Production and purification of a novel antibiotic peptide, adenoregulin, from a recombinant Escherichia coli

Biotechnol Lett. 2005 May;27(10):725-30. doi: 10.1007/s10529-005-5361-2.


Adenoregulin is a member of dermaseptin family which are vertebrate antibiotic peptides having lethal effects against a broad spectrum of bacteria, fungi and protozoa. The 99 bp adenoregulin gene was cloned in the expression vector pET32a and transformed into Escherichia coli BL21(DE3). In fed-batch cultivation of BL21(DE3)/pET32a-adr, an exponential feeding strategy was applied to gain 60 g dry cells l-1. The recombinant fusion protein Trx-ADR was expressed in a soluble form. The fusion protein was isolated by Ni2+-chelating chromatography, cleaved with CNBr and purified to homogeneity through reverse phase-HPLC and size exclusion-HPLC. The purified recombinant adenoregulin had antibacterial activity against Escherichia coli K12D31 with apparent Mr of 3.4 kDa, identical to the anticipated value.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amphibian Proteins / biosynthesis
  • Amphibian Proteins / genetics
  • Animals
  • Antimicrobial Cationic Peptides / biosynthesis*
  • Antimicrobial Cationic Peptides / genetics
  • Anura
  • Bioreactors / microbiology
  • Cell Division / drug effects
  • Chromatography, High Pressure Liquid / methods
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / cytology
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / pharmacology
  • Thioredoxins / genetics


  • Amphibian Proteins
  • Antimicrobial Cationic Peptides
  • Recombinant Fusion Proteins
  • dermaseptin B2, Phyllomedusa bicolor
  • Thioredoxins