Umami peptide synergy unveiled: A comprehensive study from molecular simulation to practical validation of sensing strategy.

Synergistic effect is one of the main properties of umami substances, as a new natural umami agent, umami peptide synergy has not been systematically explored. Presently, conventional methods relying on human sensory evaluation and intelligent instrument analysis pose challenges due to their time-consuming and lack of high throughput. This research provides a detailed molecular-level understanding of multiple umami peptides interact with T1R1-VFT simultaneous, revealing that multiple umami peptides promotes stronger binding affinity and more effective receptor activation (from -7.3 kcal mol to -11.19 kcal mol). The kinetic simulations demonstrated a significant reduction in the average fluctuation of protein amino acid residues during the binding process. Moreover, the hydrophobic regions on the protein surface were diminished following binding, and the resultant complex structure was more tightly packed, these phenomena may collectively represent the manifestation of synergistic effects. To validate the simulation results, biolayer interferometry sensing strategies were developed to measure the interaction process, indicating that umami peptides and T1R1-VFT could association and dissociation in solution without significant interactions with other proteins. When multiple umami peptides interacted with T1R1-VFT, the kinetic equilibrium constant decreased and affinity increased (from 1.2 e M to 8.3 e M), showing significant synergistic effect. Furthermore, the practical application ability of this sensing strategy was verified in a complex matrix with multiple real samples. Overall, this comprehensive study combined micro-molecular simulation and biological experiment verification, offering a deeper understanding of umami peptide synergy and paving the way for innovative approaches in flavor science and food product development.