FIG. 4.
HPLC chromatograms of the reaction product from OCA catalyzed by PraD. The reaction mixture contained the PraC reaction mixture, 500 μM MgSO4, and crude PraD (25 μg of protein/ml) in 50 mM Tris-HCl buffer (pH 7.3) in a total volume of 1 ml. The PraC reaction mixture was prepared as described in Materials and Methods. The reaction mixtures were analyzed by HPLC at the start (A to C) and after 10 min (D to F) of incubation. Compounds were monitored at 228 nm (A and D), 272 nm (B and E), and 303 nm (C and F).