applied and Environmental Microbiology, 69 (6). pp. 3600-3606. ISSN 0099-2240

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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.