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To investigate immune activity of pine nut peptides treated by PEF technology and mechanism of targeting immunoactive sitesits, immune regulatory active was evaluated by RAW 264.7 cells model and the structures of pine nut peptides were researched by fluorescence, CD, and 1D/2D NMR spectrum. These consequences showed the ability of macrophages to phagocytosis neutral red and the production of nitric oxide (NO) were improved after PEF treatment. KWFCT treated by PEF treatment with 40 kV/cm obtained the best immunocompetence. The CD spectroscopy showed that PEF could transform the secondary structures of pine nut peptides. The short‐range correlation between CγH (1.65 ppm) and CαH (3.35 ppm), and long‐range correlation between CαH (3.37 ppm) and NαH (8.07 ppm) were enhanced by PEF treatment. PEF treatment of tryptophan in the pine nut peptides enhanced the immunological activity of the pine nut peptides. Practical applications Bioactive peptides derived from food proteins have been extensively studied in recent years. However, little research has been done on the immunoactive peptide of pine nut source. PEF treatment is promising for improving certain properties of foods while maintaining the flavor, color, taste, and nutritional value of the food. This research demonstrated that PEF treatment increased the immunological activity of KWFCT and KWFM. The primary structure of KWFCT and KWFM did not change after PEF treatment, but the secondary structure was transformed into each other. A new perspective on the PEF action site is proposed, which is beneficial to the application of PEF technology.
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