Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 6.096
Filtrar
1.
J Microbiol Biotechnol ; 29(8): 1299-1309, 2019 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-31387340

RESUMO

As an opportunistic bacterial pathogen, Pseudomonas aeruginosa PAO1 contains two phenazineproducing gene operons, phzA1B1C1D1E1F1G1 (phz1) and phzA2B2C2D2E2F2G2 (phz2), each of which is independently capable of encoding all enzymes for biosynthesizing phenazines, including phenazine-1-carboxylic acid and its derivatives. Other previous study reported that the RpoS-deficient mutant SS24 overproduced pyocyanin, a derivative of phenazine-1- carboxylic acid. However, it is not known how RpoS mediates the expression of two phz operons and regulates pyocyanin biosynthesis in detail. In this study, with deletion of the rpoS gene in the PAΔphz1 mutant and the PAΔphz2 mutant respectively, we demonstrated that RpoS exerted opposite regulatory roles on the expression of the phz1and phz2 operons. We also confirmed that the phz1 operon played a critical role and especially biosynthesized much more phenazines than the phz2 operon when the rpoS gene was knocked out in P. aeruginosa. By constructing the translational reporter fusion vector lasR'-'lacZ and the chromosomal fusion mutant PAΔlasR::lacZ, we verified that RpoS deficiency caused increased expression of lasR, a transcription regulator gene in a first quorum sensing system (las) that activates overexpression of the phz1 operon, suggesting that in the absence of RpoS, LasR might act as an intermediate in overproduction of phenazine biosynthesis mediated by the phz1 operon in P. aeruginosa.


Assuntos
Proteínas de Bactérias/metabolismo , Fenazinas/metabolismo , Pseudomonas aeruginosa/metabolismo , Fator sigma/metabolismo , Transativadores/metabolismo , Proteínas de Bactérias/genética , Regulação Bacteriana da Expressão Gênica , Fusão Gênica , Óperon , Pseudomonas aeruginosa/genética , Piocianina/biossíntese , Fator sigma/genética
2.
World J Microbiol Biotechnol ; 35(9): 137, 2019 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-31432268

RESUMO

The presence of very high concentrations of organic pollutants, phenols, tannins and heavy metals mainly chromium in wastewater discharged from leather industries, tags it as one of the most polluting industries. The phenolic syntans discharged from tanning units have an adverse effect on living organisms and cause serious environmental pollution, thereby making it very imperative to remove it. Among various treatment methods available for removal of phenols, biodegradation is environment friendly. The present study aims at the remediation of phenolic syntan used in the leather industry employing individual as well as co-culture of Bacillus cereus and Pseudomonas aeruginosa at varying syntan concentration in the medium. Parameters such as chemical oxygen demand (COD), total organic carbon (TOC), total phenol content (TPC) and Fourier Transform Infrared Spectroscopy (FTIR) indicating biodegradation were analyzed. Promising results were observed with P. aeruginosa, which exhibited a reduction in TPC by 62-72% in all the concentrations of syntan tested just within 12 h of inoculation, whereas about 67 and 83% reduction in COD and TOC respectively was observed for 2000 ppm concentration at the end of 5 days. B. cereus also demonstrated very good reduction in the above parameters however; percentage was less as compared to P. aeruginosa. In the case of co-culture, the TPC reduction was higher than B. cereus but lesser than P. aeruginosa. The percentage reduction in TOC and COD was highest for 500 ppm which eventually decreased for subsequent concentrations.


Assuntos
Bacillus cereus/metabolismo , Resíduos Industriais , Fenóis/metabolismo , Pseudomonas aeruginosa/metabolismo , Poluentes Químicos da Água/metabolismo , Bacillus cereus/crescimento & desenvolvimento , Análise da Demanda Biológica de Oxigênio , Biotransformação , Carbono/análise , Técnicas de Cocultura , Fenóis/análise , Pseudomonas aeruginosa/crescimento & desenvolvimento , Espectroscopia de Infravermelho com Transformada de Fourier , Água/química
3.
Microbiol Res ; 228: 126301, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31422232

RESUMO

The in vitro inhibition of quorum sensing signal, xanthan gum secretion, biofilm formation in different Xanthomonas pathovars and biological control of bacterial blight of rice by the two bioactive extrolites produced by Pseudomonas aeruginosa strain CGK-KS-1 were explored. These extrolites were extracted from Diaion HP-20 resin with methanol and purified by preparative-thin layer chromatography. Further, spectroscopic structural elucidation revealed the tentative identity of these extrolites to be (R,3E,5E,9Z,11E)-13-((3S,5R)-5-acetyl-2,6-dimethylheptan-3-yl)-10-hydroxy-4-methyl-1,8-diazabicyclo[9.3.1]pentadeca-3,5,9,11(15),13-pentaen-2-one and (R,3E,5E,8E,11E)-13-((3S,5R)-5-acetyl-2,6-dimethylheptan-3-yl)-4-methyl-1,8-diazabicyclo[9.3.1]pentadeca-3,5,8,11(15),13-pentaene-2,10-dione, named as Chumacin-1 and Chumacin-2, respectively. Antimicrobial assay showed Chumacin-1 and Chumacin-2 exhibited a strong in vitro growth inhibition against various Xanthomonas pathovars. Quorum sensing overlay assay using a reporter strain Chromobacterium violaceum strain CV026 showed that Chumacin-1 and Chumacin-2 inhibited quorum sensing signaling. The mechanistic studies revealed that these extrolites inhibited the production of quorum sensing signaling factor, cis-11-methyl-2-dodecenoic acid; suppressed the xanthan gum secretion and also inhibited the biofilms formed by various Xanthomonas pathovars. Both Chumacin-1 and Chumacin-2 showed ROS generation in the test Xanthomonas strains, resulting in in vitro cell membrane damage was revealed through CSLM and FE-SEM micrographs. Further, greenhouse experiments using Samba Mashuri (BPT-5204) revealed that seed treatment with Chumacin-1 and Chumacin-2 along with foliar spray groups showed up to ˜80% reduction in bacterial blight disease in rice. To the best of our knowledge, this is the first report on new quorum sensing inhibitors, Chumacin-1 and Chumacin-2 produced by Pseudomonas aeruginosa strain CGK-KS-1 exhibiting DSF inhibition activity in Xanthomonas oryzae pv. oryzae.


Assuntos
Agentes de Controle Biológico/isolamento & purificação , Agentes de Controle Biológico/farmacologia , Oryza/microbiologia , Doenças das Plantas/prevenção & controle , Pseudomonas aeruginosa/metabolismo , Percepção de Quorum/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Xanthomonas/efeitos dos fármacos , Anti-Infecciosos/química , Anti-Infecciosos/isolamento & purificação , Anti-Infecciosos/metabolismo , Anti-Infecciosos/farmacologia , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Agentes de Controle Biológico/química , Chromobacterium/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Simulação de Acoplamento Molecular , Doenças das Plantas/microbiologia , Polissacarídeos Bacterianos/metabolismo , Poliestirenos , Xanthomonas/metabolismo
4.
J Agric Food Chem ; 67(29): 8191-8196, 2019 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-31282662

RESUMO

Conversion of free fatty acids into monoacylglycerol gives rise to new structural properties, particularly amphipathic property. Therefore, monoacylglycerols are widely used in pharmaceutical and food industries and are also reported to facilitate better absorption into the human body. A functional fatty acid when transformed into a monoacylglycerol will possibly conserve both the original functionality and amphipathic property. The compound 7,10-dihydroxy-8(E)-octadecenoic acid (DOD) was generated from oleic acid by Pseudomonas aeruginosa PR3 and was known to contain antimicrobial activities against a broad range of food-borne and plant pathogenic bacteria. Here, we attempted to convert DOD into its monoacylglycerol form using lipase for producing an amphipathic antibacterial agent. Consequently, the monoacylglycerol of DOD (DOD-MAG) was successfully produced by coincubating DOD, glycerol, and lipase at 30 °C. The maximum conversion yield reached 70% after 12 h of incubation. Antibacterial activity of DOD-MAG was enhanced by 8 times from the original activity of DOD against food-borne bacteria.


Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Monoglicerídeos/química , Ácidos Oleicos/química , Ácidos Oleicos/farmacologia , Pseudomonas aeruginosa/química , Antibacterianos/metabolismo , Microbiologia de Alimentos , Ácidos Oleicos/metabolismo , Pseudomonas aeruginosa/metabolismo , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/crescimento & desenvolvimento
5.
Nat Commun ; 10(1): 2931, 2019 07 03.
Artigo em Inglês | MEDLINE | ID: mdl-31270321

RESUMO

The virulence of Pseudomonas aeruginosa, a Gram-negative opportunistic pathogen, is regulated by many transcriptional factors (TFs) that control the expression of quorum sensing and protein secretion systems. Here, we report a genome-wide, network-based approach to dissect the crosstalk between 20 key virulence-related TFs. Using chromatin immunoprecipitation coupled with high-throughput sequencing (ChIP-seq), as well as RNA-seq, we identify 1200 TF-bound genes and 4775 differentially expressed genes. We experimentally validate 347 of these genes as functional target genes, and describe the regulatory relationships of the 20 TFs with their targets in a network that we call 'Pseudomonas aeruginosa genomic regulatory network' (PAGnet). Analysis of the network led to the identification of novel functions for two TFs (ExsA and GacA) in quorum sensing and nitrogen metabolism. Furthermore, we present an online platform and R package based on PAGnet to facilitate updating and user-customised analyses.


Assuntos
Proteínas de Bactérias/genética , Pseudomonas aeruginosa/genética , Fatores de Transcrição/genética , Proteínas de Bactérias/metabolismo , Imunoprecipitação da Cromatina , Regulação Bacteriana da Expressão Gênica , Genoma Bacteriano , Genômica , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/metabolismo , Pseudomonas aeruginosa/patogenicidade , Fatores de Transcrição/metabolismo , Virulência , Fatores de Virulência/genética , Fatores de Virulência/metabolismo
6.
Environ Sci Pollut Res Int ; 26(24): 24695-24706, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31240645

RESUMO

The biotoxicity of heavy metals in sediments toward benthic organisms has evoked great concern for the health of freshwater ecosystems. This study applied a sediment toxicity testing protocol to investigate the single and joint toxicity of cadmium (Cd) and lead (Pb) on Bellamya aeruginosa. B. aeruginosa were exposed to different concentrations of Cd (5, 25, and 100 mg/kg), Pb (20, 100, and 400 mg/kg), and their different concentration combinations. A suite of biomarkers, including superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), metallothionein (MT), malondialdehyde (MDA), and acetylcholinesterase (AChE), were measured after 7, 14, 21, and 28 days of exposure to evaluate their oxidative stress status. Cell apoptosis of soft tissue was also determined after exposure. Results revealed that these endpoints represented sensitive biomarkers for the characterization of the oxidative stress response induced by these metals. Specifically, a decrease of SOD and GPx and an increase of MDA were indicative of the potential failure of the antioxidant defense system in neutralizing the reactive oxygen species (ROS) generated in the exposure of the Pb-treated group. The integrated biomarker response (IBR) index revealed the most significant sub-lethal toxicity for Pb-spiked sediments, leading to the highest rate of cell apoptosis (70.8%). Exposure to Cd resulted in a time- and dose-dependent effect on MT levels, which suggested active detoxification of this metal. Exposure to the mixture resulted in amelioration of Pb toxicity, likely due to the competitive binding of Cd to active enzyme, with the result of an observed antagonistic interaction. This study indicated that B. aeruginosa represents a good biomonitor for assessing Cd and Pb contamination of sediments, and laid the foundation for their potential risk assessments in freshwater ecosystems.


Assuntos
Cádmio/toxicidade , Chumbo/toxicidade , Metalotioneína/metabolismo , Metais Pesados/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Pseudomonas aeruginosa/metabolismo , Animais , Biomarcadores/metabolismo , Catalase/metabolismo , Ecossistema , Água Doce , Glutationa Peroxidase/metabolismo , Malondialdeído/metabolismo , Metalotioneína/química , Pseudomonas aeruginosa/química , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismo
7.
Nat Commun ; 10(1): 2853, 2019 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-31253808

RESUMO

Plant innate immunity restricts growth of bacterial pathogens that threaten global food security. However, the mechanisms by which plant immunity suppresses bacterial growth remain enigmatic. Here we show that Arabidopsis thaliana secreted aspartic protease 1 and 2 (SAP1 and SAP2) cleave the evolutionarily conserved bacterial protein MucD to redundantly inhibit the growth of the bacterial pathogen Pseudomonas syringae. Antibacterial activity of SAP1 requires its protease activity in planta and in vitro. Plants overexpressing SAP1 exhibit enhanced MucD cleavage and resistance but incur no penalties in growth and reproduction, while sap1 sap2 double mutant plants exhibit compromised MucD cleavage and resistance against P. syringae. P. syringae lacking mucD shows compromised growth in planta and in vitro. Notably, growth of ΔmucD complemented with the non-cleavable MucDF106Y is not affected by SAP activity in planta and in vitro. Our findings identify the genetic factors and biochemical process underlying an antibacterial mechanism in plants.


Assuntos
Arabidopsis/metabolismo , Arabidopsis/microbiologia , Proteínas de Bactérias/metabolismo , Peptídeo Hidrolases/metabolismo , Doenças das Plantas/microbiologia , Serina Endopeptidases/metabolismo , Arabidopsis/imunologia , Proteínas de Bactérias/genética , Evolução Molecular , Regulação Enzimológica da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Filogenia , Doenças das Plantas/imunologia , Plantas Geneticamente Modificadas , Pseudomonas aeruginosa/enzimologia , Pseudomonas aeruginosa/metabolismo , Serina Endopeptidases/genética
8.
Microb Pathog ; 132: 230-242, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31082528

RESUMO

Virulence pathways in gram-negative pathogenic bacteria are regulated by quorum sensing mechanisms, through the production and sensing of N-acylhomoserine lactone (AHL) signal molecules. Enzymatic degradation to disrupt quorum-sensing in these bacteria could pave the way for the new development in decreasing resistance strains and are of significant interest for clinical, agricultural, and industrial applications. Isolated endophytic Bacillus thuringiensis strain KMCL07 showing quorum quenching activity on Pseudomonas aeruginosa PAO1 has been studied. AiiA lactonase KMMI17 identified belongs to metallo- ß-lactamase superfamily preserving conserved regions of 106HXDH-59 amino acids-H169-21 amino acids-D191 motif, significantly inhibits the biofilm formation and attenuates virulence factor pyocyanin production of PAO1. Insilico molecular docking analysis of lactonase KMMI17 using alternative catalytic site (PDB entry: 3DHA) with the AHL-based QS system regulators of PAO-1, C4 AHL, C6 AHL and 3-oxo-C12 AHL molecules showed good binding affinity between the protein and ligands, Phe111 and Tyr198 residues plays an important role in binding them. Crude enzyme extract was found to have Km value for C6-HSL: 134.2702 ±â€¯34.83 µM-1, C4-HSL: 308.217 ±â€¯139.9 µM-1 and 3-oxo-C12-HSL: 760.463 ±â€¯251.3 µM-1. LCMS analysis confirms the degradation activity of lactonase KMMI17 on AHL molecules and its hydrolytic process, which indicates the potential application of lactonase KMMI17 as a biocontrol agent or an anti-pathogenic drug.


Assuntos
Bacillus thuringiensis/metabolismo , Proteínas de Bactérias/antagonistas & inibidores , Lactonas/metabolismo , Metaloendopeptidases/antagonistas & inibidores , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/metabolismo , Percepção de Quorum/fisiologia , Proteínas de Bactérias/metabolismo , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Hidrolases de Éster Carboxílico/metabolismo , Índia , Madhuca/microbiologia , Metaloendopeptidases/metabolismo , Simulação de Acoplamento Molecular , Fenótipo , Pseudomonas aeruginosa/crescimento & desenvolvimento , Pseudomonas aeruginosa/patogenicidade , Piocianina/metabolismo , Fatores de Virulência
9.
Lett Appl Microbiol ; 69(1): 79-86, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31077423

RESUMO

Considering that bacterial biosurfactants (BSFs) are released as secondary metabolites involved in biotic relations within mixed bacterial assemblages, the hypothesis that the co-cultivation of BSF producing bacteria with biofilm-forming strains would enhance BSF synthesis was tested. Environmental BSF producing strains of Bacillus licheniformis and Pseudomonas sp. were cultivated with reference biofilm-forming strains (Pseudomonas aeruginosa and Listeria innocua). BSF production and quorum-quenching effects were tested in solid media. Tensioactive and anionic BSFs were also quantified in cell-free extracts (CFEs). BSF production increased in co-cultures with inducer strains although this was not demonstrated by all screening methods. Increased concentrations of anionic BSF were detected in CFEs of co-cultures in which Pseudomonas aeruginosa was included as inducer, which is in accordance with the observation of larger halos in cetyl trimethylammonium bromide-methylene blue agar. The results demonstrate that co-cultivation positively affects the efficiency of BSF production and that higher production yields may be attained by selecting convenient inducer partners in designed consortia. SIGNIFICANCE AND IMPACT OF THE STUDY: The high production cost of biosurfactants (BSFs) still represents a major limitation to the industrial use of these otherwise advantageous alternatives to chemical surfactants. This work demonstrates that the co-cultivation of consortia of biosurfactant-producer and biofilm-forming bacteria enhances BSF production and may contribute to the cost-effectiveness of biosurfactant-based products.


Assuntos
Bacillus licheniformis/metabolismo , Biofilmes/crescimento & desenvolvimento , Listeria/metabolismo , Pseudomonas aeruginosa/metabolismo , Tensoativos/metabolismo , Ágar/metabolismo , Técnicas de Cocultura , Percepção de Quorum
10.
Talanta ; 200: 537-546, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31036220

RESUMO

Liquid chromatography-mass spectrometry based profiling of microbial metabolites has been a challenging task due to their diverse physicochemical properties and wide concentration ranges. This study is aimed to develop a systematic platform for the broad-scale profiling of microbial metabolites by integrating aqueous-lipophilic biphasic extractions and chemical derivatizations with a data-dependent automatable metabolite annotation algorithm. This complementary strategy of detection will not only largely expand the metabolite coverage, but also facilitate the drawing out of interested submetabolome using designed chemical derivatizations. Then, the data-dependent metabolite annotation algorithm is able to automatically match the raw MS/MS data with those of compounds in the self-collected databases. The performance of this platform is illustrated through the analysis of two representative bacteria (Escherichia coli and Pseudomonas aeruginosa) and intestinal contents samples from experimental colitis mice. As a result, 292 metabolites corresponding to 875 annotated features distributing over 25 chemical families were putatively annotated in a short time. Of these metabolites, 197 and 218 are respectively from the bacteria and intestinal contents, and 107 are identified in all three biological samples. This systematic platform could be used to accomplete high-coverage detection and high-quality data processing of microbial metabolites. At the same time, chemical derivatization design and the establishment of self-collected databases will facilitate self-driven untargeted analysis.


Assuntos
Colite/metabolismo , Escherichia coli/metabolismo , Pseudomonas aeruginosa/metabolismo , Animais , Cromatografia Líquida de Alta Pressão , Colite/induzido quimicamente , Colite/microbiologia , Sulfato de Dextrana , Espectrometria de Massas , Camundongos
11.
Can J Microbiol ; 65(8): 563-574, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31009577

RESUMO

Pseudomonas aeruginosa is a virulent bacterium that secretes a variety of virulence factors that aid in establishing infections in individuals. Allicin, derived from garlic, has been shown to inhibit virulence factor production and biofilm formation in P. aeruginosa. However, the mechanisms underlying the allicin-mediated regulation of P. aeruginosa virulence remain unclear. In this study, we investigated the possible mechanisms underlying allicin-mediated virulence regulation in P. aeruginosa. The results showed that allicin attenuates the production of P. aeruginosa virulence-associated factors, such as elastase, pyocyanin, pyoverdine, and rhamnolipids, by inhibiting the rhl and pqs quorum-sensing systems. Further analysis revealed that the rhl and pqs systems play different roles during the allicin-mediated regulation process. Taken together, these results support the potential use of allicin as a therapeutic agent in controlling P. aeruginosa infection and associated mechanisms.


Assuntos
Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/patogenicidade , Percepção de Quorum/efeitos dos fármacos , Ácidos Sulfínicos/farmacologia , Fatores de Virulência/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Glicolipídeos/metabolismo , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/metabolismo , Piocianina/metabolismo , Virulência/efeitos dos fármacos , Fatores de Virulência/genética
12.
J Ind Microbiol Biotechnol ; 46(7): 1025-1038, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30989356

RESUMO

Coenzyme Q (ubiquinone) is a redox-active isoprenylated benzoquinone commonly found in living organisms. The biosynthetic pathway for this lipid has been extensively studied in Escherichia coli and Saccharomyces cerevisiae; however, little is known in Pseudomonas aeruginosa. In this study, we observed that CoQ9 is the predominant coenzyme Q synthesized by the Shenqinmycin-producing strain M18. BLASTP and domain organization analyses identified 15 putative genes for CoQ biosynthesis in M18. The roles of 5 of these genes were genetically and biochemically investigated. PAM18_4662 encodes a nonaprenyl diphosphate synthase (Nds) and determines the number of isoprenoid units of CoQ9 in M18. PAM18_0636 (coq7PA) and PAM18_5179 (ubiJPA) are essential for aerobic growth and CoQ9 biosynthesis. Deletion of ubiJPA, ubiBPA and ubiKPA led to reduced CoQ biosynthesis and an accumulation of the CoQ9 biosynthetic intermediate 3-nonaprenylphenol (NPP). Moreover, we also provide evidence that the truncated UbiJPA interacts with UbiBPA and UbiKPA to affect CoQ9 biosynthesis by forming a regulatory complex. The genetic diversity of coenzyme Q biosynthesis may provide targets for the future design of specific drugs to prevent P. aeruginosa-related infections.


Assuntos
Agentes de Controle Biológico/metabolismo , Pseudomonas aeruginosa/metabolismo , Ubiquinona/biossíntese , Vias Biossintéticas/genética , Pseudomonas aeruginosa/genética
13.
Int J Biol Macromol ; 132: 1221-1234, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-30946905

RESUMO

In the present investigation functional chitosan/silver nanocomposites (CS/Ag NCs) were successfully synthesized and found to possess favorable antibacterial activity against extended spectrum beta-lactasame (ESBL) producing Pseudomonas aeruginosa. Powder X-ray diffraction showed that the obtained CS/Ag NCs are constituted of highly crystalline Ag nanoparticles (NPs) embedded in an amorphous CS matrix material. Transmission electron microscopy (TEM) analysis provided structural information about CS/Ag NCs, revealing the formation of spherical cluster structures constituted of Ag NPs with size ranging from 6 to 18 nm embedded in the amorphous CS matrix. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Ag NPs and CS/Ag NCs were found to inhibit the ESBL producing P. aeruginosa at 80 µg/mL (76%) and 50 µg/mL (92%), respectively. Confocal laser scanning microscope (CLSM) and scanning electron microscopy (SEM) images revealed that P. aeruginosa experienced reduced cell viability and morphological cell membrane damage at desired MIC. The in-vivo toxicity effect of Ag NPs and CS/Ag NCs suggested an increased mortality rate when Artemia franciscana were exposed for 24 h to increasing concentrations of Ag NPs and CS/Ag NCs. Anti-ESBL activity and toxicity effect of CS/Ag NCs revealed that these NCs possess promising antibacterial properties to overcome numerous communicable bacterial strains.


Assuntos
Carbapenêmicos/farmacologia , Quitosana/química , Farmacorresistência Bacteriana/efeitos dos fármacos , Nanocompostos/química , Pseudomonas aeruginosa/efeitos dos fármacos , Prata/química , beta-Lactamas/metabolismo , Antibacterianos/química , Antibacterianos/farmacologia , Nanopartículas Metálicas/química , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa/metabolismo
14.
Molecules ; 24(7)2019 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-30979013

RESUMO

Rhamnolipids are a mixture of the homologs species due to variations in the rhamnose units and ß-hydroxy fatty acid moieties, mainly including Rha-C10-C10, Rha-Rha-C10-C10, and Rha-C10. In this study, strain P. aeruginosa YM4 was selected for its capacity to efficiently produce di-rhamnolipid (Rha-Rha-C10-C10) as the predominant component with soybean oil and glycerol as carbon source, accounting for 64.8% and 85.7% of total products, respectively. The critical micelle concentration (CMC) of rhamnolipid products varies with the content of di-rhamnolipid, whereby lower CMC values corresponding to higher di-rhamnolipid contents. The rhamnolipids containing 85.7% di-rhamnolipid had the lowest CMC value of 50 mg/L. Accordingly the viscosity-reducing efficiency and oil-washing efficiency of rhamnolipids increased with higher di-rhamnolipid component. At a concentration of 500 mg/L, the rhamnolipids containing 85.7% di-rhamnolipid worked best and showed 82.5% oil-washing efficiency, which offered great promise for applications in enhanced oil recovery. The results showed the variation of structure and composition of rhamnolipids had a significant effect on their application.


Assuntos
Glicolipídeos/biossíntese , Poluição por Petróleo/prevenção & controle , Pseudomonas aeruginosa/metabolismo , Ramnose/biossíntese , Carbono/química , Ácidos Graxos/química , Glicerol/química , Glicolipídeos/química , Humanos , Pseudomonas aeruginosa/química , Pseudomonas aeruginosa/genética , Ramnose/química , Óleo de Soja/química , Tensoativos/química
15.
Environ Pollut ; 250: 262-273, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30999203

RESUMO

Dibenzofuran (DBF) derivatives have caused serious environmental problems, especially those produced by paper pulp bleaching and incineration processes. Prominent for its resilient mutagenicity and toxicity, DBF poses a major challenge to human health. In the present study, a new strain of Pseudomonas aeruginosa, FA-HZ1, with high DBF-degrading activity was isolated and identified. The determined optimum conditions for cell growth of strain FA-HZ1 were a temperature of 30 °C, pH 5.0, rotation rate of 200 rpm and 0.1 mM DBF as a carbon source. The biochemical and physiological features as well as usage of different carbon sources by FA-HZ1 were studied. The new strain was positive for arginine double hydrolase, gelatinase and citric acid, while it was negative for urease and lysine decarboxylase. It could utilize citric acid as its sole carbon source, but was negative for indole and H2S production. Intermediates of DBF 1,2-dihydroxy-1,2-dihydrodibenzofuran, 1,2-dihydroxydibenzofuran, 2-hydroxy-4-(3'-oxo-3'H-benzofuran-2'-yliden)but-2-enoic acid, 2,3-dihydroxybenzofuran, 2-oxo-2-(2'-hydrophenyl)lactic acid, and 2-hydroxy-2-(2'-hydroxyphenyl)acetic acid were detected and identified through liquid chromatography-mass analyses. FA-HZ1 metabolizes DBF by both the angular and lateral dioxygenation pathways. The genomic study identified 158 genes that were involved in the catabolism of aromatic compounds. To identify the key genes responsible for DBF degradation, a proteomic study was performed. A total of 1459 proteins were identified in strain FA-HZ1, of which 100 were up-regulated and 104 were down-regulated. A novel enzyme "HZ6359 dioxygenase", was amplified and expressed in pET-28a in E. coli BL21(DE3). The recombinant plasmid was successfully constructed, and was used for further experiments to verify its function. In addition, the strain FA-HZ1 can also degrade halogenated analogues such as 2, 8-dibromo dibenzofuran and 4-(4-bromophenyl) dibenzofuran. Undoubtedly, the isolation and characterization of new strain and the designed pathways is significant, as it could lead to the development of cost-effective and alternative remediation strategies. The degradation pathway of DBF by P. aeruginosa FA-HZ1 is a promising tool of biotechnological and environmental significance.


Assuntos
Benzofuranos/metabolismo , Poluentes Ambientais/metabolismo , Pseudomonas aeruginosa/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biodegradação Ambiental , Dioxigenases/genética , Dioxigenases/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Resíduos Industriais , Redes e Vias Metabólicas/genética , Proteômica , Pseudomonas aeruginosa/química , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/crescimento & desenvolvimento
16.
Int J Mol Sci ; 20(8)2019 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-31018518

RESUMO

Anthropogenic activities have increased the amount of urban wastewater discharged into natural aquatic reservoirs containing a high amount of nutrients such as phosphorus (Pi and PO 4 - 3 ), nitrogen (NH 3 and NO 3 - ) and organic contaminants. Most of the urban wastewater in Mexico do not receive any treatment to remove nutrients. Several studies have reported that an alternative to reduce those contaminants is using consortiums of microalgae and endogenous bacteria. In this research, a genome-scale biochemical reaction network is reconstructed for the co-culture between the microalga Chlorella vulgaris and the bacterium Pseudomonas aeruginosa. Metabolic Pathway Analysis (MPA), is applied to understand the metabolic capabilities of the co-culture and to elucidate the best conditions in removing nutrients. Theoretical yields for phosphorus removal under photoheterotrophic conditions are calculated, determining their values as 0.042 mmol of PO 4 - 3 per g DW of C. vulgaris, 19.43 mmol of phosphorus (Pi) per g DW of C. vulgaris and 4.90 mmol of phosphorus (Pi) per g DW of P. aeruginosa. Similarly, according to the genome-scale biochemical reaction network the theoretical yields for nitrogen removal are 10.3 mmol of NH 3 per g DW of P. aeruginosa and 7.19 mmol of NO 3 - per g DW of C. vulgaris. Thus, this research proves the metabolic capacity of these microorganisms in removing nutrients and their theoretical yields are calculated.


Assuntos
Chlorella vulgaris/metabolismo , Redes e Vias Metabólicas , Nitrogênio/metabolismo , Fósforo/metabolismo , Pseudomonas aeruginosa/metabolismo , Técnicas de Cocultura , Águas Residuárias/microbiologia , Purificação da Água
17.
Nat Commun ; 10(1): 1520, 2019 04 03.
Artigo em Inglês | MEDLINE | ID: mdl-30944318

RESUMO

In Pseudomonas aeruginosa, MexAB-OprM plays a central role in multidrug resistance by ejecting various drug compounds, which is one of the causes of serious nosocomial infections. Although the structures of the components of MexAB-OprM have been solved individually by X-ray crystallography, no structural information for fully assembled pumps from P. aeruginosa were previously available. In this study, we present the structure of wild-type MexAB-OprM in the presence or absence of drugs at near-atomic resolution. The structure reveals that OprM does not interact with MexB directly, and that it opens its periplasmic gate by forming a complex. Furthermore, we confirm the residues essential for complex formation and observed a movement of the drug entrance gate. Based on these results, we propose mechanisms for complex formation and drug efflux.


Assuntos
Proteínas da Membrana Bacteriana Externa/química , Proteínas de Membrana Transportadoras/química , Antibacterianos/farmacocinética , Antibacterianos/farmacologia , Proteínas da Membrana Bacteriana Externa/metabolismo , Cristalografia por Raios X , Farmacorresistência Bacteriana Múltipla , Proteínas de Membrana Transportadoras/metabolismo , Modelos Moleculares , Conformação Proteica , Domínios e Motivos de Interação entre Proteínas , Subunidades Proteicas/química , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/metabolismo , Relação Estrutura-Atividade
18.
MBio ; 10(2)2019 04 16.
Artigo em Inglês | MEDLINE | ID: mdl-30992347

RESUMO

Biogenesis of the outer membrane of Gram-negative bacteria depends on dedicated macromolecular transport systems. The LolABCDE proteins make up the machinery for lipoprotein trafficking from the inner membrane (IM) across the periplasm to the outer membrane (OM). The Lol apparatus is additionally responsible for differentiating OM lipoproteins from those for the IM. In Enterobacteriaceae, a default sorting mechanism has been proposed whereby an aspartic acid at position +2 of the mature lipoproteins prevents Lol recognition and leads to their IM retention. In other bacteria, the conservation of sequences immediately following the acylated cysteine is variable. Here we show that in Pseudomonas aeruginosa, the three essential Lol proteins (LolCDE) can be replaced with those from Escherichia coli The P. aeruginosa lipoproteins MexA, OprM, PscJ, and FlgH, with different sequences at their N termini, were correctly sorted by either the E. coli or P. aeruginosa LolCDE. We further demonstrate that an inhibitor of E. coli LolCDE is active against P. aeruginosa only when expressing the E. coli orthologues. Our work shows that Lol proteins recognize a wide range of signals, consisting of an acylated cysteine and a specific conformation of the adjacent domain, determining IM retention or transport to the OM.IMPORTANCE Gram-negative bacteria build their outer membranes (OM) from components that are initially located in the inner membrane (IM). A fraction of lipoproteins is transferred to the OM by the transport machinery consisting of LolABCDE proteins. Our work demonstrates that the LolCDE complexes of the transport pathways of Escherichia coli and Pseudomonas aeruginosa are interchangeable, with the E. coli orthologues correctly sorting the P. aeruginosa lipoproteins while retaining their sensitivity to a small-molecule inhibitor. These findings question the nature of IM retention signals, identified in E. coli as aspartate at position +2 of mature lipoproteins. We propose an alternative model for the sorting of IM and OM lipoproteins based on their relative affinities for the IM and the ability of the promiscuous sorting machinery to deliver lipoproteins to their functional sites in the OM.


Assuntos
Proteínas da Membrana Bacteriana Externa/metabolismo , Membrana Celular/metabolismo , Escherichia coli/genética , Lipoproteínas/metabolismo , Transporte Proteico , Pseudomonas aeruginosa/metabolismo , Transportadores de Cassetes de Ligação de ATP/genética , Ácido Aspártico/química , Proteínas da Membrana Bacteriana Externa/genética , Proteínas de Escherichia coli/genética , Lipoproteínas/genética , Pseudomonas aeruginosa/genética
19.
Science ; 364(6436): 178-181, 2019 04 12.
Artigo em Inglês | MEDLINE | ID: mdl-30975887

RESUMO

In plants, cell-surface immune receptors sense molecular non-self-signatures. Lipid A of Gram-negative bacterial lipopolysaccharide is considered such a non-self-signature. The receptor kinase LIPOOLIGOSACCHARIDE-SPECIFIC REDUCED ELICITATION (LORE) mediates plant immune responses to Pseudomonas and Xanthomonas but not enterobacterial lipid A or lipopolysaccharide preparations. Here, we demonstrate that synthetic and bacterial lipopolysaccharide-copurified medium-chain 3-hydroxy fatty acid (mc-3-OH-FA) metabolites elicit LORE-dependent immunity. The mc-3-OH-FAs are sensed in a chain length- and hydroxylation-specific manner, with free (R)-3-hydroxydecanoic acid [(R)-3-OH-C10:0] representing the strongest immune elicitor. By contrast, bacterial compounds comprising mc-3-OH-acyl building blocks but devoid of free mc-3-OH-FAs-including lipid A or lipopolysaccharide, rhamnolipids, lipopeptides, and acyl-homoserine-lactones-do not trigger LORE-dependent responses. Hence, plants sense low-complexity bacterial metabolites to trigger immune responses.


Assuntos
Arabidopsis/imunologia , Arabidopsis/microbiologia , Ácidos Decanoicos/metabolismo , Pseudomonas aeruginosa/metabolismo , Acil-Butirolactonas/metabolismo , Ácidos Decanoicos/química , Glicolipídeos/metabolismo , Lipídeo A/metabolismo , Lipopeptídeos/metabolismo
20.
PLoS Comput Biol ; 15(4): e1006507, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30973869

RESUMO

The identification of genes essential for bacterial growth and survival represents a promising strategy for the discovery of antimicrobial targets. Essential genes can be identified on a genome-scale using transposon mutagenesis approaches; however, variability between screens and challenges with interpretation of essentiality data hinder the identification of both condition-independent and condition-dependent essential genes. To illustrate the scope of these challenges, we perform a large-scale comparison of multiple published Pseudomonas aeruginosa gene essentiality datasets, revealing substantial differences between the screens. We then contextualize essentiality using genome-scale metabolic network reconstructions and demonstrate the utility of this approach in providing functional explanations for essentiality and reconciling differences between screens. Genome-scale metabolic network reconstructions also enable a high-throughput, quantitative analysis to assess the impact of media conditions on the identification of condition-independent essential genes. Our computational model-driven analysis provides mechanistic insight into essentiality and contributes novel insights for design of future gene essentiality screens and the identification of core metabolic processes.


Assuntos
Genes Essenciais , Redes e Vias Metabólicas/genética , Biologia Computacional , Meios de Cultura , Elementos de DNA Transponíveis , Bases de Dados Genéticas/estatística & dados numéricos , Genoma Bacteriano , Sequenciamento de Nucleotídeos em Larga Escala/estatística & dados numéricos , Humanos , Modelos Genéticos , Mutagênese , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/crescimento & desenvolvimento , Pseudomonas aeruginosa/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA