Heterologous expression of a Photorhabdus luminescens syrbactin-like gene cluster results in production of the potent proteasome inhibitor glidobactin A
| Autor: | Robert Dudler, Laurent Bigler, Alexey Dudnik |
|---|---|
| Přispěvatelé: | University of Zurich, Dudler, Robert |
| Rok vydání: | 2013 |
| Předmět: |
Proteasome Endopeptidase Complex
580 Plants (Botany) Peptides Cyclic Microbiology 03 medical and health sciences 10126 Department of Plant and Microbial Biology Photorhabdus luminescens Polyketide synthase Gene cluster Pseudomonas syringae Cloning Molecular Peptide Synthases Gene 030304 developmental biology 0303 health sciences biology Pseudomonas putida 030306 microbiology 2404 Microbiology biology.organism_classification Biochemistry Multigene Family biology.protein Heterologous expression Photorhabdus Proteasome Inhibitors |
| Zdroj: | MICROBIOLOGICAL RESEARCH |
| ISSN: | 0944-5013 |
| DOI: | 10.1016/j.micres.2012.09.006 |
| Popis: | Syrbactins are cyclic peptide derivatives which are known to inhibit the eukaryotic proteasome by irreversible covalent binding to its catalytic sites. The only two members of this family characterized to date, syringolin A and glidobactin A, are secreted by certain strains of Pseudomonas syringae pv. syringae and strain K481-B101 from the order Burkholderiales, respectively. Syrbactins are the products of mixed non-ribosomal peptide/polyketide synthases encoded by gene clusters with a characteristic architecture. Similar, but not identical gene clusters are present in several other bacterial genomes, including that of Photorhabdus luminescens subsp. laumondii TT01, which is therefore hypothesized to be able to produce a syrbactin-type proteasome inhibitor. Here we report the cloning of the putative syrbactins synthetase encoding gene cluster of Ph. luminescens into a cosmid vector and its heterologous expression in Pseudomonas putida. Analysis of culture supernatants of transformed Ps. putida by HPLC and mass spectrometry revealed the presence of glidobactin A, indicating that the syrbactins-like gene cluster of Ph. luminescens encodes a glidobactin A synthetase and that this organism has the capacity to synthesize glidobactin A. |
| Databáze: | OpenAIRE |
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