ArticleACS synthetic biology2016
Cell-Free Mixing of Escherichia coli Crude Extracts to Prototype and Rationally Engineer High-Titer Mevalonate Synthesis.
Article in ACS synthetic biology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 60 papers.
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Who cites it
60 citing papers in PubMed, 148 citations in OpenAlex.
- A cell-free system for 1,6-hexanediamine synthesis using bulk chemical caprolactam as the feedstock.Synthetic and systems biotechnology · 2026Article
- Heterologous expression of NRPS and PKS pathways in Escherichia coli: from bottlenecks to biosynthetic platforms.Essays in biochemistry · 2026Review
- Timed batch inputs unlock substantially higher yields for enzymatic cascades.Nature chemistry · 2026Article
- Reprogramming the Rossmann Fold Signature Motif Creates Orthogonal Redox Biocatalysts.bioRxiv : the preprint server for biology · 2025Article
- Cell-Free-Based Thermophilic Biocatalyst for the Synthesis of Amino Acids from One-Carbon Feedstocks.ACS synthetic biology · 2025Article
- Automated and Programmable Cell-Free Systems for Scalable Synthetic Biology with a Focus on Biofoundry Integration.Journal of microbiology and biotechnology · 2025Review
- Glycosylation of Structured Protein Domains in Cell-Free Reaction Environments.ACS synthetic biology · 2025Article
- The knowledge driven DBTL cycle provides mechanistic insights while optimising dopamine production in Escherichia coli.Microbial cell factories · 2025Article
- Cell-Free Gene Expression: Methods and Applications.Chemical reviews · 2025Review
- Carbon Negative Synthesis of Amino Acids Using a Cell-Free-Based Biocatalyst.ACS synthetic biology · 2024Article
- Cell-Free Systems Biology: Characterizing Central Metabolism ofACS synthetic biology · 2024Article
- Cell-Free Synthesis: Expediting Biomanufacturing of Chemical and Biological Molecules.Molecules (Basel, Switzerland) · 2024Review
- Exploring Emergent Properties in Enzymatic Reaction Networks: Design and Control of Dynamic Functional Systems.Chemical reviews · 2024Review
- Cell-Free Expression of a Therapeutic Protein Serratiopeptidase.Molecules (Basel, Switzerland) · 2023Article
- Investigating ethanol production using the Zymomonas mobilis crude extract.Scientific reports · 2023Article
- Utilizing a cell-free protein synthesis platform for the biosynthesis of a natural product, caffeine.Synthetic biology (Oxford, England) · 2023Article
- Rewiring cell-free metabolic flux inSynthetic biology (Oxford, England) · 2023Article
- Bottom-Up Synthetic Biology Using Cell-Free Protein Synthesis.Advances in biochemical engineering/biotechnology · 2023Review
- Compartmentalized Cell-Free Expression Systems for Building Synthetic Cells.Advances in biochemical engineering/biotechnology · 2023Article
- Advancing synthetic biology through cell-free protein synthesis.Computational and structural biotechnology journal · 2023Review
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3 authors at 1 institution in 1 country.
Funding
Abstract
Cell-free metabolic engineering (CFME) is advancing a powerful paradigm for accelerating the design and synthesis of biosynthetic pathways. However, as most cell-free biomolecule synthesis systems to date use purified enzymes, energy and cofactor balance can be limiting. To address this challenge, we report a new CFME framework for building biosynthetic pathways by mixing multiple crude lysates, or extracts. In our modular approach, cell-free lysates, each selectively enriched with an overexpressed enzyme, are generated in parallel and then combinatorically mixed to construct a full biosynthetic pathway. Endogenous enzymes in the cell-free extract fuel high-level energy and cofactor regeneration. As a model, we apply our framework to synthesize mevalonate, an intermediate in isoprenoid synthesis. We use our approach to rapidly screen enzyme variants, optimize enzyme ratios, and explore cofactor landscapes for improving pathway performance. Further, we show that genomic deletions in the source strain redirect metabolic flux in resultant lysates. In an optimized system, mevalonate was synthesized at 17.6 g·L
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