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Current postexposure prophylaxis of rabies includes vaccines, human rabies immunoglobulin (RIG), equine RIG, and recombinant monoclonal antibodies (mAb). In the manufacturing of rabies recombinant mAb, charge variants are the most common source of heterogeneity. Charge variants of rabies mAb were isolated by salt gradient cation exchange chromatography (CEX) to separate acidic and basic and main charge variants. Separated variants were further extensively characterized using orthogonal analytical techniques, which include secondary and tertiary structure determination by far and near ultraviolet circular dichroism spectroscopy. Charge and size heterogeneity were evaluated using CEX, isoelectric focusing (IEF), capillary-IEF, size exclusion chromatography, sodium dodecyl sulfate polyacrylamide gel electrophoresis, and western blotting. Antigen binding affinity was assessed by enzyme linked immuno-sorbent assay and rapid florescence foci inhibition test. Results from structural and physicochemical characterizations concluded that charge variants are formed due to posttranslational modification demonstrating that the charge heterogeneity, these charge variants did neither show any considerable physicochemical change nor affect its biological function. This study shows that charge variants are effective components of mAb and there is no need of deliberate removal, until biological functions of rabies mAb will get affected.
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Quorum sensing (QS), the communication signaling network, regulates biofilm formation and several virulence factors in Pseudomonas aeruginosa PAO1, a nosocomial opportunistic pathogen. QS is considered to be a challenging target for compounds antagonistic to virulent factors. Biologically synthesized silver nanoparticles (AgNPs) are reported as anti-QS and anti-biofilm drugs against bacterial infections. The present study reports on the synthesis and characterization of Piper betle (Pb) mediated AgNPs (Pb-AgNPs). The anti-QS activity of Pb-AgNPs against Chromobacterium violaceum and the potential effect of Pb-AgNPs on QS-regulated phenotypes in PAO1 were studied. FTIR analysis exhibited that Pb-AgNPs had been capped by phytochemical constituents of Pb. Eugenol is one of the active phenolic phytochemicals in Pb leaves, therefore molecular docking of eugenol-conjugated AgNPs on QS regulator proteins (LasR, LasI and MvfR) was performed. Eugenol-conjugated AgNPs showed considerable binding interactions with QS-associated proteins. These results provide novel insights into the development of phytochemically conjugated nanoparticles as promising anti-infective candidates.
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Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Nanopartículas Metálicas/química , Pseudomonas aeruginosa/efeitos dos fármacos , Percepção de Quorum/efeitos dos fármacos , Prata/química , Biofilmes/crescimento & desenvolvimento , Chromobacterium/fisiologia , Infecção Hospitalar/microbiologia , Violeta Genciana/química , Testes de Sensibilidade Microbiana , Simulação de Acoplamento Molecular , Elastase Pancreática/química , Compostos Fitoquímicos/farmacologia , Piper betle/química , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Folhas de Planta/química , Fatores de Virulência/metabolismoRESUMO
Previously, we reported an arsenic resistant bacterium Lysinibacillus sphaericus B1-CDA, isolated from an arsenic contaminated lands. Here, we have investigated its genetic composition and evolutionary history by using massively parallel sequencing and comparative analysis with other known Lysinibacillus genomes. Assembly of the sequencing reads revealed a genome of ~4.5 Mb in size encompassing ~80% of the chromosomal DNA. We found that the set of ordered contigs contains abundant regions of similarity with other Lysinibacillus genomes and clearly identifiable genome rearrangements. Furthermore, all genes of B1-CDA that were predicted be involved in its resistance to arsenic and/or other heavy metals were annotated. The presence of arsenic responsive genes was verified by PCR in vitro conditions. The findings of this study highlight the significance of this bacterium in removing arsenics and other toxic metals from the contaminated sources. The genetic mechanisms of the isolate could be used to cope with arsenic toxicity.
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Arsênio/metabolismo , Bacillaceae/genética , Genoma Bacteriano/genética , Genômica/métodos , Arsênio/farmacologia , Bacillaceae/classificação , Bacillaceae/metabolismo , Biodegradação Ambiental , Cromossomos Bacterianos/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Farmacorresistência Bacteriana/genética , Variação Genética , Filogenia , Análise de Sequência de DNA , Especificidade da EspécieRESUMO
Chromium and chromium containing compounds are discharged into the nature as waste from anthropogenic activities, such as industries, agriculture, forest farming, mining and metallurgy. Continued disposal of these compounds to the environment leads to development of various lethal diseases in both humans and animals. In this paper, we report a soil borne bacterium, B2-DHA that can be used as a vehicle to effectively remove chromium from the contaminated sources. B2-DHA is resistant to chromium with a MIC value of 1000 µg mL(-1) potassium chromate. The bacterium has been identified as a Gram negative, Enterobacter cloacae based on biochemical characteristics and 16S rRNA gene analysis. TOF-SIMS and ICP-MS analyses confirmed intracellular accumulation of chromium and thus its removal from the contaminated liquid medium. Chromium accumulation in cells was 320 µg/g of cells dry biomass after 120-h exposure, and thus it reduced the chromium concentration in the liquid medium by as much as 81%. Environmental scanning electron micrograph revealed the effect of metals on cellular morphology of the isolates. Altogether, our results indicate that B2-DHA has the potential to reduce chromium significantly to safe levels from the contaminated environments and suggest the potential use of this bacterium in reducing human exposure to chromium, hence avoiding poisoning.
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Cromo/metabolismo , Enterobacter cloacae/genética , Enterobacter cloacae/metabolismo , Resíduos Industriais/análise , Poluentes do Solo/isolamento & purificação , Bangladesh , Biodegradação Ambiental , Enterobacter cloacae/efeitos dos fármacos , Filogenia , Microbiologia do Solo , CurtumeRESUMO
Salinity and heavy metal stress are challenging problems in agriculture. Here we report the plant growth promoting ability of three moderate halophiles, Halobacillus sp. ADN1, Halomonas sp. MAN5, and Halobacillus sp. MAN6, in presence of both salinity and heavy metal stress. Halobacillus sp. ADN1, Halomonas sp. MAN5, and Halobacillus sp. MAN6 can tolerate 25, 21, and 29% NaCl, respectively and grow in presence of 1 mM cobalt, cadmium, and nickel and 0.04 mM mercury and 0.03 mM silver. Halobacillus sp. ADN1, Halomonas sp. MAN5, and Halobacillus sp. MAN6 produced 152.5, 95.3, and 167.3 µg/ml indole acetic acid (IAA) and could solubilize 61, 53, and 75 parts per million (ppm) phosphate, respectively in the presence of 15% NaCl. The production of IAA and solubilization of phosphate was well retained in the presence of salinity and heavy metals like 1 mM cadmium, 0.7 mM nickel, 0.04 mM mercury, and 0.03 mM silver. Besides, the strains showed amylase and protease activities and could produce hydrogen cyanide and ammonia in presence of salinity and heavy metals. A mixture of three strains enhanced the root growth of Sesuvium portulacastrum under saline and heavy metal stress, where the root length increased nearly 4.5 ± 0.6 times and root dry weight increased 5.4 ± 0.5 times as compared to control. These strains can thus be useful in microbial assisted phytoremediation of polluted saline soils.
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Aizoaceae/crescimento & desenvolvimento , Halobacillus/metabolismo , Halomonas/metabolismo , Metais Pesados/farmacologia , Cloreto de Sódio/farmacologia , Aizoaceae/microbiologia , Amônia/metabolismo , Amilases/metabolismo , Técnicas de Tipagem Bacteriana , Halobacillus/genética , Halomonas/genética , Cianeto de Hidrogênio/metabolismo , Ácidos Indolacéticos/metabolismo , Peptídeo Hidrolases/metabolismo , Fosfatos/química , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/microbiologia , RNA Ribossômico 16S/genética , Salinidade , Solo , Poluentes do SoloRESUMO
The main objective of this study was to identify and isolate arsenic resistant bacteria that can be used for removing arsenic from the contaminated environment. Here we report a soil borne bacterium, B1-CDA that can serve this purpose. B1-CDA was isolated from the soil of a cultivated land in Chuadanga district located in the southwest region of Bangladesh. The morphological, biochemical and 16S rRNA analysis suggested that the isolate belongs to Lysinibacillus sphaericus. The minimum inhibitory concentration (MIC) value of the isolate is 500 mM (As) as arsenate. TOF-SIMS and ICP-MS analysis confirmed intracellular accumulation and removal of arsenics. Arsenic accumulation in cells amounted to 5.0 mg g(-1) of the cells dry biomass and thus reduced the arsenic concentration in the contaminated liquid medium by as much as 50%. These results indicate that B1-CDA has the potential for remediation of arsenic from the contaminated water. We believe the benefits of implementing this bacterium to efficiently reduce arsenic exposure will not only help to remove one aspect of human arsenic poisoning but will also benefit livestock and native animal species. Therefore, the outcome of this research will be highly significant for people in the affected area and also for human populations in other countries that have credible health concerns as a consequence of arsenic-contaminated water.
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Arsênio/metabolismo , Bacillaceae/isolamento & purificação , Bacillaceae/metabolismo , Poluentes Químicos da Água/metabolismo , Arsênio/análise , Bacillaceae/classificação , Bacillaceae/genética , Bangladesh , Biodegradação Ambiental , Humanos , Dados de Sequência Molecular , Filogenia , Microbiologia do Solo , Poluentes Químicos da Água/análiseRESUMO
The nosocomial infection outbreak caused by Pseudomonas aeruginosa remains a public health concern. Multi-drug resistant (MDR) strains of P. aeruginosa are rapidly spreading leading to a huge mortality rate because of the unavailability of promising antimicrobials. MurG glycotransferase [UDP-N-acetylglucosamine-N-acetylmuramyl (pentapeptide) pyrophosphoryl-undecaprenol N-acetylglucosamine transferase] is located at the plasma membrane and plays a key role in murein (peptidoglycan) biosynthesis in bacteria. Since MurG is required for bacterial cell wall synthesis and is non-homologous to Homo sapiens; it can be a potential target for the antagonist to treat P. aeruginosa infection. The discovery of high-resolution crystal structure of P. aeruginosa MurG offers an opportunity for the computational identification of its prospective inhibitors. Therefore, in the present study, the crystal structure of MurG (PDB ID: 3S2U) from P. aeruginosa was selected, and computational docking analyses were performed to search for functional inhibitors of MurG. IMPPAT (Indian medicinal plants, phytochemicals and therapeutic) phytomolecule database was screened by computational methods with MurG catalytic site. Docking results identified Theobromine (-8.881 kcal/mol), demethoxycurcumin (-8.850 kcal/mol), 2-alpha-hydroxycostic acid (-8.791 kcal/mol), aurantiamide (-8.779 kcal/mol) and petasiphenol (-8.685 kcal/mol) as a potential inhibitor of the MurG activity. Further, theobromine and demethoxycurcumin were subjected to MDS (molecular dynamics simulation) and free energy (MM/GBSA) analysis to comprehend the physiological state and structural stability of MurG-phytomolecules complexes. The outcomes suggested that these two phytomolecules could act as most favorable natural hit compounds for impeding the enzymatic action of MurG in P. aeruginosa, and thus it needs further validation by both in vitro and in vivo analysis. HIGHLIGHTSThe top phytomolecules such as theobromine, demethoxycurcumin, 2-alpha-hydroxycostic acid, aurantiamide and petasiphenol displayed promising binding with MurG catalytic domain.MurG complexed with theobromine and demethoxycurcumin showed the best interaction and stable by MD simulation at 100 ns.The outcome of MurG binding phytomolecules has expanded the possibility of hit phytomolecules validation.Communicated by Ramaswamy H. Sarma.
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Infecção Hospitalar , Pseudomonas aeruginosa , Humanos , Teobromina , Simulação de Acoplamento Molecular , Simulação de Dinâmica MolecularRESUMO
Vaginal health is essential to a woman's overall well-being, as abnormalities in vaginal health can lead to a variety of gynaecological disorders, such as urinary tract infections, yeast infections, and bacterial vaginosis. The vaginal microbiome is essential for the prevention of these infections. Disruptions in this microbial ecosystem can significantly impact vaginal health. The concept of utilizing probiotics and prebiotics to stimulate the growth of protective vaginal microbiota has gathered substantial interest in recent years. Probiotics are live micro-organisms that strengthen and restore vaginal microbial balance by lowering pH levels, production of bacteriocins, biofilm disruption, modulation of immune response, and production of hydrogen peroxide (H2O2), consequently combating the development of pathogens. Prebiotics are oligosaccharides that encourage the development of probiotics such as lactobacilli species. Probiotics and prebiotics also have some broader implications for vaginal health, including their role in minimizing the incidence of premature birth, optimizing fertility, managing menopausal symptoms, and preventing vaginal infections. Synbiotics are a combination of probiotics and prebiotics that deliver additional benefits by encouraging the development and activity of beneficial microbes. Furthermore, postbiotics are bioactive compounds derived from probiotic bacteria during fermentation that have immunomodulatory actions and provide an additional layer of protection against vaginal infections. The present study highlights the most prevalent vaginal infections and limitations of existing therapies that influence the vaginal microbiota. The profound consequences of probiotics and prebiotics in women's health, including their role in minimizing the prevalence of vaginal infections and promoting overall vaginal health, as well as advanced therapeutic strategies such as synbiotics and postbiotics, are also discussed. The literature offers significant insights into the mechanism, efficacy, and safety of probiotics and prebiotics to healthcare providers and researchers.
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Microbiota , Prebióticos , Probióticos , Vagina , Humanos , Feminino , Probióticos/administração & dosagem , Vagina/microbiologia , Vaginose Bacteriana/prevenção & controle , Vaginose Bacteriana/microbiologia , Vaginose Bacteriana/terapia , Lactobacillus , Simbióticos/administração & dosagemRESUMO
Gold nanoparticles (Au-NPs) were synthesized from chloroauric acid using cell free supernatant of Streptomyces sp. NK52 grown in nutrient broth. These nanoparticles were synthesized by varying pH and temperature of the reaction mixture and chloroauric acid concentration. The nanoparticles were characterized by spectrometry, X-ray diffraction, Scanning electron microscopy and energy dispersive spectrometry. Au-NP ranged from 10-100 nm in size and exhibited a polydispersive nature with various shapes like rods, hexagons, triangles, spheres. The diffraction peaks at 2theta = 38.1 degrees and 44.5 degrees could be assigned to the (1 1 1) and (2 0 0) planes of a faced centre cubic (fcc) lattice of gold. Au-NP showed 47% inhibition of lipid peroxidation in vitro. To the best of our knowledge, this is the first report on the rapid biosynthesis of Au-NP using cell free supernatant of Streptomyces sp. and its evaluation for anti-lipid peroxidation.
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Ouro/química , Peroxidação de Lipídeos/efeitos dos fármacos , Nanopartículas Metálicas , Streptomyces/metabolismo , Microscopia Eletrônica de Varredura , Espectrometria por Raios X , Difração de Raios XRESUMO
Acinetobacter baumannii is a notorious multidrug resistant bacterium responsible for several hospital acquired infections assisted by its capacity to develop biofilms. A. baumannii BfmR (RstA), a response regulator from the BfmR/S two-component signal transduction system, is the major controller of A. baumannii biofilm development and formation. As a result, BfmR represents a novel target for anti-biofilm treatment against A. baumannii. The discovery of the high-resolution crystal structure of BfmR provides a good chance for computational screening of its probable inhibitors. Therefore, in this study we aim to search new, less toxic, and natural BfmR inhibitors from 8450 phytomolecules available in the Indian Medicinal Plants, Phytochemistry and Therapeutic (IMPPAT) database by analyzing molecular docking against BfmR (PDB ID: 6BR7). Out of these 8450 phytomolecules 6742 molecules were successfully docked with BfmR with the docking score range -6.305 kcal/mol to +5.120 kcal/mol. Structure based-molecular docking (SB-MD) and ADMET (absorption, distribution, metabolism, excretion, & toxicity) profile examination revealed that Norepinephrine, Australine, Calystegine B3, 7,7 A-Diepialexine, and Alpha-Methylnoradrenaline phytocompounds strongly binds to the active site residues of BfmR. Furthermore, molecular dynamics simulation (MDS) studies for 100 ns and the binding free energy (MM/GBSA) analysis elucidated the binding mechanism of Calystegine B3, 7,7 A-Diepialexine, and Alpha-Methylnoradrenaline to BfmR. In summary, these phytocompounds seems to have the promising molecules against BfmR, and thus necessitates further verification by both in vitro and in vivo experiments. HighlightsBfmR plays a key role in biofilm development and exopolysaccharide (EPS) synthesis in A. baumannii.Computational approach to search for promising BfmR inhibitors from IMPAAT database.The lead phytomolecules such as Calystegine B3, 7,7 A-Diepialexine, and Alpha-Methylnoradrenaline displayed significant binding with BfmR active site.The outcome of BfmR binding phytomolecules has broadened the scope of hit molecules validation.Communicated by Ramaswamy H. Sarma.
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Acinetobacter baumannii , Simulação de Dinâmica Molecular , Simulação de Acoplamento Molecular , Acinetobacter baumannii/metabolismo , Nordefrin/metabolismo , Desenvolvimento de MedicamentosRESUMO
The rapid development of multi-drug resistant (MDR) pathogens adds urgency to search for novel and safe drugs having promising action on new and re-emerging infectious pathogens. Serratia marcescens is an MDR pathogen that causes several-healthcare associated infections. Curbing bacterial virulence, rather than inhibiting its growth, is a promising strategy to diminish the pathogenesis of infectious bacteria, reduce the development of antimicrobial resistance, and boost the host immune power to eradicate infections. Bergamot essential oil (BEO) is a remarkable source of promising therapeutics against pathogens. Therefore, the present investigation aimed to analyze the major phytocompounds from BEO against S. marcescens virulent proteins using in silico studies. The analysis of BEO phytocompounds was achieved by Gas chromatography-mass spectrometry (GC-MS) method. The molecular docking was carried out using the SP and XP docking protocol of the Glide program. The drug-likeness and pharmacokinetics properties (ADMET properties) were analyzed with SwissADME and pkCSM server. The results revealed that the major compounds present in BEO are Linalool (8.17%), D-Limonene (21.26%), and Linalyl acetate (26.91%). Molecular docking analysis revealed that these compounds docked strongly within the binding cavities of Serratia protease and FabI model which in turn curb the pathogenesis of this bacteria. Linalool interacted with the Serratia protease and FabI with a binding energy of - 3.130 kcal/mol and - 3.939 kcal/mol, respectively. Based on the pharmacokinetics findings all lead BEO phytocompounds appear to be promising drug candidates. Overall, these results represent a significant step in the development of plant-based compounds as a promising inhibitor of the virulent proteins of the MDR S. marcescens.
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Óleos Voláteis , Óleos Voláteis/farmacologia , Óleos Voláteis/química , Serratia marcescens , Peptídeo Hidrolases , Simulação de Acoplamento MolecularRESUMO
COVID-19 shook the world during the pandemic, where the climax it reached was vaccine manufacturing at an unfathomable pace. Alternative promising solutions to prevent infection from SARS-CoV-2 and its variants will remain crucial in the years to come. Due to its key role in viral replication, the major protease (Mpro) enzyme of SARS-CoV-2 can be an attractive therapeutic target. In the present work, natural terpenoids from mangrove medicinal plant Xylocarpus moluccensis (Lam.) M. Roem. were screened using computational methods for inhibition of Mpro protein. Out of sixty-seven terpenoids, Angolensic acid methyl ester, Moluccensin V, Thaixylomolin F, Godavarin J, and Xylomexicanolide A were shortlisted based on their docking scores and interaction affinities (- 13.502 to - 15.52 kcal/mol). The efficacy was validated by the 100 ns molecular dynamics study. Lead terpenoids were within the acceptable range of RMSD and RMSF with a mean value of 2.5 Å and 1.5 Å, respectively indicating that they bound tightly within Mpro and there was minimal fluctuation and stability of Mpro upon binding of these terpenoids. The utmost favorable binding strengths as calculated by MM-GBSA, were of Angolensic acid methyl ester and Moluccensin V with binding free energies (ΔGbind) of - 39.084, and - 43.160 kcal/mol, respectively. The terpenoids showed no violations in terms of Drug Likeliness and ADMET predictions. Overall, the findings indicate that Angolensic acid methyl ester and Moluccensin V are effective terpenoids having strong binding interaction with Mpro protein, which must be tested in vitro as an effective anti-SARS-CoV-2 drug.
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Antivirais , Magnoliopsida , Terpenos , Simulação por Computador , Magnoliopsida/química , Terpenos/química , SARS-CoV-2 , Proteases 3C de Coronavírus/antagonistas & inibidores , Simulação de Acoplamento Molecular , Tratamento Farmacológico da COVID-19 , Antivirais/química , TermodinâmicaRESUMO
The recent outbreak of SARS-CoV-2 has quickly become a worldwide pandemic and generated panic threats for both the human population and the global economy. The unavailability of effective vaccines or drugs has enforced researchers to hunt for a potential drug to combat this virus. Plant-derived phytocompounds are of applicable interest in the search for novel drugs. Bioflavonoids from Rhus succedanea are already reported to exert antiviral activity against RNA viruses. SARS-CoV-2 Mpro protease plays a vital role in viral replication and therefore can be considered as a promising target for drug development. A computational approach has been employed to search for promising potent bioflavonoids from Rhus succedanea against SARS-CoV-2 Mpro protease. Binding affinities and binding modes between the biflavonoids and Mpro enzyme suggest that all six biflavonoids exhibit possible interaction with the Mpro catalytic site (-19.47 to -27.04 kcal/mol). However, Amentoflavone (-27.04 kcal/mol) and Agathisflavone (-25.87 kcal/mol) interact strongly with the catalytic residues. Molecular dynamic simulations (100 ns) further revealed that these two biflavonoids complexes with the Mpro enzyme are highly stable and are of less conformational fluctuations. Also, the hydrophobic and hydrophilic surface mapping on the Mpro structure as well as biflavonoids were utilized for the further lead optimization process. Altogether, our findings showed that these natural biflavonoids can be utilized as promising SARS-CoV-2 Mpro inhibitors and thus, the computational approach provides an initial footstep towards experimental studies in in vitro and in vivo, which is necessary for the therapeutic development of novel and safe drugs to control SARS-CoV-2. Communicated by Ramaswamy H. SarmaResearch highlightsRhus succedanea biflavonoids have antiviral activity.The molecular interactions and molecular dynamics displayed that all six biflavonoids bound with a good affinity to the same catalytic site of Mpro.The compound Amentoflavone has a strong binding affinity (-27.0441 kcal/mol) towards Mpro.The binding site properties of SARS-CoV-2-Mpro can be utilized in a novel discovery and lead optimization of the SARS-CoV-2-Mpro inhibitor.
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Biflavonoides , Tratamento Farmacológico da COVID-19 , Rhus , Antivirais/química , Antivirais/farmacologia , Biflavonoides/farmacologia , Proteases 3C de Coronavírus , Cisteína Endopeptidases/química , Humanos , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Inibidores de Proteases/química , Inibidores de Proteases/farmacologia , Rhus/metabolismo , SARS-CoV-2 , Proteínas não Estruturais Virais/químicaRESUMO
In this study we aim to investigate the computational docking approach of biofabricated silver nanoparticles against P. aeruginosa virulent exoenzymes, such as ExoS and ExoY. Therefore, the synthesis and characterization of biofabricated silver nanoparticles using Piper betle leaves (Pb-AgNPs) were carried out. The surface topology and functional group attachment on the surface of Pb-AgNPs were analyzed using UV-visible spectroscopy, Scanning Electron Microscopy, Fourier Transformed Infrared Spectroscopy (FTIR), and X-Ray Diffraction. The FTIR analysis revealed that the synthesized silver nanoparticles were capped with P. betle phytochemicals importantly Eugenol and Hydroxychavicol. These are the major bioactive compounds present in P. betle leaves; therefore, computational docking of Eugenol-conjugated AgNPs (PbEu-AgNPs) and Hydroxychavicol-conjugated AgNPs (PbHy-AgNPs) against ExoS and ExoY was performed. The active residues of PbEu-AgNPs and PbHy-AgNPs interacted with the active site of ExoS and ExoY exoenzymes. Biofabricated AgNP-mediated inhibition of these virulent exoenzymes blocked the adverse effect of P. aeruginosa on the host cell. The computational analysis provides new approach into the design of biofabricated AgNPs as promising anti-infective nanomedicine agents. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03367-0.
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The linking of polysaccharide in glycoconjugate vaccine with carrier protein is an imperative step to develop a strong memory response. The excessive use of similar carrier protein known to result in bystander immunity warrants an urgent need for new carrier protein. The preparation of the glycoconjugate vaccine using cyanylation chemistry is to link the active cyanate ester site of polysaccharide with the carrier protein. In the present study, transferrin binding protein-B (Tbp-B) has been explored as a new carrier protein to develop in silico pneumococcal polysaccharide serotype-5 (PnPs-5) conjugate vaccine. The homology model of Tbp-B was constructed using the Prime module and stereochemically validated using ProSA, PDBsum and ProQ. The selected model revealed a Z-score of -5.6 within the X-ray region in ProSA analysis, LGscore: 9.776, and MaxSub: 0.8 in protein quality predictor suggesting its preferred use. Loop modeling and active site analysis followed by in silico PnPs-5 activation with cyanalyting agent CDAP was docked with Tbp-B using Glide module. The complex stability of cyanate esters with Tbp-B, analyzed by molecular dynamics (MD) simulation, revealed an average RMSD of 2.49 Å for its binding to the receptor. The RMSF values of cyanate ester-1, -2, and -3 were observed to be 1.06, 1.39 and 0.79 Å, respectively. The higher RMSF of 1.39 Å of cyanate ester-2 was further found unstable which corroborates its non-binding to the protein and also incurring conformational changes to a carrier protein. Molecular simulations revealed that cyanate ester-1 and cyanate ester-3 formed stable conjugates with carrier protein Tbp-B. Communicated by Ramaswamy H. Sarma.
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Proteínas de Transporte , Neisseria meningitidis , Proteínas de Transporte/metabolismo , Proteína B de Ligação a Transferrina/metabolismo , Antígenos/metabolismo , Neisseria meningitidis/metabolismo , Glicoconjugados/metabolismo , Simulação de Dinâmica Molecular , Simulação de Acoplamento MolecularRESUMO
The interest in naturally occurring essential oils from medicinal plants has increased extremely over the last decade markedly because they possess antimicrobial and antioxidant protective properties against different chronic diseases. Extensive survival of drug-resistant infectious bacteria depends on quorum sensing (QS) signaling network which raises the need for alternative antibacterial compounds. The aim of this study was to examine the phytochemical compounds of patchouli essential oil (PEO) and to assess its antioxidant activity. Antioxidant studies estimated by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method showed that the PEO has effective antioxidant activity (IC50 19.53 µg/mL). QS inhibitory activity of PEO was examined by employing the biosensor strain, Chromobacterium violaceum CV12472. At sub-lethal concentrations, PEO potentially reduced the QS regulated violacein synthesis in CV12472 without inhibiting its cell proliferation. Moreover, it also effectively reduced the production of some QS regulated virulence factors and biofilm development in P. aeruginosa PAO1 without hindering its growth. Phytochemical analysis of PEO was done by GC/MS technique. Molecular docking of PEO major compounds with QS (LasR and FabI) and biofilm regulator proteins (MvfR and Sialidase) of PAO1 was evaluated. These phytocompounds showed potential hydrogen binding interactions with these proteins. The overall results, in vitro and in silico, suggest that PEO could be applied as biocontrol agent against antibiotic resistance pathogens. Communicated by Ramaswamy H. Sarma.
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Óleos Voláteis , Pogostemon , Antibacterianos/farmacologia , Antioxidantes/farmacologia , Biofilmes , Simulação de Acoplamento Molecular , Óleos Voláteis/farmacologia , Extratos Vegetais/farmacologia , Percepção de Quorum , Fatores de VirulênciaRESUMO
Biological methods offer eco-friendly and cost-effective alternatives for the synthesis of silver nanoparticles (AgNPs). The present study highlights a green process where AgNPs were synthesized and optimized by using silver nitrate (AgNO3) and the aqueous extract of Piper betle (Pbet) leaf as the reducing and capping agent. The stable and optimized process for the synthesis of Pbet-AgNPs was exposure of reaction mixture into the sunlight for 40 min, pH 9.0, and 2 mM AgNO3 using 1:4 diluted Pbet leaf aqueous extract. The optimized Pbet-AgNPs were characterized by UV-visible spectroscopy, high-resolution field emission scanning electron microscopy (FE-SEM), X-ray diffractometry (XRD), and Fourier-transform infrared spectroscopy (FTIR). The prepared Pbet-AgNPs were spherical in shape with size in the range of 6-14 nm. These nanoparticles were stable for 6 months in aqueous solution at room temperature under dark conditions. The biogenic synthesized Pbet-AgNPs are found to have significant antifungal activity against plant pathogenic fungi, Alternaria brassicae and Fusarium solani. Synthesized Pbet-AgNPs potentially reduced the fungal growth in a dose-dependent manner. Microscopic observation of treated mycelium showed that Pbet-AgNPs could disrupt the mycelium cell wall and induce cellular permeability. Protein leakage assay supports these findings. Overall, this study revealed the efficacy of green synthesized AgNPs to control the plant fungal pathogens. Pbet leaves are a rich source of phenolic biomolecule(s). It was hypothesized that these biomolecule(s) mediated metal reduction reactions. In this context, the present work investigates the phytobiomolecule(s) of the aqueous extract of Pbet leaves using high-resolution liquid chromatography-mass spectroscopy (HR-LCMS) method. The analysis revealed that eugenol, chavicol, and hydroxychavicol were present in the Pbet aqueous extract.
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Nanopartículas Metálicas , Piper betle , Extratos Vegetais , Folhas de Planta , Prata , Nitrato de Prata , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
Polluted waters are an important reservoir for antibiotic resistance genes and multidrug-resistant bacteria. This report describes the microbial community, antibiotic resistance genes, and the genetic profile of extended spectrum ß-lactamase strains isolated from rivers at, Pune, India. ESBL-producing bacteria isolated from diverse river water catchments running through Pune City were characterized for their antibiotic resistance. The microbial community and types of genes which confer antibiotic resistance were identified followed by the isolation of antibiotic-resistant bacteria on selective media and their genome analysis. Four representative isolates were sequenced using next generation sequencing for genomic analysis. They were identified as Pseudomonas aeruginosa, Escherichia coli, and two isolates were Enterobacter cloacae. The genes associated with the multidrug efflux pumps, such as tolC, macA, macB, adeL, and rosB, were detected in the isolates. As MacAB-TolC is an ABC type efflux pump responsible for conferring resistance in bacteria to several antibiotics, potential efflux pump inhibitors were identified by molecular docking. The homology model of their MacB protein with that from Escherichia coli K12 demonstrated structural changes in different motifs of MacB. Molecular docking of reported efflux pump inhibitors revealed the highest binding affinity of compound MC207-110 against MacB. It also details the potential efflux pump inhibitors that can serve as possible drug targets in drug development and discovery.
Assuntos
Proteínas de Bactérias , Rios , Antibacterianos/farmacologia , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Índia , Testes de Sensibilidade Microbiana , Simulação de Acoplamento MolecularRESUMO
We have cloned, characterized and transformed the AtACR2 gene (arsenic reductase 2) of Arabidopsis thaliana into the genome of tobacco (Nicotiana tabacum, var Sumsun). Our results revealed that the transgenic tobacco plants are more tolerant to arsenic than the wild type ones. These plants can grow on culture medium containing 200µM arsenate, whereas the wild type can barely survive under this condition. Furthermore, when exposed to 100µM arsenate for 35days the amount of arsenic accumulated in the shoots of transgenic plants was significantly lower (28µg/g d wt.) than that found in the shoots of non-transgenic controls (40µg/g d wt.). However, the arsenic content in the roots of transgenic plants was significantly higher (2400µg/g d. wt.) than that (2100µg/g d. wt.) observed in roots of wild type plants. We have demonstrated that Arabidopsis thaliana AtACR2 gene is a potential candidate for genetic engineering of plants to develop new crop cultivars that can be grown on arsenic contaminated fields to reduce arsenic content of the soil and can become a source of food containing no arsenic or exhibiting substantially reduced amount of this metalloid.