Figure 6.

Demonstration of nonspecific partnering with diverse combinations of P450_cam, reductases, auxiliary redox partners, substrates and electron donors: All the reactions were carried out at 27 ± 1°C in 100 mM potassium phosphate buffer (pH 7.4). The initial concentration of substrate employed was 200 μM. Enzyme concentrations were as follows: [P450cam] = 0.5 μM and [Pdr] = 0.6 μM. Electron donors: [NADH] and [H₂O₂] = 1 mM, [O${}_2^{.-}$] = 50 μM. [Cyt. c] and [Vit. C] = 1 μM. (A) P450_cam reactions with pNP (a CYP2E1 marker substrate), employing Pdr and NADH as redox equivalent generating agents. The control had only pNP, P450_cam, Pdr, and NADH. (B) Schematic of HPLC profiles obtained with P450_cam and coumarin (a CYP2B6 substrate), employing superoxide as the sole redox agent. The inset to the right shows the original area under curve (AUC) values obtained.