Characterization of fluoroalcohols-induced intermediates of Mucor miehei lipase at low pH

We have previously characterized an acid-unfolded (U-A) state of Mucor miehei lipase at pH 2. The effect of 2,2,2-trifluoroethanol (TFE) and 1,1,1,3,3,3-hexafluoroisopropanol (HFIP) resulted in characterization of molten-globule (MG) like states with sheet secondary structure at 15% (v/v) TFE and 6% (v/v) HFIP. Helical states accumulate at 80% (v/v) TFE and 30% (v/v) HFIP.