ArticleACS synthetic biology2017
In Vitro Reconstruction of Nonribosomal Peptide Biosynthesis Directly from DNA Using Cell-Free Protein Synthesis.
Article in ACS synthetic biology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 50 papers.
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Who cites it
50 citing papers in PubMed, 103 citations in OpenAlex.
- A Simple and Versatile Cell-Free Expression Method for Producing Secondary Metabolites.ACS synthetic biology · 2026Article
- Online monitoring of the mitochondrial respiration activity and protein formation in the Almost Living Cell-free Expression (ALiCE) system.BMC biotechnology · 2025Article
- Cyanobacterial Peptides in Anticancer Therapy: A Comprehensive Review of Mechanisms, Clinical Advances, and Biotechnological Innovation.Marine drugs · 2025Review
- A Design of Experiments Approach for Enhancing Room Temperature Stability of a Lyophilised and Paper-Based Bacterial Cell-Free System.Bioengineering (Basel, Switzerland) · 2025Article
- BeyondChemical reviews · 2025Review
- Cell-Free Systems: Ideal Platforms for Accelerating the Discovery and Production of Peptide-Based Antibiotics.International journal of molecular sciences · 2024Review
- Cell-free biosynthesis and engineering of ribosomally synthesized lanthipeptides.Nature communications · 2024Article
- Cell-free protein synthesis with technical additives - expanding the parameter space of in vitro gene expression.Beilstein journal of organic chemistry · 2024Article
- Present and future outlooks on environmental DNA-based methods for antibiotic discovery.Current opinion in microbiology · 2023Review
- Cell-free metabolic engineering enables selective biotransformation of fatty acids to value-added chemicals.Metabolic engineering communications · 2023Article
- Investigating and Optimizing the Lysate-Based Expression of Nonribosomal Peptide Synthetases Using a Reporter System.ACS synthetic biology · 2023Article
- Cell-Free Expression of a Therapeutic Protein Serratiopeptidase.Molecules (Basel, Switzerland) · 2023Article
- Natural and engineered cyclodipeptides: Biosynthesis, chemical diversity, and engineering strategies for diversification and high-yield bioproduction.Engineering microbiology · 2023Review
- Cell-free Biosynthesis of Peptidomimetics.Biotechnology and bioprocess engineering : BBE · 2023Review
- Cell-Free Display Techniques for Protein Evolution.Advances in biochemical engineering/biotechnology · 2023Article
- Utilizing a cell-free protein synthesis platform for the biosynthesis of a natural product, caffeine.Synthetic biology (Oxford, England) · 2023Article
- Ice Cores as a Source for Antimicrobials: From Bioprospecting to Biodesign.Biodesign research · 2023Review
- One-pot synthesis of 2,5-diketopiperazine with high titer and versatility using adenylation enzyme.Applied microbiology and biotechnology · 2022Article
- A High-Yield Streptomyces Transcription-Translation Toolkit for Synthetic Biology and Natural Product Applications.Journal of visualized experiments : JoVE · 2021Article
- Review
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
Abstract
Genome sequencing has revealed that a far greater number of natural product biosynthetic pathways exist than there are known natural products. To access these molecules directly and deterministically, a new generation of heterologous expression methods is needed. Cell-free protein synthesis has not previously been used to study nonribosomal peptide biosynthesis, and provides a tunable platform with advantages over conventional methods for protein expression. Here, we demonstrate the use of cell-free protein synthesis to biosynthesize a cyclic dipeptide with correct absolute stereochemistry. From a single-pot reaction, we measured the expression of two nonribosomal peptide synthetases larger than 100 kDa, and detected high-level production of a diketopiperazine. Using quantitative LC-MS and synthetically prepared standard, we observed production of this metabolite at levels higher than previously reported from cell-based recombinant expression, approximately 12 mg/L. Overall, this work represents a first step to apply cell-free protein synthesis to discover and characterize new natural products.
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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.