Molecular Cloning, Prokaryotic Expression, and Immunological Characterization of β-Enolase from Grass Carp...

β-Enolase is a cross-allergen commonly found in fungi, plants, and aquatic products. Although studies on the allergenicity of fish enolase have been reported in recent years, they are still limited to a few species of marine fish. Therefore, the detection of freshwater fish in the food industry requires more studies of the molecular characterization as well as the allergenicity of enolase. In this study, the nucleotide sequence of β-enolase from grass carp was obtained by molecular cloning technology. Structural domain analysis showed that it contained the characteristic structural domains of the enolase superfamily, and homology analysis indicated that enolases are highly conserved evolutionarily. Recombinant β-enolase was obtained by prokaryotic expression, and its allergenicity was assessed by β-enolase-sensitized mice, which confirmed the ability of β-enolase to trigger an allergic response and cause a rise in Th1 and Th2 immune responses in mice. These results suggest that β-enolase could be used as a characterizing substance for the detection of fish allergens in the food industry as well as the preparation of drugs for allergy-related studies.