Biochemical and Biophysical Research Communications
Antibiotic activity and synergistic effect of antimicrobial peptide against pathogens from a patient with gallstones
Section snippets
Materials and methods
Peptide synthesis. The peptides were synthesized by the solid phase method using Fmoc(9-fluorenyl-methoxycarbonyl)-chemistry [22]. Rink amide 4-methyl benzhydrylamine (MBHA) resin (0.55 mmol/g) was used as the support to obtain a C-terminal amidate peptide. The coupling of Fmoc-amino acids was performed with N-hydroxybenzotriazole (HOBt) and dicyclohexylcarbodiimide (DCC). Amino acid side chains were protected as follows: tert-butyl (Asp), trityl (Gln), tert-butyloxycarbonyl (Lys). Deprotection
Peptide synthesis, antibacterial, and hemolytic activities
A peptide corresponding to HP (2–20)-amide was chemically synthesized, purified to homogeneity, and subjected to MALDI-mass spectroscopic analysis (Table 1). We previously showed that HP (2–20) inhibits the growth of Escherichia coli and Bacillus megaterium[20]. Melittin, a toxin from honeybee venom, has also been reported to possess potent antimicrobial activity over a broader spectrum [12]. When we compared the antibacterial MIC [25] of HP (2–20) and melittin, we found that bacteria were
Discussion
A peptide corresponding to HP (2–20) was chemically synthesized, purified to homogeneity, and subjected to MALDI-mass spectroscopy. It was found to have a molecular ion peak of 2319.3 Da, which agrees well with the calculated molecular weight (2320.0 Da). This synthetic HP (2–20) had been shown to possess potent antibacterial activity [15], [22]. When we tested it against various kinds of Gram-positive and Gram-negative bacteria, we found that HP (2–20) exhibits potent, broad-spectrum
Acknowledgments
This work was supported, in part, by grants from the Ministry of Science and Technology, Korea, and the Korea Science and Engineering Foundation through the Research Center for Proteineous Materials, and from the National R&D Program for Fusion strategy of Advanced Technologies.
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