Preparation of human metabolites of propranolol using laboratory-evolved bacterial cytochromes P450.
Testing the toxicities and biological activities of the human metabolites of drugs is important for development of safe and effective pharmaceuticals. Producing these metabolites using human cytochrome P450s is difficult, however, because the human enzymes are costly, poorly stable, and slow. We have used directed evolution to generate variants of P450 BM3 from Bacillus megaterium that function via the "peroxide shunt" pathway, using hydrogen peroxide in place of the reductase domain, oxygen and NADPH. Here, we report further evolution of the P450 BM3 heme domain peroxygenase to enhance production of the authentic human metabolites of propranolol by this biocatalytic route. This system offers a versatile, cost-effective, and scaleable route to the synthesis of drug metabolites.
Otey
Christopher R
CR
Biochemistry and Molecular Biophysics, California Institute of Technology, Pasadena, 91125, USA.
Bandara
Geethani
G
Lalonde
James
J
Takahashi
Katsuyuki
K
Arnold
Frances H
FH
D001410:Bacillus megaterium / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001711:Biotransformation; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D019020:Directed Molecular Evolution; D005798:Genes, Bacterial; D006418:Heme / Q000378:metabolism; D006801:Humans; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D011433:Propranolol / Q000378:metabolism*; D015202:Protein Engineering
D001426:Bacterial Proteins; D006418:Heme; D003577:Cytochrome P-450 Enzyme System; D011433:Propranolol; D006899:Mixed Function Oxygenases; C022333:peroxygenase