Expression of Rat Liver Cytochrome P-450MC cDNA in <i>Saccharomyces cerevisiae</i>

Abstract

Rat liver cytochrome P-450MC cDNA was inserted between the ADH1 promoter and terminator regions of the yeast expression vector pAAH5. On introduction of the resulting recombinant plasmid pAMC1, Saccharomyces cerevisiae cells synthesized up to 8 X 10(5) molecules per 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.