Protective Effect of EDC/NHS Cross-Linking Against Urea-Induced Collagen Destabilization in Ready-to-Eat Sea Cucumber...

Ready-to-eat sea cucumbers (RSC) cannot be preserved at room temperature due to autolysis, which is closely related to the instability of collagen resulting from the disruption of hydrogen bonds. To investigate the protective effect of N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS) cross-linking against disruption of hydrogen bonds and its role in stabilizing RSC quality at room temperature, this study designed comparative experiments involving EDC/NHS cross-linking treatments with varying sequences of hydrogen bonds disruption. The results indicated that EDC/NHS positively affects the stabilization of the collagen structure in RSC. The various quality parameters of both groups of RSC that underwent cross-linking treatment before and after hydrogen bonds disruption were significantly better than those of the control group, which only experienced the breaking of hydrogen bonds. Notably, the Eb group, which underwent EDC/NHS cross-linking treatment prior to the disruption of the hydrogen bonds network, yielded even more favorable results. Preliminary analyses of textural properties and moisture content suggested that EDC/NHS helps delay the deterioration of RSC quality. The levels of soluble components and carbonyl groups indicated that prior cross-linking treatment is more effective in mitigating collagen degradation and oxidation. Differential scanning calorimetry revealed that the reduction in ΔH for the Eb group was only 2.4%. Furthermore, fluorescence spectroscopy, Fourier transform infrared spectroscopy, and circular dichroism spectroscopy, examined from the perspectives of secondary and tertiary structures respectively, indicated that the cross-linking mechanism of EDC/NHS involves the formation of a more robust network of amide bonds, thereby preventing the disruption of hydrogen bonds and enhancing collagen stability, enabling it to better resist the cleavage of hydrogen bonds due to urea. The scanning electron microscope and Van Gieson's staining techniques offer a clearer illustration of this point from a microscopic perspective. Moreover, molecular docking simulations have indicated the cross-linking mechanism of EDC/NHS at the atomic level, thereby establishing a scientific foundation for the potential application and development of EDC/NHS in room-temperature storage technologies for RSC.