Electron-spin-resonance evidence for enzymic reduction of oxygen to a free radical, the superoxide ion.
1. An electron-spin-resonance signal with g( parallel)2.08 and g( perpendicular)2.00 is observed by the rapid-freezing technique during the oxidation of substrates by molecular oxygen catalysed by xanthine oxidase at pH10. 2. The intensity of this signal is shown to depend on oxygen rather than on enzyme concentration, indicating that it is due to an oxygen free radical and not to the enzyme. 3. The same species is shown to be produced in the reaction at pH10 between hydrogen peroxide and periodate ions. Studies with this system have facilitated comparison of the properties of the oxygen radical with data in the literature on the products of pulse radiolysis of oxygenated water over a wide pH range. 4. It is concluded that the species observed is the superoxide ion, O(2) (-), and that the stability of this ion is greatly increased in alkaline solution. A mechanism explaining the alkaline stability is proposed. 5. The importance of O(2) (-) in the enzymic reaction is discussed.
D000468:Alkalies; D055598:Chemical Phenomena; D002621:Chemistry; D004578:Electron Spin Resonance Spectroscopy; D005609:Free Radicals / Q000096:biosynthesis; D006861:Hydrogen Peroxide; D006863:Hydrogen-Ion Concentration; D010087:Oxides / Q000096:biosynthesis*; D010100:Oxygen / Q000378:metabolism*; D010504:Periodic Acid; D012996:Solutions; D014867:Water; D014969:Xanthine Oxidase / Q000378:metabolism*
D000468:Alkalies; D005609:Free Radicals; D010087:Oxides; D012996:Solutions; D014867:Water; D010504:Periodic Acid; D006861:Hydrogen Peroxide; D014969:Xanthine Oxidase; D010100:Oxygen