ArticleMetabolic engineering2020
Total in vitro biosynthesis of the nonribosomal macrolactone peptide valinomycin.
Article in Metabolic engineering, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 49 papers.
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Who cites it
49 citing papers in PubMed, 96 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
- Tailor-made poly-γ-glutamic acid for property diversity and biological function.World journal of microbiology & biotechnology · 2026Review
- De Novo Biosynthesis of Valinomycin From Glucose Using In Vitro Reconstituted Hybrid Pathways.Chembiochem : a European journal of chemical biology · 2026Article
- Recent Advances in Beta-Alanine Production via Enzymatic Catalysis and Microbial Whole-Cell Catalysis.Biology · 2026Review
- Machine learning-driving optimization and spatial assembly of a cell-free system for high-yield liquiritigenin production.Advanced biotechnology · 2026Article
- A Simple and Versatile Cell-Free Expression Method for Producing Secondary Metabolites.ACS synthetic biology · 2026Article
- Reconstituting transcription-translation-coupled DNA replication within complex in vitro biological systems.Nature communications · 2025Article
- Protein Engineering of Biosynthetic Enzymes Unlocks Libraries of Bioactive Tilimycin Analogs.ACS catalysis · 2025Article
- Online monitoring of the mitochondrial respiration activity and protein formation in the Almost Living Cell-free Expression (ALiCE) system.BMC biotechnology · 2025Article
- Microbial secondary metabolites: advancements to accelerate discovery towards application.Nature reviews. Microbiology · 2025Review
- An efficient lysate-based approach for biosynthesis of the pyrrolobenzodiazepine natural product tilimycin.Journal of biotechnology · 2025Article
- Cyanobacterial Peptides in Anticancer Therapy: A Comprehensive Review of Mechanisms, Clinical Advances, and Biotechnological Innovation.Marine drugs · 2025Review
- BeyondChemical reviews · 2025Review
- Cell-Free Gene Expression: Methods and Applications.Chemical 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 Synthesis: Expediting Biomanufacturing of Chemical and Biological Molecules.Molecules (Basel, Switzerland) · 2024Review
- Biocompatible strategies for peptide macrocyclisation.Chemical science · 2024Review
- Development and Challenges of Cyclic Peptides for Immunomodulation.Current protein & peptide science · 2024Review
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 2 countries.
Funding
Abstract
Natural products are important because of their significant pharmaceutical properties such as antiviral, antimicrobial, and anticancer activity. Recent breakthroughs in DNA sequencing reveal that a great number of cryptic natural product biosynthetic gene clusters are encoded in microbial genomes, for example, those of Streptomyces species. However, it is still challenging to access compounds from these clusters because many source organisms are uncultivable or the genes are silent during laboratory cultivation. To address this challenge, we develop an efficient cell-free platform for the rapid, in vitro total biosynthesis of the nonribosomal peptide valinomycin as a model. We achieve this goal in two ways. First, we used a cell-free protein synthesis (CFPS) system to express the entire valinomycin biosynthetic gene cluster (>19 kb) in a single-pot reaction, giving rise to approximately 37 μg/L of valinomycin after optimization. Second, we coupled CFPS with cell-free metabolic engineering system by mixing two enzyme-enriched cell lysates to perform a two-stage biosynthesis. This strategy improved valinomycin production ~5000-fold to nearly 30 mg/L. We expect that cell-free biosynthetic systems will provide a new avenue to express, discover, and characterize natural product gene clusters of interest in vitro.
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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.