Functional Characterization of Galectin-8 from Golden Pompano Trachinotus ovatus Reveals Its Broad-Spectrum Antimicrobial Activity

Abstract

Galectins exhibit a variety of biological functions through interactions with their ligands, including galactose and its derivatives. Tandem-repeat galectins, such as Galectin-8, can act as pattern recognition receptors to aggregate and neutralize bacterial pathogens. In this study, Galectin-8 was identified in Trachinotus ovatus (golden pompano). Galectin-8 consists of two carbohydrate recognition domains (CRDs) connected by a linker region. Furthermore, molecular docking analysis suggests that the C-terminal CRD can bind galactose, mannose, and N-acetylglucosamine at similar binding sites. ToGal-8 expression levels were highest in the brain and blood of healthy T. ovatus. However, following infection with Streptococcus agalactiae, expression levels in the spleen and head kidney surged at 48 h, while liver expression significantly decreased by 96 h. Cytoplasmic Galectin-8 expression was upregulated after stimulation by peptidoglycan compared with lipopolysaccharide. Recombinant ToGal-8 (rToGal-8) was produced using a prokaryotic expression system. This protein could agglutinate red blood cells from rabbits, carp, and T. ovatus independently of Ca²⁺. Moreover, it was also effective in aggregating and eliminating several bacterial strains, such as Staphylococcus aureus, Bacillus subtilis, Vibrio vulnificus, S. agalactiae, Pseudomonas aeruginosa, and Aeromonas hydrophila. Therefore, this study provides an in-depth analysis of the function of T. ovatus Galectin-8 for the first time, offering guidance for the healthy aquaculture of T. ovatus.