RESUMEN
PURPOSE: The study aimed at isolating and identifying potential anti-quorum sensing (QS) compounds from Cinnamomum verum leaves against Pseudomonas aeruginosa. METHODOLOGY: Isolation of anti-QS compounds from C. verum leaf ethanol extract was carried out by column chromatography. The bioactive fraction was analysed by UV, IR, and GCMS spectroscopy. Various virulence assays were performed to assess the QS quenching ability of the purified compounds. In vivo toxicity of the purified compounds was examined in zebrafish model. The expression of the virulence genes was evaluated by qPCR analysis and in silico assessment was accomplished to check the binding ability of the compounds with the autoinducer molecule. KEY FINDINGS: The QS inhibitors isolated and identified showed a remarkable ability in reducing the production of elastase, pyocyanin, swarming motility and biofilm formation in P. aeruginosa. In the presence of the characterized compounds, the expression of virulence genes of P. aeruginosa was significantly reduced. Toxicity studies in zebrafish model indicated no effects on development and organogenesis at a concentration below 100 mg/l. Further, in silico analysis demonstrated the binding efficiency of the anti-QS compounds to AHL molecules, thus proving the QS quenching ability of the isolated compounds. SIGNIFICANCE: To the best of our knowledge this is the first report of isolation of anti-QS compounds from C. verum leaves against P. aeruginosa. The identified compounds qualify as potential QS antagonists. Further studies on these compounds can pave way for an effective and attractive anti-pathogenic therapy, to overcome the emergence of antibiotic resistance in bacteria.
Asunto(s)
Cinnamomum zeylanicum/metabolismo , Extractos Vegetales/farmacología , Percepción de Quorum/efectos de los fármacos , Antibacterianos/farmacología , Biopelículas/efectos de los fármacos , Cinnamomum zeylanicum/química , Cinnamomum zeylanicum/enzimología , Flavonoides/farmacología , Hojas de la Planta/química , Pseudomonas aeruginosa/efectos de los fármacos , Percepción de Quorum/fisiología , Virulencia/genética , Factores de VirulenciaRESUMEN
Carotenoids are widely used in functional foods, cosmetics, and health supplements, and their importance and scope of use are continuously expanding. Here, we characterized carotenoid biosynthetic genes of the plant-pathogenic bacterium Pantoea ananatis, which carries a carotenoid biosynthetic gene cluster (including crtE, X, Y, I, B, and Z) on a plasmid. Reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed that the crtEXYIB gene cluster is transcribed as a single transcript and crtZ is independently transcribed in the opposite direction. Using splicing by overlap extension with polymerase chain reaction (SOE by PCR) based on asymmetric amplification, we reassembled crtE-B, crtE-B-I, and crtE-B-I-Y. High-performance liquid chromatography confirmed that Escherichia coli expressing the reassembled crtE-B, crtE-B-I, and crtE-B-I-Y operons produced phytoene, lycopene, and ß-carotene, respectively. We found that the carotenoids conferred tolerance to UV radiation and toxoflavin. Pantoea ananatis shares rice environments with the toxoflavin producer Burkholderia glumae and is considered to be the first reported example of producing and using carotenoids to withstand toxoflavin. We confirmed that carotenoid production by P. ananatis depends on RpoS, which is positively regulated by Hfq/ArcZ and negatively regulated by ClpP, similar to an important regulatory network of E. coli (HfqArcZ âRpoS Í° ClpXP). We also demonstrated that Hfq-controlled quorum signaling de-represses EanR to activate RpoS, thereby initiating carotenoid production. Survival genes such as those responsible for the production of carotenoids of the plant-pathogenic P. ananatis must be expressed promptly to overcome stressful environments and compete with other microorganisms. This mechanism is likely maintained by a brake with excellent performance, such as EanR.
Asunto(s)
Carotenoides/metabolismo , Proteína de Factor 1 del Huésped/metabolismo , Pantoea/efectos de los fármacos , Pantoea/metabolismo , Pirimidinonas/farmacología , Percepción de Quorum/fisiología , Triazinas/farmacología , Proteínas Bacterianas/metabolismo , Endopeptidasa Clp/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Escherichia coli/metabolismo , Familia de Multigenes/genética , Plásmidos/genética , Factor sigma/metabolismo , Rayos UltravioletaRESUMEN
Quorum sensing is a communication system among bacteria to sense the proper time to express their virulence factors. Quorum sensing inhibition is a therapeutic strategy to block bacterial mechanisms of virulence. The aim of this study was to synthesize and evaluate new bioisosteres of N-acyl homoserine lactones as Quorum sensing inhibitors in Chromobacterium violaceum CV026 by quantifying the specific production of violacein. Five series of compounds with different heterocyclic scaffolds were synthesized in good yields: thiazoles, 16a-c, thiazolines 17a-c, benzimidazoles 18a-c, pyridines 19a-c and imidazolines 32a-c. All 15 compounds showed activity as Quorum sensing inhibitors except 16a. Compounds 16b, 17a-c, 18a, 18c, 19c and 32b exhibited activity at concentrations of 10 µM and 100 µM, highlighting the activity of benzimidazole 18a (IC50 = 36.67 µM) and 32b (IC50 = 85.03 µM). Pyridine 19c displayed the best quorum sensing inhibition activity (IC50 = 9.66 µM). Molecular docking simulations were conducted for all test compounds on the Chromobacterium violaceum CviR protein to gain insight into the process of quorum sensing inhibition. The in-silico data reveal that all 15 the compounds have higher affinity for the protein than the native AHL ligand (1). A strong correlation was found between the theoretical and experimental results.
Asunto(s)
Percepción de Quorum/fisiología , Acil-Butirolactonas/metabolismo , Chromobacterium/química , Indoles/metabolismo , Simulación del Acoplamiento Molecular , Extractos Vegetales/química , Percepción de Quorum/genéticaRESUMEN
Given the major threat of phytopathogenic bacteria to food production and ecosystem stability worldwide, novel alternatives to conventional chemicals-based agricultural practices are needed to combat these bacteria. The objective of this study is to evaluate the ability of Pseudomonas segetis strain P6, which was isolated from the Salicornia europaea rhizosphere, to act as a potential biocontrol agent given its plant growth-promoting (PGP) and quorum quenching (QQ) activities. Seed biopriming and in vivo assays of tomato plants inoculated with strain P6 resulted in an increase in seedling height and weight. We detected QQ activity, involving enzymatic degradation of signal molecules in quorum sensing communication systems, against a broad range of N-acylhomoserine lactones (AHLs). HPLC-MRM data and phylogenetic analysis indicated that the QQ enzyme was an acylase. The QQ activity of strain P6 reduced soft rot symptoms caused by Dickeya solani, Pectobacterium atrosepticum and P. carotovorum on potato and carrot. In vivo assays showed that the PGP and QQ activities of strain P6 protect tomato plants against Pseudomonas syringae pv. tomato, indicating that strain P6 could have biotechnological applications. To our knowledge, this is the first report to show PGP and QQ activities in an indigenous Pseudomonas strain from Salicornia plants.
Asunto(s)
Chenopodiaceae/química , Pseudomonas/patogenicidad , Cromatografía Líquida de Alta Presión , Daucus carota/microbiología , Dickeya , Gammaproteobacteria/patogenicidad , Pectobacterium/patogenicidad , Pectobacterium carotovorum/patogenicidad , Pseudomonas syringae/patogenicidad , Percepción de Quorum/fisiología , Solanum tuberosum/microbiologíaRESUMEN
The geocaulosphere is home to microbes that establish communication between themselves and others that disrupt them. These cell-to-cell communication systems are based on the synthesis and perception of signaling molecules, of which the best known belong to the N-acyl-homoserine lactone (AHL) family. Among indigenous bacteria, certain Gram-positive actinobacteria can sense AHLs produced by soft-rot Gram-negative phytopathogens and can degrade the quorum-sensing AHL signals to impair the expression of virulence factors. We mimicked this interaction by introducing dual-color reporter strains suitable for monitoring both the location of the cells and their quorum-sensing and -quenching activities, in potato tubers. The exchange of AHL signals within the pathogen's cell quorum was clearly detected by the presence of bright green fluorescence instead of blue in a portion of Pectobacterium-tagged cells. This phenomenon in Rhodococcus cells was accompanied by a change from red fluorescence to orange, showing that the disappearance of signaling molecules is due to rhodococcal AHL degradation rather than the inhibition of AHL production. Rhodococci are victorious in this fight for the control of AHL-based communication, as their jamming activity is powerful enough to prevent the onset of disease symptoms.
Asunto(s)
Percepción de Quorum/fisiología , Acil-Butirolactonas/metabolismo , Control Biológico de Vectores , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , Rhodococcus/genética , Rhodococcus/metabolismo , Rhodococcus/fisiología , Solanum tuberosum/microbiología , Factores de Virulencia/metabolismoRESUMEN
The Burkholderia cepacia complex (Bcc) is a family of closely related bacterial pathogens that are the causative agent of deadly human infections. Virulence in Bcc species has been shown to be controlled by the CepI/CepR quorum sensing (QS) system, which is mediated by an N-acyl L-homoserine lactone (AHL) signal (C8-AHL) and its cognate LuxR-type receptor (CepR). Chemical strategies to block QS in Bcc members would represent an approach to intercept this bacterial communication process and further delineate its role in infection. In the current study, we sought to identify non-native AHLs capable of agonizing or antagonizing CepR, and thereby QS, in a Bcc member. We screened a library of AHL analogs in cell-based reporters for CepR, and identified numerous highly potent CepR agonists and antagonists. These compounds remain active in a Bcc member, B. multivorans, with one agonist 250-fold more potent than the native ligand C8-AHL, and can affect QS-controlled motility. Further, the CepR antagonists prolong C. elegans survival in an infection model. These AHL analogs are the first reported non-native molecules that both directly modulate CepR and impact QS-controlled phenotypes in a Bcc member, and represent valuable chemical tools to assess the role of QS in Bcc infections.
Asunto(s)
Proteínas Bacterianas/antagonistas & inhibidores , Complejo Burkholderia cepacia/efectos de los fármacos , Complejo Burkholderia cepacia/patogenicidad , Percepción de Quorum/efectos de los fármacos , Acil-Butirolactonas/metabolismo , Animales , Proteínas Bacterianas/agonistas , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Infecciones por Burkholderia/microbiología , Caenorhabditis elegans/efectos de los fármacos , Evaluación Preclínica de Medicamentos , Escherichia coli/genética , Genes Reporteros , Ligandos , Percepción de Quorum/fisiología , beta-Galactosidasa/genéticaRESUMEN
BACKGROUND: Globally, people utilize plants as the main source of remedy to heal various ailments. Medicinal plants have been utilized to treat ailments since the invention of modern scientific systems of medicine. The common remedy of infectious diseases mainly depends on the inhibition capacity of compounds or killing potential. The issue may give a clue for the development of a novel antimicrobial agent. METHODS: Currently, microorganisms which are resistant towards antibiotics are probably a matter of serious concern for the overall well-being of health. At the moment, new therapeutic targets aside from the microorganism wall-based activities are in progress. For instance, the autoinducer molecules produced by the quorum sensing system are used to control antibiotic resistance and biofilm formation. RESULTS: This therapeutic target is well-studied worldwide, however, the scientific data are not updated and only current studies started to gain insight into its perspective as a target to struggle against infectious diseases. Microbial resistance against antimicrobial compounds is a topic of serious concern in recent time. CONCLUSION: Hence, this paper aims to confer a current overview of the novel compounds, quorum sensing, quorum quenching, biofilm formation in the development of antibiotic resistance and an update on their importance as a potential target for natural substances.
Asunto(s)
Antibacterianos/química , Farmacorresistencia Bacteriana Múltiple/efectos de los fármacos , Extractos Vegetales/química , Plantas Medicinales/química , Animales , Antibacterianos/farmacología , Antiinfecciosos/química , Antiinfecciosos/farmacología , Bacterias/efectos de los fármacos , Bacterias/crecimiento & desarrollo , Biopelículas/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Farmacorresistencia Bacteriana Múltiple/fisiología , Humanos , Extractos Vegetales/farmacología , Percepción de Quorum/efectos de los fármacos , Percepción de Quorum/fisiologíaRESUMEN
Ochrobactrum spp. are ubiquitous bacteria attracting growing attention as important members of microbiomes of plants and nematodes and as a source of enzymes for biotechnology. Strain Ochrobactrum sp. A44T was isolated from the rhizosphere of a field-grown potato in Gelderland, the Netherlands. The strain can interfere with quorum sensing (QS) of Gram-negative bacteria through inactivation of N-acyl homoserine lactones (AHLs) and protect plant tissue against soft rot pathogens, the virulence of which is governed by QS. Phylogenetic analysis based on 16S rRNA gene alone and concatenation of 16S rRNA gene and MLSA genes (groEL and gyrB) revealed that the closest relatives of A44T are O. grignonense OgA9aT, O. thiophenivorans DSM 7216T, O. pseudogrignonense CCUG 30717T, O. pituitosum CCUG 50899T, and O. rhizosphaerae PR17T. Genomes of all six type strains were sequenced, significantly expanding the possibility of genome-based analyses in Ochrobactrum spp. Average nucleotide identity (ANIb) and genome-to-genome distance (GGDC) values for A44T and the related strains were below the single species thresholds (95% and 70%, respectively), with the highest scores obtained for O. pituitosum CCUG 50899T (87.31%; 35.6%), O. rhizosphaerae PR17T (86.80%; 34.3%), and O. grignonense OgA9aT (86.30%; 33.6%). Distinction of A44T from the related type strains was supported by chemotaxonomic and biochemical analyses. Comparative genomics revealed that the core genome for the newly sequenced strains comprises 2731 genes, constituting 50-66% of each individual genome. Through phenotype-to-genotype study, we found that the non-motile strain O. thiophenivorans DSM 7216T lacks a cluster of genes related to flagella formation. Moreover, we explored the genetic background of distinct urease activity among the strains. Here, we propose to establish a novel species Ochrobactrum quorumnocens, with A44T as the type strain (= LMG 30544T = PCM 2957T).
Asunto(s)
Ochrobactrum/genética , Solanum tuberosum/microbiología , Proteínas Bacterianas/genética , ADN Bacteriano/genética , ADN Ribosómico/genética , Flagelos/genética , Genoma Bacteriano , Familia de Multigenes , Países Bajos , Ochrobactrum/clasificación , Ochrobactrum/fisiología , Filogenia , Percepción de Quorum/genética , Percepción de Quorum/fisiología , ARN Ribosómico 16S/genética , Rizosfera , Especificidad de la Especie , Ureasa/genéticaRESUMEN
Following the increasing antibiotic resistance of pathogenic bacteria, the use of medicinal herbs as antibacterial agents has attracted growing attention. Pseudomonas aeruginosa is a human opportunistic pathogen that uses quorum sensing for regulating virulence gene expression (pyocyanin, protease, and elastase production and biofilm formation). This study examined the anti-quorum sensing activity of Quercus infectoria, Zataria multiflora and Trachyspermum copticum extracts on standard P. aeruginosa strain. The minimum inhibitory concentration (MIC) of Q. infectoria, Z. multiflora and T. copticum extracts for standard P. aeruginosa strain was determined through micro dilution. Microtiter plates were used to evaluate the anti-quorum sensing effects of the three extracts (at a sub-MIC concentration) on pyocyanin, protease, and elastase production and biofilm formation. The acetone extract of Q. infectoria showed the highest anti-quorum sensing activity and reduced the pyocyanin, protease, and elastase production and biofilm formation by 89.1%, 78%, 73.3%, and 70.1%, respectively. The corresponding values were 88.2%, 72.1%, 69%, and 61.1% for the methanol extract of Z. multiflora and 70.6%, 63.42%, 60.1%, and 59.1% for the methanol extract of T. copticum. Considering the high anti-quorum sensing activity of the studied extracts, especially the acetone extract of Q. infectoria, these herbs can be used as antipathogenic drugs.
Asunto(s)
Biopelículas/efectos de los fármacos , Extractos Vegetales/farmacología , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/metabolismo , Percepción de Quorum/fisiología , Factores de Virulencia/metabolismo , Carum/química , Lamiaceae/química , Pruebas de Sensibilidad Microbiana , Elastasa Pancreática/metabolismo , Péptido Hidrolasas/metabolismo , Extractos Vegetales/química , Plantas Medicinales/química , Piocianina/metabolismo , Quercus/química , Percepción de Quorum/efectos de los fármacosRESUMEN
Quorum sensing (QS) is a population density-dependent regulatory system in bacteria that couples gene expression to cell density through accumulation of diffusible signaling molecules. Pectobacteria are causal agents of soft rot disease in a range of economically important crops. They rely on QS to coordinate their main virulence factor, production of plant cell wall degrading enzymes (PCWDEs). Plants have evolved an array of antimicrobial compounds to anticipate and cope with pathogens, of which essential oils (EOs) are widely recognized. Here, volatile EOs, carvacrol and eugenol, were shown to specifically interfere with QS, the master regulator of virulence in pectobacteria, resulting in strong inhibition of QS genes, biofilm formation and PCWDEs, thereby leading to impaired infection. Accumulation of the signal molecule N-acylhomoserine lactone declined upon treatment with EOs, suggesting direct interaction of EOs with either homoserine lactone synthase (ExpI) or with the regulatory protein (ExpR). Homology models of both proteins were constructed and docking simulations were performed to test the above hypotheses. The resulting binding modes and docking scores of carvacrol and eugenol support potential binding to ExpI/ExpR, with stronger interactions than previously known inhibitors of both proteins. The results demonstrate the potential involvement of phytochemicals in the control of Pectobacterium.
Asunto(s)
Pectobacterium/efectos de los fármacos , Aceites de Plantas/farmacología , Percepción de Quorum/efectos de los fármacos , 4-Butirolactona/análogos & derivados , 4-Butirolactona/farmacología , Secuencia de Aminoácidos , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Proteínas Bacterianas/fisiología , Biopelículas/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Cimenos , Eugenol/farmacología , Expresión Génica/efectos de los fármacos , Genes Bacterianos , Modelos Moleculares , Monoterpenos/farmacología , Aceites Volátiles/farmacología , Pectobacterium/patogenicidad , Pectobacterium/fisiología , Fenoles/farmacología , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , Poligalacturonasa/antagonistas & inhibidores , Polisacárido Liasas/antagonistas & inhibidores , Percepción de Quorum/genética , Percepción de Quorum/fisiología , Homología de Secuencia de Aminoácido , Homología Estructural de Proteína , Virulencia/efectos de los fármacos , Virulencia/genética , Virulencia/fisiologíaRESUMEN
Thermophilic campylobacters are a major cause of bacterial food-borne diarrhoeal disease. Adherence and biofilm formation are key elements of Campylobacter jejuni persistence in unfavourable environmental conditions. The phytochemical analysis of Euodia ruticarpa fruit ethanol solution extract (EREE) indicated that the major compounds were evodiamine (1), rutaecarpine (2) and evocarpine (9). E. ruticarpa fruit ethanol solution extract, compounds 1 and 2 as well as a mixture of quinolinone alkaloids with 41.7% of 9 were tested for antibacterial, antibiofilm and antiquorum sensing activities against C. jejuni. Minimal inhibitory concentrations varied from 64 to 1024 µg/mL. A mutant strain that lacks the functional gene coding for the CmeB efflux pump protein was the most susceptible. Interestingly, in addition to the wild-type (NCTC 11168) and cmeB mutant, also a mutant that lacks autoinducer-2 production (luxS) was able to adhere (1 h) and to produce a biofilm (24, 48 and 72 h). The subinhibitory concentrations of all preparations at least partly inhibited C. jejuni adhesion and biofilm formation with the most visible effect of the quinolinone alkaloid fraction. Using a Vibrio harveyi luminescence assay, the inhibition of autoinducer-2 production was observed in the wild-type and cmeB mutant after 48 h with the most visible effect of EREE and its fraction Q. Copyright © 2016 John Wiley & Sons, Ltd.
Asunto(s)
Biopelículas/efectos de los fármacos , Campylobacter jejuni/efectos de los fármacos , Evodia/química , Frutas/química , Percepción de Quorum/fisiología , Antibacterianos/farmacologíaRESUMEN
Quorum sensing (QS) is bacterial cell-to-cell communication with small signal molecules such as acyl-homoserine lactones (AHL) that control a number of phenotypes including the regulation of virulence determinants in pathogenic bacteria. Therefore, quorum sensing degrader has been suggested as one of the biocontrol strategies to fight bacterial infections. In the present study, different bacterial QS degrader strains were isolated from Artemia and screened using Chromobacterium violaceum CV026 bioassay. The results showed that six bacterial strains (four Gram-positive and two Gram-negative) isolated from Artemia were able to degrade AHL in two different in vitro assays. All the strains were later identified through 16S rRNA gene sequencing as Rhodococcus opacus, Strepsporangium roseum, Streptomyces alboniger, Enterobacter clocae and Bacillus litoralis. Highest bacterial AHL degrader, Bacillus litoralis BP-ART/6 fully degraded 10 ppm AHL in 9 hrs. The present study showed that bacterial strains isolated from Artemia can act as a QS degrader. ?
Asunto(s)
Artemisia/microbiología , Bacterias/clasificación , Bacterias/aislamiento & purificación , Percepción de Quorum/fisiología , AnimalesRESUMEN
Membrane fouling and energy consumption are interconnected and considered as a bottleneck in membrane bioreactor (MBR) applications. This study investigated synergistic combinations of quorum quenching (QQ) and physical cleaning under different cleaning conditions and aeration intensities with respect to fouling control and energy saving. The MBR operated with periodic air backpulsing had a lower fouling tendency compared with the reactor operated with relaxation. Frequent physical cleanings mitigated the membrane fouling, but irreversible fouling inevitably occurred over time. A significant improvement in fouling control was accomplished when QQ was coupled with physical cleanings, particularly in the filtration/relaxation mode. The submerged QQ vessel helped operate the MBRs stably even at the lowest end of aeration intensity (51 s(-1) in G value), without any significant loss of membrane permeability. The specific membrane filtration energy of the QQMBR remained low and independent of aeration intensities tested, whereas that of the normal MBR sharply increased with decreased aeration rates. The QQMBR with low aeration intensity (51 s(-1)) reduced approximately 27% of the specific aeration energy required for the MBR operated at high aeration intensity (103 s(-1)). QQ bacteria should hamper the formation of a biofilm on the membrane surface, but mixed liquor properties and treatment performances were not affected by the QQ activity.
Asunto(s)
Incrustaciones Biológicas/prevención & control , Reactores Biológicos/microbiología , Metabolismo Energético , Membranas Artificiales , Percepción de Quorum/fisiología , Carbono/análisis , Metabolismo Energético/fisiología , Modelos Biológicos , Fósforo/análisis , Espectroscopía Infrarroja por Transformada de Fourier , Aguas Residuales/químicaRESUMEN
BACKGROUND & OBJECTIVES: Emergence of antimicrobial resistance in Pseudomonas aeruginosa has led to the search for alternative agents for infections control. Natural products have been a good alternative to present antibiotics. The present study was undertaken to evaluate the effectiveness of cranberry in attenuation of virulence of P. aeruginosa in experimental urinary tract infection (UTI) in mouse model. Efforts were also directed to explore the action of cranberry towards virulence of P. aeruginosa through quorum sensing (QS) inhibition. METHODS: Efficacy of cranberry was evaluated in an experimental UTI mouse model and on production of QS signals, alginate, pyochelin, haemolysin, phospholipase-C, cell-surface hydrophobicity, uroepithelial cell-adhesion assay and biofilm formation by already standardized methods. RESULTS: Presence of cranberry showed significant decline in the production of QS signals, biofilm formation and virulence factors of P. aeruginosa in vitro (P<0.001). Further, cranberry was found to be useful in prevention of experimental UTI in mouse model as indicated by reduced renal bacterial colonization and kidney tissues destruction. INTERPRETATION & CONCLUSIONS: The findings of the present study indicated that cranberry inhibited QS and hence elaboration of virulence factors of P. aeruginosa. It also affected the adherence ability of this pathogen. This approach can lead to the discovery of new category of safe anti-bacterial drugs from dietary sources such as cranberry with reduced toxicity without the risk of antibiotic resistance.
Asunto(s)
Frutas/química , Extractos Vegetales/farmacología , Infecciones por Pseudomonas/prevención & control , Pseudomonas aeruginosa/efectos de los fármacos , Percepción de Quorum/efectos de los fármacos , Infecciones Urinarias/microbiología , Vaccinium macrocarpon/química , Animales , Biopelículas/efectos de los fármacos , Proteínas Hemolisinas/metabolismo , Ratones , Microscopía Electrónica de Rastreo , Fenoles/metabolismo , Percepción de Quorum/fisiología , Tiazoles/metabolismo , Virulencia , Factores de Virulencia/metabolismoRESUMEN
Essential oils (EOs) form an important part of traditional medicine so their anti-microbial and, in the recent past, antiquorum sensing activity has been well studied. However it is likely that due to their hydrophobic nature and reduced solubility in aqueous environments full potential of their activity cannot be realized. hence it is only rational to formulate a process to make these molecules more polar in nature. The present paper reports synthesis of sophorolipids using 12 different essential oils as substrates, thus providing surfactant-like properties to these EOs. The synthesis protocol makes the use of Candida bombicola ATCC 22214 as producer organism. The production process required 7 days of incubation at 28°C and 180 rpm. Preliminary characterization of the synthesized essential oil sophorolipids (EOSLs) was performed using thin layer chromatography (TLC) and Fourier transform infrared spectroscopy (FTIR). Additionally, essential oils that were incapable of mediating quorum sensing inhibition (QSI) on their own became potent quorum sensing inhibitors upon conversion into their corresponding EOSLs. Antibiofilm potential of these EOSLs was also demonstrated using V. cholerae as test organism. Use of essential oils as substrates for glycolipid synthesis has not been attempted previously, and hence this is the first report.
Asunto(s)
Biopelículas/efectos de los fármacos , Glucolípidos/síntesis química , Aceites Volátiles/síntesis química , Percepción de Quorum/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Candida/fisiología , Glucolípidos/farmacología , Aceites Volátiles/farmacología , Percepción de Quorum/fisiología , Espectroscopía Infrarroja por Transformada de Fourier/métodosRESUMEN
BACKGROUND: Quorum sensing (QS), as the basis of bacterial cell-to-cell communication, is a promising approach to reduce the incidence of multidrug resistance. The objective of this study was to search for novel quorum sensing inhibitors from plants and control detrimental infections. METHODS: The crude extracts of Ficus carica and Perilla frutescens were examined for their anti-QS properties. Powdered plant samples were treated sequentially with organic solvents of increasing polarity. The extracts of each solvent were concentrated in vacuo to give crude extracts and tested against Chromobacterium violaceum CV026 and Pseudomonas aeruginosa PA01 especially. Anti-QS activity was measured by quantifying violacein production and swarming motility. RESULTS: All extracts of these two plants display anti-QS ability. Interestingly, the extract of F. carica with dichloromethane and of P. frutescens with MeOH exhibited the most pronounced inhibition of QS activity. CONCLUSIONS: These two plants can offer bioactive natural products with potential for attenuating pathogens.
Asunto(s)
Antibacterianos/farmacología , Medicamentos Herbarios Chinos/farmacología , Ficus , Perilla frutescens , Percepción de Quorum/efectos de los fármacos , Antibacterianos/aislamiento & purificación , Chromobacterium/efectos de los fármacos , Chromobacterium/fisiología , Medicamentos Herbarios Chinos/aislamiento & purificación , Humanos , Hojas de la Planta , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/fisiología , Percepción de Quorum/fisiologíaRESUMEN
Extracts of Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum, inhibited quorum sensing in Chromobacterium violaceum CV026. G. lucidum fruiting bodies were milled and extracted with ethyl acetate. The crude extract was dissolved in an appropriate concentration of methanol, sterilized by filtration through a 0.22-µm membrane filter, and added to Ch. Violaceum CV026 cultures, which were used as an indicator to monitor quorum sensing inhibition. Inhibitory activity was measured by quantifying violacein production using a microplate reader. Methanol-soluble compounds extracted from G. lucidum significantly inhibited quorum sensing-controlled behavior in Ch. Violaceum in a concentration-dependent manner. The results suggest that compounds in G. lucidum might be useful to control and handle detrimental infections caused by human, animal, and plant pathogens. Further studies are in progress in our lab to isolate the specific compounds from G. lucidum extract, evaluate them as quorum sensing inhibitors, and analyze their mechanism of action.
Asunto(s)
Chromobacterium/efectos de los fármacos , Chromobacterium/fisiología , Cuerpos Fructíferos de los Hongos/química , Ganoderma/química , Percepción de Quorum/fisiologíaRESUMEN
In recent years, antibiotic resistance of bacteria has become a global health crisis. Especially, the new class of "superbug" was found in South Asia, which is resistant to almost known antibiotics and causes worldwide alarm. Through the underlying mechanisms of bacterial pathogenecity, the expression of many pathogen virulence factors is regulated by the process of quorum sensing. Screening efficient quorum sensing inhibitors is an especially compelling approach to the future treatment of bacterial infections and antibiotic resistance. This article focuses on bacterial quorum sensing system, quorum sensing screening model for in vitro and evaluation of animal models in vivo, recent research of quorum sensing inhibitors and so on.
Asunto(s)
Antibacterianos/farmacología , Infecciones Bacterianas , Farmacorresistencia Bacteriana , Pseudomonas aeruginosa/fisiología , Percepción de Quorum/efectos de los fármacos , Animales , Antibacterianos/uso terapéutico , Infecciones Bacterianas/tratamiento farmacológico , Modelos Animales de Enfermedad , Medicamentos Herbarios Chinos/farmacología , Humanos , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/patogenicidad , Percepción de Quorum/fisiología , Virulencia/efectos de los fármacos , Factores de Virulencia/metabolismoRESUMEN
Quorum sensing (QS) is a process by which bacteria use small molecules or peptidic signals to assess their local population densities. At sufficiently high density, bacteria can alter gene expression levels to regulate group behaviors involved in a range of important and diverse phenotypes, including virulence factor production, biofilm formation, root nodulation, and bioluminescence. Gram-negative bacteria most commonly use N-acylated l-homoserine lactones (AHLs) as their QS signals. The AHL lactone ring is hydrolyzed relatively rapidly at biological pH, and the ring-opened product is QS inactive. We seek to identify AHL analogues with heightened hydrolytic stability, and thereby potentially heightened activity, for use as non-native modulators of bacterial QS. As part of this effort, we probed the utility of thiolactone analogues in the current study as QS agonists and antagonists in Gram-negative bacteria. A focused library of thiolactone analogs was designed and rapidly synthesized in solution. We examined the activity of the library as agonists and antagonists of LuxR-type QS receptors in Pseudomonas aeruginosa (LasR), Vibrio fischeri (LuxR), and Agrobacterium tumefaciens (TraR) using bacterial reporter strains. The thiolactone library contained several highly active compounds, including some of the most active LuxR inhibitors and the most active synthetic TraR agonist reported to date. Analysis of a representative thiolactone analog revealed that its hydrolysis half-life was almost double that of its parent AHL in bacterial growth medium.
Asunto(s)
Acil-Butirolactonas/síntesis química , Bacterias Gramnegativas/efectos de los fármacos , Pseudomonas aeruginosa/efectos de los fármacos , Percepción de Quorum/efectos de los fármacos , Proteínas Represoras/efectos de los fármacos , Transactivadores/efectos de los fármacos , Acetilación , Acil-Butirolactonas/química , Acil-Butirolactonas/farmacocinética , Acil-Butirolactonas/farmacología , Relación Dosis-Respuesta a Droga , Diseño de Fármacos , Evaluación Preclínica de Medicamentos , Genes Reporteros/efectos de los fármacos , Bacterias Gramnegativas/genética , Bacterias Gramnegativas/metabolismo , Hidrólisis , Terapia Molecular Dirigida , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/metabolismo , Percepción de Quorum/fisiología , Proteínas Represoras/agonistas , Proteínas Represoras/antagonistas & inhibidores , Proteínas Represoras/fisiología , Transactivadores/agonistas , Transactivadores/antagonistas & inhibidores , Transactivadores/fisiologíaRESUMEN
AIMS: This study aimed to search for novel quorum sensing (QS) inhibitors from mushroom and to analyze their inhibitory activity, with a view to their possible use in controlling detrimental infections. METHODS: The bioactive metabolites produced by mushroom cultivation were tested for their abilities to inhibit QS-regulated behavior. All mushroom strains were cultivated in potato-dextrose medium by large-scale submerged fermentation. The culture supernatant was condensed into 0.2 vol by freeze-drying. The condensed supernatant was sterilized by filtration through a 0.22-µm membrane filter and added to Chromobacterium violaceum CV026 cultures, which were used to monitor QS inhibition. Inhibitory activity was measured by quantifying violacein production using a microplate reader. RESULTS: The results have revealed that, of 102 mushroom strains, the bioactive metabolites produced by 14 basidiomycetes were found to inhibit violacein production, a QS-regulated behavior in C. violaceum. CONCLUSIONS: Higher fungi can produce QS-inhibitory compounds.