A cryptic PKS–NRPS gene locus in the plant commensal Pseudomonas fluorescens Pf-5 codes for the biosynthesis of an antimitotic rhizoxin complex

Brendel, Nicole; Partida‐Martínez, Laila P.; Scherlach, Kirstin; Hertweck, Christian

doi:10.1039/b707762a

Cited by 61 publications

(46 citation statements)

References 14 publications

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“…(12) and Pseudomonas sp. (13). Further downstream, the PKS genes have resemblances to gene clusters reported from various Nostocales or Oscillatoriales.…”

Section: Resultsmentioning

confidence: 88%

“…S6-S9). This method allowed collection of highquality 1 H-and 13 C-spectra, as well as COSY-, HSQC-, and HMBC-2D-NMR data from microgram amounts of the 13 The NMR data fully supported the identity of the compound as a member of the pederin group and allowed elucidation of its constitution. In combination with bioinformatic analysis it was possible to predict most of the stereogenic elements, except the configuration at C12 and C14 ( Fig.…”

Section: Resultsmentioning

confidence: 93%

“…was cultured in 25 L of BG-11 supplemented with 13 C-labeled NaHCO 3 , and after 5 wk, cyanobacterial biomass from 10 L of culture was freeze-dried and extracted. HPLC-electrospray ionization (ESI)-MS analysis confirmed that most components in the extracts had multiple 13 C atoms incorporated into individual molecules. To obtain insights into structural features of these compounds, the crude extract was subjected to repetitive HPLC-solid phase extraction (SPE) purification with subsequent NMR analysis of target molecules eluted with fully deuterated solvent (SI Appendix, Figs.…”

Section: Resultsmentioning

confidence: 94%

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Metagenomic natural product discovery in lichen provides evidence for a family of biosynthetic pathways in diverse symbioses

Kampa

Gagunashvili

Gulder

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

125

127

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Bacteria are a major source of natural products that provide rich opportunities for both chemical and biological investigation. Although the vast majority of known bacterial metabolites derive from free-living organisms, increasing evidence supports the widespread existence of chemically prolific bacteria living in symbioses. A strategy based on bioinformatic prediction, symbiont cultivation, isotopic enrichment, and advanced analytics was used to characterize a unique polyketide, nosperin, from a lichen-associated Nostoc sp. cyanobacterium. The biosynthetic gene cluster and the structure of nosperin, determined from 30 μg of compound, are related to those of the pederin group previously known only from nonphotosynthetic bacteria associated with beetles and marine sponges. The presence of this natural product family in such highly dissimilar associations suggests that some bacterial metabolites may be specific to symbioses with eukaryotes and encourages exploration of other symbioses for drug discovery and better understanding of ecological interactions mediated by complex bacterial metabolites.biosynthesis | Peltigera membranacea | trans-acyltransferase polyketide synthase | 13 C nuclear magnetic resonance

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“…(12) and Pseudomonas sp. (13). Further downstream, the PKS genes have resemblances to gene clusters reported from various Nostocales or Oscillatoriales.…”

Section: Resultsmentioning

confidence: 88%

Section: Resultsmentioning

confidence: 93%

Section: Resultsmentioning

confidence: 94%

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Metagenomic natural product discovery in lichen provides evidence for a family of biosynthetic pathways in diverse symbioses

Kampa

Gagunashvili

Gulder

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

125

127

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“…However, WF-1360 B and the new compound 22Z-WF-1360 F, which were detected in Pf-5 culture supernatants in this study, were not reported previously. Instead, Brendel et al (4) isolated from cultures of Pf-5 several seco-rhizoxins, derivatives with an open ␦-lactone ring. These seco-rhizoxins included rhizoxin D3, S1, S2, and Z1, which are the corresponding seco forms of WF-1360 C, WF-1360B, WF-1360 F, and 22Z-WF-1360 F, respectively.…”

Section: Discussionmentioning

confidence: 99%

Isolation and Identification of Rhizoxin Analogs from Pseudomonas fluorescens Pf-5 by Using a Genomic Mining Strategy

Loper

Henkels

Shaffer

et al. 2008

Appl Environ Microbiol

113

101

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The products synthesized from a hybrid polyketide synthase/nonribosomal peptide synthetase gene cluster in the genome of Pseudomonas fluorescens Pf-5 were identified using a genomics-guided strategy involving insertional mutagenesis and subsequent metabolite profiling. Five analogs of rhizoxin, a 16-member macrolide with antifungal, phytotoxic, and antitumor activities, were produced by Pf-5, but not by a mutant with an insertion in the gene cluster. The five rhizoxin analogs, one of which had not been described previously, were differentially toxic to two agriculturally important plant pathogens, Botrytis cinerea and Phytophthora ramorum. The rhizoxin analogs also caused swelling of rice roots, a symptom characteristic of rhizoxin itself, but were less toxic to pea and cucumber roots. Of the rhizoxin analogs produced by Pf-5, the predominant compound, WF-1360 F, and the newly described compound 22Z-WF-1360 F were most toxic against the two plant pathogens and three plant species. These rhizoxin analogs were tested against a panel of human cancer lines, and they exhibited potent but nonselective cytotoxicity. This study highlights the value of the genomic sequence of the soil bacterium P. fluorescens Pf-5 in providing leads for the discovery of novel metabolites with significant biological properties.Secondary-metabolite production is a striking characteristic of Pseudomonas spp. (5, 28), and the current availability of genomic sequence data for several Pseudomonas spp. further highlights the capacity for secondary-metabolite production in this group of bacteria. For example, at least 6% of the genome of Pseudomonas fluorescens Pf-5, a rhizosphere bacterium that suppresses plant diseases, is devoted to secondary metabolism, with gene clusters for the biosynthesis of two siderophores, hydrogen cyanide, and several antibiotics (pyrrolnitrin, 2,4-diacetylphloroglucinol, and pyoluteorin) distributed throughout the genome (30). In addition to the secondary metabolites known to be produced by Pf-5 prior to genomic sequencing, three orphan gene clusters were identified in the genome of this bacterium (41). These three orphan genetic loci contain sequences that are characteristic of polyketide synthases (PKS) or nonribosomal peptide synthetases (NRPS) (51). One of the orphan metabolites has since been identified as orfamide A, the founder of a new group of bioactive cyclic lipopeptides that lyses zoospores of an oomycete plant pathogen and functions in the swarming motility of Pf-5 (11). In our continuing effort to identify the products of orphan pathways from the P. fluorescens Pf-5 genome, we describe here the products of a cluster containing genes with characteristic sequences of both PKS and NRPS. Using a screening strategy comparing Pf-5 to a derivative with a mutation in this cluster (12), we isolated and identified several metabolites structurally related to rhizoxin (Fig.

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“…Because few growing chains with both ␣-and ␤-keto groups have been examined on PKS assembly-line modules, it is not yet known how many dual-functioning ␣/␤ KR domains exist. Blastp searches of the dual-function PksJ-KR revealed numerous orthologs predominantly in trans-AT clusters, including those responsible for mupirocin, bryostatin, and rhizoxin biosynthesis (19,20), with 44%, 44%, and 43% sequence identity, respectively. Notably, many of these possible dual-function ␣/␤ KRs are predicted to act on C 2 -extended ␣-ketoacyl-Gly-S-T thioesters, perhaps reflecting a preference for conformationally flexible substrates (21).…”

Section: Discussionmentioning

confidence: 99%

A ketoreductase domain in the PksJ protein of the bacillaene assembly line carries out both α- and β-ketone reduction during chain growth

Calderone

Bumpus

Kelleher

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

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A cryptic PKS–NRPS gene locus in the plant commensal Pseudomonas fluorescens Pf-5 codes for the biosynthesis of an antimitotic rhizoxin complex

Abstract: Targeted gene inactivation and metabolic profiling revealed that the cryptic PKS-NRPS gene cluster in the genome of the plant commensal Pseudomonas fluorescens Pf-5 codes for the biosynthesis of antiproliferative and antifungal rhizoxin derivatives.

Cited by 61 publications

References 14 publications

Metagenomic natural product discovery in lichen provides evidence for a family of biosynthetic pathways in diverse symbioses

Metagenomic natural product discovery in lichen provides evidence for a family of biosynthetic pathways in diverse symbioses

Isolation and Identification of Rhizoxin Analogs from Pseudomonas fluorescens Pf-5 by Using a Genomic Mining Strategy

A ketoreductase domain in the PksJ protein of the bacillaene assembly line carries out both α- and β-ketone reduction during chain growth

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