title: Production of human cytochrome P450 2D6 drug metabolites with recombinant microbes--a comparative study.
abstract: The processes of drug development require efficient strategies to produce the respective drug metabolites, which are often difficult to obtain. Biotransformations employing recombinant microorganisms as whole-cell biocatalysts have become an attractive alternative to the chemical syntheses of such metabolites. For the first time, the potential of four different microbial systems expressing the human cytochrome P450 2D6 (CYP2D6), which is one of the most important drug-metabolizing enzymes, were compared and evaluated for such applications. The microbial host Pichia pastoris was the most efficient at expressing CYP2D6. Without additional over-expression of chaperons, the achieved yield of CYP2D6 was the highest of microbial hosts reported so far. Therefore, the system described in this study outperformed the previously reported expression of the N-terminally modified enzyme. It was also shown that the activities of the whole-cell conversions of bufuralol in recombinant P. pastoris were significantly higher than the Escherichia coli catalyst, which expressed the same unmodified gene.
authors.affiliation: Institute of Molecular Biotechnology, Graz University of Technology, Graz, Austria.
authors.forename: Martina
authors.initials: M
authors.lastname: Geier
authors.forename: Andreas
authors.initials: A
authors.lastname: Braun
authors.forename: Anita
authors.initials: A
authors.lastname: Emmerstorfer
authors.forename: Harald
authors.initials: H
authors.lastname: Pichler
authors.forename: Anton
authors.initials: A
authors.lastname: Glieder
mesh_terms: D019149:Bioreactors / Q000382:microbiology*; D001709:Biotechnology / Q000379:methods; D001711:Biotransformation; D002462:Cell Membrane; D003001:Cloning, Molecular; D019389:Cytochrome P-450 CYP2D6 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D004983:Ethanolamines / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006801:Humans; D009251:NADPH-Ferrihemoprotein Reductase / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004718:Saccharomycetales / Q000235:genetics / Q000378:metabolism*; D013696:Temperature
chemical_list: D004983:Ethanolamines; D011994:Recombinant Proteins; C010831:bufuralol; D019389:Cytochrome P-450 CYP2D6; D009251:NADPH-Ferrihemoprotein Reductase; D005947:Glucose