Acidic peptide-mediated expression of the antimicrobial peptide buforin II as tandem repeats in Escherichia coli

Jae H. Lee, Il Minn, Chan B. Park, Sun C. Kim

Research output: Contribution to journalArticlepeer-review

Abstract

Antimicrobial peptides have received increasing attention as a new pharmaceutical substance, because of their broad spectrum of antimicrobial activities and the rapid development of multidrug-resistant pathogenic microorganisms. The main obstacle to the wide application of antimicrobial peptides has been the lack of a cost-effective, mass-production method. A novel mass-production method for an antimicrobial peptide of 21 amino acids, buforin II, which was isolated from the stomach of the amphibian Bufo bufo gargarizans, has been developed. This method is based on the neutralization of the positive charges of buforin II by fusing to an acidic peptide to avoid the lethal effect of the expressed antimicrobial peptide on the host cells. The fusion peptide was expressed in Escherichia coli as tandem repeats to increase the product yield. Multimers of the acidic peptide-buforin II fusion peptide were expressed at high levels without causing damage to the cells. The presence of cysteine residues in the acidic peptide was critical for the high level expression of the fusion peptide multimers. Multimers of this fusion peptide were expressed as inclusion bodies, and about 107 mg of pure buforin II was obtained from 1 L of E. coli culture by cleaving the multimers with CNBr. Recombinant buforin II had an antimicrobial activity identical to that of natural buforin II. These results may lead to a general, cost-effective solution to the mass production of antimicrobial peptides and other basic peptides which are lethal to the host strain.

Original languageEnglish (US)
Pages (from-to)53-60
Number of pages8
JournalProtein Expression and Purification
Volume12
Issue number1
DOIs
StatePublished - Feb 1998
Externally publishedYes

Keywords

  • Acidic peptide
  • Antimicrobial peptide
  • Escherichia coli
  • Expression
  • Tandem multimer

ASJC Scopus subject areas

  • Biotechnology

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