ArticleMicrobial cell factories2017
A high-throughput expression screening platform to optimize the production of antimicrobial peptides.
Article in Microbial cell factories, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 58 citations in OpenAlex.
- Antimicrobial peptides: emerging next-generation strategy for sustainable plant disease management.Frontiers in antibiotics · 2026Review
- A High-Throughput Multi Well Plate-Based Approach for the Combined Expression, Export, and Assay of Recombinant Proteins.Current protocols · 2025Article
- Synthesis and Evaluation of Aquatic Antimicrobial Peptides Derived from Marine Metagenomes Using a High-Throughput Screening Approach.Marine drugs · 2025Article
- Unlocking the power of antimicrobial peptides: advances in production, optimization, and therapeutics.Frontiers in cellular and infection microbiology · 2025Review
- Current achievements, strategies, obstacles, and overcoming the challenges of the protein engineering in Pichia pastoris expression system.World journal of microbiology & biotechnology · 2023Review
- Antimicrobial peptides from Bacillus spp. and strategies to enhance their yield.Applied microbiology and biotechnology · 2023Review
- Understanding the Necessity of Regulatory Protein Machinery in Heterologous Expression of Class-III Type of Ocins.The protein journal · 2023Review
- Efficient recombinant production of mouse-derived cryptdin family peptides by a novel facilitation strategy for inclusion body formation.Microbial cell factories · 2023Article
- Application of antimicrobial peptides in plant protection: making use of the overlooked merits.Frontiers in plant science · 2023Review
- Efficient Production of Broad-Spectrum Antimicrobials byFrontiers in bioengineering and biotechnology · 2022Article
- Rapid screening of high expressing Escherichia coli colonies using a novel dicistronic-autoinducible system.Microbial cell factories · 2021Article
- CRISPR-Cas9-mediated genomic multiloci integration in Pichia pastoris.Microbial cell factories · 2019Article
- Selection of High Producers From Combinatorial Libraries for the Production of Recombinant Proteins inFrontiers in bioengineering and biotechnology · 2019Article
- Process Intensification for an Insect Antimicrobial Peptide Elastin-Like Polypeptide Fusion Produced in Redox-EngineeredFrontiers in bioengineering and biotechnology · 2019Article
- Single-cell cloning enables the selection of more productiveBiotechnology reports (Amsterdam, Netherlands) · 2018Article
- GoldenPiCS: a Golden Gate-derived modular cloning system for applied synthetic biology in the yeast Pichia pastoris.BMC systems biology · 2017Article
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Corrections and comments
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Authors and funding
8 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundAntimicrobial peptides (AMPs) are promising candidates for the development of novel antibiotics, but it is difficult to produce sufficient quantities for preclinical and clinical studies due to their toxicity towards microbial expression hosts. To avoid laborious trial-and-error testing for the identification of suitable expression constructs, we have developed a small-scale expression screening platform based on a combinatorial plasmid library.
resultsThe combinatorial library is based on the Golden Gate cloning system. In each reaction, six donor plasmids (each containing one component: a promoter, fusion partner 1, fusion partner 2, protease cleavage site, gene of interest, or transcriptional terminator) were combined with one acceptor plasmid to yield the final expression construct. As a proof of concept, screening was carried out in Escherichia coli and Pichia pastoris to study the expression of three different model AMPs with challenging characteristics, such as host toxicity or multiple disulfide bonds. The corresponding genes were successfully cloned in 27 E. coli and 18 P. pastoris expression plasmids, each in a one-step Golden Gate reaction. After transformation, small-scale expression screening in microtiter plates was followed by AMP quantification using a His
conclusionsThe optimization of recombinant protein production is best achieved from first principles by initially optimizing the genetic construct. The unrestricted combination of multiple plasmid features yields a comprehensive library of expression strains that can be screened for optimal productivity. The availability of such a platform could benefit all laboratories working in the field of recombinant protein expression.
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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.