Purification and characterization of mycoferritin from Aspergillus parasiticus (255).
As intracellular iron storage molecules, only hydroxymate type siderophores have been reported in ascomycetes and basidiomycetes. This is the first report documenting the presence of mycoferritin in ascomycetes. The fungus, Aspergillus parasiticus (255), is capable of producing mycoferritin only upon induction with iron in yeast extract sucrose (YES) medium. The same has been purified from Aspergillus sps by application of conventional biochemical techniques. The molecular mass, yield, iron and carbohydrate contents of the HPLC purified protein were 460kDa, 0.012mg/g of wet mycelia, 1.6% and 6.0%, respectively. The iron content was much lower than Mortierella alpina mycoferritin (17%). Native PAGE revealed the presence of trimeric and monomeric forms of ferritin. Subunit analysis by SDS-PAGE showed a single protein subunit of approximately 20kDa suggesting structural simplicity of the apoferritin shell. Variation in amino acid composition was noted upon comparison with ferritins of other species. Interestingly, no phenylalanine could be detected in the mycoferritin of Aspergillus sps. The acidic amino acid content was 1.5-1.6 fold higher than mammalian and fish ferritins. The spectral characteristics (UV/VIS and fluorescence) of mycoferritin were akin to equine spleen ferritin. However, circular dichroic spectra revealed a lower degree of helicity.
Shashidhar
J
J
Department of Biochemistry, University College of Science, Osmania University, Hyderabad 500 007, Andhra Pradesh, India.
Sashidhar
R B
RB
Deshpande
Vijay
V
D000596:Amino Acids / Q000032:analysis; D024342:Amino Acids, Acidic / Q000032:analysis; D001230:Aspergillus / Q000737:chemistry*; D002241:Carbohydrates / Q000032:analysis; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D005293:Ferritins / Q000737:chemistry* / Q000302:isolation & purification*; D007501:Iron / Q000032:analysis; D008970:Molecular Weight; D010649:Phenylalanine / Q000032:analysis; D017433:Protein Structure, Secondary; D021122:Protein Subunits / Q000737:chemistry / Q000302:isolation & purification; D013057:Spectrum Analysis
D000596:Amino Acids; D024342:Amino Acids, Acidic; D002241:Carbohydrates; D021122:Protein Subunits; D010649:Phenylalanine; D005293:Ferritins; D007501:Iron