title: Genetically engineered yeast cells and their applications.
abstract: The first generation of yeast expression systems relies on inducible expression cassettes borne by multicopy plasmids for production of unmodified human P450s and on the endogenous NADPH-P450 reductase to support activities. A second generation of engineered yeast involved targeted genomic modifications allowing overexpression of the yeast reductase and coexpression of human cytochrome b5 and of a phase II enzyme such as epoxide hydrolase. These features allow improved P450 turnover numbers and simulation of some phase I-phase II couplings. In the third generation, the human reductase was substituted for the yeast reductase by genome engineering. Simultaneously, induction procedures were optimized to reach high P450 specific contents. Dramatic improvements (1000-fold) of yeast-expressed P450 activities have thus been obtained. To get more insight into complex metabolic events, such as that of a typical pollutant: benzo[a]pyrene, an approach was designed which involves a complementary use of yeast expression and computer simulations.
authors.affiliation: Centre de Génétique Moléculaire du CNRS, UPR 2420, Gifu-sur-Yvette, France.
authors.forename: D
authors.initials: D
authors.lastname: Pompon
authors.forename: A
authors.initials: A
authors.lastname: Perret
authors.forename: A
authors.initials: A
authors.lastname: Bellamine
authors.forename: R
authors.initials: R
authors.lastname: Laine
authors.forename: J C
authors.initials: JC
authors.lastname: Gautier
authors.forename: P
authors.initials: P
authors.lastname: Urban
mesh_terms: D001564:Benzo(a)pyrene / Q000378:metabolism; D003198:Computer Simulation; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis*; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis*; D015003:Yeasts / Q000235:genetics*
chemical_list: D011994:Recombinant Proteins; D001564:Benzo(a)pyrene; D003577:Cytochrome P-450 Enzyme System