Isolation and Characterization of the Gene Cluster for Biosynthesis of the Thiopeptide Antibiotic TP-1161

• Published in: Applied and Environmental Microbiology Vol. 76; no. 21; pp. 7093 - 7101
• Main Authors: Engelhardt, Kerstin, Degnes, Kristin F, Zotchev, Sergey B
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01-11-2010; American Society for Microbiology (ASM)
• Subjects: Actinomycetales - genetics; Actinomycetales - metabolism; Actinomycetes; Anti-Bacterial Agents - biosynthesis; Antibiotics; Biological and medical sciences; Biosynthesis; Chromosome Mapping; Cloning, Molecular; Cosmids - genetics; Fermentation; Fundamental and applied biological sciences. Psychology; Gene expression; Gene Expression - genetics; Gene loci; Genes, Bacterial - genetics; Genetics and Molecular Biology; Microbiology; Molecular Sequence Data; Multigene Family - genetics; Nocardiopsis; Peptides; Peptides - genetics; Peptides, Cyclic - biosynthesis; Sequence Alignment; Sequence Analysis, DNA; Streptomyces coelicolor; Thiazoles; Characterization; Gene cluster; Isolation; Antibiotic; Biosynthesis
• ISSN: 0099-2240; 1098-5336; 1098-6596
• DOI: 10.1128/aem.01442-10

Recently, we isolated a new thiopeptide antibiotic, TP-1161, from the fermentation broth of a marine actinomycete typed as a member of the genus NOCARDIOPSIS: Here we report the identification, isolation, and analysis of the TP-1161 biosynthetic gene cluster from this species. The gene cluster was identified by mining a draft genome sequence using the predicted structural peptide sequence of TP-1161. Functional assignment of a ~16-kb genomic region revealed 13 open reading frames proposed to constitute the TP-1161 biosynthetic locus. While the typical core set of thiopeptide modification enzymes contains one cyclodehydratase/dehydrogenase pair, paralogous genes predicted to encode additional cyclodehydratases and dehydrogenases were identified. Although attempts at heterologous expression of the TP-1161 gene cluster in Streptomyces coelicolor failed, its identity was confirmed through the targeted gene inactivation in the original host.