Expression of Rat Liver Cytochrome P-450MC cDNA inSaccharomyces cerevisiae

Abstract

Rat liver cytochrome P-450MC complementaryDNA was inserted between the ADH1 promoter and terminator regions of the yeast expression vector pAAH5. On introduction of the resulting recombinant plasmid pAMC1, S. cerevisiae cells synthesized up to 8 .times. 105 molecules/cell of the cytochrome P-450MC protein, most of which was localized in yeast microsomes. Approximately half of the synthesized cytochrome contained heme in the enzyme molecule. These formed a functional electron-transport chain in the microsomes which exhibited aryl hydrocarbon hydroxylase activity toward benzo[a]pyrene.