Fig 6. Evaluation of secondary structure for SEA and SEE after heating and re-cooling.

Transition curves upon heating (40°C—95°C, dark grey) and cooling (40°C, light grey) were recorded by CD spectroscopy at 220 nm for (A) SEA and (D) SEE at pH 5.0 in presence of 0.1 mM ZnCl₂. Far-UV CD spectra (195–250 nm) for (B) SEA and (E) SEE at pH 5.0 in presence of 0.1 mM ZnCl₂ recorded at 40°C (blue), 95°C (red), and after subsequent cooling to 40°C (dark blue). Quantification of secondary structural elements using the far-UV CD spectra for (C) SEA and (F) SEE at 40°C (blue), upon heating to 95°C (red) and cooling back to 40°C (dark blue). The RMSD between experimental and fitted data for SEA is: 0.235, 0.615 and 0.152 for initial 40°C, 95°C and after subsequent cooling to 40°C, respectively. The RMSD values for SEE are 0.157, 0.179 and 0.153, respectively, for the same conditions.