ArticleApplied and environmental microbiology2014
Using transcriptional control to increase titers of secreted heterologous proteins by the type III secretion system.
Article in Applied and environmental microbiology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed.
- Tumor-specific cytokine therapy mediated by engineered Salmonella with a synthetic protein delivery system.Scientific reports · 2026Article
- A high throughput assay for measuring secreted protein based on a de novo fluorescent reporter reveals regulatory and structural insights in Salmonella type three secretion system.Protein science : a publication of the Protein Society · 2025Article
- Experimental Approaches to Visualize Effector Protein Translocation During Host-Pathogen Interactions.BioEssays : news and reviews in molecular, cellular and developmental biology · 2025Review
- Evaluation of theFrontiers in bioengineering and biotechnology · 2025Article
- Article
- Construction of a constitutively active type III secretion system for heterologous protein secretion.Applied microbiology and biotechnology · 2023Article
- Recruitment of heterologous substrates by bacterial secretion systems for transkingdom translocation.Frontiers in cellular and infection microbiology · 2023Review
- An optimized growth medium for increased recombinant protein secretion titer via the type III secretion system.Microbial cell factories · 2021Article
- Delivery of Heterologous Proteins, Enzymes, and Antigens via the Bacterial Type III Secretion System.Microorganisms · 2020Review
- Developing Gram-negative bacteria for the secretion of heterologous proteins.Microbial cell factories · 2018Review
- An estimate is worth about a thousand experiments: using order-of-magnitude estimates to identify cellular engineering targets.Microbial cell factories · 2018Review
- Proteins adopt functionally active conformations after type III secretion.Microbial cell factories · 2016Article
- YscU/FlhB of Yersinia pseudotuberculosis Harbors a C-terminal Type III Secretion Signal.The Journal of biological chemistry · 2015Article
- Article
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Authors and funding
5 authors.
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Abstract
The type III secretion system (T3SS) encoded at the Salmonella pathogenicity island 1 (SPI-1) locus secretes protein directly from the cytosol to the culture media in a concerted, one-step process, bypassing the periplasm. While this approach is attractive for heterologous protein production, product titers are too low for many applications. In addition, the expression of the SPI-1 gene cluster is subject to native regulation, which requires culturing conditions that are not ideal for high-density growth. We used transcriptional control to increase the amount of protein that is secreted into the extracellular space by the T3SS of Salmonella enterica. The controlled expression of the gene encoding SPI-1 transcription factor HilA circumvents the requirement of endogenous induction conditions and allows for synthetic induction of the secretion system. This strategy increases the number of cells that express SPI-1 genes, as measured by promoter activity. In addition, protein secretion titer is sensitive to the time of addition and the concentration of inducer for the protein to be secreted and SPI-1 gene cluster. Overexpression of hilA increases secreted protein titer by >10-fold and enables recovery of up to 28±9 mg/liter of secreted protein from an 8-h culture. We also demonstrate that the protein beta-lactamase is able to adopt an active conformation after secretion, and the increase in secreted titer from hilA overexpression also correlates to increased enzyme activity in the culture supernatant.
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