The stability of the molybdenum-azotochelin complex and its effect on siderophore production in Azotobacter vinelandii

A. K. Duhme, R. C. Hider, M. J. Naldrett, R. N. Pau

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Azotobacter vinelandii produces five siderophores with different metal binding properties, depending on the concentrations of Fe(III) and molybdate in the growth medium. The three lower protonation constants of the unusual bis(catecholamide) siderophore azotochelin (L) were determined by a simultaneous spectrophotometric and potentiometric titration as log K5=3.65(5), log K4=7.41(3) and log K3=8.54(4). The metal-ligand equilibrium constant for [MoO2(L)]3- was obtained from analysis of the absorbance concentration data: at 20°C and pH 6.6, log K(eq) =4(1). Based on an average log K(a) value of 12.1 for the two basic phenolic oxygens of azotochelin, the equilibrium formation constant was converted into the conventional formation constant K(f(MoL)) = [MoO2L3-]/[MoO22+][L5-] = 1035 M-1. To assess the influence of molybdenum-siderophore interactions on metal uptake in A. vinelandii, the dose-response effect of molybdate in the growth medium on siderophorc biosynthesis was followed by UV-vis spectroscopy and HPLC. It could be shown that the formation of molybdenum siderophore complexes clearly reduces the concentration of free siderophores available for iron solubilization. Furthermore, in media with initial molybdate concentrations up to 100 μM, the molybdenum azotochelin complex is the predominant molybdenum species, suggesting that azotochelin might also possess sequestering activity towards molybdenum. Even higher molybdate levels result in a complete repression of the synthesis of the tetradentate siderophore azotochelin, while they initiate the alternative release of the more efficient iron chelator, the hexadentate siderophore protochelin.

Original languageEnglish (US)
Pages (from-to)520-526
Number of pages7
JournalJournal of Biological Inorganic Chemistry
Volume3
Issue number5
DOIs
StatePublished - Oct 1 1998

Fingerprint

Azotobacter vinelandii
Siderophores
Molybdenum
Metals
Iron
Protonation
Biosynthesis
Equilibrium constants
Chelating Agents
Growth
azotochelin
Ultraviolet spectroscopy
Titration
Spectrum Analysis
High Pressure Liquid Chromatography
Oxygen
Ligands
molybdate

Keywords

  • Azotobacter vinelandii
  • Azotochelin
  • Metal uptake
  • Molybdenum
  • Siderophore

ASJC Scopus subject areas

  • Biochemistry
  • Inorganic Chemistry

Cite this

The stability of the molybdenum-azotochelin complex and its effect on siderophore production in Azotobacter vinelandii. / Duhme, A. K.; Hider, R. C.; Naldrett, M. J.; Pau, R. N.

In: Journal of Biological Inorganic Chemistry, Vol. 3, No. 5, 01.10.1998, p. 520-526.

Research output: Contribution to journalArticle

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AU - Pau, R. N.

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N2 - Azotobacter vinelandii produces five siderophores with different metal binding properties, depending on the concentrations of Fe(III) and molybdate in the growth medium. The three lower protonation constants of the unusual bis(catecholamide) siderophore azotochelin (L) were determined by a simultaneous spectrophotometric and potentiometric titration as log K5=3.65(5), log K4=7.41(3) and log K3=8.54(4). The metal-ligand equilibrium constant for [MoO2(L)]3- was obtained from analysis of the absorbance concentration data: at 20°C and pH 6.6, log K(eq) =4(1). Based on an average log K(a) value of 12.1 for the two basic phenolic oxygens of azotochelin, the equilibrium formation constant was converted into the conventional formation constant K(f(MoL)) = [MoO2L3-]/[MoO22+][L5-] = 1035 M-1. To assess the influence of molybdenum-siderophore interactions on metal uptake in A. vinelandii, the dose-response effect of molybdate in the growth medium on siderophorc biosynthesis was followed by UV-vis spectroscopy and HPLC. It could be shown that the formation of molybdenum siderophore complexes clearly reduces the concentration of free siderophores available for iron solubilization. Furthermore, in media with initial molybdate concentrations up to 100 μM, the molybdenum azotochelin complex is the predominant molybdenum species, suggesting that azotochelin might also possess sequestering activity towards molybdenum. Even higher molybdate levels result in a complete repression of the synthesis of the tetradentate siderophore azotochelin, while they initiate the alternative release of the more efficient iron chelator, the hexadentate siderophore protochelin.

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