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Design of a fluorometric substrate for determination of Michaelis-Menten kinetics of VapCPAE2754. (A) MALDI-TOF spectra of RNA oligonucleotide cleavage and (B) RNA oligonucleotide sequence showing only one cleavage site. (C) Reaction velocities were determined from the slopes of the fluorescent data sets and the calibration plot. The Michaelis-Menten equation was fit to the data for substrate concentrations 0.25–7.5 μM. (D) From the curve fit Vmax, KM and kcat were calculated.
