ArticleProceedings of the National Academy of Sciences of the United States of America2011
A system for the targeted amplification of bacterial gene clusters multiplies antibiotic yield in Streptomyces coelicolor.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed, 74 citations in OpenAlex.
- Overproduction of endusamycin inSynthetic and systems biotechnology · 2025Article
- A tunable and reversible thermo-inducible bio-switch for streptomycetes.Nucleic acids research · 2025Article
- Metabolic engineering of Streptomyces roseosporus for increased production of clinically important antibiotic daptomycin.Microbial biotechnology · 2024Article
- An efficient method for targeted cloning of large DNA fragments from Streptomyces.Applied microbiology and biotechnology · 2023Article
- Multiplying the heterologous production of spinosad through tandem amplification of its biosynthetic gene cluster in Streptomyces coelicolor.Microbial biotechnology · 2022Article
- Multiscale engineering of microbial cell factories: A step forward towards sustainable natural products industry.Synthetic and systems biotechnology · 2022Review
- Improving the Yield and Quality of Daptomycin inFrontiers in microbiology · 2022Article
- Biosynthetic Potential ofAntibiotics (Basel, Switzerland) · 2021Article
- Review
- Use of Permanent Wall-Deficient Cells as a System for the Discovery of New-to-Nature Metabolites.Microorganisms · 2020Article
- Biosynthesis of aromatic polyketides in microorganisms using type II polyketide synthases.Microbial cell factories · 2020Review
- Cloning and Overexpression of theFrontiers in microbiology · 2020Article
- Heterologous expression-facilitated natural products' discovery in actinomycetes.Journal of industrial microbiology & biotechnology · 2019Review
- Recent Advances in Synthetic Biology Approaches to Optimize Production of Bioactive Natural Products in Actinobacteria.Frontiers in microbiology · 2019Review
- A Review of the Microbial Production of Bioactive Natural Products and Biologics.Frontiers in microbiology · 2019Review
- Regulation of antibiotic biosynthesis in actinomycetes: Perspectives and challenges.Synthetic and systems biotechnology · 2018Review
- Biosynthesis and Metabolic Engineering of Pseudo-oligosaccharides.Emerging topics in life sciences · 2018Article
- Engineering microbial hosts for production of bacterial natural products.Natural product reports · 2016Review
- Genetic manipulation of secondary metabolite biosynthesis for improved production in Streptomyces and other actinomycetes.Journal of industrial microbiology & biotechnology · 2016Review
- Activity-Independent Discovery of Secondary Metabolites Using Chemical Elicitation and Cheminformatic Inference.ACS chemical biology · 2015Article
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Authors and funding
5 authors at 2 institutions in 2 countries.
Funding
Abstract
Gene clusters found in bacterial species classified as Streptomyces encode the majority of known antibiotics as well as many pharmaceutically active compounds. A site-specific recombination system similar to those that mediate plasmid conjugation was engineered to catalyze tandem amplification of one of these gene clusters in a heterologous Streptomyces species. Three genetic elements were known to be required for DNA amplification in S. kanamyceticus: the oriT-like recombination sites RsA and RsB, and ZouA, a site-specific relaxase similar to TraA proteins that catalyze plasmid transfer. We inserted RsA and RsB sequences into the S. coelicolor genome flanking a cluster of 22 genes (act) responsible for biosynthesis of the polyketide antibiotic actinorhodin. Recombination between RsA and RsB generated zouA-dependent DNA amplification resulting in 4-12 tandem copies of the act gene cluster averaging nine repeats per genome. This resulted in a 20-fold increase in actinorhodin production compared with the parental strain. To determine whether the recombination event required taxon-specific genetic effectors or generalized bacterial recombination (recA), it was also analyzed in the heterologous host Escherichia coli. zouA was expressed under the control of an inducible promoter in wild-type and recA mutant strains. A plasmid was constructed with recombination sites RsA and RsB bordering a drug resistance marker. Induction of zouA expression generated hybrid RsB/RsA sites, evidence of site-specific recombination that occurred independently of recA. ZouA-mediated DNA amplification promises to be a valuable tool for increasing the activities of commercially important biosynthetic, degradative, and photosynthetic pathways in a wide variety of organisms.
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