In vivo metal substitution in bacteroides superoxide dismutase
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The effect of various growth conditions on the type of superoxide dismutase (SGD) formed anaerobically in three Bacteriodes species was studied. B. fragilis, B. distasonis, and B. thetaiotaomicron were grown in ironâ restricted media with or without manganese supplementation. Iron availability was decreased by treatment of the media with chelex-100, a metal-chelating resin, and addition of desferrioxamine mesylate (desferal, Ciba-Geigy), an iron chelator. Mn-containing (MnSOD) and Fe-containing superoxide dismutase (EeSOD) activities in cell extracts were differentiated by inhibition with azide and inactivation by H202. The amount of Mn-containing superoxide dismutase was estimated by the fraction of azide- and H202, -resistant activity. Cells grown in untreated media contained approximately 90% FeSOD and 10% MnSOD. Cells grown in Fe-restricted media supplemented with graded amounts of manganese synthesized a progressively larger fraction of MnSOD. Hemin, added to the Fe-restricted media, did not serve as an iron source for FeSOD formation. Superoxide dismutase specific activities varied (3-6 U/mg) in each extract but not as a function of manganese concentration.
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