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Parvalbumin (PV) induces IgE‐mediated allergic reactions and is an important allergen in fish species. The effects of high hydrostatic pressure (HHP) treatment (300, 450, 600 MPa; 10 min) on the structural characteristics of PV in cultured large yellow croakers (Larimichthys crocea) were investigated. The effects of HHP treatment were analyzed by free sulfhydryl content, surface hydrophobicity, circular dichroism spectroscopy (CD), laser Raman spectroscopy, and endogenous fluorescence spectroscopy. The results showed that after HHP treatment, the free sulfhydryl content in PV decreased slightly and the surface hydrophobicity increased, indicating changes in the tertiary structure of PV. CD showed that with increase in pressure, the content of α‐helix in the secondary structure of PV decreased slightly. This indicated that the stability of the secondary structure of PV was affected by HHP treatment. The secondary structure of PV was dominated by α‐helix and that the α‐helix content in PV also increased. The redshift of endogenous fluorescence spectroscopy decreases the fluorescence intensity. Practical applications: Parvalbumin (PV) in the large yellow croaker is the main allergen with a molecular weight of 12 kDa. It is an allergy risk to consumers. The application of high hydrostatic pressure (HHP) treatment of 200 to 400 MPa could be successfully applied to changes of the tertiary and secondary structure of PV. This suggests a direct and important relationship between the structural characteristics and allergenicity of PV. The application of HHP treatment effectively reduced the allergenicity of PV. It is useful to desensitization of the large yellow croaker before processing. [ABSTRACT FROM AUTHOR]