Increasingly, studies are using ultrasound to elevate the functional properties of proteins, so the interaction between phenolic compounds and proteins induced by ultrasound needs to be further understood. β-Lactoglobulin (β-LG) at pH 8.1, which exists mainly as monomers, was ultrasound treated at 20 kHz ultrasonic intensity and 30% amplitude for 0-5 min and subsequently interacted with resveratrol. Fluorescence data showed that ultrasound pretreatment improved binding constant (Ka ) from (1.62 ± 0.45) × 105 to (9.43 ± 0.55) × 105 M-1 and binding number from 1.13 ± 0.09 to 1.28 ± 0.11 in a static quenching mode. Fluorescence resonance energy transfer (FRET) analysis indicated that resveratrol bound to the surface hydrophobic pocket of native and treated proteins with no obvious changes in energy transfer efficiency (E) and Föster's distance (r). Thermodynamic parameters indicated that ultrasonication shifted the main driving force from the hydrophobic force for native and 1-min treated β-LG to van der Waals forces and hydrogen bonding for both 3-min and 5-min treated proteins. Ultrasonication and resveratrol addition generated significant differences in surface hydrophobicity and the surface charge of the protein (P < 0.05), whereas they had little influence on the secondary structure of β-LG. Compared with the native β-LG/resveratrol complex, ultrasound-treated protein complexes showed significantly stronger 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) scavenging capacity (P < 0.05), and kept relatively stable after 180-min irradiation. Data provided by this study can lead to a better comprehension of the structure and molecular events occurring during the complexing process between an ultrasound-pretreated protein with polyphenol.
Keywords: fluorescence spectra; resveratrol; ultrasonication; β-lactoglobulin.
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