α naphtyl acetate

MZ0003 deacetylase activity was detected using the Acetate Colorimetric Assay Kit (Sigma-Aldrich, St Louis, MO, USA). The substrates used, purchased from Sigma-Aldrich, were: 4-methylumbelliferyl acetate and 7-aminocephalosporanic acid (dissolved in DMSO); α-naphtyl acetate, β-D-glucose pentaacetate and β-D-galactose pentaacetate (dissolved in methanol); 5-bromo-4-chloro-3-indoyl acetate (dissolved in ethanol); p-NP acetate and cellulose acetate (dissolved in acetonitrile). The reaction mixtures, each containing 10 μg of protein and substrate, were pre-incubated in 5 mM Na-phosphate buffer pH 7.3 for 30 min at 35°C. Thereafter the assay was developed according to the manufacturer’s instructions and the absorbance was read at 450 nm. Values for each substrate were taken as the mean of three independent measurements after subtraction of a blank sample, without MZ0003, to correct for background degradation. The amount of acetate liberated was calculated in nM μl^-1 from a standard curve according to the manufacturer’s instructions.