ArticleThe EMBO journal2003
Directed evolution of an extremely fast phosphotriesterase by in vitro compartmentalization.
Article in The EMBO journal, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 76 papers.
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Who cites it
76 citing papers in PubMed, 318 citations in OpenAlex.
- Directed evolution of enzymes at the crossroads of tradition and innovation.FEBS open bio · 2026Review
- Peptide ligase-mediated display: A cell-free platform for tunable selection of affinity peptides.PNAS nexus · 2026Article
- AI-Driven Enzyme Engineering: Emerging Models and Next-Generation Biotechnological Applications.Molecules (Basel, Switzerland) · 2025Review
- Autonomous ribosome biogenesis in vitro.Nature communications · 2025Article
- Cell-Free Gene Expression: Methods and Applications.Chemical reviews · 2025Review
- A sticky situation - simple method for rapid poissonian encapsulation of highly aggregation-prone microbeads in polydisperse emulsions.Frontiers in bioengineering and biotechnology · 2025Article
- Applications of Microbial Organophosphate-Degrading Enzymes to Detoxification of Organophosphorous Compounds for Medical Countermeasures against Poisoning and Environmental Remediation.International journal of molecular sciences · 2024Review
- Thermostable in vitro transcription-translation compatible with microfluidic droplets.Microbial cell factories · 2024Article
- Engineering Enzymes for Environmental Sustainability.Angewandte Chemie (International ed. in English) · 2023Review
- Engineering Enzymes for Environmental Sustainability.Angewandte Chemie (Weinheim an der Bergstrasse, Germany) · 2023Review
- Article
- Ultrahigh-Throughput Enzyme Engineering and Discovery inChemical reviews · 2023Review
- Ultrahigh-Throughput Directed Evolution of a Metal-Free α/β-Hydrolase with a Cys-His-Asp Triad into an Efficient Phosphotriesterase.Journal of the American Chemical Society · 2023Article
- Progresses in Cell-Free In Vitro Evolution.Advances in biochemical engineering/biotechnology · 2023Review
- Engineering Ag43 Signal Peptides with Bacterial Display and Selection.Methods and protocols · 2022Article
- Fungi-on-a-Chip: microfluidic platforms for single-cell studies on fungi.FEMS microbiology reviews · 2022Review
- Directed Evolution in Drops: Molecular Aspects and Applications.ACS synthetic biology · 2021Review
- Droplet Microfluidics and Directed Evolution of Enzymes: An Intertwined Journey.Angewandte Chemie (International ed. in English) · 2021Review
- Directed Evolution Methods for Enzyme Engineering.Molecules (Basel, Switzerland) · 2021Review
- Pollen-inspired enzymatic microparticles to reduce organophosphate toxicity in managed pollinators.Nature food · 2021Article
16 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We describe the selection of a phosphotriesterase with a very fast k(cat) (over 10(5) s(-1)), 63 times higher than the already very efficient wild-type enzyme. The enzyme was selected from a library of 3.4 x 10(7) mutated phosphotriesterase genes using a novel strategy based on linking genotype and phenotype by in vitro compartmentalization (IVC) using water-in-oil emulsions. First, microbeads, each displaying a single gene and multiple copies of the encoded protein, are formed by compartmentalized in vitro translation. These microbeads can then be selected for catalysis or binding. To select for catalysis the microbeads are re-emulsified in a reaction buffer of choice with a soluble substrate. The product and any unreacted substrate are coupled to the beads when the reaction is finished. Product-coated beads, displaying active enzymes and the genes that encode them, are detected with anti-product antibodies and selected using flow cytometry. This completely in vitro process selects for all enzymatic features simultaneously (substrate recognition, product formation, rate acceleration and turnover) and single enzyme molecules can be detected.
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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.