Interaction of Different Polyphenols with Bovine Serum Albumin (BSA) and Human Salivary α-Amylase (HSA) by Fluorescence Quenching

Phenolic compounds are responsible for major organoleptic characteristics of plant-derived food and beverages; these substances have received much attention, given that the major function of these compounds is their antioxidant ability. In the context of this study, our major aim was study the binding of several phenolic compounds such as (+)-catechin, (−)-epicatechin, (−)-epicatechin gallate, malvidin-3-glucoside, tannic acid, procyanidin B4, procyanidin B2 gallate, and procyanidin oligomers to different proteins (bovine serum albumin and human α-amylase) by fluorescence quenching of protein intrinsic fluorescence. From the spectra obtained, the Stern−Volmer, the apparent static, and the bimolecular quenching constants were calculated. The structure of polyphenols revealed to significantly affect the binding/quenching process; in general, the binding affinity increased with the molecular weight of polyphenol compounds and in the presence of galloyl groups. For catechin monomer and procyanidin dimer B4, the KSV was 14100 and 13800 M-1, respectively, and for galloyl derivatives, the KSV was 19500 and 21900 M-1, respectively. Tannic acid was shown to be the major quenching molecule for both proteins. However, comparing different proteins, the same polyphenol showed different quenching effects, which are suggested to be related to the three-dimensional structure of the proteins studied. For (+)-catechin and BSA, the KSV was 8700 M-1, and with α-amylase, it was 14100 M-1; for tannic acid, the KSV was 100548 and 110674 M-1, respectively. From the results obtained, besides the main binding analysis performed, we conclude that this technique is more sensitive than thought because we can detect several interactions that have not been proven by other methods, namely, nephelometry. Overall, fluorescence quenching has proven to be a very sensitive technique with many potentialities to analyze the interaction between polyphenols and proteins. Keywords: Polyphenols; bovine serum albumin; human salivary α-amylase; fluorescence quenching