Metabolomics and Dereplication Study of the Endophytic Fungus Aspergillus chevalieriin Search of Bioactive Natural Compounds (original) (raw)
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Journal of Advanced Pharmacy Research
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 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. 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 chevalieri 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 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. The pure compounds were tested for the anticancer and antimicrobial activity, compound (1) exhibited significant antitrypanosomal 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.
Journal of Chromatography B
This study aims to identify bioactive anticancer and anti-trypanosome secondary metabolites from the fermentation culture of Aspergillus flocculus endophyte assisted by modern metabolomics technologies. The endophyte was isolated from the stem of the medicinal plant Markhamia platycalyx and identified using phylogenetics. Principle component analysis was employed to screen for the optimum growth endophyte culturing conditions and revealing that the 30-days rice culture (RC-30d) provided the highest levels of the bioactive agents. To pinpoint for active chemicals in endophyte crude extracts and successive fractions, a new application of molecular interaction network is implemented to correlate the chemical and biological profiles of the anti-trypanosome active fractions to highlight the metabolites mediating for bioactivity prior to purification trials. Multivariate data analysis (MVDA), with the aid of dereplication studies, efficiently annotated the putatively active anticancer molecules. The small-scale RC-30d fungal culture was purified using high-throughput chromatographic techniques to yield compound 1, a novel polyketide molecule though inactive. Whereas, active fractions revealed from the bioactivity guided fractionation of medium scale RC-30d culture were further purified to yield 7 metabolites, 5 of which namely cis-4-hydroxymellein, 5-hydroxymellein, diorcinol, botryoisocoumarin A and mellein, inhibited the growth of chronic myelogenous leukaemia cell line K562 at 30 μM. 3hydroxymellein and diorcinol exhibited a respective inhibition of 56% and 97% to the sleeping sickness causing parasite Trypanosoma brucei brucei. More interestingly, the antitrypanosomal activity of A. flocculus extract appeared to be mediated by the synergistic effect of the active steroidal compounds i.e. ergosterol peroxide, ergosterol and campesterol. The isolated structures were elucidated by using 1D, 2D NMR and HR-ESIMS.
2013
During the research for bioactive secondary metabolites from microorganisms, the endophytic fungi Aspergillus fumigatus sp. isolate R7 was found to produce a set of promising bioactive compounds (1-10) after its large scale fermentation, working up and purification using a series of chromatographic techniques. Structural elucidation of the yielded compounds using intensive studies of their NMR ( 1 H, 13 C& 2D NMR) and mass (EI MS, ESI MS) spectrometry confirmed them as linoleic acid (1), R(-)-glycerol monolinoleate (2), bis-dethio-(bis-methyl-thio)-gliotoxin (3), fumiquinazoline-F (4), fumiquinazoline-D (5), (Z,Z)-N,N’-[1-[(4-Hydroxy-phenyl)methylene]-2-[(4-methoxy-phenyl)-methylene]-1,2-ethanediyl]-bis-formamide (6), pyrazoline-3-one trimer (7), Tricho-9-ene-2a,3a,11a,16-tetraol (8), 2’-deoxy-thymidine (9), and cerebroside A (10). In this article, taxonomical characterization, fermentation, structural characterization of the obtained metabolites were reported together with their an...
International Journal of Pharmacy and Pharmaceutical Sciences, 2020
Objective: Aspergillus fungus is a rich source of natural products with broad biological activities. This study was conducted to identify secondary metabolites from the rice culture of Aspergillus species isolated from Melaleuca subulata leaves and evaluated their anticancer activity. Methods: Ethyl acetate extract was fractionated on silica gel and Sephadex columns. Structures of the compounds were established using physical and chemical methods. Cytotoxic activities of the extract and pure compounds against two human cancer cell lines (Mcf-7and Hep G2) were evaluated using microculture tetrazolium assay as well as the mode of the cytotoxicity was evaluated. Molecular docking studies have been performed using the Hsp 90 enzyme as an anticancer target. Results: Methyl linoleate (1), arugosin C (2), ergosterol (3), sterigmatocystin (4), diorcinol (5), alternariol-5-O-methyl ether (6), averufin (7), averufanin (8), and alternariol (9) were identified from ethyl acetate extract. All tested compounds exhibit week activity against MCF-7 and Hep G2 cell lines but a mixture of compounds 7 and 8 is considered to be more active towards both MCF-7 and Hep G 2 in comparison to other compounds. Compound 4 exhibits moderate activity against Hep G2 only as well as the ethyl acetate extract exerts moderate activity against MCF-7 cell line Moreover, compound 4 and a mixture of 7 and 8 caused a decrease in the number of Hep G2 cancer cells due to apoptotic and necrotic processes. Most active anticancer candidates 7 and 8 showed binding to the active site similar to geldanamycin reference ligand. Conclusion: Secondary metabolites identified from Aspergillus sp. and their anticancer activity were evaluated. Molecular docking suggested active candidates as Hsp 90 inhibitors.
ANTIFUNGAL POTENTIAL OF BACTERIAL METABOLITES: ISOLATION, SCREENING AND CHARACTERIZATION
The increased use of antimicrobial agents resulted in the development of resistance against most of the commonly used antimicrobial drugs. Therefore the need for safe and effective antimicrobial agent have increased with time. This study was designed in an attempt to search for such natural antimicrobial agents against fungi. A total of fifty five indigenous bacterial strains were collected from different clinical specimens. These strains were identified by conventional methods as Staphylococcus aureus, S. epidermidis, Bacillus subtilis, Pseudomonas aeruginosa, Proteus mirabilis and P. vulgaris. The cell free culture supernatant (CFS) of these strains were used in antifungal screening by using agar well diffusion assay against different fungal strains. Of all the bacterial strains screened, 36.4% (20/55) showed antifungal potential. However, S. aureus, B. subtilis and P. mirabilis demonstrated bioactivity against some fungal strains but 71.4% (15/21) strains of P.aeruginosa demonstrated significant antifungal activity. The most potent antifungal strains P. aeruginosa strains MS 9 and MS 10 exhibited antifungal activity against Rhizopus sp., Aspergillus spp., Penicillium sp., Microsporum sp., Trichophyton spp. and Saccharomyces species. However, no activity was recorded against Candida spp. The physico-chemical characterization of these two strains MS 9 and MS 10 revealed retention of bioactivity of these metabolites both at low temperature (00C and 40C) and at high temperature up till 700C. Similarly exposure to organic solvents such as ethanol, methanol and formaldehyde also had no effect on their antifungal potential. The findings suggest further purification and possible application of these metabolites as antifungal agent in future. Keywords: Antifungal, Ps. aeruginosa, phytopathogenic fungi, dermatophytes.
Analytical techniques for discovery of bioactive compounds from marine fungi
TrAC Trends in Analytical Chemistry, 2012
Marine fungi are a promising source of novel bioactive compounds as lead structures for medicine and for plant protection. The current analytical techniques and future perspectives of analytical methodologies are reviewed from the point of view of the discovery and the characterization of bioactive compounds isolated from marine fungi. This critical overview also includes a general assessment of sampling and preparation of extracts, the comparison of different methods used for separation and isolation, as well as different strategies used for structural characterization of the bioactive compounds. The evolution of the application of the bioassays for discovery of bioactive compounds is also ascertained. Finally, this review addresses the advantages and disadvantages of such techniques, and comments on future applications and potential research interest within this field.
Isolation of Bioactive Compounds from Aspergillus terreus LS07
Indonesian Journal of Chemistry, 2014
This study aims to search for the active components from Aspergillus terreus LS07 which isolated from an Indonesian soil. Bioassay-guided fractionations of the ethyl acetate (EtOAc) extract against α-glucosidase and DPPH free radical to give four isolated compounds: oleic acid (1), ergosterol (2), butyrolactone I (3), and butyrolactone II (4). The structures of these metabolites were assigned on the basis of detailed spectroscopic analysis. Oleic acid (1) was showed significant activity toward α-glucosidase with IC50 value of 8.54 μM, but not for antioxidant. Butyrolactone I (3) and II (4) were showed significant activities against the α-glucosidase with their IC50 values at 52.17 and 96.01 μM, and those against DPPH free radicals at 51.39 and 17.64 μM, respectively. On the other hand, ergosterol (2) did not show any activities.
Frontiers in Microbiology
Endophytic microorganisms are an important source of bioactive secondary metabolites. In this study, fungal endophytes obtained from A*STAR’s Natural Product Library (NPL) and previously isolated from different habitats of Singapore were investigated for their diversity, antimicrobial, and cytotoxic activities. A total of 222 fungal strains were identified on the basis of sequence analysis of ITS region of the rDNA gene. The identified fungal strains belong to 59 genera distributed in 20 orders. Majority of the identified strains (99%; 219 strains) belong to the phylum Ascomycota, while two strains belonged to the phylum Basidiomycota, and only one strain was from Mucoromycota phylum. The most dominant genus was Colletotrichum accounting for 27% of all the identified strains. Chemical elicitation using 5-azacytidine and suberoylanilide hydroxamic acid (SAHA) and variation of fermentation media resulted in the discovery of more bioactive strains. Bioassay-guided isolation and structu...
Pharmacognosy Journal
Endophytic fungal isolation Endophytic fungi were isolated from plant parts according to the method of Kyeremeh et al. (2017). 18 To make ensure that, isolated fungi are endophytes, the plant part surfaces were sterilized by rinsing with sterile distilled water (SDW), followed by dipping in to 70% ethanol for 1 min. the pieces were again rinsed ABSTRACT Background: Endophytes are the richest sources of natural compounds, showing biological varieties and pharmacological activities. Objective: This study aims to isolate bioactive secondary metabolites from endophytic Aspergillus tubenginses with study the biological activity of the isolated bioactive compounds. Materials and Methods: Anofinic acid were obtained from Aspergillus tubenginses crude extract using chromatographic techniques and characterized by spectral analysis. Results: Nine endophytic fungi were isolated from Hyoscyamus muticus plant. The most efficient isolate was AF3 identified as Aspergillus tubenginses ASH4 by 18S rRNA gene sequencing. Anofinic acid is an isolated active metabolite biosynthesized by A. tubenginsis was extracted from ethyl acetate with UPAC name of 2,2-dimethyl-2H-1-benzopyran-6-carboxylic acid. It shows a strong antimicrobial activity against human pathogenic bacteria such as Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, Candida albicans and Bacillus subtilis. Moreover, anofinic acid inhibits biofilm formation and has antioxidant activity, with strong activity against some carcinoma cells such as HCT-116, Hep-G2 and MCF-7. Conclusion: Anofinic acid was purified from the endophytic Aspergillus tubenginses crude extract and showed antimicrobial, antibiofilm, antioxidant, anticancer activities.
Secondary metabolites and bioactivity of two fungal strains
Purpose The investigation of two fungal strains isolated from Egyptian habitats, namely, the endophytic Fusarium poae FUN1 and the terrestrial Penicillium italicum FUN2 to illustrate their chemical constituents and their bioactivities. Materials and methods See General instrumental procedures. Results Linoleic acid (1), indole-3-acetic acid methyl ester (2) and Nb-acetyltryptamine (3) were produced by F. poae FUN1, whereas P. italicum FUN2 also delivered linoleic acid (1) in addition to cis-cyclo-(prolyl,valyl) (4). The structures of compounds (1)–(4) were elucidated by 1D and 2D NMR, MS data and through comparison with literature reports. In this article, the taxonomical characterization of both fungal strains, their upscale fermentation and the antimicrobial and cytotoxic activities tested have been described. Conclusion Two different fungal strains, endophytic F. poae FUN1 and terrestrial P. italicum FUN2, were intensively studied biologically and chemically. Four bioactive compounds (1)–(4) were isolated, and structurally confirmed by intensive studies of NMR and MS. The antimicrobial and cytotoxic activities of the fungal extracts and their delivered compounds were studied. This might be helpful for the cure of recent diseases, and drug-resistant phenomena as well as in the development of pharmaceutical, agrochemical and biochemical agents and their lead compounds.