title: Properties of 2-nitropropane dioxygenase of Hansenula mrakii. Formation and participation of superoxide.
abstract: 2-Nitropropane dioxygenase, purified to homogeneity from a yeast, Hansenula mrakii, is significantly inhibited by superoxide dismutase and various scavengers for superoxide anion such as cytochrome c, epinephrine, NADH, thiols, and polyhydric phenols. The reduction of cytochrome c and the oxidation of epinephrine and NADH are concomitant with the inhibition of enzymatic oxygenation. Neither the oxidation nor the reduction occursin the presence of superoxide dismutase or in the absence of 2-nitropropane or oxygen. Superoxide anion added externally induces the oxygenation. These findings indicate the generation of superoxide anion and its participation in the oxygenation of 2-nitropropane. The difference spectrum of the binding of NADH to 2-nitropropane dioxygenase exhibits a negative peak at 353 nm. One mole of NADH is bound to 1 mol of the enzyme and the pro-R hydrogen of the nicotinamide moiety of bound NADH predominantly is transferred to superoxide anion formed enzymatically or given externally. Thus, the diastereotopic hydrogen of NADH is discriminated by the enzyme, although not completely.
mesh_terms: D001203:Ascomycota / Q000201:enzymology*; D003574:Cytochrome c Group / Q000378:metabolism; D049308:Dioxygenases; D004837:Epinephrine / Q000378:metabolism; D007700:Kinetics; D009243:NAD / Q000378:metabolism; D009574:Nitro Compounds; D010100:Oxygen / Q000378:metabolism*; D010105:Oxygenases / Q000378:metabolism*; D010843:Pichia / Q000201:enzymology*; D013482:Superoxide Dismutase / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism*
chemical_list: D003574:Cytochrome c Group; D009574:Nitro Compounds; D009243:NAD; D013481:Superoxides; D010105:Oxygenases; D049308:Dioxygenases; C021876:2-nitropropane dioxygenase; D013482:Superoxide Dismutase; D010100:Oxygen; D004837:Epinephrine