IDENTIFICATION AND CHARACTERIZATION OF POTENT ANTIMICROBIAL SECONDARY METABOLITE GENERATED FROM ENDOPHYTIC FUNGI ISOLATED FROM JAMBLANG PLANT ( Eugenia cumini L)

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

Jurnal MIPA ◽  
2019 ◽  
Vol 8 (2) ◽  
pp. 42
Author(s):  
Florentin Natalia Melsadalam ◽  
Dewa Gede Katja ◽  
Meiske S. Sangi

Telah dilakukan penelitian yang bertujuan untuk mengidentifikasi senyawa metabolit sekunder  dari ekstrak daun kaf menggunakan tiga jenis pelarut yakni n-heksan, etil asetat, dan metanol. Ekstrak yang diperoleh diuji aktivitas antibakterinya dengan metode difusi yang menggunakan bakteri Staphylococcusaureus dan Escherichia coli. Dari hasil pengujian fitokimia, ketiga ekstrak tersebut menunjukan bahwa sampel mengandung senyawa; alkaloid, saponin, steroid, flavonoid dan tanin. Pengujian aktivitas antibakteri  dengan metode difusi menunjukan hasil yang paling tinggi aktivitas antibakteri adalah  estrak etil asetat dengan zona hambat 35,75 mm terhadap bakteri  Staphylococcus aureus, sedangkan pada ekstrak metanol memiliki zona hambat 23,75 mm terhadap bakteri Escherichia coliResearch has been carried out aimed at identifying secondary metabolites of kaf leaf extract using three types of solvents, namely n-hexane, ethyl acetate, and methanol.The obtained extracts were tested for antibacterial activity by diffusion method using Staphylococcus aureus and Escherichia coli bacteria. From the results of phytochemical testing, the three extracts showed that the sample contained compounds; alkaloids, saponins, steroids, flavonoids and tannins. Antibacterial activity testing with diffusion method showed the highest antibacterial activity was estracethyl ethyl acetate with 35.75 mm inhibition zone against Staphylococcus aureus bacteria, whereas methanol had 23.75 mm inhibitory zone against Escherichia coli bacteria


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.


PHARMACON ◽  
2019 ◽  
Vol 8 (3) ◽  
pp. 525
Author(s):  
Ayu Natasya Paputungan ◽  
Widya Astuty Lolo ◽  
Imam Jayanto

Mangosteen leaves have flavonoid compounds, tannins, and saponins that can be efficacious as antibacterial. The aim of this study was to determine the fraction of mangosteen leaves having an antibacterial effect and knowing the class of compounds identified as having antibacterial activity after TLC- Bioautography testing was carried out. The samples were extracted using 96% maceratarion method and fractioned using liquid-liquid fractionation method with methanol, n-hexane and ethyl acetate solvents, antibacterial activity using agar diffusion method (Kirby and Bauer) with 3 concetrations namely 10%, 20% and 30%. Thin Layer Chromatography (TLC) uses n-hexane and chloroform solvens. TLC-Bioautography uses contact bioautography methods. The resultd showed that mangosteen leaves in methanol fraction with a concentration of 30% had a very large inhibitory activity again Staphylococcus aureus and ethyl acetate fraction with a concentration of 30% had the gratest antibacterial activity against  Escherichia coli. The results of the TLC- Bioautography study showed that the flavonoids compounds after spraying with AlCl3 and the mangosteen leaf Biosutography test had inhibitory zone activity against the bacteria Staphylococcus aureus dan Escherichia coli. Keywords: Mangosteen Leaves. Antibacterial, TLC Bioautography.  ABSTRAK Daun manggis mempunyai senyawa flavonoid, tanin, dan saponin yang dapat berkhasiat sebagai antibakteri. Penelitian ini bertujuan untuk mengetahui fraksi daun manggis memiliki efek antibakteri dan mengetahui golongan senyawa yang teridentifikasi memiliki aktivitas antibakteri setelah dilakukan pengujian KLT Bioautografi. Sampel diektrak dengan metode maserasi dengan pelarut 96% dan difraksinasi dengan metode  fraksinasi cair-cair dengan pelarut metanol, n-heksan dan etil asetat, aktivitas antibakteri menggunakan metode difusi agar (Kirby and Bauer) dengan 3 kosentrasi yaitu 10%, 20% dan 30%. Kromatografi Lapis Tipis (KLT) menggunakan pelarut n-heksan dan klorofom. KLT-Bioautografi menggunakan metode bioautografi kontak. Hasil penelitian menunjukan daun manggis pada fraksi metanol  dengan kosentrasi 30% memiliki aktivitas zona hambat ppaling besar terhadapat bakteri Staphylococcus aureus dan fraksi etil asetat dengan kosentrasi 30% memiliki aktivitas antibakteri paling besar terhadap  Escherichia coli. Hasil penelitian KLT-Bioautografi menunjukan golongan senyawa flavonoid setelah disemprotkan dengan AlCl3 dan uji Bioautografi daun manggis memiliki aktivitas zona hambat terhadap bakteri Staphylococcus aureus dan Escherichia coli. Kata kunci : Daun Manggis, Antibakteri, KLT- Bioautografi.


2018 ◽  
Vol 1 (1) ◽  
Author(s):  
Vera Nurviana

<p class="Default"><em>Limus (Mangifera foetida Lour.) Is one of the local Indonesian fruits of the anacardiaceae family. Limus contain secondary metabolites that have pharmacological effects, including the kernel waste of the seeds of the fruit. This study aims to determine the antibacterial activity against Staphylococcus aureus and Escherichia coli on the fraction of kernel seed of limus fruits ethanolic extract in invitro using solid diffusion method with positive control Tetracycline HCl. The extraction process was carried out by maceration method, then fractionation was obtained to obtain the fraction of n-hexane, ethyl acetate fraction and water fraction. Based on the result of the research, the fraction of n-hexane, ethyl acetate fraction, and water fraction of kernel ethanol extract limus fruit seed have antibacterial activity against Staphylococcus aureus and Escherichia coli. The highest activity was shown by fraction of ethyl acetate with minimum killing concentration (KBM) to Staphylococcus aureus was 5% with diameter equal to 4,33 ± 1.17 mm equal to 30,27 μg / mL Tetracycline HCl, and KBM value to Escherichia coli.4% (7.60 ± 0.14) mm equivalent to 26, 04 μg / mL Tetracycline HCl.<br /> <br /> <strong></strong></em></p><p class="Default"><strong><em>Keywords: </em></strong><em>Antibacterial, Staphylococcus aureus, Escherichia coli, Kernel, M. foetida.</em></p>


Author(s):  
ZAMHARIRA MUSLIM ◽  
YONANIKO DEPHINTO

Objective: This research aims to analyze the ability of robusta coffee leaves fraction extract to inhibit the growth of Staphylococcus aureus and Escherichia coli and also determine the minimum inhibitory concentration (MIC). Methods: Antibacterial activity evaluated by the disc diffusion method observed in four types of fraction of extract robusta coffee leaves (n-hexane, ethyl acetate, ethanol, and water). Each extract divided into three various concentrations, 5%, 10%, and 15%. Determination of antimicrobial activity in vitro by the disk diffusion method. Results: Ethyl acetate fraction of coffee leaves extract produced the largest diameter zone of inhibition of bacterial growth compared to other extraction fractions of 17.28 mm in E. coli and 18.58 mm in S. aureus. The MIC of coffee leaves extract fraction water, ethyl acetate, and n-hexane on E. coli and S. aureus is 5%, while the fraction ethanol MIC is 10%. Conclusion: The antibacterial effect of ethyl acetate fraction of coffee leaves extract showed an antibacterial effect that was better than the fraction of n-hexane, ethanol, and water.


2019 ◽  
Vol 18 (2) ◽  
pp. 223-232
Author(s):  
AO Shittu ◽  
A Aliyu ◽  
MS David ◽  
NS Njinga ◽  
HI Ishaq

In order to authenticate and ascertain the various claim by the rural dwellers that depend on chewing sticks for their oral hygiene and protection against innumerable diseases, the antibacterial activity of the roots of Fagara zanthoxyloides and Distemonanthus benthamianus were evaluated. Preliminary phytochemical screening was carried out using standard methods. In-vitro antibacterial activity of the methanol extracts of both plants was also carried out using the agar well diffusion method against standard strains of Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922, Salmonella typhii ATCC 14028, Citrobacter freundii ATCC 8090 and also some clinical isolates of Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis at different concentrations and ratios. Preliminary phytochemical screening showed that the extracts contained alkaloids, saponin, tannins, steroids, flavonoids, anthraquinones, cardiac glycosides and phenol. All the test microorganisms were susceptible to the inhibitory effect of the extracts at concentrations of 200 mg/ml and 500 mg/ml. F. zanthoxyloides had better antimicrobial activity with zones of inhibition ranging from 21.0 to 26.0 mm at 200 mg/ml and 24.3 to 29.3 mm at 500 mg/ml while D. benthamianus had zones of inhibition ranging from 17.7 to 26.7 mm at 200 mg/mL and 19.0 to 26.7 mm at 500 mg/mL. Some of the test microorganisms were resistant to the standard antibiotics (cefuroxime, ciprofloxacin, ofloxacin and gentamicin). Combinations of the extracts in ratios 50:50 and 75:25 yielded no increase in activity. The MIC and MBC for D. benthamianus ranged from 6.25 mg/ml to 100 mg/mL and 25 to 200 mg/ml, respectively while that of F. zanthoxyloides ranged from 1.56 mg/ml to 12.5 mg/mL and 50 to 200 mg/ml, respectively. The antimicrobial activity demonstrated by F. zanthoxyloides and D. benthamianus indicates that they would be valuable in the management of urinary, respiratory and gastrointestinal tracts infections while at the same time helping to mitigate the problem of antimicrobial resistance. Phenolic compounds of plants, among them flavonoids are the chief constituents, which have potent antioxidant activities Dhaka Univ. J. Pharm. Sci. 18(2): 223-232, 2019 (December)


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.


KOVALEN ◽  
2019 ◽  
Vol 5 (3) ◽  
pp. 233-239
Author(s):  
Ni Luh Sumitriasih ◽  
Ahmad Ridhay ◽  
Indriani

A study was conducted on the antibacterial activity test on n-hexane, ethyl acetate, and ethanol extracts of ebony bark (Diospyros celebica Bakh.) using a diffusion method. The purpose of this research is to know the antibacterial activity of ebony bark against Staphylococcus aureus and Escherichia coli bacteria using n-hexane, ethyl acetate dan ethanol. The method of extraction used in this research was maceration method with three polarity levels of solvent. The antibacterial activity test was performed by diffusion-well test. The results showed that ethyl acetate extract gave the highest inhibition of 26.14 mm in gram-positive bacteria (Staphylococcus aureus) and ethanol extract of 25.97 mm in gram-negative bacteria (Escherichia coli). Keywords: Ebony (Diospyros celebica. Bakh), Tree Bark, Inhibitory, Antibacterial


Author(s):  
Melisa Selly Liswandari

UJI AKTIVITAS ANTIBAKTERI ALGA HIJAU (Ulva sp.) DARI PANTAI SORIDO BIAK TERHADAP BAKTERI Escherichia coli  DAN Staphylococcus aureus Melisa Selly Liswandari1), Daniel Lantang2), Septriyanto Dirgantara1)1)Program Studi Farmasi, Fakultas MIPA Universitas Cenderawasih, Jayapura2)Jurusan Biologi, Fakultas MIPA Universitas Cenderawasih, Jayapura, Kampus UNCEN Waena. Jl Perumnas III Waena Jayapura 99351 PapuaEmail*: [email protected]  ABSTRACT The green algae (Ulva sp.) are potentially as antibacterial agent because it contains triterpenoids, flavonoids, and saponins. The objective of this research are to know antibacterial activity of green algae (Ulva sp.) extracts using different levels of solvent polarity and the most effective concentration inhibits the growth of bacterial Escherichia coli and Staphylococcus aureus. Green algae (Ulva sp.) extracted by stratified soxletation method using solvent, eq diethyl ether, ethyl acetate, and ethanol 96%. Test antibacterial activity using the disc diffusion method. This research using the of CRD (Completely Randomize Design) with 7 treatments was concentration of 100 ppm, 250 ppm, 500 ppm, 750 ppm, 1000 ppm, positive control (ciprofloxacin 5µg) and negative control (aquades steril) with 3 repetitions. The data was analyzed using ANOVA and continued by HSD (Honestly Significant Difference) test with 95% confidence level. The result was demonstrated that extract green algae (Ulva sp.) the most effective to inhibits the growth of bacterial Escherichia coli is ethyl acetate extract with concentration 250 ppm (9,26 mm) and bacterial Staphylococcus aureus is ethanol 96% extract with concentration 750 ppm (9,57 mm). Keywords : Ulva sp., antibacterial agent, Escherichia coli, Staphylococcus aureus. ABSTRAK             Alga hijau (Ulva sp.) berpotensi sebagai antibakteri karena mengandung triterpenoid, flavonoid, dan saponin. Tujuan dari penelitian ini adalah untuk mengetahui aktivitas antibakteri ekstrak alga hijau (Ulva sp.) yang menggunakan tingkat kepolaran pelarut berbeda dan konsentrasi yang paling efektif dalam menghambat pertumbuhan bakteri Escherichia coli dan Staphylococcus aureus. Alga hijau (Ulva sp.) diekstraksi dengan metode sokletasi bertingkat menggunakan pelarut dietil eter, etil asetat, dan etanol 96%. Uji aktivitas antibakteri menggunakan metode difusi cakram. Penelitian ini menggunakan RAL dengan 7 perlakuan yaitu konsentrasi 100 ppm, 250 ppm, 500 ppm, 750 ppm, 1000 ppm, kontrol positif (ciprofloxasin 5µg) dan kontrol negatif (akuades steril) dengan 3 pengulangan. Data analisis menggunakan ANOVA dan dilakukan uji lanjut BNJ dengan tingkat kepercayaan 95%. Hasil penelitian menunjukkan bahwa ekstrak alga hijau (Ulva sp.) yang paling efektif untuk menghambat pertumbuhan bakteri Escherichia coli adalah ekstrak etil asetat dengan konsentrasi 250 ppm (9,26 mm) dan bakteri Staphylococcus aureus adalah ekstrak etanol 96% dengan konsentrasi 750 ppm (9,57 mm). Kata kunci : Ulva sp., antibakteri, Escherichia coli, Staphylococcus aureus. 


2010 ◽  
Vol 53 (3) ◽  
pp. 629-632 ◽  
Author(s):  
Henrique Pereira Ramos ◽  
Glaucia Hollander Braun ◽  
Mônica Tallarico Pupo ◽  
Suraia Said

Papulaspora immersa and Arthrinium state of Apiospora montagnei Sacc. were isolated from the roots of Smallanthus sonchifolius (yacón). The crude extracts from their cultures inhibited the growth of Staphylococcus aureus, Kocuria rhizophila, Pseudomonas aeruginosa and Escherichia coli. The more relevant results were observed in the ethyl acetate extract from P immersa against P aeruginosa (90 µg/mL) and ethyl acetate extract from Arthrinium state of A montagnei Sacc. against P aeruginosa (160 µg/mL). The two endophytic fungi isolated from yacón roots as well as their antimicrobial activity detected in the crude extracts cultures were being reported for the first time.


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