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

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.

Three genomes differentially contribute to the biosynthesis of benzoxazinones in hexaploid wheat

“…The TaBX2-TaBX5 enzymes encoding membrane-bound cytochrome P450s were expressed in yeast (Saccharomyces cerevisiae). The coding sequences of the three homoeologs of TaBx2-TaBx5 were separately inserted into HindIII site of the vector pAAH5N (26), and the vector was introduced into S. cerevisiae AH22 strain. The culture of transformed yeast cells, preparation of recombinant microsome fractions, and determination of P450 contents were performed as described in ref.…”

Downstream reactions and engineering in the microbially reconstituted pathway for Taxol

“…This can be mitigated by the native capabilities to complement reductase activity when using S. cerevisiae, but the same level of convenience may not be provided with E. coli (Barnes et al 1991;Jennewein et al 2005;Oeda et al 1985). Again thanks to work by Rodney Croteau's group, a reductase has been isolated from Taxus cuspidata (Jennewein et al 2005).…”

Expression of cytochrome P‐450d by Saccharomyces cerevisiae

“…Oeda et al [8] reported a high expression of rat liver P-450c in the yeast under the control of a yeast alcohol dehydrogenase promoter. The efficiency of the expression of P-450d in the yeast of our system is as high as that of P-450c by using the alcohol dehydrogenase promoter [8]. We attempted to express P-450a by using the yeast alcohol dehydrogenase promoter, but did not succeed in the high expression of P-450a for unknown reasons.…”