Characterization of antimicrobial properties of TroH2A-29 peptide from golden pompano (Trachinotus ovatus).

Antimicrobial peptides (AMPs) are small, potent molecules that serve as a crucial first line of defense across a wide range of organisms, including fish. In this study, we investigated the antimicrobial properties of a novel peptide, spanning residues 52 to 80 of the full-length histone H2A protein, comprising a total of 29 amino acids. This peptide, designated as Histone H2A-29 (TroH2A-29), was derived from the golden pompano (Trachinotus ovatus) and evaluated for its activity against both Gram-positive bacteria, Lactococcus garvieae and Staphylococcus epidermidis, and Gram-negative bacteria, Vibrio alginolyticus and Vibrio harveyi. The expression of TroH2A in the intestines, liver, and gills of T. ovatus was significantly upregulated after bacterial infections with L. garvieae and V. harveyi. The highest expression levels were observed at 48 h post-infection in the intestines and at different time points in the liver and gills. TroH2A-29 exhibited a high hydrophobic ratio (51 %) and formed an α-helical structure, suggesting its potential as an antimicrobial agent. Notably, TroH2A-29 induced significant agglutination of all four bacterial species in the presence of Ca⁺. TroH2A-29 demonstrated bactericidal effects against L. garvieae, V. harveyi, and V. alginolyticus, with a MIC of 60 μM. However, it showed no antibacterial activity against S. epidermidis. Transmission electron microscopy (TEM) revealed that TroH2A-29 caused morphological damage to the bacterial cells, including cell collapse in L. garvieae and shrinkage in V. alginolyticus and V. harveyi. No morphological changes were observed in S. epidermidis. Membrane permeability assays showed that TroH2A-29 increased membrane disruption in L. garvieae, V. harveyi, and V. alginolyticus, but had little effect on S. epidermidis. Additionally, TroH2A-29 caused membrane depolarization in all tested bacterial strains. These findings highlight the potential of TroH2A-29 as a novel antimicrobial peptide with selective bactericidal properties.