This study aimed to explore the influence of gardenia yellow on in vitro wheat starch digestion. The influence of gardenia yellow on the digestion properties of starch was determined through in vitro digestion, and its action mechanism on slow starch digestion was revealed by laser scanning confocal microscopy, enzymatic inhibition dynamics, and other means of characterization. Results showed that gardenia yellow could inhibit starch digestion, significantly increase the resistant starch and slowly digestible starch contents in starch (P < 0.05), and trigger the decrease in glycemic and hydrolysis indices. Furthermore, gardenia yellow could spontaneously bind to the catalytic sites of α-amylase and α-glucosidase, affect their secondary structures through vdW force and hydrophobic interaction, and reduce their catalytic abilities to inhibit the digestion process of wheat starch. Therefore, the interactions of gardenia yellow with starch and digestive enzymes jointly promote the slow digestion of starch.
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