Binding studies of crocin to β-Lactoglobulin and its impacts on both components

Light susceptibility of crocin and high degree of unsaturation limit its application in food systems. This study aimed to investigate the impact of molecular binding of crocin to beta-lactoglobulin (beta-LG) on cmcin stability as well as the conformational stability of beta-LG. The overall binding affinity was estimated about 1.74 x 10(3) M-1 through following UV spectral changes of beta-LG at different concentrations of crocin. A 1:1 binding stoichiometry for the complex of crocin and beta-LG was recognized using Job's method. The results of molecular docking and molecular dynamic simulation indicated that in addition to a single hydrogen bond with an oxygen linker attached to polyene chain, hydrophobic interactions play the major role in binding of crocin's polyene chain to beta-LG surface. Three-dimensional emission and CD spectra showed conformational alterations in both tertiary and secondary structure of beta-LG varying with crocin concentration. Depending on the temperature, crocin showed a dual effect on the thermodynamic stability of beta-LG. Crocin destabilized beta-LG up to room temperature, but stabilized at higher temperatures. The CD spectra of crocin in the Soret region and simulation results proved the bending of polyene chain and changes in the angle between sugar groups and polyene chain in combination with the protein. beta-LG complexation enhanced the chemical stability of crocin by 20% during a 30-day storage, while its degradation was accelerated under UV irradiation. According to the results, it is predicted crocin functionalities to be affected by beta-LG association and vice versa.