ArticleApplied and environmental microbiology2010
Product-induced gene expression, a product-responsive reporter assay used to screen metagenomic libraries for enzyme-encoding genes.
Article in Applied and environmental microbiology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 41 papers.
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Who cites it
41 citing papers in PubMed, 122 citations in OpenAlex.
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- Microbial Transcription Factor-Based Biosensors: Innovations from Design to Applications in Synthetic Biology.Biosensors · 2025Review
- Combining biosensor and metabolic network optimization strategies for enhanced L-threonine production in Escherichia coli.Biotechnology for biofuels and bioproducts · 2025Article
- Microbial amidases: Characterization, advances and biotechnological applications.Biotechnology notes (Amsterdam, Netherlands) · 2025Review
- Genetically encoded biosensors for the circular plastics bioeconomy.Metabolic engineering communications · 2024Article
- Marine Bioprospecting, Biocatalysis and Process Development.Microorganisms · 2022Review
- Transcription Factor-Based Biosensors for Detecting Pathogens.Biosensors · 2022Review
- Strategies for Natural Products Discovery from Uncultured Microorganisms.Molecules (Basel, Switzerland) · 2021Review
- A genome-wide approach for identification and characterisation of metabolite-inducible systems.Nature communications · 2020Article
- Antagonistic Control of Genetic Circuit Performance for Rapid Analysis of Targeted Enzyme Activity in Living Cells.Frontiers in molecular biosciences · 2020Article
- Transcription Factor Engineering for High-Throughput Strain Evolution and Organic Acid Bioproduction: A Review.Frontiers in bioengineering and biotechnology · 2020Review
- De novo design of programmable inducible promoters.Nucleic acids research · 2019Article
- Biochemical Characteristics of Microbial Enzymes and Their Significance from Industrial Perspectives.Molecular biotechnology · 2019Review
- 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
- The evolving interface between synthetic biology and functional metagenomics.Nature chemical biology · 2018Review
Corrections and comments
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Authors and funding
2 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A reporter assay-based screening method for enzymes, which we named product-induced gene expression (PIGEX), was developed and used to screen a metagenomic library for amidases. A benzoate-responsive transcriptional activator, BenR, was placed upstream of the gene encoding green fluorescent protein and used as a sensor. Escherichia coli sensor cells carrying the benR-gfp gene cassette fluoresced in response to benzoate concentrations as low as 10 μM but were completely unresponsive to the substrate benzamide. An E. coli metagenomic library consisting of 96,000 clones was grown in 96-well format in LB medium containing benzamide. The library cells were then cocultivated with sensor cells. Eleven amidase genes were recovered from 143 fluorescent wells; eight of these genes were homologous to known bacterial amidase genes while three were novel genes. In addition to their activity toward benzamide, the enzymes were active toward various substrates, including d- and l-amino acid amides, and displayed enantioselectivity. Thus, we demonstrated that PIGEX is an effective approach for screening novel enzymes based on product detection.
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Registered trials
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.