Molecular Biotechnology

, Volume 18, Issue 3, pp 193–198

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

  • Sung-Wook Hwang
  • Jae-Hyun Lee
  • Heung-Bok Park
  • Sang-Hyun Pyo
  • Jin-Eon So
  • Hyun-Soo Lee
  • Seung-Suh Hong
  • Jin-Hyun Kim
Research

DOI: 10.1385/MB:18:3:193

Cite this article as:
Hwang, SW., Lee, JH., Park, HB. 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 peptide magainin derivative fusion protein purification hydroxylamine treatment chromatography 

Copyright information

© Humana Press Inc. 2001

Authors and Affiliations

  • Sung-Wook Hwang
  • Jae-Hyun Lee
  • Heung-Bok Park
  • Sang-Hyun Pyo
  • Jin-Eon So
  • Hyun-Soo Lee
  • Seung-Suh Hong
  • Jin-Hyun Kim
    • 1
  1. 1.Department of Chemical EngineeringKongju National UniversityChungnamSouth Korea

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