Exploring the inhibitory effect of small molecules on pancreatic lipase (PL) has become an important method for screening active ingredients with anti-obesity effect in vitro. The binding reaction and mechanism of genistein (Gen) and naringenin (Nar) with PL were studied by fluorescence spectroscopy， non-radiative energy transfer theory and high performance liquid chromatography. At the same time， three-dimensional fluorescence， surface hydrophobicity， X-ray diffraction， infrared spectroscopy， particle size， differential scanning calorimetry， and scanning electron microscopy were used to characterize the structure of the composites from different angles， and their influence on the structure and properties of PL were comprehensively clarified. The results showed that the quenching of PL by Gen and Nar was static quenching， and the binding constants were 42.39 L/mg and 22.57 L/mg， respectively. The addition of both reduced the α-helix content of PL， increased the content of β-turn， β-sheet and random coil， and increased the crystallinity from 25.60% to 32.77% and 38.00%， respectively. The surface hydrophobicity decreased and the thermal stability decreased (the denaturation temperature decreased from 76.04 ℃ to 65.00 ℃ and 65.05 ℃， respectively)， and the PL changed from a smooth sheet structure to a rod-like particle. These results indicate that Gen and Nar significantly affect the structure and properties of PL， providing new ideas for the development of new PL inhibitors.

multispectral methods  /  genistein  /  naringenin  /  pancreatic lipase  /  inhibition mechanism