ArticleMetabolic engineering communications2021
A lysate proteome engineering strategy for enhancing cell-free metabolite production.
Article in Metabolic engineering communications, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 19 citations in OpenAlex.
- Cell-Free-Based Thermophilic Biocatalyst for the Synthesis of Amino Acids from One-Carbon Feedstocks.ACS synthetic biology · 2025Article
- Cell-Free Gene Expression: Methods and Applications.Chemical reviews · 2025Review
- Profiling expression strategies for a type III polyketide synthase in a lysate-based, cell-free system.Scientific reports · 2024Article
- Engineering cell-free systems by chemoproteomic-assisted phenotypic screening.RSC chemical biology · 2024Article
- Designing of an extract production protocol for industrial application of cell-free protein synthesis technology: Building from a current best practice to a quality by design approach.Engineering biology · 2023Review
- Immobilised-enzyme microreactors for the identification and synthesis of conjugated drug metabolites.RSC advances · 2023Article
- Investigating and Optimizing the Lysate-Based Expression of Nonribosomal Peptide Synthetases Using a Reporter System.ACS synthetic biology · 2023Article
- Rewiring cell-free metabolic flux inSynthetic biology (Oxford, England) · 2023Article
- Cell-Free Protein Synthesis for High-Throughput Biosynthetic Pathway Prototyping.Methods in molecular biology (Clifton, N.J.) · 2022Article
- An integrated in vivo/in vitro framework to enhance cell-free biosynthesis with metabolically rewired yeast extracts.Nature communications · 2021Article
- Biofoundry-assisted expression and characterization of plant proteins.Synthetic biology (Oxford, England) · 2021Article
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Cell-free systems present a significant opportunity to harness the metabolic potential of diverse organisms. Removing the cellular context provides the ability to produce biological products without the need to maintain cell viability and enables metabolic engineers to explore novel chemical transformation systems. Crude extracts maintain much of a cell's capabilities. However, only limited tools are available for engineering the contents of the extracts used for cell-free systems. Thus, our ability to take full advantage of the potential of crude extracts for cell-free metabolic engineering is constrained. Here, we employ Multiplex Automated Genomic Engineering (MAGE) to tag proteins for selective depletion from crude extracts so as to specifically direct chemical production. Specific edits to central metabolism are possible without significantly impacting cell growth. Selective removal of pyruvate degrading enzymes resulted in engineered crude lysates that are capable of up to 40-fold increases in pyruvate production when compared to the non-engineered extract. The described approach melds the tools of systems and synthetic biology to showcase the effectiveness of cell-free metabolic engineering for applications like bioprototyping and bioproduction.
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