The partial characterization of purified nitrite reductase and hydroxylamine oxidase from Nitrosomonas europaea.
Nitrite reductase has been separated from cell-free extracts of Nitrosomonas and partially purified from hydroxylamine oxidase by polyacrylamide-gel electrophoresis. In its oxidized state the enzyme, which did not contain haem, had an extinction maximum at 590nm, which was abolished on reduction. Sodium diethyldithiocarbamate was a potent inhibitor of nitrite reductase. Enzyme activity was stimulated 2.5-fold when remixed with hydroxylamine oxidase, but was unaffected by mammalian cytochrome c. The enzyme also exhibited a low hydroxylamine-dependent nitrite reductase activity. The results suggest that this enzyme is similar to the copper-containing ;denitrifying enzyme' of Pseudomonas denitrificans. A dithionite-reduced, 465nm-absorbing haemoprotein was associated with homogeneous preparations of hydroxylamine oxidase. The band at 465nm maximum was not reduced during the oxidation of hydroxylamine although the extinction was abolished on addition of hydroxylamine, NO(2) (-) or CO. These last-named compounds when added to the oxidized enzyme precluded the appearance of the 465nm-absorption band on addition of dithionite. Several properties of 465nm-absorbing haemoprotein are described.
D000645:Ammonium Sulfate; D002248:Carbon Monoxide; D055598:Chemical Phenomena; D011232:Chemical Precipitation; D002621:Chemistry; D002848:Chromatography, DEAE-Cellulose; D003574:Cytochrome c Group / Q000494:pharmacology; D003956:Dialysis; D004591:Electrophoresis, Polyacrylamide Gel; D006420:Hemeproteins; D006898:Hydroxylamines; D009247:NADH, NADPH Oxidoreductases / Q000302:isolation & purification*; D009572:Nitrite Reductases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D009606:Nitrosomonas / Q000201:enzymology*; D010088:Oxidoreductases / Q000302:isolation & purification*; D012967:Sodium Dodecyl Sulfate; D013053:Spectrophotometry; D013447:Sulfites; D013859:Thiocarbamates / Q000494:pharmacology
D003574:Cytochrome c Group; D006420:Hemeproteins; D006898:Hydroxylamines; D013447:Sulfites; D013859:Thiocarbamates; D012967:Sodium Dodecyl Sulfate; D002248:Carbon Monoxide; D010088:Oxidoreductases; D009247:NADH, NADPH Oxidoreductases; D009572:Nitrite Reductases; D000645:Ammonium Sulfate