[HTML][HTML] Directed evolution of mammalian cytochrome P450 2B1: mutations outside of the active site enhance the metabolism of several substrates, including the …
Cytochrome P450 2B1 has been subjected to directed evolution to investigate the role of
amino acid residues outside of the active site and to engineer novel, more active P450
catalysts. A high throughput screening system was developed to measure H 2 O 2-
supported oxidation of the marker fluorogenic substrate 7-ethoxy-4-trifluoromethylcoumarin
(7-EFC). Random mutagenesis by error-prone polymerase chain reaction and activity
screening were optimized using the L209A mutant of P450 2B1 in an N-terminally modified …
amino acid residues outside of the active site and to engineer novel, more active P450
catalysts. A high throughput screening system was developed to measure H 2 O 2-
supported oxidation of the marker fluorogenic substrate 7-ethoxy-4-trifluoromethylcoumarin
(7-EFC). Random mutagenesis by error-prone polymerase chain reaction and activity
screening were optimized using the L209A mutant of P450 2B1 in an N-terminally modified …