ArticleFrontiers in bioengineering and biotechnology2019
Selection of High Producers From Combinatorial Libraries for the Production of Recombinant Proteins in
Article in Frontiers in bioengineering and biotechnology, 2019. 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, 36 citations in OpenAlex.
- Is faster always better? A critical assessment of Vibrio natriegens as microbial host for recombinant protein production.Bioprocess and biosystems engineering · 2026Review
- Evaluation of Vibrio natriegens as a fast-growing alternative host for plasmid DNA production.Microbial cell factories · 2026Article
- Efficient Separation of a Novel Microbial Chassis,Membranes · 2025Article
- Vibrio natriegens: Application of a Fast-Growing Halophilic Bacterium.Advances in biochemical engineering/biotechnology · 2025Review
- Heterologous Production of Antimicrobial Peptides: Notes to Consider.The protein journal · 2024Article
- Vibrio natriegens as a superior host for the production of c-type cytochromes and difficult-to-express redox proteins.Scientific reports · 2024Article
- Identification and Cross-Characterisation of Artificial Promoters and 5' Untranslated Regions inFrontiers in bioengineering and biotechnology · 2022Article
- Rapid screening of high expressing Escherichia coli colonies using a novel dicistronic-autoinducible system.Microbial cell factories · 2021Article
- Intelligent host engineering for metabolic flux optimisation in biotechnology.The Biochemical journal · 2021Review
- High-cell-density fed-batch cultivations of Vibrio natriegens.Biotechnology letters · 2021Article
- Optimization of Cephalosporin C Acylase Expression in Escherichia coli by High-Throughput Screening a Constitutive Promoter Mutant library.Applied biochemistry and biotechnology · 2021Article
- Strategies to Enhance Periplasmic Recombinant Protein Production Yields inFrontiers in bioengineering and biotechnology · 2021Review
- Article
- Changes inFrontiers in microbiology · 2020Article
Corrections and comments
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Authors and funding
5 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The optimization of recombinant protein production in bacteria is an important stage of process development, especially for difficult-to-express proteins that are particularly sensitive or recalcitrant. The optimal expression level must be neither too low, which would limit yields, nor too high, which would promote the formation of insoluble inclusion bodies. Expression can be optimized by testing different combinations of elements such as ribosome binding sites and N-terminal affinity tags, but the rate of protein synthesis is strongly dependent on mRNA secondary structures so the combined effects of these elements must be taken into account. This substantially increases the complexity of high-throughput expression screening. To address this limitation, we generated libraries of constructs systematically combining different ribosome binding sites, N-terminal affinity tags, and periplasmic translocation sequences representing two secretion pathways. Each construct also contained a green fluorescent protein (GFP) tag to allow the identification of high producers and a thrombin cleavage site enabling the removal of fusion tags. To achieve proof of principle, we generated libraries of 200 different combinations of elements for the expression of an antimicrobial peptide (AMPs), an antifungal peptide, and the enzyme urate oxidase (uricase) in
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Registered trials
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