Intein-mediated expression of cecropin in Escherichia coli

Mauricio Díaz, Elena Venturini, Stefano Marchetti, Gloria Arenas, Sergio H. Marshall

Research output: Contribution to journalArticlepeer-review

3 Scopus citations


Different strategies have been used to overcome the difficulties to produce antimicrobial peptides. Here we used Intein Mediated Purification with an Affinity Chitin-binding Tag (IMPACT-System, New England Biolabs) for the expression of the antimicrobial peptide cecropin to reduce its sensitivity to intracellular proteases and use its inducible self-cleaving capability to remove the carrier. Cecropin was cloned into suitable expression vector pTYB11, and expression induced by IPTG in Escherichia coli ER2566. The use of 22°C induction allowed the expression of cecropin with its intein carrier in soluble form. Cell extracts were purified by chitin affinity chromatography and intein-mediated splicing of the target protein was achieved by thiol addition, obtaining a final yield of 2.5 mg cecropin/l. Cecropin cleaved from the intein had its proper biologically active form, showing a micromolar antimicrobial activity against Vibrio ordalii, Vibrio alginolyticus and Escherichia coli.

Original languageEnglish
Pages (from-to)30-38
Number of pages9
JournalElectronic Journal of Biotechnology
Issue number2
StatePublished - 15 Mar 2012


  • Antimicrobial
  • Cecropin
  • Fusion
  • Intein
  • Peptide
  • Soluble


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