Differential induction of peroxygenase-dependent microsomal aniline hydroxylase by chronic ethanol ingestion.

The liver microsomal-mediated hydroxylation of aniline, which is selectively induced by chronic (EtOH) ingestion, has been studied as a function of NADPH plus dioxygen (O2)- or hydroperoxide-dependent reactions. Consistent with the well-documented induction of aniline hydroxylase following chronic ethanol -ingestion, the results showed selectivity towards aniline hydroxylase by the NADPH plus O2- and tert-butyl hydroperoxide (t-BuOOH)-dependent reactions with microsomes from EtOH-fed rats. On the other hand, the cumene hydroperoxide (CumOOH)-dependent aniline hydroxylase activity was not discriminated between microsomes from EtOH- and pair-fed rats. In parallel experiments with positive controls, CumOOH did show selectivity for phenobarbital (PB)-induced microsomal aniline hydroxylase compared to chow-fed rats. The Kcat/KM values, which indicate the efficiency of enzyme catalysis, for NADPH plus O2-, t-BuOOH, and CumOOH-dependent aniline hydroxylase from EtOH-fed rats were 102, 37, and 5 and from pair-fed rats were 68, 4, and 4 (nmol p-aminophenol/min/nmol cytochrome P-450)/mM aniline, respectively. The relative Kcat/KM ratio for EtOH-fed to that of pair-fed microsomal aniline hydroxylase from NADPH plus O2-, t-BuOOH-, and CumOOH-dependent reactions were 1.5, 7.4, and 1.2, respectively. The present preliminary studies indicate that the catalytic efficiency of EtOH-induced aniline hydroxylase is significantly greater for the t-BuOOH-dependent reaction.