Dataset Information

Insight into Unprecedented Diversity of Cyanopeptides in Eutrophic Ponds Using an MS/MS Networking Approach.

ABSTRACT: Man-made shallow fishponds in the Czech Republic have been facing high eutrophication since the 1950s. Anthropogenic eutrophication and feeding of fish have strongly affected the physicochemical properties of water and its aquatic community composition, leading to harmful algal bloom formation. In our current study, we characterized the phytoplankton community across three eutrophic ponds to assess the phytoplankton dynamics during the vegetation season. We microscopically identified and quantified 29 cyanobacterial taxa comprising non-toxigenic and toxigenic species. Further, a detailed cyanopeptides (CNPs) profiling was performed using molecular networking analysis of liquid chromatography-tandem mass spectrometry (LC-MS/MS) data coupled with a dereplication strategy. This MS networking approach, coupled with dereplication, on the online global natural product social networking (GNPS) web platform led us to putatively identify forty CNPs: fourteen anabaenopeptins, ten microcystins, five cyanopeptolins, six microginins, two cyanobactins, a dipeptide radiosumin, a cyclooctapeptide planktocyclin, and epidolastatin 12. We applied the binary logistic regression to estimate the CNPs producers by correlating the GNPS data with the species abundance. The usage of the GNPS web platform proved a valuable approach for the rapid and simultaneous detection of a large number of peptides and rapid risk assessments for harmful blooms.

SUBMITTER: Kust A

PROVIDER: S-EPMC7551678 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature

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Insight into Unprecedented Diversity of Cyanopeptides in Eutrophic Ponds Using an MS/MS Networking Approach.

Kust Andreja A Řeháková Klára K Vrba Jaroslav J Maicher Vincent V Mareš Jan J Hrouzek Pavel P Chiriac Maria-Cecilia MC Benedová Zdeňka Z Tesařová Blanka B Saurav Kumar K

Toxins 20200831 9

Man-made shallow fishponds in the Czech Republic have been facing high eutrophication since the 1950s. Anthropogenic eutrophication and feeding of fish have strongly affected the physicochemical properties of water and its aquatic community composition, leading to harmful algal bloom formation. In our current study, we characterized the phytoplankton community across three eutrophic ponds to assess the phytoplankton dynamics during the vegetation season. We microscopically identified and quantif ...[more]

PMID: 32878042

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Project description:Cucurbitaceae comprises 800 species, the majority of which are known for their nutritive, economic, and health-promoting effects. This study aims at the metabolome profiling of cucumber (Cucumis sativus) and bottle gourd (Lagenaria siceraria) fruits in a comparative manner for the first time, considering that both species are reported to exhibit several in-common phytochemical classes and bioactivities. Nevertheless, bottle gourd is far less known and/or consumed than cucumber, which is famous worldwide. A multiplex approach, including HR-UPLC/MS/MS, GNPS networking, SPME, and GC/MS, was employed to profile primary and secondary metabolites in both species that could mediate for new health and nutritive aspects, in addition to their aroma profiling, which affects the consumers' preferences. Spectroscopic datasets were analyzed using multivariate data analyses (PCA and OPLS) for assigning biomarkers that distinguish each fruit. Herein, 107 metabolites were annotated in cucumber and bottle gourd fruits via HR-UPLC/MS/MS analysis in both modes, aided by GNPS networking. Metabolites belong to amino acids, organic acids, cinnamates, alkaloids, flavonoids, pterocarpans, alkyl glycosides, sesquiterpenes, saponins, lignans, fatty acids/amides, and lysophospholipids, including several first-time reported metabolites and classes in Cucurbitaceae. Aroma profiling detected 93 volatiles presented at comparable levels in both species, from which it can be inferred that bottle gourds possess a consumer-pleasant aroma, although data analyses detected further enrichment of bottle gourd with ketones and esters versus aldehydes in cucumber. GC/MS analysis of silylated compounds detected 49 peaks in both species, including alcohols, amino acids, fatty acids/esters, nitrogenous compounds, organic acids, phenolic acids, steroids, and sugars, from which data analyses recognized that the bottle gourd was further enriched with fatty acids in contrast to higher sugar levels in cucumber. This study provides new possible attributes for both species in nutrition and health-care fields based on the newly detected metabolites, and further highlights the potential of the less famous fruit "bottle gourd", recommending its propagation.

Project description:BACKGROUND:Herbicidin F has an undecose tricyclic furano-pyrano-pyran structure with post-decorations. It was detected from Streptomyces mobaraensis US-43 fermentation broth as a trace component by HPLC-MS analysis. As herbicidins exhibit herbicidal, antibacterial, antifungal and antiparasitic activities, we are attracted to explore more analogues for further development. RESULTS:The genome of S. mobaraensis US-43 was sequenced and a herbicidin biosynthetic gene cluster (hcd) was localized. The cluster contains structural genes, one transporter and three potential transcription regulatory genes. Overexpression of the three regulators respectively showed that only hcdR2 overexpression significantly improved the production of herbicidin F, and obviously increased the transcripts of 7 structural genes as well as the transporter gene. After performing homology searches using BLASTP in the GenBank database, 14 hcd-like clusters were found with a cluster-situated hcdR2 homologue. These HcdR2 orthologues showed overall structural similarity, especially in the C-terminal DNA binding domain. Based on bioinformatics analysis, a 21-bp consensus binding motif of HcdR2 was detected within 30 promoter regions in these genome-mined clusters. EMSA results verified that HcdR2 bound to the predicted consensus sequence. Additionally, we employed molecular networking to explore novel herbicidin analogues in hcdR2 overexpression strain. As a result, ten herbicidin analogues including six new compounds were identified based on MS/MS fragments. Herbicidin O was further purified and confirmed by 1H NMR spectrum. CONCLUSIONS:A herbicidin biosynthetic gene cluster (hcd) was identified in S. mobaraensis US-43. HcdR2, a member of LuxR family, was identified as the pathway-specific positive regulator, and the production of herbicidin F was dramatically increased by overexpression of hcdR2. Combined with molecular networking, ten herbicidin congeners including six novel herbicidin analogues were picked out from the secondary metabolites of hcdR2 overexpression strain. The orthologues of herbicidin F pathway-specific regulator HcdR2 were present in most of the genome-mined homologous biosynthetic gene clusters, which possessed at least one consensus binding motif with LuxR family characteristic. These results indicated that the combination of overexpression of hcdR2 orthologous regulator and molecular networking might be an effective way to exploit the "cryptic" herbicidin-related biosynthetic gene clusters for discovery of novel herbicidin analogues.

Dataset Information

Insight into Unprecedented Diversity of Cyanopeptides in Eutrophic Ponds Using an MS/MS Networking Approach.

Publications

Insight into Unprecedented Diversity of Cyanopeptides in Eutrophic Ponds Using an MS/MS Networking Approach.

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