scholarly journals In vitro Inhibitory Activity of Ethyl Acetate Extract of Symbiotic Bacteria Isolated from the Marine Sponge Haliclona fascigera against Multidrug Resistant Organism (MDRO)

Author(s):  
Dian Handayani ◽  
Murniati Murniati ◽  
Rustini Rustini
Author(s):  
Sneha Arora ◽  
Shoma Paul Nandi

  Objective: Screening of medicinal plants for the presence of beta-lactamase inhibitor identified three plants; Terminalia chebula, Terminalia bellirica, and Ocimum tenuiflorum, extracts of which inhibit beta-lactamase enzyme in vitro. The objective of this study was to evaluate and compare beta-lactamase inhibiting potential of these plant extracts.Methods: Extracts of these plants were prepared with 6 solvents of different polarity. Beta-lactamase inhibition study was performed using antibiotic-resistant bacteria in bioassay and by micro-iodometric assay. Multidrug-resistant clinical strains of Escherichia coli and laboratory strain with plasmid carrying beta-lactamase gene as positive control were used.Results: Our results from bioassay, as well as micro-iodometric assay for enzyme activity, confirmed the presence of beta-lactamase inhibitor in these plant extracts. Among the extracts made by different solvents, hexane and ethyl acetate extract of T. chebula, hexane extract of T. bellirica, and all extracts of O. tenuiflorum except dichloromethane, possessed beta-lactamase inhibitor. Multidrug-resistant clinical isolate of E. coli AIIMS-1 could be reverted by applying 50 μg/μl of extract of all the medicinal plants. The micro-iodometric result showed highest beta-lactamase inhibition with O. tenuiflorum extracts. Comparative evaluation of the O. tenuiflorum extracts with increasing concentration of inhibitor suggests that ethyl acetate extract of O. tenuiflorum contains the highest inhibition potential, which is comparable with clavulanic acid.Conclusion: The results demonstrated that the ethyl acetate extract of O. tenuiflorum contain the highest level of beta-lactamase inhibitor, which in the future can be used as an alternative to synthetic beta-lactamase inhibitors that are presently being used to control beta-lactam antibiotic resistance


Author(s):  
Bina Lohita Sari ◽  
Abdul Mun’im ◽  
Arry Yanuar ◽  
Rezi Riadhi

<p><strong>Objective: </strong><em>Terminalia catappa</em> L. (<em>T. catappa</em> L.) fruit has inhibitory activity on α-glucosidase, therefore, can be a potential natural source for the treatment of type II diabetes mellitus. Inhibitory activity of ethanol fruit extract with IC<sub>50</sub> 3.02 µg/ml was the strongest inhibition when compared with 54 medicinal plants used as an antidiabetic agent in Indonesia. This project was aimed to find the active compound from <em>T</em><em>.</em><em> catappa</em> L. fruit using molecular docking, identification ethyl acetate subfraction using TLC and GC-MC, determine <em>in vitro</em> test on α-glucosidase inhibitory activity from ethyl acetate extract and subfraction.</p><p><strong>Methods: </strong>Molecular docking using AutoDock 4.2 was performed to predict the binding modes of<strong> </strong>α-glucosidase enzyme from <em>Saccharomyces cereviciae</em> with 13 chemical constituents of <em>T. catappa</em>. α-Glucosidase enzyme was obtained from Protein Data Bank (PDB code: 3A4A). Acarbose, voglibose and miglitol were used as standards. Docking result determines the highest binding energy (ΔG) and inhibition constants (Ki) as an active compound. Visualization of amino acid residues around the active compound was identified with PyMOL and LigPlot. Screening of active compound was carried out by <em>T</em><em>.</em><em> catappa</em> L. fruit remaceration extraction use hexane and ethyl acetate. Ethyl acetate extract was separated on silica gel column chromatography using n-hexane, ethyl acetate and methanol sequentially based on polarity of each solvent. Identification of an active compound from ethyl acetate sub fractions using TLC and GC-MS method. The inhibitory activity of the active compound of α-glucosidase was determined with <em>in vitro</em> test using α-glucosidase enzyme.</p><p><strong>Results: </strong>The highest binding energy and inhibition constant is β–sitosterol with ΔG-10.61 kcal/mol and Ki 0.02 µM. The ligand was situated around of 18 amino acid residues. Ethyl acetate subfractions A, B and C showed that subfraction B contains similar spot characteristic and Rf value (0.42) with β-Sitosterol standard. Identification with GC-MS gave β–sitosterol acetate and sitostenone. Redocking process of β–sitosterol acetate and sitostenone showed ΔG-11.14 kcal/mol and-9.79 kcal/mol with Ki 0.01 μM and 0.07 μM respectively. <em>In vitro</em> test of acarbose, ethyl acetate extract and subfraction B gave IC<sub>50</sub> 17.52; 192.51 and 296.28 µg/ml.</p><p><strong>Conclusion: </strong>Three steroids that are β-sitosterol, β-sitosterol acetate and sitostenone were<strong> t</strong>he active compounds responsible for α-glucosidase inhibitory activity of <em>T</em><em>.</em><em> catappa </em>L. fruit. According to the <em>in vitro</em> test, ethyl acetate extract has stronger α-glucosidase inhibitory activity than ethyl acetate subfraction B.</p>


Author(s):  
Shubhaisi Das ◽  
Sunanda Burman ◽  
Goutam Chandra

Background: The only remedy for up surging problem of antibiotic resistance is the discovery of antibacterial agents of natural origin. Objective: The present study was aimed at finding antibacterial potential of crude and solvent extracts of mature leaves of Plumeria pudica. Methods: Antibacterial activity of three different solvent extracts were evaluated in four human and four fish pathogenic bacteria by measuring the zone of inhibition and determining Minimum Inhibitory Concentration and Minimum Bactericidal Concentration values. Standard antibiotics were used as positive control. Preliminary phytochemical screening of most effective extract i.e., ethyl acetate extract, Fourier Transform Infra Red analysis and GC-MS analysis of the Thin Layer Chromatographic (TLC) fraction of ethyl acetate extract were done meticulously. All experiments were done thrice and analyzed statistically. Results: Crude leaf extracts and solvent extracts caused good inhibition of bacterial growth in all selected bacteria. Ethyl acetate extract showed highest inhibition zones in all tested strains with maximum inhibition (19.50±0.29 mm) in Escherichia coli (MTCC 739). MBC/MIC of the extracts indicated that all three solvent extracts were bactericidal. Preliminary phytochemical tests revealed the presence of tannins, steroids and alkaloids and FT-IR analysis revealed presence of many functional groups namely alcoholic, amide, amine salt and aldehyde groups. From the GC-MS analysis of TLC fraction of ethyl acetate extract five different bioactive compounds e.g., 2,4-ditert –butylphenyl 5-hydroxypentanoate, Oxalic acid; allyl nonyl ester, 7,9-Ditert-butyl-1-oxaspiro(4,5)deca-6,9-diene-2,8-dione, Dibutyl phthalate and 2,3,5,8-tetramethyl-decane were identified. Conclusion: Leaf extracts of P. pudica contain bioactive compounds that can be used as broad spectrum bactericidal agent.


2018 ◽  
Vol 34 (11) ◽  
pp. 1586-1590 ◽  
Author(s):  
Wassila Benabderrahmane ◽  
Amel Amrani ◽  
Ouahiba Benaissa ◽  
Marta Lores ◽  
J. Pablo Lamas ◽  
...  

Author(s):  
Abdul Mun'im ◽  
Muhammad Ashar Munadhil ◽  
Nuraini Puspitasari ◽  
Azminah . ◽  
Arry Yanuar

ABSTRACTObjectives: To evaluate the angiotensin converting enzyme (ACE) inhibitory activity of melinjo (Gnetum gnemon) seed extract and to study moleculardocking of stilbene contained in melinjo seeds.Methods: Melinjo seed powders were extracted with n-hexane, dichloromethane, ethyl acetate, methanol, and water successively. The extracts wereevaluated ACE inhibitory activities using ACE kit-Wist and the phenolic content using Folin–Ciocalteu method. The extract demonstrated the highestACE inhibitory activity was subjected to liquid chromatography-mass spectrometry (LC-MS) to know its stilbene constituent. The stilbene constituentsin melinjo seed were performed molecular docking using AutoDock Vina, and ligand-receptor Interactions were processed using Ligand Scout.Results: The ethyl acetate extract demonstrated the highest ACE inhibition activity with inhibitory concentration 50% value of 9.77 × 10−8 μg/mLand the highest total phenolic content (575.9 mg gallic acid equivalent/g). Ultra-performance LC-MS analysis of ethyl acetate extract has detected theexistency of resveratrol, gnetin C, ε-viniferin, and gnemonoside A/B. These compounds displayed similar physiochemical properties to lisinopril (ACEinhibitor), as in silico molecular docking studies demonstrated that they fit into the lisinopril receptors.Conclusion: In vitro analysis ethyl acetate extract from melinjo seeds demonstrated the highest ACE inhibitory activity. Molecular docking analysisindicated that resveratrol dimers, gnetin C and gnemonoside A can be considered ACE inhibitor.Keywords: Angiotensin converting enzyme inhibitor, Gnetum gnemon, Melinjo, Total phenolic, Antihypertension, Molecular docking.


2018 ◽  
Vol 18 (3) ◽  
pp. 753-767 ◽  
Author(s):  
Amina Boussaada ◽  
Rabah Arhab ◽  
Serena Calabrò ◽  
Raffaella Grazioli ◽  
Maria Ferrara ◽  
...  

Abstract The aim of the research was to evaluate the effect of three Eucalyptus globulus extracts rich in phenolic compounds, especially flavonoids, on rumen fermentation, methane (CH4) production, organic matter degradability and protozoa population using an in vitro gas production technique. Four concentrations (0, 50, 75 and 100 mg) of three Eucalyptus extracts (ethyl acetate, n-butanol and aqueous) were added to a diet of ruminants (forage: concentrate ratio 60:40) and incubated at 39°C under anaerobiosis with buffered rumen fluid. After 24 h, the fermentation fluid was analysed for ammonia-N and volatile fatty acids (VFA). Organic matter degradability (OMD) and protozoa were also determined; in vitro gas production was also recorded and CH4 concentration was measured. Compared to the control, CH4 production was significantly lower for ethyl acetate extract (P<0.05), but higher for n-butanol and aqueous extracts. Production of ammonia- N was lower in all Eucalyptus extracts (P<0.05). Propionate production (P<0.05) increased for ethyl acetate and n-butanol extracts, whereas no effect was registered for VFA, for all Eucalyptus extracts. Ethyl acetate extract decreased in vitro OMD (P<0.05), whereas n-butanol and aqueous extracts were comparable to the control. Protozoa population decreased (P<0.05) for all extracts in comparison with the control. Eucalyptus ethyl acetate extract might be promising to be used as a potent anti-methanogenic additive. Moreover, the assessment of the right dosage seems to be important to decrease methane production, without reducing feed nutritional value.


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