Reduced oxy intermediate observed in D251N cytochrome P450cam

Kinetic measurements coupled with UV-visible spectrophotometry, electron paramagnetic resonance measurements and gas chromatograpy analysis were used to determine the nature of D251N cytochrome P450cam intermediate. The UV-visible spectral methods showed that the UV-visible spectrum of the spectral intermediate was significantly higher from that of oxy-P450. A newly observed catalytic intermediate was formed from the D251N mutant of P450cam.