Lipoxygenase-catalyzed oxidation of chlorpromazine by hydrogen peroxide at acidic pH.
The hydroperoxidase activity of soybean lipoxygenase, a non-heme protein, oxidizes chlorpromazine using H2O2 at acidic pHs ranging from 3.0 to 4.0. The enzyme is assayed at pH 3.5, at which the half-life is 2 h (lower pHs cause higher inactivation rates). This oxidation is enzymatical since boiled enzyme or even iron ions both with H2O2 failed to produce any increase in absorbance. In addition, the concentration of CPZ radical cation formed and the concomitant enzyme activity directly depends on the enzyme concentration up to 0.23 microM. The Vmax value is 125 mumol/min per mg protein and the Km for chlorpromazine and H2O2 are 2.1 mM and 0.25 mM, respectively. Similar results were obtained when linoleic acid hydroperoxide was used instead of H2O2 with a Km value of 95 microM. The radical cation obtained in the oxidation of chlorpromazine by lipoxygenase decays by a disproportionation reaction. This permits to consider the overall reaction as a sum of an enzymatic reaction coupled with a chemical second order reaction with substrate regeneration, similar to those produced by peroxidases from different sources.
Pérez-Gilabert
M
M
Departamento de Bioquímica y Biología Molecular, Facultad de Biología, Universidad de Murcia, Spain.
Sánchez-Ferrer
A
A
García-Carmona
F
F
D002746:Chlorpromazine / Q000737:chemistry / Q000378:metabolism*; D004795:Enzyme Stability; D006861:Hydrogen Peroxide / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008084:Lipoxygenase / Q000378:metabolism*; D010084:Oxidation-Reduction; D013025:Glycine max
D006861:Hydrogen Peroxide; D008084:Lipoxygenase; D002746:Chlorpromazine