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A representative time course of pNPA hydrolysis by WT CES1. The reaction was performed in a 1 ml volume containing Tris–HCl, pH 7.5, absolute ethanol, 100 mM pNPA and 20 μl purified WT CES1 at 37 °C. Hydrolysis was determined after incubating the reaction at 37 °C for 10 min, and the formation of pNP was followed at 405 nm for 10 min. A blank reaction was performed by replacing the enzyme with buffer.
