Universal Chemical Assay for the Detection and Determination of Siderophores

Anal Biochem. 1987 Jan;160(1):47-56. doi: 10.1016/0003-2697(87)90612-9.

Abstract

A universal method to detect and determine siderophores was developed by using their high affinity for iron(III). The ternary complex chrome azurol S/iron(III)/hexadecyltrimethylammonium bromide, with an extinction coefficient of approximately 100,000 M-1 cm-1 at 630 nm, serves as an indicator. When a strong chelator removes the iron from the dye, its color turns from blue to orange. Because of the high sensitivity, determination of siderophores in solution and their characterization by paper electrophoresis chromatography can be performed directly on supernatants of culture fluids. The method is also applicable to agar plates. Orange halos around the colonies on blue agar are indicative of siderophore excretion. It was demonstrated with Escherichia coli strains that biosynthetic, transport, and regulatory mutations in the enterobactin system are clearly distinguishable. The method was successfully used to screen mutants in the iron uptake system of two Rhizobium meliloti strains, DM5 and 1021.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Colorimetry / methods
  • Electrophoresis, Paper
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Indicators and Reagents
  • Iron / metabolism*
  • Iron Chelating Agents / analysis*
  • Mutation
  • Rhizobium / genetics
  • Rhizobium / metabolism
  • Siderophores

Substances

  • Indicators and Reagents
  • Iron Chelating Agents
  • Siderophores
  • Iron