Molecular Cloning and Heterologous Expression of the Dehydrophos Biosynthetic Gene Cluster
| Autor: | Wilfred A. van der Donk, Benjamin T. Circello, William W. Metcalf, Andrew C. Eliot, Jin Hee Lee |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Clinical Biochemistry
Organophosphonates Streptomyces luridus 010402 general chemistry 01 natural sciences Streptomyces Biochemistry Article 03 medical and health sciences Phosphoenolpyruvate mutase Gene cluster Drug Discovery Cloning Molecular ORFS Molecular Biology 030304 developmental biology Genetics Pharmacology 0303 health sciences biology Genomics General Medicine biology.organism_classification Anti-Bacterial Agents 0104 chemical sciences Fosmid Open reading frame CHEMBIO Genes Bacterial Phosphotransferases (Phosphomutases) Multigene Family Mutation Molecular Medicine Heterologous expression Oligopeptides |
| Zdroj: | Chemistry & Biology. 17(4):402-411 |
| ISSN: | 1074-5521 |
| DOI: | 10.1016/j.chembiol.2010.03.007 |
| Popis: | Dehydrophos is a vinyl phosphonate tripeptide produced by Streptomyces luridus with demonstrated broad spectrum antibiotic activity. To identify genes necessary for biosynthesis of this unusual compound we screened a fosmid library of S. luridus for the presence of the phosphoenolpyruvate mutase gene, which is required for biosynthesis of most phosphonates. Integration of one such fosmid clone into the chromosome of Streptomyces lividans led to heterologous production of dehydrophos. Deletion analysis of this clone allowed identification of the minimal contiguous dehydrophos cluster, which contained 17 open reading frames (ORFs). Bioinformatic analyses of these ORFs are consistent with a proposed biosynthetic pathway that generates dehydrophos from phosphoenolpyruvate. The early steps of this pathway are supported by analysis of intermediates accumulated by blocked mutants and in vitro biochemical experiments. |
| Databáze: | OpenAIRE |
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