Abstract：Numerous antimicrobial peptides have been isolated from frog skin secretions. In this study, we purified an antibacterial peptide of 33 amino acids, brevinin2GHa1, from the skin secretion of Hylarana guentheri. The skin secretion from the dorsal skin glands of young adult frogs was obtained by 10-Volt electrical stimulation. Sephadex G-50 size exclusion gel filtration, followed with reverse-phase high performance liquid chromatography (RP-HPLC) was used for purification. Brevinin2GHa1 inhibited the growth of both Gram-positive and Gram-negative bacteria. The minimum inhibitory concentrations (MICs) against Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Salmonella were 7.8, 3.9, 2.0, and 250 μg/mL. The secondary structure of brevinin2GHa1 was analyzed using circular dichroism spectroscopy. High level of random coils was indicated when brevinin-2GHa1 was dissolved in deionized water, whereas increased ordered α-helix conformation in SDS (10 mmol/L) or different concentrations of TFE was observed. The results provided useful clues to elucidate the antibacterial mechanism of brevinin-2GHal.
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