background: Parvalbumin (PV) can be subdivided into two phylogenetic lineages, αPV and βPV. The bony fish βPV is considered a major fish allergen. However, there is no available report on the immunological property and epitope mapping of bony fish αPV. results: To characterize the allergenic property of bony fish αPV and investigate the difference in allergenic property of bony fish αPV and βPV, turbot (Scophthalmus maximus) αPV and βPV were identified by mass spectrometry and were expressed in Escherichia coli system in this study. Spectra analysis and three-dimensional (3D) modeling showed the similar structure between αPV and βPV. However, αPV exhibited lower immunoglobulin E/immunoglobulin G (IgE/IgG) binding capacity than βPV. Three identified βPV epitopes possessed higher IgE reactivity and more hydrophobic residues than three identified αPV epitopes. In addition, less similarity in sequence homology of αPV epitopes was observed with allergen sequences in database. conclusions: These finding expanded information on fish PV epitopes and substantiated the difference in allergenicity and epitope mapping between fish αPV and βPV, which will improve the epitope-based detection tools of PV and diagnostic of PV induced fish allergy. © 2023 Society of Chemical Industry.