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.
D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004355:Drug Stability; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005037:Ethylmorphine-N-Demethylase / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D005965:Glucuronates / Q000378:metabolism; D014453:Glucuronosyltransferase / Q000378:metabolism*; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism; D009575:Nitroanisole O-Demethylase / Q000378:metabolism; D009578:Nitrobenzenes / Q000378:metabolism; D011817:Rabbits
D004800:Enzymes, Immobilized; D005965:Glucuronates; D009578:Nitrobenzenes; C002730:4-nitrophenylglucuronide; D003577:Cytochrome P-450 Enzyme System; D009575:Nitroanisole O-Demethylase; D005037:Ethylmorphine-N-Demethylase; D009251:NADPH-Ferrihemoprotein Reductase; D014453:Glucuronosyltransferase