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Substrate vs. velocity curve with PNPP. Equal amounts of purified enzyme were incubated with various concentrations of PNPP and the amount of PNP formed was monitored at OD400. All three exhibit Michaelis–Menten kinetics, and protein from a mock purification exhibited a velocity of ∼0.2 mmol PNP released/min × mg protein regardless of substrate concentration (data not shown).
