Abstract
Lactobacillus acidophilus is an important probiotic. The β-glucosidase produced by L. acidophilus GIM1.208 can transform quercetin glycosides of Rosa roxburghii Tratt to release quercetin and improve the functional activity of raw materials. Understanding the interaction and the characteristics of the two will lay a theoretical foundation for the site-directed transformation and functional application of the catalytic active site of enzymes. In our study, using the heterologously expressed and highly stable, purified L. acidophilus GIM1.208 BGL as the strain, the representative quercetin in β-glucosidase and Rosa roxburghii Tratt was preliminarily predicted and explored using ultraviolet-visible absorption spectroscopy. Fluorescence spectroscopy combined with molecular docking was used to determine the interaction characteristics of the glycoside substrates, rutin (Rut) and isoquercitrin (Iso). Results from molecular docking showed that Asp159, Arg56, Iso294, Phe292, and Gly25 were the main residues of β-glucosidase and Rut. Arg56 was found to be the most crucial residue of β-glucosidase and isoquercitrin; the interaction between Rut and Iso and β-glucosidase was mainly driven by hydrogen bonding. The combined free energy of β-glucosidase and Iso was found to be -182.10 kcal/mol, while that of β-glucosidase and Rut was -32.37 kcal/mol. The results of fluorescence spectroscopy showed that the fluorescence intensity of β-glucosidase decreased with an increase in Rut and Iso concentrations. This interaction made β-glucosidase quench endogenous fluorescence, which was static quenching. The binding constants of Rut and Iso with β-glucosidase were determined to be 0.50×107 and 0.31×107 L/mol, respectively, indicating that rutin had a stronger affinity when interacting with β-glucosidase. These findings were consistent with our prediction results determined using molecular docking studies.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
