title: Heterologous expression of the thiopeptide antibiotic GE2270 from Planobispora rosea ATCC 53733 in Streptomyces coelicolor requires deletion of ribosomal genes from the expression construct.
abstract: GE2270 is a thiopeptide antibiotic generated by extensive posttranslational modifications of a ribosomally generated precursor peptide. Thiopeptides are especially active against Gram-positive bacteria, including methicillin resistant Staphylococcus aureus (MRSA). In this study the GE2270 biosynthetic gene cluster (pbt) from Planobispora rosea ATCC 53733 was successfully expressed in the heterologous host strain Streptomyces coelicolor M1146. Notably, exconjugants containing the pbt gene cluster could only be obtained after deletion of the major part of the ribosomal genes flanking the gene cluster. This is a striking example that genes belonging to primary metabolism can prevent the successful conjugative transfer of DNA from phylogenetic distant species and thus complicate heterologous expression of secondary metabolite gene clusters. GE2270 production in the heterologous producer strain increased after introduction of the constitutive ermE* promoter upstream of the GE2270 resistance gene tuf from P. rosea. Insertion of the inducible tcp830 promoter resulted in inducible GE2270 production. When the regulatory gene pbtR was deleted, the resulting strain ceased to produce GE2270, suggesting an essential role of PbtR as a putative transcriptional activator of GE2270 expression.
authors.affiliation: Pharmaceutical Institute, University of Tübingen, Tübingen, Germany; German Centre for Infection Research (DZIF), partner site Tübingen, Tübingen, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Flinspach
authors.affiliation: Pharmaceutical Institute, University of Tübingen, Tübingen, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Kapitzke
authors.affiliation: KtedoGen Srl, Milan, Italy.
authors.forename: Arianna
authors.initials: A
authors.lastname: Tocchetti
authors.affiliation: KtedoGen Srl, Milan, Italy; NAICONS Srl, Milan, Italy.
authors.forename: Margherita
authors.initials: M
authors.lastname: Sosio
authors.affiliation: Pharmaceutical Institute, University of Tübingen, Tübingen, Germany; German Centre for Infection Research (DZIF), partner site Tübingen, Tübingen, Germany.
authors.forename: Alexander K
authors.initials: AK
authors.lastname: Apel
mesh_terms: D000192:Actinomycetales / Q000235:genetics*; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000494:pharmacology; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D003001:Cloning, Molecular; D003360:Cosmids / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics*; D052978:Disk Diffusion Antimicrobial Tests; D024881:Drug Resistance, Bacterial; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D058889:Molecular Typing; D005810:Multigene Family; D010456:Peptides, Cyclic / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D010802:Phylogeny; D011401:Promoter Regions, Genetic; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D048372:Streptomyces coelicolor / Q000187:drug effects / Q000235:genetics*; D013844:Thiazoles / Q000494:pharmacology; D015533:Transcriptional Activation
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004275:DNA, Ribosomal; D010456:Peptides, Cyclic; D012336:RNA, Ribosomal, 16S; D013844:Thiazoles; C068122:GE 2270 A