Cyclic enterobacterial common antigen: Potential contaminant of bacterially expressed protein preparations

Paul J A Erbel, Ronald Seidel, Scott E. Macintosh, Lisa N. Gentile, Juan C. Amor, Richard A. Kahn, James H. Prestegard, Lawrence P. McIntosh, Kevin H. Gardner

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

We have previously reported the identification of the cyclic enterobacterial common antigen (ECACYC) polysaccharide in E. coli strains commonly used for heterologous protein expression (PJA Erbel et al., J. Bacteriol. 185 (2003): 1995). Following this initial report, interactions among several NMR groups established that characteristic N-acetyl signals of ECACYC have been observed in 15N-1H HSQC spectra of samples of various bacterially-expressed proteins suggesting that this water-soluble carbohydrate is a common contaminant. We provide NMR spectroscopic tools to recognize ECACYC in protein samples, as well as several methods to remove this contaminant. Early recognition of ECA-based NMR signals will prevent time-consuming analyses of this copurifying carbohydrate.

Original languageEnglish (US)
Pages (from-to)199-204
Number of pages6
JournalJournal of biomolecular NMR
Volume29
Issue number2
DOIs
StatePublished - Jun 1 2004

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Keywords

  • Enterobacterial common antigen
  • Heterologous protein expression
  • N-acetylated carbohydrate
  • Protein purification

ASJC Scopus subject areas

  • Biochemistry
  • Spectroscopy

Cite this

Erbel, P. J. A., Seidel, R., Macintosh, S. E., Gentile, L. N., Amor, J. C., Kahn, R. A., Prestegard, J. H., McIntosh, L. P., & Gardner, K. H. (2004). Cyclic enterobacterial common antigen: Potential contaminant of bacterially expressed protein preparations. Journal of biomolecular NMR, 29(2), 199-204. https://doi.org/10.1023/B:JNMR.0000019252.65073.24