ArticleApplied and environmental microbiology2011
Engineering Escherichia coli BL21(DE3) derivative strains to minimize E. coli protein contamination after purification by immobilized metal affinity chromatography.
Article in Applied and environmental microbiology, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 74 papers.
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Who cites it
74 citing papers in PubMed, 129 citations in OpenAlex.
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- IMACulate(DE3), an E. coli Strain for High Purity His-Tagged Protein Purifications.Biotechnology and bioengineering · 2026Article
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- International multisite implementation of distributed cell-free protein biomanufacturing to advance health and research equity.Science advances · 2026Article
- Coordination chemistry-enabled drug delivery systems: metal-ligand platforms for controlled release and targeted therapeutics.Journal of nanobiotechnology · 2026Review
- Cryo-EM reveals ArnA contamination during purification of a ciliary protein complex.Acta crystallographica. Section D, Structural biology · 2026Article
- A katE katG double-knockout E. coli strain eliminates the risk of catalase contamination in recombinant proteins.Applied microbiology and biotechnology · 2026Article
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- Serendipitous high-resolution structure of Escherichia coli carbonic anhydrase 2.Acta crystallographica. Section F, Structural biology communications · 2025Article
- Changing expression system alters oligomerization and proinflammatory activity of recombinant human S100A9.bioRxiv : the preprint server for biology · 2024Article
- Recombinant and Synthetic Affibodies Function Comparably for Modulating Protein Release.Cellular and molecular bioengineering · 2024Article
- Producing recombinant proteins in Vibrio natriegens.Microbial cell factories · 2024Article
- Bacterial biofilm growth and perturbation by serine protease from Bacillus sp.Archives of microbiology · 2024Article
- Optimized expression and purification of a human adenosine deaminase in E. coli and characterization of its Asp8Asn variant.Protein expression and purification · 2024Article
- Substrate Sequestration and Chain Flipping in Human Mitochondrial Acyl Carrier Protein.Biochemistry · 2023Article
14 more citing papers are in PubMed but not listed here.
Corrections and comments
- Erratum issued
Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Recombinant His-tagged proteins expressed in Escherichia coli and purified by immobilized metal affinity chromatography (IMAC) are commonly coeluted with native E. coli proteins, especially if the recombinant protein is expressed at a low level. The E. coli contaminants display high affinity to divalent nickel or cobalt ions, mainly due to the presence of clustered histidine residues or biologically relevant metal binding sites. To improve the final purity of expressed His-tagged protein, we engineered E. coli BL21(DE3) expression strains in which the most recurring contaminants are either expressed with an alternative tag or mutated to decrease their affinity to divalent cations. The current study presents the design, engineering, and characterization of two E. coli BL21(DE3) derivatives, NiCo21(DE3) and NiCo22(DE3), which express the endogenous proteins SlyD, Can, ArnA, and (optionally) AceE fused at their C terminus to a chitin binding domain (CBD) and the protein GlmS, with six surface histidines replaced by alanines. We show that each E. coli CBD-tagged protein remains active and can be efficiently eliminated from an IMAC elution fraction using a chitin column flowthrough step, while the modification of GlmS results in loss of affinity for nickel-containing resin. The "NiCo" strains uniquely complement existing methods for improving the purity of recombinant His-tagged protein.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.