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Antimicrobial proteins and peptides had been found from a wide varietyof organisms in the last few years. These molecules have attracted muchresearch interest because of their biochemical diversity, broadspecificity on anti-viral, anti-bacterial, anti-fungi, anti-protozoanparasites, anti-tumoural, and wound-healing effects. Antimicrobialproteins and peptides play key roles in innate immunity. They interactdirectly with bacteria and kill them. The brown-spotted grouper,Epinephelus fario , is an important marine fish cultured in southernChina. Recently, bacteria and virus have caused high mortality in E.fario cultures, but its endogenous antimicrobial peptides and proteinshave not been explored. An antimicrobial component was found from theskin homogenate of E. fario. After the skin homogenate was digestedwith trypsin, its antimicrobial activity was lost, which showed thatthe antimicrobial component is a protein. The antimicrobial protein(Efap) was purified from the skin homogenate of E. fario by successiveion-exchange and gel filtration chromatography. Efap was demonstratedto be single protein band by SDS-PAGE, with the apparent molecularweight of 41 kD. Efap exhibited antimicrobial activity both for theGram-positive bacteria, Staphylococcus aureus, Micrococcus luteus andBacillus subtilis, and for the Gram-negative bacteria, Vibrioalginolyticus , Vibrio parahaemolyticus , Vibrio fluvialis ,Pasteurella multocida , Aeromonas hydrophila , Eschrrichiu coli ,and Pseudomonas aeruginosa . Except A. hydrophila, P. aeruginosa, andE. coli (MIC>20 mol/L), most of the tested Gram-negative bacteriawere sensitive to Efap (MIC<20 mol/L). Interestingly, Efap showedpotent antimicrobial activity against Gram-positive bacteria S. aureus(MIC 5-10 mol/L) but comparatively weak antimicrobial activity againstM. luteus and B. subtilis. The broad antimicrobial activities of Efapsuggest that it contributes to the innate host defence of E. fario.

Purification and Antimicrobial Activity of Antimicrobial Protein from Brown-spotted Grouper, <I>Epinephelus fario</I>