Role of the siderophore azotobactin in the bacterial acquisition of nitrogenase metal cofactors

Fixation of dinitrogen by soil bacteria is catalyzed by the enzyme nitrogenase which requires iron, molybdenum, and/or vanadium as metal cofactors. Under conditions of iron deficiency, the ubiquitous N₂-fixing bacterium Azotobacter vinelandii produces azotobactin, a fluorescent pyoverdine-like compound which serves as a siderophore. Azotobatin's hydroxamate, catechol, and α-hydroxy-acid moieties endow it with a very high affinity for Fe^III, and the Fe complex is taken up by the bacterium. Here we show that azotobactin also serves for the uptake of Mo and V. Azotobactin forms strong complexes with molybdate and vanadate and the complexes are taken up by regulated transport systems. The kinetics of complexation of molybdate and vanadate by azotobactin are faster than the complexation of Fe^III, which is either precipitated or bound to strong complexing agents. As a result of this kinetic advantage, the Mo and V complexes of azotobactin form despite the higher affinity of the compound for Fe, which is present in large excess in the environment. The results obtained here for azotobactin and previous data for the bisand tris-catechols produced by A. vinelandii show that those "siderophores" are really "metallophores" that promote the bacterial acquisition of Mo and V in addition to Fe.