ArticleNucleic acids research2014
Engineering an Acinetobacter regulon for biosensing and high-throughput enzyme screening in E. coli via flow cytometry.
Article in Nucleic acids research, 2014. 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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Who cites it
30 citing papers in PubMed, 67 citations in OpenAlex.
- Advancing lignocellulosic conversion though biosensor-enabled metabolic engineering.Green chemistry : an international journal and green chemistry resource : GC · 2025Review
- Biological Switches: Past and Future Milestones of Transcription Factor-Based Biosensors.ACS synthetic biology · 2025Review
- Tuning the performance of a TphR-based terephthalate biosensor with a design of experiments approach.Metabolic engineering communications · 2024Article
- Getting the Right Clones in an Automated Manner: An Alternative to Sophisticated Colony-Picking Robotics.Bioengineering (Basel, Switzerland) · 2024Article
- A hybrid RNA-protein biosensor for high-throughput screening of adenosylcobalamin biosynthesis.Synthetic and systems biotechnology · 2024Article
- Enhancing the erythritol production of Yarrowia lipolytica by high-throughput screening based on highly sensitive artificial sensor and anchor protein cwp2.Journal of industrial microbiology & biotechnology · 2024Article
- Biosensors for the detection of chorismate and cis,cis-muconic acid in Corynebacterium glutamicum.Journal of industrial microbiology & biotechnology · 2024Article
- Have you tried turning it off and on again? Oscillating selection to enhance fitness-landscape traversal in adaptive laboratory evolution experiments.Metabolic engineering communications · 2023Review
- A Genetically Encoded Biosensor for the Detection of Levulinic Acid.Journal of microbiology and biotechnology · 2023Article
- Targeted mutagenesis and high-throughput screening of diversified gene and promoter libraries for isolating gain-of-function mutations.Frontiers in bioengineering and biotechnology · 2023Article
- Identification of acetic acid sensitive strains through biosensor-based screening of a Saccharomyces cerevisiae CRISPRi library.Microbial cell factories · 2022Article
- Directed Evolution of Transcription Factor-Based Biosensors for Altered Effector Specificity.Methods in molecular biology (Clifton, N.J.) · 2022Article
- A unified design allows fine-tuning of biosensor parameters and application across bacterial species.Metabolic engineering communications · 2020Article
- Biosensors and Sensing Systems for Rapid Analysis of Phenolic Compounds from Plants: A Comprehensive Review.Biosensors · 2020Review
- Review
- Displaced by Deceivers: Prevention of Biosensor Cross-Talk Is Pivotal for Successful Biosensor-Based High-Throughput Screening Campaigns.ACS synthetic biology · 2019Article
- Directed evolution of the PcaV allosteric transcription factor to generate a biosensor for aromatic aldehydes.Journal of biological engineering · 2019Article
- Genetically encoded biosensors for lignocellulose valorization.Biotechnology for biofuels · 2019Review
- An E. coli biosensor for screening of cDNA libraries for isochorismate pyruvate lyase-encoding cDNAs.Molecular genetics and genomics : MGG · 2018Article
- In vivo biosensors: mechanisms, development, and applications.Journal of industrial microbiology & biotechnology · 2018Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We created a single cell sorting system to screen for enzyme activity in Escherichia coli producing 3,4 dihydroxy benzoate (34DHB). To do so, we engineered a transcription factor regulon controlling the expression of green fluorescent protein (GFP) for induction by 34DHB. An autoregulated transcription factor, pcaU, was borrowed from Acinetobacter sp ADP1 to E. coli and its promoter region adapted for activity in E. Coli. The engineered pcaU regulon was inducible at >5 μM exogenous 34DHB, making it a sensitive biosensor for this industrially significant nylon precursor. Addition of a second plasmid provided IPTG inducible expression of dehydroshikimate dehydratase enzyme (AsbF), which converts endogenous dehydroshikimate to 34DHB. This system produced GFP fluorescence in an IPTG dose-dependent manner, and was easily detected in single cell on flow cytometer despite a moderate catalytic efficiency of AsbF. Using fluorescence-activated cell sorting (FACS), individual cells carrying the active AsbF could be isolated even when diluted into a decoy population of cells carrying a mutant (inactivated) AsbF variant at one part in a million. The same biosensor was also effective for further optimization of itself. FACS on E. coli carrying randomized loci in the promoter showed several variants with enhanced response to 34DHB.
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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.