Abstract
Objectives: This study aimed to point the significant rule of metabolomics tools to assess the chemistry of the bioactive metabolites produced by endophytic fungus Aspergillus chevelieri isolated from Lagerostromia tomentosa C. presl. The anticancer and antimicrobial activity of crude extracts, fractions and pure compounds were investigated as part of this study. Methods: An endophytic fungus (Aspergillus chevelieri) was isolated from the tissues of the stem of Lagerostromia tomentosa C. presl and identified through molecular biological procedure by DNA isolation, PCR, DNA sequencing and through searching the Gene Bank. Metabolomics profiling and dereplication studies were employed to choose the optimum growth medium and conditions that yield the most significant metabolites. The crude extract of the 30-days rice culture of Aspergillus chevelieri was subjected to bioactivity and metabolomics guided isolation approach. The structure of the isolated compounds was determined on the basis of 1D, 2D NMR and mass spectrometry (HR-ESIMS) analysis. Results: four bioactive fractions were further purified to produce five pure compounds, which are Ergosterol (1), Ergosterol peroxide (2), Campesterol (3), Flavoglaucin (4) and 3-O- methyl caffeic acid (5). Multivariate data analysis highlighted the most significant metabolites contributed to the bioactivity. Only 30 days RC extract of A. chevelieri was active among all fungal extracts. The pure compounds were tested for the anticancer and antimicrobial activity, compound (1) exhibited significant anti-trypanosomal activity, while compounds (2, 3, 4 and 5) effectively inhibited the growth of Escherichia coli, Staphylococcus aureus and Candida albicans. Conclusion: A combination of metabolomic- and bioassay-guided approaches gives an access to a shorter and faster route to highlight the active metabolites, which are highly correlated to the bioactivity during the first stage of fractionation.
Highlights
MHz), 13C-NMR (DMSO, 100 MHz) data presented in table 1; ESIHRMS(pos): m/z [M-H]- 395.3309
The heat map of all extracts showed that Aspergillus chevalieri extract and Alternaria alternate fungal extracts were the richest in metabolites of different mass range as shown in figure 1
The cytotoxicity assay showed that Aspergillus chevalieri was active against A278O cell line and non-toxic for the normal cells PNT2A (Figure 2), implying that Aspergillus chevalieri could have a unique chemical and biological fingerprints
Summary
This study aimed to point the significant rule of metabolomics tools to assess the chemistry of the bioactive metabolites produced by endophytic fungus Aspergillus chevalieri isolated from Lagerostromia tomentosa C. presl. The anticancer of crude extracts, fractions and pure compounds and antimicrobial of pure compounds were investigated as part of this study. Methods: An endophytic fungus (Aspergillus chevalieri) was isolated from the tissues of the stem of Lagerostromia tomentosa C. The crude extract of the 30-days rice culture of Aspergillus chevalieri was subjected to bioactivity and metabolomics guided isolation approach. Multivariate data analysis highlighted the most significant metabolites contributed to the bioactivity. Conclusion: A combination of metabolomic- and bioassay-guided approaches gives an access to a shorter and faster route to highlight the active metabolites, which are highly correlated to the bioactivity during the first stage of fractionation
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