Purification and characterization of a manganese-containing superoxide dismutase from a carboxydobacterium, Pseudomonas carboxydohydrogena.
Superoxide dismutase from Pseudomonas carboxydohydrogena, a carboxydobacterium, grown on carbon monoxide was purified 37.6-fold in seven steps to homogeneity, with a yield of 1.4%. The final specific activity was 2,396 units per mg protein as determined by an assay based on a 50% decrease in the rate of cytochrome c reduction. The molecular weight of the native enzyme was determined to be 42,500. Sodium dodecyl sulfate gel electrophoresis revealed two identical subunits of molecular weight 21,700. The optimal pH for enzyme activity was found to be 9.0. The enzyme was stable to heat treatment. The isoelectric point of the native enzyme was found to be 7.1. The enzyme showed an absorption peak at 280 nm with a shoulder at around 289 nm. Sodium azide, but not sodium cyanide and hydrogen peroxide, was found to inhibit the enzyme activity. One mol of native enzyme was found to contain 1.09 g-atom of manganese. Analysis of amino acid composition revealed that the enzyme contains cysteine. The superoxide dismutase of P. carboxydohydrogena was found to have antigenic sites identical to those of Oligotropha carboxydovorans. The enzyme showed partial identity to Hydrogenophaga pseudoflava and Escherichia coli enzymes, but no identity to Acinetobacter sp. strain JC1 enzyme.
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Department of Biology, Yonsei University, Seoul, Korea.
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D000596:Amino Acids / Q000032:analysis; D000942:Antigens, Bacterial / Q000276:immunology; D019281:Dimerization; D004791:Enzyme Inhibitors; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008970:Molecular Weight; D011487:Protein Conformation; D011549:Pseudomonas / Q000201:enzymology*; D019810:Sodium Azide / Q000494:pharmacology; D013482:Superoxide Dismutase / Q000737:chemistry*; D013696:Temperature
D000596:Amino Acids; D000942:Antigens, Bacterial; D004791:Enzyme Inhibitors; D019810:Sodium Azide; D013482:Superoxide Dismutase