Endophytic fungi are now recognized as sources of pharmacologically beneficial, novel bioactive compounds. This study was carried out to evaluate antiproliferative and antioxidative potential of a seaweed endophytic fungus Talaromyces purpureogenus. Extracts with different solvents of the fungus grown on different liquid media were assayed for the antiproliferative and antioxidative activities. Tested 6 cancer cell lines, the highest antiproliferative activity was observed in ethyl acetate extract of total culture grown in Potato Dextrose Broth for 28 days in a dose-dependent manner. The highest antioxidative activity was observed in hexane extract of fungal culture grown in Malt Extract Broth for 21 days. Analyzed for secondary metabolites, the extract revealed the presence of phenolics, alkaloids, flavonoids, steroids and terpenoids. Further, Gas Chromatography Mass Spectroscopy (GCMS) analysis of the extract revealed the presence of several compounds including 3-nitropropanoic acid, 4H-pyran-4-one 5-hydroxy-2-(hydroxymethyl), hexadecanoic acid, and octadecanoic acid, known to be cytotoxic or antioxidative. Among different cell lines tested, HeLa cells were the most vulnerable to the treatment of the fungal extract with an IC50 value of 101 ± 1 μg/mL. The extract showed no significant cytotoxicity to the normal human embryonic kidney cell line (HEK 293 T) in the MTT assay. The ethyl acetate extract induced membrane damage and mitochondrial depolarization and thereby apoptosis and cytotoxicity in HeLa cells. The study marks marine-derived endophytes as potential sources for discovery of novel drugs.
Endophytes are an unexploited source of pharmacologically relevant compounds owing to their species richness and diversity. In this study, a total of 26 endophytic fungi were isolated and identified from 10 marine algal samples collected from the Konkan coast, Goa, India. Eighteen of the fungal isolates belonged to phylum Ascomycota while one belonged to phylum Basidiomycota based on ITS sequencing. Further, the genus Aspergillus sp. was the most common and abundant endophyte found in the sampled algal species. A significant antibacterial activity against five pathogenic bacteria was exhibited by the extracts of fungal isolates AG1.1, AG1.1 (G) and VG2.6 (agar diffusion assay). The extracts of fungal endophytes VB1.1, PG1.2 and VG2.6 demonstrated good antioxidant activity (DPPH scavenging assay). Further, cytotoxicity of all the endophytic extracts on human cancer cell lines was determined by MTT and resazurin assay. The crude extract of Aspergillus unguis (AG 1.2) showed the highest cytotoxic potential on cervical cancer (HeLa), breast cancer (MCF-7), lung cancer (A549), and skin cancer (A431) cell lines in a concentration dependent manner. Moreover, Gas Chromatography-Mass Spectroscopy analysis of the extract of A. unguis (AG 1.2) confirmed the presence of several bioactive metabolites including azelaic acid, azetidine, and furopyrans. The extract of A. unguis (AG 1.2) demonstrated G1 phase cell cycle arrest, reactive oxygen species (ROS)-dependent MMP loss and apoptosis-dependent cell death in A431 cells. The algae-derived fungal endophytes of Konkan coast are a rich source of novel pharmaceutically active compounds as indicated by this work.
The endophytic fungal community in the marine ecosystem has been demonstrated to be relevant source of novel and pharmacologically active secondary metabolites. The current study focused on the evaluation of cytotoxic and apoptosis induction potential in the culture extracts of endophytic fungi associated with Sargassum muticum, a marine brown alga. The cytotoxicity of the four marine endophytes, Aspergillus sp., Nigrospora sphaerica, Talaromyces purpureogenus, and Talaromyces stipitatus, was evaluated by the MTT assay on HeLa cells. Further, several physicochemical parameters, including growth curve, culture media, and organic solvents, were optimized for enhanced cytotoxic activity of the selected extract. The Aspergillus sp. ethyl acetate extract (ASE) showed maximum cytotoxicity on multiple cancer cell lines. Chemical investigation of the metabolites by gas chromatography–mass spectroscopy (GC-MS) showed the presence of several compounds, including quinoline, indole, 2,4-bis(1,1-dimethylethyl) phenol, and hexadecenoic acid, known to be cytotoxic in ASE. The ASE was then tested for cytotoxicity in vitro on a panel of six human cancer cell lines, namely, HeLa (cervical adenocarcinoma), MCF-7 (breast adenocarcinoma), Hep G2 (hepatocellular carcinoma), A-549 (lung carcinoma), A-431 (skin/epidermis carcinoma), and LN-229 (glioblastoma). HeLa cells were most vulnerable to ASE treatment with an IC50 value of 24 ± 2 μg/ml. The mechanism of cytotoxicity exhibited by the ASE was further investigated on Hela cells. The results showed that the ASE was capable of inducing apoptosis in HeLa cells through production of reactive oxygen species, depolarization of mitochondrial membrane, and activation of the caspase-3 pathway, which shows a possible activation of the intrinsic apoptosis pathway. It also arrested the HeLa cells at the G2/M phase of the cell cycle, eventually leading to apoptosis. Through this study, we add to the knowledge about the marine algae associated with fungal endophytes and report its potential for purifying specific compounds responsible for cytotoxicity.
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