scholarly journals Evaluation of antimicrobial, antioxidant and cytotoxic properties of bioactive compounds produced from endophytic fungi of Himalayan yew (Taxus wallichiana) in Nepal

F1000Research ◽  
2020 ◽  
Vol 9 ◽  
pp. 379 ◽  
Author(s):  
Dhurva Prasad Gauchan ◽  
Pratistha Kandel ◽  
Astha Tuladhar ◽  
Ashesh Acharya ◽  
Upendra Kadel ◽  
...  

Background: Endophytic fungi are largely underexplored in the discovery of natural bioactive products though being rich sources of novel compounds with promising pharmaceutical potential. In this study, Taxus wallichiana, which has huge medicinal value, was investigated for its endophytic diversity and capability to produce bioactive secondary metabolites by analyzing antioxidant, antimicrobial and cytotoxic properties. Methods: The endophytes were identified by ITS-PCR using genomic DNA samples. The secondary metabolites were extracted by solvent extraction method using ethyl acetate. The antioxidant activity was analyzed by Thin Layer Chromatography, Total Phenol Content (TPC), Total Flavonoid Content (TFC) and DPPH assay, and the antimicrobial activity was analyzed by agar-well diffusion method. Brine shrimp lethality assay was used to analyze the cytotoxicity of the fungal extracts. Results: Out of 16 different Taxus trees sampled from different locations of Dhorpatan, 13 distinctive endophytic fungi were isolated and grouped into 9 different genera: Bjerkandera, Trichoderma, Preussia, Botrytis, Arthrinium, Alternaria, Cladosporium, Sporormiella and Daldinia. The ethyl acetate extracts isolated from three endophytic fungi: Alternaria alternata, Cladosporium cladosporioides and Alternaria brassicae showed significant TPC values of 204±6.144, 312.3±2.147 and 152.7±4.958µg GAE/mg of dry extract, respectively, and TFC values of 177.9±2.911, 644.1±4.202 and 96.38±3.851µg RE/mg of dry extract, respectively. Furthermore, these three extracts showed a dose dependent radical scavenging activity with IC50 concentration of 22.85, 22.15 and 23.001 µg/ml, respectively. The extracts of C. cladosporioides and A. brassicae also showed promising antimicrobial activity against Escherichia coli, Staphylococcus aureus and Bacillus subtilis with a minimum inhibitory concentration of 250μg/ml for all bacteria. Both the samples showed cytotoxic property against shrimp nauplii with LC50 of 104.2 and 125.9µg/ml, respectively. Conclusions: The crude fungal extracts obtained from endophytes: A. alternata, C. cladosporioides and A. brassicae upon purification and further identification of the bioactive compounds can be a fascinating source for novel pharmaceutical agents.

F1000Research ◽  
2020 ◽  
Vol 9 ◽  
pp. 379
Author(s):  
Dhurva Prasad Gauchan ◽  
Pratistha Kandel ◽  
Astha Tuladhar ◽  
Ashesh Acharya ◽  
Upendra Kadel ◽  
...  

Background: Endophytic fungi are largely underexplored in the discovery of natural bioactive products though being rich sources of novel compounds with promising pharmaceutical potential. In this study, Taxus wallichiana, which has huge medicinal value, was investigated for its endophytic diversity and capability to produce bioactive secondary metabolites by analyzing antioxidant, antimicrobial and cytotoxic properties. Methods: The endophytes were identified by ITS-PCR using genomic DNA samples. The secondary metabolites were extracted by solvent extraction method using ethyl acetate. The antioxidant activity was analyzed by Thin Layer Chromatography, Total Phenol Content (TPC), Total Flavonoid Content (TFC) and DPPH assay, and the antimicrobial activity was analyzed by agar-well diffusion method. Brine shrimp lethality assay was used to analyze the cytotoxicity of the fungal extracts. Results: Out of 16 different Taxus trees sampled from different locations of Dhorpatan, 13 distinctive endophytic fungi were isolated and grouped into 9 different genera: Bjerkandera, Trichoderma, Preussia, Botrytis, Arthrinium, Alternaria, Cladosporium, Sporormiella and Daldinia. The ethyl acetate extracts isolated from three endophytic fungi: Alternaria alternata, Cladosporium cladosporioides and Alternaria brassicae showed significant TPC values of 204±6.144, 312.3±2.147 and 152.7±4.958µg GAE/mg of dry extract, respectively, and TFC values of 177.9±2.911, 644.1±4.202 and 96.38±3.851µg RE/mg of dry extract, respectively. Furthermore, these three extracts showed a dose dependent radical scavenging activity with IC50 concentration of 22.85, 22.15 and 23.001 µg/ml, respectively. The extracts of C. cladosporioides and A. brassicae also showed promising antimicrobial activity against Escherichia coli, Staphylococcus aureus and Bacillus subtilis with a minimum inhibitory concentration of 250μg/ml for all bacteria. Both the samples showed cytotoxic property against shrimp nauplii with LC50 of 104.2 and 125.9µg/ml, respectively. Conclusions: The crude fungal extracts obtained from endophytes: A. alternata, C. cladosporioides and A. brassicae upon purification and further identification of the bioactive compounds can be a fascinating source for novel pharmaceutical agents.


Author(s):  
Shirly Kumala ◽  
Ng Vini Aprilia ◽  
Partomuan Simanjuntak

Objective: Colletotrichium capsici endophytic fungi isolated from Jamblang plant (Eugenia cumini  L).  The secondary metabolites of this plant has the potent antibacterial efficacy  as well as diarrheal and anti-diabetic. This research focussed on isolation of the endophytic microbes from branches of Jamblang plants and their secondary metabolites.Methods: Isolation of endophytes were performed in PDA(potato dextrose agar) using direct seed plant. Endophytic fungi isolates with strongest antimicrobial activity against the bacteria Staphylococcus aureus, Escherichia coli were fermented in Potato Dextrose Yeast (PDY) to produce  large scale of the metabolites.  Supernatant was extracted with ethyl acetate solvent. Ethyl acetate extract fractionated by column chromatography (SiO3, n-hexane- ethyl acetate = 50:1 ~ 1:1) and obtained three fractions. Further, agar diffusion method was performed to assess their anti-microbial activity.Results:  Antibacterial  test  results  indicated that fraction III had  the antibacterial  activity Staphylococcus aureus with  inhibition zone diameter  of 10.7 mm but no observed antibacterial activity against Escherichia coli.  Furthermore, identification  by GC-MS showed that compounds present in fraction III was mainly fatty acid and phenolic compounds.Conclusion:  In conclusion, secondary metabolites isolated from Jamblang plants branches contained predominantly fatty acid and phenol related compounds that could be responsible for its potent anti microbial activity. Keywords : Endophytic fungi, Jamblang (Eugenia cumini L.), antimicrobial activity


2016 ◽  
Vol 6 (01) ◽  
pp. 1494 ◽  
Author(s):  
Ananda Danagoudar ◽  
Chandrashekhar G. Joshi* ◽  
M.T. Nivya ◽  
H.M. Manjunath ◽  
Jagadeesha Poyya ◽  
...  

Endophytic fungi are one of the untapped resources of therapeutic compounds for various diseases. The present study focused on the antimicrobial as well as larvicidal activity of ethyl acetate extract of endophytic fungi isolated from Tragia involucrata. The ethyl acetate extract of Penicillium citrinum CGJ-C1 (GenBank No.KT780618), Penicillium citrinum CGJ-C2 (KP739821), Cladosporium sp. (KP739822), and Cryptendoxyla hypophloia CGJ-D2 (KT780619) was subjected to antimicrobial activity against a panel of microorganisms by disc diffusion method, larvicidal activity against Culex quinquefasciatus. All the extracts showed significant antimicrobial activity against the tested organisms ranging from 8±0.32 to 13±2.11. The extent of activity was comparable to the standard drugs. The larvicidal potential of the endophytes was superior to T.involucrata extract. The larvicidal activity was found to be dose and time dependent with LC50 value ranging from 4.25- 158.06ppm after 24hrs of treatment. This is the first report on the bioactivity of the endophytes isolated from T.involucrata. Further studies on the bio-guided isolation of lead compound will benefit the people suffering from microbe’s related diseases.


2014 ◽  
Vol 5 (2) ◽  
Author(s):  
Widya Lestari

Endophytic fungi are a group of fungi whose part or whole life is in living plant tissue and usually does not harm the host. Endophytic fungi generally produce secondary metabolites that have beneficial biological activities such as anti-cancer compounds, antifungi or antibacterials. Control of plant diseases caused by Rigidoporus microporus and Ganoderma boninense can be done by using biological agents, namely by using endophytic fungi. This fungus has metabolite compounds that can inhibit the growth of Rigidoporus microporus and Ganoderma boninense. This study aims to determine what content is found in endophytic mushroom extracts isolated from rubber plants (Hevea brasiliensis). Endophytic fungi are extracted by maceration using methanol and ethyl acetate solvents. The extract was tested for antifungal activity using diffusion method. The content of secondary metabolites was tested by phytochemical test. The results of endophytic fungal extraction with methanol and ethyl acetate solvents resulted in antifungal activity with inhibitory zones in a row namely WL01: 2.19 and 2.15 mm Fungi Isolates on Ganoderma boninense while 3.09 mm WL02 against Rigidoporus microporus Based on the phytochemical test results, endophytic fungal extract with methanol and ethyl acetate solvents showed the presence of alkaloid, flavonoid, phenol and saponin. Keywords: endofit, fungi, hevea, secondary metabolites


2019 ◽  
Vol 3 (1) ◽  
pp. 10-18 ◽  
Author(s):  
Peter M. Eze ◽  
Joy C. Nnanna ◽  
Ugochukwu Okezie ◽  
Happiness S. Buzugbe ◽  
Chika C. Abba ◽  
...  

AbstractEndophytic fungi associated with Nigerian plants have recently generated significant interest in drug discovery programmes due to their immense potential to contribute to the discovery of new bioactive compounds. This study was carried out to investigate the secondary metabolites of endophytic fungi isolated from leaves ofNewbouldia laevis, Ocimum gratissimum, andCarica papayaThe plants were collected from Agulu, Anambra State, South-East Nigeria. Endophytic fungal isolation, fungal fermentation; and extraction of secondary metabolites were carried out using standard methods. The crude extracts were screened for antimicrobial activities using the agar well diffusion method, and were also subjected to high performance liquid chromatography (HPLC) analysis to identify their constituents. A total of five endophytic fungi was isolated, two fromN. laevis(NL-L1 and NL-L2), one fromO. gratissimum(SL-L1), and two fromC. papaya(PPL-LAC and PPL-LE2). In the antimicrobial assay, the extracts of NL-L2, SL-L1, and PPL-LE2 displayed mild antibacterial activity against both Gram negative and Gram positive test bacteria. PPL-LAC extract showed mild activity only againstS. aureus, while no antimicrobial activity was recorded for NL-L1 extract. All the endophytic fungal extracts showed no activity against the test fungiC. albicansandA. fumigatusHPLC analysis of the fungal extracts revealed the presence of ethyl 4-hydroxyphenyl acetate and ferulic acid in NL-L1; ruspolinone in NL-L2; protocatechuic acid, scytalone, and cladosporin in SL-L1; indole-3-acetic acid and indole-3-carbaldehyde in PPL-LE2; and indole-3-acetic acid in PPL-LAC. The findings of this study revealed the potentials possessed by these plants as source of endophytes that express biological active compounds. These endophytes hold key of possibilities to the discovery of novel molecules for pharmaceutical, agricultural and industrial applications.


2016 ◽  
Vol 2016 ◽  
pp. 1-8 ◽  
Author(s):  
José Henrique Faleiro ◽  
Randys Caldeira Gonçalves ◽  
Mara Núbia Guimarães dos Santos ◽  
Diego Pereira da Silva ◽  
Plínio Lázaro Faleiro Naves ◽  
...  

It is known that the phytochemical identification and assessment of biological effects caused by the constituent species Psidium cattleianum, which belongs to family Myrtaceae, are poorly held in the literature. The aim of the current study is to investigate the composition of secondary metabolites, the toxicity, and the antimicrobial activity of P. cattleianum leaves. The crude ethanolic extract of the plant was obtained through maceration and fractionated with hexane, dichloromethane, and ethyl acetate. The crude ethanol extract and the fractions were subjected to phytochemical screening and tested against the microcrustacean Artemia salina for toxicological assessment. Antimicrobial tests with crude ethanol extract and the fractions were carried out through the agar diffusion method using broth microdilution against Staphylococcus aureus, S. epidermidis, Burkholderia cepacia, and Escherichia coli strains. A variety of secondary metabolite groups such as catechins, steroids, phenolic compounds, flavonoids, and saponins was detected. Regarding toxicity, hexane and dichloromethane fractions were considered nontoxic, whereas the crude ethanol extract and the ethyl acetate fraction showed low toxicity. The crude ethanol extract and the fractions, except for the hexane fraction, showed activity against the tested strains. Therefore, the composition of the secondary metabolites, the low toxicity, and the antimicrobial activity suggest that this species is promising in the search and development of new drugs.


Author(s):  
Susan Kurian ◽  
V. S. Josekumar

<p><strong>Objective: </strong>To evaluate the antimicrobial activity and cytotoxicity of hexane, ethyl acetate and methanol extracts of <em>Alysicarpus vaginalis </em>var<em>. nummularifolius </em>(DC) along with phytochemical analysis.</p><p><strong>Methods: </strong>The crude extract of hexane (AVH), ethyl acetate (AVE) and methanol (AVM) of <em>Alysicarpus vaginalis </em>var<em>. nummularifolius </em>(DC) were prepared and analysed for phytochemical constituents using standard methods. The cytotoxicity activity of the plant extracts was predicted using brine shrimp lethality assay (BSLA). The antimicrobial activity and the minimal inhibitory concentration (MIC) of the plant extracts were examined against 5 bacterial and 2 fungal strains using agar well diffusion method, and two fold serial dilution method, respectively.</p><p><strong>Results: </strong>The phytochemical screening studies showed a higher concentration of saponins, alkaloids, flavonoids, phenols, quinones and terpenoids in AVM than the other two extracts. The LC<sub>50</sub> value of AVH and AVE were found to be 900.05 µg/ml and 754.35 µg/ml respectively using BSLA while that of AVM was&gt;1000 ug/ml. All the extracts of the plant showed antimicrobial activity against most of the test organisms. The MIC values of AVM were lower than AVE for all the microbial strains except for <em>Pseudomonas aeroginosa</em> where AVE (107.87µg/ml) exhibited higher value than AVM (51µg/ml).</p><p><strong>Conclusion: </strong>The present study concluded AVM with a high presence of phytochemicals. The AVE and AVM were found to possess promising antimicrobial activity when compared with the standards. The AVM exhibited lesser toxicity when compared with AVH and AVE.<strong></strong></p><p> </p>


2021 ◽  
Vol 22 (10) ◽  
Author(s):  
Syarifah Syarifah ◽  
Elfita Elfita ◽  
HARY WIDJAJANTI ◽  
ARUM SETIAWAN ◽  
ALFIA R. KURNIAWATI

Abstract. Syarifah, Elfita, Widjajanti H, Setiawan A, Kurniawati AR. 2021. Diversity of endophytic fungi from the root bark of Syzygium zeylanicum, and the antibacterial activity of fungal extracts, and secondary metabolite. Biodiversitas 22: 4572-4582. The decoction of the root bark of Syzygium zeylanicum has been used as traditional medicine, such as for treating pathogenic bacterial infections. Endophytic fungi that live in medicinal plant tissues have a high species diversity and biological activities correlate with their host. Therefore, this study aimed to explore the diversity of endophytic fungi from the root bark of S. zeylanicum and to determine the antibacterial activity of endophytic fungi and their secondary metabolites. In this study, we isolate and identify the endophytic fungi from the root bark of S. zeylanicum, continued by screening their antibacterial activity against two Gram-negative bacteria (Escherichia coli InaCCB5 and Salmonella thypi ATCC1048 and two Gram-positive bacteria (Staphylococcus aureus InaCCB4 and Bacillus subtilis InaCCB1204) by the Kirby-Bauer method. The fungal extract with the highest antibacterial activity proceeded with the isolation and determination of the structure of their bioactive compounds. The isolates were morphologically identified. Isolates that showed strong antibacterial activity were identified by molecular identification. Isolation of bioactive compounds was carried out by chromatographic techniques and the determination of the structure of pure chemical compounds was performed by the spectroscopic analysis. In total, there were 8 isolates of endophytic fungi were obtained from the root bark of S. zeylanicum, namely SZR1 – SZR8. SZR2 isolate has the highest antibacterial activity. Molecular identification through phylogenetic analysis showed that SZR2 isolate had high similarity with Penicillium brefeldianum. Isolation of bioactive compounds from SZR2 produced compound 1 in the form of light yellow crystals which showed strong antibacterial activity against S. typhi, E. coli, and B. subtilis with MIC values of 64 g/mL. Compound 1 was identified as p-hydroxybenzaldehyde, which was also obtained in its host. In conclusion, the endophytic fungus Penicillium brefeldianum produces similar secondary metabolites and antibacterial activity as its host plant.


2020 ◽  
Vol 36 (2) ◽  
Author(s):  
Ana Flávia da Silva ◽  
Marisa de Oliveira Lopes ◽  
Cláudio Daniel Cerdeira ◽  
Ingridy Simone Ribeiro ◽  
Isael Aparecido Rosa ◽  
...  

The radish (Raphanus sativus L.) is a vegetable of the Brassicaceae family cultivated worldwide and has several medicinal properties. Its biological activities are related to various secondary metabolites present in the species, especially phenolics. Thus, the objectives of this study were the chemical analysis and evaluation of the antioxidant and antimicrobial activities of the dry extract and fractions of the fodder turnip leaves (R. sativus var. oleiferus Metzg.). Samples were analyzed by mass spectrometry and the antioxidant activity was evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical method and the reducing power method. Antimicrobial activity was determined by the agar diffusion and microdilution methods. The total phenols were concentrated in the butanol fraction (121.27 mg GAE/g) and the flavonoids were concentrated in the ethyl acetate fraction (98.02 mg EQ/g). The ethyl acetate fraction showed the best antioxidants results, with 83.45% of free radical scavenging and 11.34% of ferric ions reduction. The analysis of antimicrobial activity showed that the dry extract had the highest average zone of inhibition against Bacillus subtilis (18.67 mm). Smaller values of the minimum inhibitory concentration for Micrococcus luteus were, and the ethyl acetate fraction showed a lower minimum inhibitory concentration (0.1 mg/ml) for that microorganism. There was a strong correlation between the antioxidant activity and the content of phenols and flavonoids. The results showed the potential antioxidant and antimicrobial activities of this extract with the ethyl acetate fraction being most promising for further studies.


INDIAN DRUGS ◽  
2015 ◽  
Vol 52 (04) ◽  
pp. 37-41
Author(s):  
S. M Pawaskar ◽  
◽  
K. C. Sasangan

The present study was undertaken to evaluate in vitro antimicrobial activity of the successive leaf extracts of Cynodon dactylon in petroleum ether, ethyl acetate, acetone, ethanol, methanol and water, against various gram positive & gram negative bacterial strains using zone of inhibition. Both Agar well diffusion method & Agar disc diffusion method were used to evaluate the antibacterial efficacy. The microorganisms used in the test were – Escherichia coli, Proteus vulgaris, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Shigella flexneri, Salmonella typhi, Salmonella paratyphi A, Salmonella paratyphi B, Bacillus subtilis, Streptococcus pyogenes, Vibrio cholerac and Enterobacter aerogenes. Two fungal strains - Candida albicans and Saccharomyces cerevisiae were also tested to evaluate the antifungal potential of the said plant extract. The reference antibiotics chloramphenicol & ampicillin (antibacterial); nystatin & clotrimazole and (antifungal) were also tested against these standard microorganisms used in the assay and the results were compared with that of the plant extracts.The results of the study revealed that all the seven successive extracts of the leaf powder of Cynodon dactylon ( L.) Pers. exhibited prominent antimicrobial and antifungal activity against all microorganisms used in the study. The nonpolar extracts i.e. petroleum ether, ethyl acetate and acetone showed the most significant antibacterial and antifungal activity against all tested organisms. The petroleum ether and ethyl acetate extracts showing maximum inhibition in the range of 8 mm - 15 mm. This was closely followed by, acetone extract, which showed the inhibition in the range of 9 mm - 13 mm.


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