muramidase and epicatechin-gallate

The binding of caffeic acid (CA) and/or (-)-epicatechin gallate (ECG) to lysozyme was investigated by multispectroscopic methods and molecular docking. The effects of the single and combined binding on the structure, activity and stability of lysozyme and the synergistic antioxidant activity of CA and ECG were also studied. Fluorescence quenching spectra, time-resolved fluorescence spectra, and UV-vis absorption difference spectra all ascertained the static quenching mechanism of lysozyme by CA/ECG. Thermodynamic parameters indicated that CA and ECG competitively bound to lysozyme, and CA had a stronger binding affinity, which was consistent with the results of molecular docking. Hydrogen bonding, van der Waals' force and electrostatic interaction were the main driving forces for the binding process. Synchronous fluorescence spectra displayed that the interaction of CA/ECG exposed the tryptophan residues of lysozyme to a more hydrophilic environment. Circular dichroism spectroscopy, Fourier transform infrared spectroscopy and dynamic light scattering indicated that the binding of CA and/or ECG to lysozyme resulted in the change of the secondary structure and increased the particle size of lysozyme. The binding of CA and/or ECG to lysozyme inhibited the enzyme activity and enhanced the thermal stability of lysozyme. The combined application of CA and ECG showed antioxidant synergy which was influenced by the encapsulation of lysozyme and cellular uptake. In summary, this work provides theoretical guidance for lysozyme as a carrier for the combined application of CA and ECG.

Topics: Antioxidants; Binding Sites; Caffeic Acids; Catechin; Circular Dichroism; Hydrogen Bonding; Molecular Docking Simulation; Muramidase; Protein Binding; Spectrometry, Fluorescence; Thermodynamics

Green tea polyphenols (GTPs) are found to be potent inhibitors of amyloid fibril formation. We report the effective inhibitory property of (-)-epicatechin gallate (ECG) during the alkali-salt induced fibrillogenesis of hen egg white lysozyme (HEWL) at 37 °C. Spectroscopic techniques such as fluorescence, circular dichroism and microscopic images show that (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECG), and (-)-epigallocatechin gallate (EGCG) show moderate inhibition of fibrillation with ECG as the most potent polyphenol. Aromatic interactions, hydrophobic interactions, the radical scavenging activity and autoxidation of polyphenols are likely to be the major reasons for ECG being the most effective inhibitor.

Topics: Alkalies; Animals; Benzothiazoles; Catechin; Chickens; Circular Dichroism; Guanosine Triphosphate; Hydrogen-Ion Concentration; Kinetics; Microscopy, Fluorescence; Muramidase; Nephelometry and Turbidimetry; Protein Structure, Secondary; Salts; Thiazoles; Time Factors; Tryptophan