Fig. 1
Biochemical characterization of PgPepO and TfPepO. (A) The velocity of the reaction (V) was determined for various concentrations of the substrate Mca-RPPGFSAFK(Dnp)-OH, and the Michaelis–Menten constant (Km) was calculated using a GraphPad Prism macro. (B, C) Effect of pH (B) and temperature (C) on PepO activity. Enzymatic activity was measured at various pHs (B) and temperatures (C). The pH with the highest V (B), and the V at 37 °C (C), which was regarded as 100%. (D) Thermal stability of PgPepO and TfPepO. PepOs were incubated at the indicated temperatures for 30 min, then their enzymatic activities were measured. The activity at 37 °C was set at 100%. (E, F) Effects of inhibitors and reducing agents on the activities of PgPepO and TfPepO. PepOs were incubated with the indicated compounds, and their activities were measured. PepO activity determined at 37 °C in 100 mM Tris, 150 mM NaCl, 0.02% NaN3, 0.05% Pluronic F-127 (pH 7.5) was regarded as 100%. Data are mean ± SD (N = 3).
