FIGURE 1.
Michaelis-Menten curves for the determination of apparent Km and Vmax values for ATP. The specific activity of each FadD32 was deduced from the dose-response curve for the PiColorLock-based assay with lauric acid (200 μm) as a substrate (as described under “Experimental Procedures”). The initial velocity of the FadD32 reaction was assessed spectrophotometrically in the coupled assay. We determined the steady-state apparent parameters Km (Michaelis constant) and Vmax (maximum velocity) by using non-linear regression to fit the Michaelis-Menten equation to the data. Means and standard errors of the means of two replicates are shown.