Comparative studies on the interaction of nine flavonoids with trypsin

In this study, the interaction between nine classic flavonoids (including baicalin, quercetin, myricetin, rutin, puerarin, daidzein, liquiritin and isoliquiritin) and trypsin was investigated by fluorescence spectroscopy and molecular modeling methods. The results reveal that all flavonoids can interact with trypsin to form flavonoid-trypsin complexes. The binding parameters obtained from the data at different temperatures indicate that all flavonoids can spontaneously bind with trypsin with one binding site. The binding constants of trypsin with nine classic flavonoids are in the following order as: baicalin > myricetin > rutin > isoliquiritin > hesperidin > puerarin > quercetin > daidzein > liquiritin. The interaction forces between flavonoids and trypsin may be electrostatic forces (except for rutin/puerarin/daidzein), hydrophobic interactions as well as van der Waals forces. Synchronous fluorescence spectroscopy shows that the interaction between flavonoids and trypsin changes the hydrophobicity of the microenvironment of tryptophan (Trp) residues. All flavonoids close to tyrosine (Tyr) residues but have no effect on the microenvironment around Tyr residues except for hesperidin and liquiritin. Molecular modeling displays that all flavonoids bind directly into trypsin cavity site and lead to a decrease in enzyme activity.

Keywords: Flavonoids; Fluorescence spectroscopy; Molecular modeling; Trypsin.