Daptomycin biosynthesis in Streptomyces roseosporus: cloning and analysis of the gene cluster and revision of peptide stereochemistry
| Autor: | Ian Parr, Andrew Whiting, Steven M. Martin, Renee Brost, Vivian Miao, Julia Penn, Paul Brian, Richard H. Baltz, Robert Ford, Mario Bouchard, Stephen K. Wrigley, Marie-Françoise Coëffet-LeGal, Christopher J. Silva |
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| Rok vydání: | 2005 |
| Předmět: |
Streptomyces roseosporus
Stereochemistry Amino Acid Motifs Molecular Sequence Data Peptides Cyclic Microbiology Streptomyces Condensation domain Open Reading Frames Bacterial Proteins Daptomycin Gene cluster medicine Amino Acid Sequence Cloning Molecular Peptide Synthases Peptide sequence Conserved Sequence chemistry.chemical_classification Base Sequence biology Stereoisomerism Sequence Analysis DNA biology.organism_classification Anti-Bacterial Agents Amino acid Protein Subunits Biochemistry chemistry Multigene Family Streptomyces lividans Heterologous expression medicine.drug |
| Zdroj: | Microbiology. 151:1507-1523 |
| ISSN: | 1465-2080 1350-0872 |
| DOI: | 10.1099/mic.0.27757-0 |
| Popis: | Daptomycin is a 13 amino acid, cyclic lipopeptide produced by a non-ribosomal peptide synthetase (NRPS) mechanism inStreptomyces roseosporus. A 128 kb region ofS. roseosporusDNA was cloned and verified by heterologous expression inStreptomyces lividansto contain the daptomycin biosynthetic gene cluster (dpt). The cloned region was completely sequenced and three genes (dptA,dptBC,dptD) encoding the three subunits of an NRPS were identified. The catalytic domains in the subunits, predicted to couple five, six or two amino acids, respectively, included a novel activation domain and amino-acid-binding pocket for incorporating the unusual amino acidl-kynurenine (Kyn), three types of condensation domains and an extra epimerase domain (E-domain) in the second module. Novel genes (dptE,dptF) whose products likely work in conjunction with a unique condensation domain to acylate the first amino acid, as well as other genes (dptI,dptJ) probably involved in supply of the non-proteinogenic amino acidsl-3-methylglutamic acid and Kyn, were located next to the NRPS genes. The unexpected E-domain suggested that daptomycin would haved-Asn, rather thanl-Asn, as originally assigned, and this was confirmed by comparing stereospecific synthetic peptides and the natural product both chemically and microbiologically. |
| Databáze: | OpenAIRE |
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