Effects of N-Terminal and C-Terminal Polyhistidine Tag on the Stability and Function of the Thermophilic P450 CYP119

Figure 6

Changes in the UV-Visible spectra during Amplex Red oxidation by H₂O₂ in the presence of WT CYP119 (a) and N-His-CYP119 (b); the UV-visible spectra taken at 0, 2, 5, 10, 20, 30, 45, 60, and 90 minutes after addition of H₂O₂. The reaction components were 10 μM Amplex Red, 1.5 mM H₂O₂, and 1.2 μM enzyme in 50 mM sodium phosphate buffer at pH 7.4. Time course of changes in absorbance at 570 nm (c) and 415 nm (d) in the same reaction (△, N-His-CYP119; ◯, WT CYP119).

(a)

(b)

(c)

(d)