Overexpression, purification, and characterization of recombinant CamCPR.

<p>a, SDS-PAGE analyses of CamCPR (I) and tCamCPR (II). M, protein ruler; Lanes 1–4, purified enzyme; Lane 5 in I, cell lysate; Lane 5 in II, whole cell; Lane 6 in I, whole cell; Lane 6 in II, cell lysate. The target band was indicated with an arrowhead. b, the UV spectrum of tCamCPR was measured in 50 mM tris-HCl buffer (pH 7.4). c, the effects of different buffers with various pH on the cytochrome c reducing activity of tCamCPR. d–f, the steady-state kinetic constants of recombinant CamCPR. g, HPLC-DAD analyses of the reaction mixture of CamCPR supported cinnamic acid 4-hydrxoylase activity. Panel I, the authentic <i>p</i>-coumaric acid (♦) and <i>trans</i>-cinnamic acid (•); The HPLC traces of the whole reaction containing the cell lysates (panel II), the cells (panel III), and the boiled cells (panel IV) of the recombinant tCamCPR and CYP73A25 as catalyst.