ArticleMicrobial cell factories2012
The LIKE system, a novel protein expression toolbox for Bacillus subtilis based on the liaI promoter.
Article in Microbial cell factories, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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30 citing papers in PubMed.
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- Engineering Bacillus subtilis for high-value bioproduction: recent advances and applications.Microbial cell factories · 2025Review
- Delaying production with prokaryotic inducible expression systems.Microbial cell factories · 2024Review
- Secretion-Catalyzed Assembly of Protein Biomaterials on a Bacterial Membrane Surface.Angewandte Chemie (International ed. in English) · 2023Article
- The effect of manipulating glucuronic acid biosynthetic pathway in Bacillus subtilis strain on hyaluronic acid production.AMB Express · 2023Article
- Metabolic engineering of Bacillus subtilis toward the efficient and stable production of CAMB Express · 2023Article
- The Novel Synthetic Antibiotic BDTL049 Based on a Dendritic System Induces Lipid Domain Formation while Escaping the Cell Envelope Stress Resistance Determinants.Pharmaceutics · 2023Article
- Development and application of a rapid all-in-one plasmid CRISPR-Cas9 system for iterative genome editing in Bacillus subtilis.Microbial cell factories · 2022Article
- A host-vector toolbox for improved secretory protein overproduction in Bacillus subtilis.Applied microbiology and biotechnology · 2022Article
- An operator-based expression toolkit forProceedings of the National Academy of Sciences of the United States of America · 2022Article
- Bacterial two-component systems as sensors for synthetic biology applications.Current opinion in systems biology · 2021Article
- The multifunctionality of expression systems inExperimental biology and medicine (Maywood, N.J.) · 2021Review
- An overview and future prospects of recombinant protein production in Bacillus subtilis.Applied microbiology and biotechnology · 2021Review
- Efficient Genome Editing by a Miniature CRISPR-AsCas12f1 Nuclease inFrontiers in bioengineering and biotechnology · 2021Review
- Label-Free Multiple Reaction Monitoring, a Promising Method for Quantification Analyses of Specific Proteins in Bacteria.International journal of molecular sciences · 2020Article
- The Epipeptide YydF Intrinsically Triggers the Cell Envelope Stress Response ofFrontiers in microbiology · 2020Article
- Identification and Characterization of Genes Required for 5-Hydroxyuridine Synthesis in Bacillus subtilis and Escherichia coli tRNA.Journal of bacteriology · 2019Article
- An EngineeredACS synthetic biology · 2019Article
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4 authors.
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Abstract
backgroundBacillus subtilis is a very important Gram-positive model organism of high biotechnological relevance, which is widely used as a host for the production of both secreted and cytoplasmic proteins. We developed a novel and efficient expression system, based on the liaI promoter (PliaI) from B. subtilis, which is under control of the LiaRS antibiotic-inducible two-component system. In the absence of a stimulus, this promoter is kept tightly inactive. Upon induction by cell wall antibiotics, it shows an over 100-fold increase in activity within 10 min.
resultsBased on these traits of PliaI, we developed a novel LiaRS-controlled gene expression system for B. subtilis (the "LIKE" system). Two expression vectors, the integrative pLIKE-int and the replicative pLIKE-rep, were constructed. To enhance the performance of the PliaI-derived system, site-directed mutagenesis was employed to optimize the ribosome binding site and alter its spacing to the initiation codon used for the translational fusion. The impact of these genetic modifications on protein production yield was measured using GFP as a model protein. Moreover, a number of tailored B. subtilis expression strains containing different markerless chromosomal deletions of the liaIH region were constructed to circumvent undesired protein production, enhance the positive autoregulation of the LiaRS system and thereby increase target gene expression strength from the PliaI promoter.
conclusionsThe LIKE protein expression system is a novel protein expression system, which offers a number of advantages over existing systems. Its major advantages are (i) a tightly switched-off promoter during exponential growth in the absence of a stimulus, (ii) a concentration-dependent activation of PliaI in the presence of suitable inducers, (iii) a very fast but transient response with a very high dynamic range of over 100-fold (up to 1,000-fold) induction, (iv) a choice from a range of well-defined, commercially available, and affordable inducers and (v) the convenient conversion of LIKE-derived inducible expression strains into strong constitutive protein production factories.
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