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Chemotaxonomic profiling of Penicillium setosum using high-resolution mass spectrometry (LC-Q-ToF-MS).

ABSTRACT: In the present study, secondary metabolites produced by an endophytic fungus Penicillium setosum were extracted using colony agar plug and culture broth extraction methods. High resolution LC-MS was used to explore the chemical nature of the secondary metabolites, as well, compare the reliability of the methods. P. setosum was chemotaxonomically distinguished from other members of section Lanata-divaricata, by its ability to produce mycotoxin, patulin and also by the presence of certain phenol-derived compounds, like quercetin, dihydroflavonols (dihydroquercetin and dihydromyricetin), kaempferol, luteolin, while some Penicillium specific compounds such as, citromycetin and andrastin D reveal its similarity towards section Lanata-Divaricata members. For the first time, the presence of dihydroquercetin is remarkably and spectrometrically confirmed from a microbial source. In addition, a few polyketides, anthroquinone compounds, hydrocarbons, and fatty acids were also detected in the culture extract. Being the first report on the production of polyphenolic compounds by an endophytic fungus of Penicillium species, the current research is crucial, and moreover the starin itself is a novel species.

SUBMITTER: George TK

PROVIDER: S-EPMC6819834 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature

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Chemotaxonomic profiling of Penicillium setosum using high-resolution mass spectrometry (LC-Q-ToF-MS).

George Tijith K TK Devadasan Dineep D Jisha M S MS

Heliyon 20190926 9

In the present study, secondary metabolites produced by an endophytic fungus Penicillium setosum were extracted using colony agar plug and culture broth extraction methods. High resolution LC-MS was used to explore the chemical nature of the secondary metabolites, as well, compare the reliability of the methods. P. setosum was chemotaxonomically distinguished from other members of section Lanata-divaricata, by its ability to produce mycotoxin, patulin and also by the presenc ...[more]

PMID: 31687578

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Project description:IntroductionSince ancient times medicinal plants have been used as medicine in many parts of the world to promote human health and longevity. In recent years many novel secondary metabolites of plants have been isolated and reported to provide lead compounds for new drug discoveries. Solanum mauritianum Scopoli is native to South America. It is reported to be used by native South Americans during famine as a vegetable and as medicine to cure various diseases. In South Africa the plant is viewed as weed and is facing eradication, however, this plant is a valuable subject for research into its potential pharmaceutical and chemical uses. This study elucidated the metabolic profile of fungal endophytes that have promising bioactive secondary metabolites against pathogenic microorganisms, including mycobacterium species.Material and methodsFungal endophytes from a weed Solanum mauritianum Scop. were used to synthesize secondary metabolites. Gas chromatograph high-resolution time-of-flight mass spectrometry (GC-HRTOF-MS) was used to analyse volatile compounds to prove that potentially fungal endophytes could be extracted from this weed. Extracts obtained with ethyl acetate were screened for phytochemicals and analyzed using a gas chromatograph high-resolution time-of-flight mass spectrometry system. Principal component analysis was used to compare the gas chromatograph high-resolution time-of-flight mass spectrometry data for differences/similarities in their clustering. Phytochemical screening was conducted on the crude extracts of fungal endophytes obtained from different parts of Solanum mauritianum Scopoli (leaves, ripe fruit, unripe fruit and stems).ResultsPhytochemical screening indicated the presents of alkaloids, flavonoids, glycosides, phenols, quinones and saponins. Quinones were not present in the crude extracts of Fusarium sp. A total of 991 compounds were observed in the fungal endophytes, and Cladosporium sp. (23.8%) had the highest number of compounds, compared to Paracamarosporium leucadendri (1.7%) and Talaromyces sp. (1.5%). Some volatile compounds such as eicosane, 2-pentadecanone, 2-methyloctacosane, hexacosane and tridecanoic acid methyl ester with antibacterial activity were also observed.ConclusionCompositional variations between the plant and fungal endophyte phytochemicals were observed. The results of this study indicate that fungal endophytes from Solanum mauritianum Scop. contain compounds that can be exploited for numerous pharmaceutical and medicinal applications.

Project description:Canine oral tumors are relatively common neoplasms in dogs. For disease monitoring and early diagnosis, salivary biomarkers are appropriate because saliva collection is non-invasive and requires no professional skills. In the era of omics, matrix-assisted laser desorption/ionization with time-of-flight mass spectrometry (MALDI-TOF MS) coupled with liquid chromatography-tandem MS (LC-MS/MS) are suitable to identify potential disease-associated peptides and proteins. The present study aimed to use MALDI-TOF MS and LC-MS/MS to search for particular peptide mass fingerprints (PMFs) and conceivable biomarkers in saliva of dogs with early- and late-stage oral melanoma (EOM and LOM, respectively), oral squamous cell carcinoma (OSCC), benign oral tumors (BN), and periodontitis and healthy controls (CP). Pooled saliva samples in each group were used to be representative of population change. Unique PMFs were obtained and specific peptide fragments were sequenced by LC-MS/MS and BLAST-searched with mammalian protein databases. Seven peptide fragments appeared in the tumor groups (EOM, LOM, OSCC and BN) at 1096, 1208, 1322, 1794, 1864, 2354 and 2483 Da, two peptide fragments appeared in the LOM and OSCC groups at 2450 and 3492 Da, and in the CP controls at 2544 and 3026 Da. Also, protein-chemotherapy drug interaction networks were exhibited. Using western blot analysis, the expression of sentrin-specific protease 7 (SENP7), a peptide fragment at 1096 Da, in OSCC was significantly increased, as was the expression of TLR4, a peptide fragment at 3492 Da, in LOM and OSCC, compared with the CP group. The expression of nuclear factor kappa B (NF-κB), a TLR4 partner, was notably increased in OSCC compared with CP, BN and EOM. The expression was also enhanced in LOM compared with EOM. Expressed protein sequences from western blots were verified by LC-MS/MS. Western blots were then performed with individual samples in each group. The results showed the elevated expression of TLR4 in LOM and OSCC, compared with that in CP and BN, the increased expression of NF-κB in LOM and OSCC, compared with CP and in LOM compared with BN, and the enhanced expression of SENP7 in LOM and OSCC, compared with that in CP and BN. In conclusion, discrete clusters of EOM, LOM, OSCC, BN and CP groups and potential protein candidates associated with the diseases were demonstrated by salivary proteomics. Western blot analysis verified SENP7, TLR4 and NF-κB as potential salivary biomarkers of canine oral tumors.

Project description:Monitoring of marine mammal steroid hormone status using matrices alternative to blood is desirable due to the ability to remotely collect samples, which minimizes stress to the animal. However, measurement techniques in alternative matrices such as blubber described to date are limited in the number and types of hormones measured. Therefore, a new method using bead homogenization to QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) extraction, C18 post extraction cleanup and analysis by liquid chromatography tandem mass spectrometry (LC-MS/MS) was developed and applied to the measurement of hormone suites in bottlenose dolphin blubber. Validations were conducted in blubber from fresh dead stranded bottlenose dolphin. The final method consisting of two LC separations and garnet bead homogenization was tested for extraction efficiencies. Steroids were separated using a biphenyl column for reproductive hormones and C18 column for corticosteroids. Three hormones previously noted in blubber, testosterone, progesterone, and cortisol, were quantified in addition to previously unmeasured androstenedione, 17-hydroxyprogesterone, 11-deoxycortisol, 11-deoxycorticosterone, and cortisone in a single sample (0.4 g blubber). Extraction efficiencies of all hormones from blubber ranged from 84% to 112% and all RSDs were comparable to those reported using immunoassay methods (< 15%). The method was successfully applied to remote biopsied blubber samples to measure baseline hormone concentrations. Through this method, increased coverage of steroid hormone pathways from a single remotely collected sample potentially enhances the ability to interpret biological phenomena such as reproduction and stress in wild dolphin populations. Graphical abstract The steroid hormone profile is quantifiable from a single sample of bottlenose dolphin blubber using liquid chromatography tandem mass spectrometry. This profile can be applied to remotely collected dart biopsies and be used to determine reproductive or stress status of a wild-living dolphin.

Project description:BackgroundLC-MALDI-TOF/TOF analysis is a potent tool in biomarkers discovery characterized by its high sensitivity and high throughput capacity. However, methods based on MALDI-TOF/TOF for biomarkers discovery still need optimization, in particular to reduce analysis time and to evaluate their reproducibility for peak intensities measurement. The aims of this methodological study were: (i) to optimize and critically evaluate each step of urine biomarker discovery method based on Nano-LC coupled off-line to MALDI-TOF/TOF, taking full advantage of the dual decoupling between Nano-LC, MS and MS/MS to reduce the overall analysis time; (ii) to evaluate the quantitative performance and reproducibility of nano-LC-MALDI analysis in biomarker discovery; and (iii) to evaluate the robustness of biomarkers selection.ResultsA pool of urine sample spiked at increasing concentrations with a mixture of standard peptides was used as a specimen for biological samples with or without biomarkers. Extraction and nano-LC-MS variabilities were estimated by analyzing in triplicates and hexaplicates, respectively. The stability of chromatographic fractions immobilised with MALDI matrix on MALDI plates was evaluated by successive MS acquisitions after different storage times at different temperatures.Low coefficient of variation (CV%: 10-22%) and high correlation (R2 > 0.96) values were obtained for the quantification of the spiked peptides, allowing quantification of these peptides in the low fentomole range, correct group discrimination and selection of "specific" markers using principal component analysis. Excellent peptide integrity and stable signal intensity were found when MALDI plates were stored for periods of up to 2 months at +4 degrees C. This allowed storage of MALDI plates between LC separation and MS acquisition (first decoupling), and between MS and MSMS acquisitions while the selection of inter-group discriminative ions is done (second decoupling). Finally the recording of MSMS spectra to obtain structural information was focused only on discriminative ions in order to minimize analysis time.ConclusionContrary to other classical approaches with direct online coupling of chromatographic separation and on the flight MS and/or MSMS data acquisition for all detected analytes, our dual decoupling strategy allowed us to focus on the most discriminative analytes, giving us more time to acquire more replicates of the same urine samples thus increasing detection sensitivity and mass precision.

Dataset Information

Chemotaxonomic profiling of Penicillium setosum using high-resolution mass spectrometry (LC-Q-ToF-MS).

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Chemotaxonomic profiling of <i>Penicillium setosum</i> using high-resolution mass spectrometry (LC-Q-ToF-MS).

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