Design, expression, and characterization of a novel cecropin A-derived peptide with high antibacterial activity
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In recent years, antimicrobial peptides have received increased interest and are potential substitutes for antibiotics. However, natural antimicrobial peptides are always toxic to mammalian cells and usually exhibit weak antibacterial activity, which restrict their wide application. In this study, a novel antibacterial peptide named PEW300 was designed with three mutations to the parental peptide cecropin A. As predicted by bioinformatic programs, the positive charge of PEW300 increased from + 6 to + 9 compared with cecropin A, and the grand average of hydropathicity increased from − 0.084 to − 0.008. Expression of PEW300 resulted in serious inhibition of Escherichia coli BL21(DE3) cells, indicating designed PEW300 may have stronger antibacterial activity. A simple, fast, and low-cost approach without tedious protein purification steps was selected for the efficient production of PEW300 by fusion with ELK16 and about 7.38 μg/mg wet cell weight PEW300 was eventually obtained. Purified PEW300 exhibited strong antibacterial activity against various Gram-positive and Gram-negative bacteria which was enhanced four- to sevenfold compared with the parental peptide cecropin A. Besides, PEW300 had no hemolytic activity toward mammalian cells even at high concentration (224 ng/μl). PEW300 showed good stability in neutral and alkaline solutions. Moreover, PEW300 was thermally stable even at up to 100 °C and resistant to proteinase K, pepsin, snailase, and trypsin. The incubation with human serum had no effect on the antibacterial activity of PEW300. All these results demonstrated that PEW300 designed in this work may have good potential as a candidate pharmaceutical agent.
KeywordsAntimicrobial peptide Escherichia coli Cecropin A PEW300 Prokaryotic expression
This study was supported by the Innovative Program of Department of Education of Guangdong Province, China (grant number 2013KJCX0013) and Natural Science Foundation of Guangdong Province, China (grant number 2015A030310322).
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Conflict of interest
The authors declare that they have no competing interests.
This article does not contain any studies with human participants or animals performed by any of the authors.
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We state this is not applicable. The manuscript does not contain data from any individual person.
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