Simultaneous immobilization of cytochrome P-450 and glucuronyltransferase for synthesis of drug metabolites.

Cytochrome P-450, NADPH-cytochrome P-450 reductase, and glucuronyltransferase were immobilized simultaneously on cyanogen bromide-activated Sepharose from phenobarbital-induced rabbit liver microsomes. The activity of the P-450 system was demonstrated by the N-demethylation of ethylmorphine and the O-demethylation of p-nitroanisole. p-Nitrophenol produced from the oxidation of p-nitroanisole was conjugated to p-nitrophenyl glucuronide, as evidenced by isolation and characterization of the glucuronide by GC/MS.