Fluorescence of pyoverdin in response to iron and other common well water metals.

When the fluorescent siderophore pyoverdin (produced by Pseudomonas aeruginosa) binds to a metal ion the fluorescence changes. A pyoverdin solution (in 0.1 M acetate buffer, pH 5.0) was placed in the microwells of a 96-well plate and varying concentrations of the metal cations Al(3+), Ca(2+), Cu(2+), Fe(2+), Fe(3+), Mn(2+), Mg(2+), and Zn(2+) were added. The fluorescence of pyoverdin 60 sec after the addition of an equimolar concentration of metal indicated: (1) no change for Ca(2+), Fe(2+), Mn(2+), Mg(2+), and Zn(2+); (2) a small increase (109%) for Al(3+); (3) decreases in fluorescence for Cu(2+) (83%) and for Fe(3+) (66%). The fluorescence of pyoverdin 24 hr after the addition of equimolar metal indicated: 1) very little change for Ca(2+), Mn(2+), Mg(2+), and Zn(2+); 2) a very large (270%) increase in fluorescence due to Al(3+); 3) an increase (113%) due to Cu(2+); 4) large decreases in fluorescence for both Fe(2+) (15%) and Fe(3+) (0%). Thus, for an iron assay using a free solution of pyoverdin, even with a short (60 sec.) reaction time there can be interference due to Cu(2+), and interference due to high levels of Al(3+).