1887

Abstract

Iron is an essential nutrient for microbial growth and bacteria have evolved numerous routes to solubilize and scavenge this biometal, which is often present at very low concentrations in host tissue. We recently used a MOPS-based medium to induce iron limitation in K-12 during the characterization of novel siderophore-conjugated antibiotics. In this study we confirm that growth media derived from commercially available M9 salts are unsuitable for studies of iron-limited growth, probably through the contamination of the sodium phosphate buffer components with over 100 µM iron. In contrast, MOPS-based media that are treated with metal-binding Chelex resin allow the free iron concentration to be reduced to growth-limiting levels. Despite these measures a small amount of growth is still observed in these iron-depleted media. By growing in conditions that theoretically increase the demand for iron-dependent enzymes, namely by replacing the glucose carbon source for acetate and by switching to a microaerobic atmosphere, we can reduce background growth even further. Finally, we demonstrate that by adding an exogeneous siderophore to the growth media which is poorly used by , we can completely prevent growth, perhaps mimicking the situation in host tissue. In conclusion, this short study provides practical experimental insight into low iron media and how to augment the growth conditions of for extreme iron-limited growth.

Keyword(s): Escherichia coli , iron and siderophores
Funding
This study was supported by the:
  • Engineering and Physical Sciences Research Council (Award EP/L024829/1)
    • Principle Award Recipient: Anne-KathrinDuhme-Klair
  • Engineering and Physical Sciences Research Council (Award EP/T007338/1)
    • Principle Award Recipient: Anne-KathrinDuhme-Klair
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
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2024-06-03
2024-06-19
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