FIGURE 3.

Time course of substrate consumption and product formation by recombinant purified enzymes PaaZ or PaaZ-E256Q from E. coli K12 (A) and ECH-Aa and PacL from A. aromaticum (B). The reaction mixtures contained 50 mm Tris-HCl (pH 8.0), 0.25 mm oxepin-CoA, and 1.5 mm NADP⁺ (for A) or 1.5 mm NAD⁺ (for B). The reactions at 30 °C were started by the addition of PaaZ (0.5 μg), PaaZ-E256Q (0.8 μg), or ECH-Aa (46 μg). The continuous lines in A present the time course for PaaZ, and the dotted lines present the time course for PaaZ-E256Q. The continuous lines in B show the time course for ECH-Aa in the presence of 28 μg of PacL, and the dotted lines show the time course for ECH-Aa alone. The roman numerals indicate the compounds in Figs. 1 and 4.