Figure 2.

(a) Influence of different type and different percentages of organic co-solvents (50 mM Tris-HCl buffer pH 8.0 supplemented with increasing vol% of organic co-solvent) on CaOYE protein melting point T_m (°C) (as measured by Thermofluor assay). All measurements have been performed in triplicate; (b) CaOYE activity-pH profile: the specific activity (U/mg) was determined by monitoring the reduction of 2-cyclohexen-1-one in the presence of NADPH. A universal buffer of constant ionic strength, 50 mM CAPS, 50 mM Tris-HCl, 50 mM MES and 50 mM AcOH adjusted to different pH values (4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0 and 11.0) at 25 °C by 5 M NaOH or 5 M HCl, was used to determine the activity of the enzyme.