Purification and characterization of YFGAP, a GAPDH-related novel antimicrobial peptide, from the skin of yellowfin tuna, Thunnus albacares.

A 3.4 kDa of antimicrobial peptide was purified from an acidified skin extract of the yellowfin tuna, Thunnus albacares, by preparative acid-urea-polyacrylamide gel electrophoresis and C(18) reversed-phase HPLC. A comparison of the N-terminal amino acid sequence of the purified peptide with that of other known polypeptides revealed high homology with the N-terminus of glyceraldehyde-3-phosphate dehydrogenase (GAPDH); thus, this peptide was designated as the yellowfin tuna GAPDH-related antimicrobial peptide (YFGAP). YFGAP showed potent antimicrobial activity against Gram-positive bacteria, such as Bacillus subtilis, Micrococcus luteus, and Streptococcus iniae (minimal effective concentrations [MECs], 1.2-17.0 μg/mL), and Gram-negative bacteria, such as Aeromonas hydrophila, Escherichia coli D31, and Vibrio parahaemolyticus (MECs, 3.1-12.0 μg/mL) without significant hemolytic activity. According to the secondary structural prediction and the homology modeling, this peptide forms an amphipathic structure and consists of three secondary structural motifs including one α-helix and two parallel β-strands. This peptide did not show membrane permeabilization ability and its activity was bacteriostatic rather than bactericidal. This is the first report of the isolation of an antimicrobial peptide from a tuna species and the first description of the antimicrobial function of the N-terminus of GAPDH of an animal species.