Optimized E. coli expression strain LOBSTR eliminates common contaminants from His-tag purification

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  • Kasper Røjkjær Andersen
  • Nina C Leksa, Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States
  • Thomas U Schwartz, Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States

His-tag affinity purification is one of the most commonly used methods to purify recombinant proteins expressed in E. coli. One drawback of using the His-tag is the co-purification of contaminating histidine-rich E. coli proteins. We engineered a new E. coli expression strain, LOBSTR (low background strain), which eliminates the most abundant contaminants. LOBSTR is derived from the E. coli BL21(DE3) strain and carries genomically modified copies of arnA and slyD, whose protein products exhibit reduced affinities to Ni and Co resins, resulting in a much higher purity of the target protein. The use of LOBSTR enables the pursuit of challenging low-expressing protein targets by reducing background contamination with no additional purification steps, materials, or costs, and thus pushes the limits of standard His-tag purifications.

Original languageEnglish
JournalProteins - Structure Function and Bioinformatics
Volume81
Issue11
Pages (from-to)1857-1861
Number of pages5
ISSN0887-3585
DOIs
Publication statusPublished - Nov 2013

    Research areas

  • Chromatography, Affinity, Escherichia coli, Escherichia coli Proteins, Histidine, Recombinant Proteins

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