ArticleApplied and environmental microbiology2015
Multiplexed integrating plasmids for engineering of the erythromycin gene cluster for expression in Streptomyces spp. and combinatorial biosynthesis.
Article in Applied and environmental microbiology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 26 citations in OpenAlex.
- Current Approaches for Genetic Manipulation ofBiotech (Basel (Switzerland)) · 2025Review
- Discovery, characterization, and engineering of an advantageous Streptomyces host for heterologous expression of natural product biosynthetic gene clusters.Microbial cell factories · 2024Article
- Use of orthogonal serine integrases to multiplex plasmid conjugation and integration fromAccess microbiology · 2021Article
- Application of serine integrases for secondary metabolite pathway assembly inSynthetic and systems biotechnology · 2020Article
- Bioprospecting Through Cloning of Whole Natural Product Biosynthetic Gene Clusters.Frontiers in bioengineering and biotechnology · 2020Review
- Genetic platforms for heterologous expression of microbial natural products.Natural product reports · 2019Review
- Integrating vectors for genetic studies in the rare Actinomycete Amycolatopsis marina.BMC biotechnology · 2019Article
- Recent advances in natural products exploitation inEngineering in life sciences · 2019Review
- Refactoring the Cryptic Streptophenazine Biosynthetic Gene Cluster Unites Phenazine, Polyketide, and Nonribosomal Peptide Biochemistry.Cell chemical biology · 2019Article
- Recent advances in the discovery and combinatorial biosynthesis of microbial 14-membered macrolides and macrolactones.Journal of industrial microbiology & biotechnology · 2019Review
- Genome Integration and Excision by a New Streptomyces Bacteriophage, ϕJoe.Applied and environmental microbiology · 2017Article
- Synthetic Biology of Natural Products.Cold Spring Harbor perspectives in biology · 2016Review
- Modern Approaches to the Genome Editing of Antibiotic Biosynthetic Clusters in Actinomycetes.Acta naturaeArticle
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Authors and funding
7 authors at 4 institutions in 2 countries.
Funding
Abstract
Bacteria in the genus Streptomyces and its close relatives are prolific producers of secondary metabolites with antibiotic activity. Genome sequencing of these bacteria has revealed a rich source of potentially new antibiotic pathways, whose products have never been observed. Moreover, these new pathways can provide novel genes that could be used in combinatorial biosynthesis approaches to generate unnatural analogues of existing antibiotics. We explore here the use of multiple orthologous integrating plasmid systems, based on the int/attP loci from phages TG1, SV1, and ϕBT1, to express the polyketide synthase (PKS) for erythromycin in a heterologous Streptomyces host. Streptomyces strains containing the three polyketide synthase genes eryAI, eryAII, and eryAIII expressed from three different integrated plasmids produced the aglycone intermediate, 6-deoxyerythronolide B (6-dEB). A further pair of integrating plasmids, both derived from the ϕC31 int/attP locus, were constructed carrying a gene cassette for glycosylation of the aglycone intermediates, with or without the tailoring gene, eryF, required for the synthesis of erythronolide B (EB). Liquid chromatography-mass spectrometry of the metabolites indicated the production of angolosaminyl-6-dEB and angolosaminyl-EB. The advantages of using multiplexed integrating plasmids for engineering expression and for combinatorial biosynthesis were demonstrated.
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