Abstract
Herein, the effect of pyridinium based ionic liquids – butyl pyridinium bromide (BPB) and octyl pyridinium bromide (OPB) on the native and denatured state of β-lactoglobulin (BLG) was studied using multi-spectroscopic techniques like steady state fluorescence spectroscopy, time resolved fluorescence, time resolved fluorescence anisotropy, circular dichroism (CD) and dynamic light scattering (DLS). Circular dichroism revealed the increase in the negative ellipticity of the native protein in presence of both the ionic liquids which confirms the increase in the secondary structural content of the protein. Time resolved fluorescence and time resolved fluorescence anisotropy revealed the presence of slow and fast decaying fluorophore populations. There was a decrease in the hydrodynamic diameter of the protein when treated with both the ionic liquids. The protein was denatured utilizing chemical denaturant Guanidinium hydrochloride (GdmCl). The denatured protein was treated with ionic liquids – BPB and OPB. The ionic liquids seem to induce a refolding effect in the denatured protein. The detailed study revealed that BPB was more capable of refolding the unfolded BLG than OPB. The results of all the techniques utilized here corroborate with each other.
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