Tributyrin assay in Petri dishes and spectroscopic analyses of Est16.

Tributyrin assays of Est16 (50 μg/mL) in Petri dishes containing 0.23% Tris-HCl (w/v), 1.2% agar (w/v) and 1% tributyrin at pH 7.0 (A), pH 8.0 (B) and pH 9.0 (C). Circular dichroism assays of Est16 at pH 7.0 (D), pH 8.0 (E) and pH 9.0 (F). The samples contained 5.64 μmol of enzyme and the spectra were recorded using a Jasco J-810 spectropolarimeter with a 10 mm path length. Similarly, the thermal denaturation of the enzyme was measured at 222 nm as a function of the temperature at pH 7.0 (G), pH 8.0 (H) and pH 9.0 (I). The melting temperature (T_m) is shown for each spectrum.