Molecular Biotechnology

, Volume 18, Issue 3, pp 193–198

A simple method for the purification of an antimicrobial peptide in recombinant Escherichia coli

Authors

  • Sung-Wook Hwang
  • Jae-Hyun Lee
  • Heung-Bok Park
  • Sang-Hyun Pyo
  • Jin-Eon So
  • Hyun-Soo Lee
  • Seung-Suh Hong
    • Department of Chemical EngineeringKongju National University
Research

DOI: 10.1385/MB:18:3:193

Cite this article as:
Hwang, S., Lee, J., Park, H. et al. Mol Biotechnol (2001) 18: 193. doi:10.1385/MB:18:3:193

Abstract

A magainin derivative, designated MSI-344, was produced in Escherichia coli as fusion protein, by utilizing a truncated amidophsphoribosyltransferase of E. coli as a fusion partner. Bacterial cells transformed with the gene encoding the fusion protein were grown to a high cell density and induced with isopropyl-1-thio-b-D-galatoside (IPTG) to initiate product expression. The fusion protein was accumulated into cytoplasmic inclusion body and recombinant MSI-344 was released from the fusion partner by hydroxylamine treatment. Following cleavage of the fusion protein with hydroxylamine, the released MSI-344 was purified to homogeneity by cationic exchange chromatography. The final purity was at least 95% by reversed-phase high performance liquid chromatography (RP-HPLC). Purified recombinant MSI-344 was found to be indistinguishable from the synthetic peptide determined by amino acid sequences and antimicrobial activity assay.

Index Entries

Antimicrobial peptidemagainin derivativefusion proteinpurificationhydroxylamine treatmentchromatography

Copyright information

© Humana Press Inc. 2001