ArticleCommunications biology2021
Tailoring the evolution of BL21(DE3) uncovers a key role for RNA stability in gene expression toxicity.
Article in Communications biology, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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19 citing papers in PubMed.
- High-cell-density Escherichia coli-based nanobody production: benchmarking of subcellular targeting, strain background and process development strategies.Microbial cell factories · 2026Article
- An engineered biofactory for efficient production of diverse recombinant superoxide dismutase isozymes loaded with specific metal ions for biochemical characterization.bioRxiv : the preprint server for biology · 2026Article
- How far can you go? Extrapolating values of catalytic activity from known protein landscapes in natural and directed evolution.Chemical Society reviews · 2026Review
- Leaky recombinant expression reveals design constraints of bicistronic synthetic operons in Escherichia coli.Scientific reports · 2026Article
- Multilayered regulation by RNA thermometers enables precise control of Cas9 expression in E. coli.Nucleic acids research · 2026Article
- A multi-dentate, cooperative interaction between endo- and exo-ribonucleases within the bacterial RNA degradosome.Nucleic acids research · 2025Article
- Optimization of CYP27A1 recombinant protein expression.Protein expression and purification · 2025Article
- Development of a cost-effective serodiagnosis for African swine fever using solubility-enhanced recombinant p54, p30, and p72.Veterinary world · 2025Article
- Innovations, Challenges and Future Directions of T7RNA Polymerase in Microbial Cell Factories.ACS synthetic biology · 2025Review
- Recombinant multiepitope proteins expressed in Escherichia coli cells and their potential for immunodiagnosis.Microbial cell factories · 2024Review
- Escherichia coli strains with precise domain deletions in the ribonuclease RNase E can achieve greatly enhanced levels of membrane protein production.Protein science : a publication of the Protein Society · 2024Article
- Tips for efficiently maintaining pET expression plasmids.Current genetics · 2023Article
- Engineering and Expression Strategies for Optimization of L-Asparaginase Development and Production.International journal of molecular sciences · 2023Review
- Dynamic feedback regulation for efficient membrane protein production using a small RNA-based genetic circuit in Escherichia coli.Microbial cell factories · 2022Article
- Efficient Bacterial Genome Engineering throughout the Central Dogma Using the Dual-Selection MarkerACS synthetic biology · 2022Article
- Cloning and Expression of Heparinase Gene from a Novel Strain Raoultella NX-TZ-3-15.Applied biochemistry and biotechnology · 2022Article
- Current trends in biopharmaceuticals production in Escherichia coli.Biotechnology letters · 2022Review
- Strategies for Successful Over-Expression of Human Membrane Transport Systems Using Bacterial Hosts: Future Perspectives.International journal of molecular sciences · 2022Review
- A standardized genome architecture for bacterial synthetic biology (SEGA).Nature communications · 2021Article
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2 authors.
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No grant is acknowledged in the PubMed record.
Abstract
Gene expression toxicity is an important biological phenomenon and a major bottleneck in biotechnology. Escherichia coli BL21(DE3) is the most popular choice for recombinant protein production, and various derivatives have been evolved or engineered to facilitate improved yield and tolerance to toxic genes. However, previous efforts to evolve BL21, such as the Walker strains C41 and C43, resulted only in decreased expression strength of the T7 system. This reveals little about the mechanisms at play and constitutes only marginal progress towards a generally higher producing cell factory. Here, we restrict the solution space for BL21(DE3) to evolve tolerance and isolate a mutant strain Evo21(DE3) with a truncation in the essential RNase E. This suggests that RNA stability plays a central role in gene expression toxicity. The evolved rne truncation is similar to a mutation previously engineered into the commercially available BL21Star(DE3), which challenges the existing assumption that this strain is unsuitable for expressing toxic proteins. We isolated another dominant mutation in a presumed substrate binding site of RNase E that improves protein production further when provided as an auxiliary plasmid. This makes it easy to improve other BL21 variants and points to RNases as prime targets for cell factory optimisation.
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