ArticleNature chemical biology2020
In vitro prototyping and rapid optimization of biosynthetic enzymes for cell design.
Article in Nature chemical biology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 98 papers.
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Who cites it
98 citing papers in PubMed, 248 citations in OpenAlex.
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- Microbial cell-free protein synthesis and its progression toward industrial use.Microbiology (Reading, England) · 2026Review
- A unifying equation for fermentation sustainability across the titer-rate-yield landscape.Nature communications · 2026Article
- Cell-Free Protein Synthesis in Porous Parylene Scaffolds.Small methods · 2026Article
- Intrinsically disordered protein droplet-enhanced oligonucleotide assembly enables rapid oligonucleotide-to-protein expression.Nucleic acids research · 2026Article
- Design-driven optimization of low-cost reagent formulations for reproducible and high-yielding cell-free gene expression.Nature communications · 2026Article
- Synthetic Biology of Plants and Microbes for Agriculture, Environment, and Future Applications.Chemical reviews · 2026Review
- A Multi-Functional Heterogeneous Biocatalyst for the Oxygen-Free Oxidative Condensation of Primary Alcohols into β‑Hydroxy Acids.ACS catalysis · 2026Article
- A Simple and Versatile Cell-Free Expression Method for Producing Secondary Metabolites.ACS synthetic biology · 2026Article
- Whole-cell and cell-free biosensor-driven metabolic engineering.Current opinion in biotechnology · 2025Review
- LDBT instead of DBTL: combining machine learning and rapid cell-free testing.Nature communications · 2025Article
- Reconstituting alternative life using the test-bed of cell-free systems.Philosophical transactions of the Royal Society of London. Series B, Biological sciences · 2025Review
- Automated and Programmable Cell-Free Systems for Scalable Synthetic Biology with a Focus on Biofoundry Integration.Journal of microbiology and biotechnology · 2025Review
- Characterizing and engineering post-translational modifications with high-throughput cell-free expression.Nature communications · 2025Article
- Microbial secondary metabolites: advancements to accelerate discovery towards application.Nature reviews. Microbiology · 2025Review
- The knowledge driven DBTL cycle provides mechanistic insights while optimising dopamine production in Escherichia coli.Microbial cell factories · 2025Article
- AI-driven high-throughput droplet screening of cell-free gene expression.Nature communications · 2025Article
- Cell-Free Systems to Mimic and Expand Metabolism.ACS synthetic biology · 2025Review
- Accelerated enzyme engineering by machine-learning guided cell-free expression.Nature communications · 2025Article
38 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
20 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The design and optimization of biosynthetic pathways for industrially relevant, non-model organisms is challenging due to transformation idiosyncrasies, reduced numbers of validated genetic parts and a lack of high-throughput workflows. Here we describe a platform for in vitro prototyping and rapid optimization of biosynthetic enzymes (iPROBE) to accelerate this process. In iPROBE, cell lysates are enriched with biosynthetic enzymes by cell-free protein synthesis and then metabolic pathways are assembled in a mix-and-match fashion to assess pathway performance. We demonstrate iPROBE by screening 54 different cell-free pathways for 3-hydroxybutyrate production and optimizing a six-step butanol pathway across 205 permutations using data-driven design. Observing a strong correlation (r = 0.79) between cell-free and cellular performance, we then scaled up our highest-performing pathway, which improved in vivo 3-HB production in Clostridium by 20-fold to 14.63 ± 0.48 g l
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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.