LC-MS/MS Method Development for the Discovery and Identification of Amphidinols Produced by Amphidinium

Wellkamp, Marvin; Camacho, F. Garcı́a; Durán-Riveroll, Lorena M.; Tebben, Jan; Tillmann, Urban; Krock, Bernd

doi:10.3390/md18100497

Cited by 20 publications

(38 citation statements)

References 37 publications

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“…The retention times for the compounds of interest ranged from 3.4-4.1 min with m/z at 1667, 1361, 1463 and 1343 during a 16 min UHPLC run (Table 1). The main fragments observed in the MS/MS fragmentation spectra of the compounds at m/z 1343, 1361 and 1463 [M + Na] + were found at m/z 1085 and 687, while the main fragments of amphidinols/luteophanols were shown to be at m/z 903 or 1105 following the numbering recently proposed by Wellkamp et al [29]. The only known derivatives with fragments at m/z 1085 (C-29/C-30) and 687 (C-29/C-30 and C-1/C-1 ) were identified as amphidinol A (m/z 1361) and B (m/z 1463) from the comprehensive analysis of Wellkamp et al [29] and Cutigano et al [28].…”

Section: Chemical Profiling Of the Bioactive Fractionsmentioning

confidence: 56%

“…The UV profiles were then used as comparative data in UHPLC-DAD-HRMS/MS to obtain the MS spectra of these major compounds. Amphidinol type compounds were detected as major compounds in the bioactive fractions G-J due to characteristic m/z and fragmentation patterns, but these compounds were absent in fraction K [29]. The retention times for the compounds of interest ranged from 3.4-4.1 min with m/z at 1667, 1361, 1463 and 1343 during a 16 min UHPLC run (Table 1).…”

Section: Chemical Profiling Of the Bioactive Fractionsmentioning

confidence: 99%

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Antibacterial Activity and Amphidinol Profiling of the Marine Dinoflagellate Amphidinium carterae (Subclade III)

Barone

Murphy

Parkes

et al. 2021

IJMS

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Microalgae have received growing interest for their capacity to produce bioactive metabolites. This study aimed at characterising the antimicrobial potential of the marine dinoflagellate Amphidinium carterae strain LACW11, isolated from the west of Ireland. Amphidinolides have been identified as cytotoxic polyoxygenated polyketides produced by several Amphidinium species. Phylogenetic inference assigned our strain to Amphidinium carterae subclade III, along with isolates interspersed in different geographic regions. A two-stage extraction and fractionation process of the biomass was carried out. Extracts obtained after stage-1 were tested for bioactivity against bacterial ATCC strains of Staphylococcus aureus, Enterococcus faecalis, Escherichia coli and Pseudomonas aeruginosa. The stage-2 solid phase extraction provided 16 fractions, which were tested against S. aureus and E. faecalis. Fractions I, J and K yielded minimum inhibitory concentrations between 16 μg/mL and 256 μg/mL for both Gram-positive. A targeted metabolomic approach using UHPLC-HRMS/MS analysis applied on fractions G to J evidenced the presence of amphidinol type compounds AM-A, AM-B, AM-22 and a new derivative dehydroAM-A, with characteristic masses of m/z 1361, 1463, 1667 and 1343, respectively. Combining the results of the biological assays with the targeted metabolomic approach, we could conclude that AM-A and the new derivative dehydroAM-A are responsible for the detected antimicrobial bioactivity.

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Section: Chemical Profiling Of the Bioactive Fractionsmentioning

confidence: 56%

Section: Chemical Profiling Of the Bioactive Fractionsmentioning

confidence: 99%

Antibacterial Activity and Amphidinol Profiling of the Marine Dinoflagellate Amphidinium carterae (Subclade III)

Barone

Murphy

Parkes

et al. 2021

IJMS

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“…is as low as 1.2 μg L −1 [ 20 ]; 400-fold times lower than that obtained in our previously analyzed supernatant of 78 L [ 6 ]. On the other hand, a recent study revealed that the AM profile and cell quotas of eight Amphidinium strains were extraordinarily diverse [ 14 ]. In that study, lingshuiol A (3 fg cell −1 ) and luteophanol D (<1 fg cell −1 ) were detected in cells of the strain ACRN03 (the same strain as in the study presented here), but at a trace level (near the limit of detection).…”

Section: Resultsmentioning

confidence: 99%

“…Despite the above difficulties, we have recently developed strategies related to the production of pilot-scale cultures of dinoflagellates of the genus Amphidinium [ 6 , 7 , 8 , 9 ]. Amphidinium species are known to produce super carbon chain compounds including amphidinols (AMs) and other related metabolites [ 10 , 11 , 12 , 13 , 14 ]. This growing family of opened long-chain polyketides, characterized by a hairpin shape constituted by a central common core delimited by two tetrahydropyran rings separating two moieties—one mainly polyhydroxilic and other polyenic—is known to elicit potent antifungal, ichthyotoxic, hemolytic, cytotoxic, antiprotozoan or antidiatom activities [ 15 ].…”

Section: Introductionmentioning

confidence: 99%

Isolation and Structural Elucidation of New Amphidinol Analogues from Amphidinium carterae Cultivated in a Pilot-Scale Photobioreactor

et al. 2021

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The demand for valuable products from dinoflagellate biotechnology has increased remarkably in recent years due to their many prospective applications. However, there remain many challenges that need to be addressed in order to make dinoflagellate bioactives a commercial reality. In this article, we describe the technical feasibility of producing and recovering amphidinol analogues (AMs) excreted into a culture broth of Amphidinium carterae ACRN03, successfully cultured in an LED-illuminated pilot-scale (80 L) bubble column photobioreactor operated in fed-batch mode with a pulse feeding strategy. We report on the isolation of new structurally related AMs, amphidinol 24 (1, AM24), amphidinol 25 (2, AM25) and amphidinol 26 (3, AM26), from a singular fraction resulting from the downstream processing. Their planar structures were elucidated by extensive NMR and HRMS analysis, whereas the relative configuration of the C-32®C-47 bis-tetrahydropyran core was confirmed to be antipodal in accord with the recently revised configuration of AM3. The hemolytic activities of the new metabolites and other related derivatives were evaluated, and structure–activity conclusions were established. Their isolation was based on a straightforward and high-performance bioprocess that could be suitable for the commercial development of AMs or other high-value compounds from shear sensitive dinoflagellates.

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“…Unfortunately, much of the early work on the natural products chemistry and structural elucidation of polyketides was based upon cultured Amphidinium isolates of uncertain or unspecified taxonomic identification at the species level. Given the uncertainty in Amphidinium species assignment, and high variability and diversity among strains in capability to biosynthesize polyketides [ 8 , 12 , 27 ], it is essential to identify and characterize the strain first. Then the biological activity of the extracts should be screened, and finally the respective secondary metabolites identified and purified for further bioactivity assessment.…”

Section: Introductionmentioning

confidence: 99%

Toxicity Bioassay and Cytotoxic Effects of the Benthic Marine Dinoflagellate Amphidinium operculatum

Mejía-Camacho

Durán-Riveroll

Cembella

2021

JoX

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Benthic dinoflagellates produce a wide array of bioactive compounds, primarily polyketides, that cause toxic effects on human consumers of seafood and perhaps mediate species interactions in the benthic microenvironment. This study assesses toxic and other bioactive effects of the benthic dinoflagellate Amphidinium operculatum (strain AA60) in two targeted bioassays. The brine shrimp (Artemia salina) bioassay revealed lethal effects of direct exposure to live dinoflagellate cells (Treatment A) and even higher potency with ethanolic extracts of lysed cells (Treatment D). There were no inimical bioactive effects of components released to the aqueous growth medium (Treatment B) or from aqueous cell lysates (Treatment C). The hypothesis that released bioactive compounds provide a chemical defense against metazoan grazers is therefore not supported by these results. The cytotoxic effect of ethanolic crude extracts of this dinoflagellate exhibited mild to high growth reduction effects on six human cancer cell lines. In particular, crude cell-free extracts proved highly growth-inhibitory activity towards breast and lung cancer cell lines MCF-7 and SKLU-1, respectively. Preliminary anti-cancer results indicate that natural bioactive compounds from Amphidinium are worthy of structural characterization and further toxicological investigation as potential therapeutants.

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LC-MS/MS Method Development for the Discovery and Identification of Amphidinols Produced by Amphidinium

Cited by 20 publications

References 37 publications

Antibacterial Activity and Amphidinol Profiling of the Marine Dinoflagellate Amphidinium carterae (Subclade III)

Antibacterial Activity and Amphidinol Profiling of the Marine Dinoflagellate Amphidinium carterae (Subclade III)

Isolation and Structural Elucidation of New Amphidinol Analogues from Amphidinium carterae Cultivated in a Pilot-Scale Photobioreactor

Toxicity Bioassay and Cytotoxic Effects of the Benthic Marine Dinoflagellate Amphidinium operculatum

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