Chemical characterization of fungal siderophores.

Siderophores of twenty fungi belonging to Zygomycotina (5 Mucorales), Ascomycotina (7 aspergilli, 6 penicillia, Neurospora crassa) and Deuteromycotina (Fusarium dimerum) were examined for their chemical nature. Siderophores produced by fungi other than Mucorales were all hydroxamates. Mucorales produced carboxylate siderophores. Catecholate type of siderophores were not detectable. Hydroxamate siderophores were mostly (9 out of 15) trihydroxamates, while six were dihydroxamates. Monohydroxamate nature was not shown by any of the 15 test fungal siderophores. In ligand properties, 12 out of 15 hydroxamate siderophores formed hexadentate ligands, while two formed tetradentates and one bidentate. There was good correlation between number of hydroxamate groups and ligand property.

Isolation and characterization of a copper containing protein from blue cell walls of Neurospora crassa.

Culturing Neurospora crassa in presence of toxic amounts of copper (0.63 mM) resulted in blue coloured mycelia and cell walls. Significant amounts (approximately 45%) of total mycelial copper were associated with cell wall isolates under conditions of copper toxicity. Hence, such blue cell walls were analysed to identify specific ligands involved in copper binding. While decuprification of the blue cell walls with 8-hydroxy quinoline (8 HQ) did not alter their copper binding abilities, similar treatment with EDTA (10 mM) decreased such abilities indicating that EDTA treatment lead to loss of copper binding ligands from cell walls. Treatment of blue cell walls with 8 HQ followed by EDTA resulted in the solubilization of a copper binding protein (relative MW approximately 14 kDa) which was associated with phosphate and carbohydrate moieties. On amino acid analysis, this protein was found to be devoid of free thiol groupings but enriched in acidic and basic amino acids, distinguishing it from classical intracellular metal binding proteins such as metallo-thioneins and phytochelatins that are inducively synthesized under conditions of metal toxicity. The biological significance of the isolated wall-bound copper binding protein, which appears to be a normal constituent of cell walls, is discussed in relation to cytoplasmic metal binding proteins and mechanism(s) adapted by fungi in countering metal toxicity.