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1.
Ecotoxicol Environ Saf ; 208: 111621, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33396141

RESUMO

The study explored the polycyclic aromatic hydrocarbon tolerance of indigenous biosurfactant producing microorganisms. Three bacterial species were isolated from crude oil contaminated sites of Haldia, West Bengal. The three species were screened for biosurfactant production and identified by 16S rRNA sequencing as Brevundimonas sp. IITISM 11, Pseudomonas sp. IITISM 19 and Pseudomonas sp. IITISM 24. The strains showed emulsification activities of 51%, 57% and 63%, respectively. The purified biosurfactants were characterised using FT-IR, GC-MS and NMR spectroscopy and found to have structural similarities to glycolipopeptides, cyclic lipopeptides and glycolipids. The biosurfactants produced were found to be stable under a wide range of temperature (0-100 °C), pH (4-12) and salinity (up to 20% NaCl). Moreover, the strains displayed tolerance to high concentrations (275 mg/L) of anthracene and fluorene and showed a good amount of cell surface hydrophobicity with different hydrocarbons. The study reports the production and characterisation of biosurfactant by Brevundimonas sp. for the first time. Additionally, the kinetic parameters of the bacterial strains grown on up to 300 mg/L concentration of anthracene and fluorene, ranged between 0.0131 and 0.0156 µmax (h-1), while the Ks(mg/L) ranged between 59.28 and 102.66 for Monod's Model. For Haldane-Andrew's model, µmax (h-1) varied between 0.0168 and 0.0198. The inhibition constant was highest for Pseudomonas sp. IITISM 19 on anthracene and Brevundimonas sp. IITISM 11 on fluorene. The findings of the study suggest that indigenous biosurfactant producing strains have tolerance to high PAH concentrations and can be exploited for bioremediation purposes.


Assuntos
Antracenos/metabolismo , Biodegradação Ambiental , Fluorenos/metabolismo , Tensoativos/metabolismo , Antracenos/química , Bactérias/metabolismo , Fluorenos/química , Glicolipídeos , Hidrocarbonetos/metabolismo , Cinética , Petróleo/metabolismo , Hidrocarbonetos Policíclicos Aromáticos/metabolismo , Pseudomonas/metabolismo , RNA Ribossômico 16S/metabolismo , Espectroscopia de Infravermelho com Transformada de Fourier , Tensoativos/química
2.
Chemosphere ; 263: 127975, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32828061

RESUMO

The capability of different strains derived from soil, activated sludge, farm sludge, and worms' excreta were investigated for biodegradation of high-density polyethylene, polystyrene foam, polypropylene and polyethylene terephthalate in unstimulated and stimulated conditions. Biodegradation using naturally occurring microbial strains examined in mixed (270 days) and individual (100 days) systems, while H2O2 stimulated strains were tested only in the mixed system (30 days). Penicillium raperi, Aspergillus flavus, Penicillium glaucoroseum and Pseudomonas sp. were isolated as the most plastic degrading microbes. Maximum weight loss was seen by incubation of polyethylene with Aspergillus flavus (5.5%) in unstimulated mix condition. Fourier Transform Infrared Spectroscopy (FT-IR) revealed formation of new functional groups as hydroxyl, carbonyl, alkene and alkoxy in the treated plastics. Visualisation of plastics by optical, atomic force (AFM) and electron microscopy (SEM) were also illustrated biodegradation. The derived by-products from microbial degradation was tested, and found no inhibition on microbial growth and performance.


Assuntos
Biodegradação Ambiental , Plásticos/metabolismo , Aspergillus flavus/metabolismo , Peróxido de Hidrogênio/metabolismo , Penicillium , Polietileno/metabolismo , Poliestirenos/metabolismo , Pseudomonas/metabolismo , Espectroscopia de Infravermelho com Transformada de Fourier
3.
Ecotoxicol Environ Saf ; 207: 111292, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-32919193

RESUMO

As a hydrophobic pollutant, benzo(a)pyrene (BaP) is difficult to be degraded by microbes due to its poor water solubility. To improve its water solubility, this study harvested a biosurfactant from swine wastewater. The role of the biosurfactant in BaP biodegradation in contaminated water and soil were investigated. The biodegradation kinetics of BaP in contaminated water and the improvement of soil properties were determined. Results showed that critical micelle concentration (CMC) of the biosurfactant was 46.8 mg/L. The biosurfactant has a high pH stability in range of 3-9 and a strong salt stability in NaCl concentration range of 0-20%. At concentrations of 1, 2, 3, 4 and 5 CMC, the biosurfactant increased BaP water solubility by 1.4, 2.6, 4.0, 5.2 and 6.6 times. BaP biodegradation in contaminated water was effectively promoted by the biosurfactant, and the concentrations of BaP in sludge phase decreased to 1.015 mg/L (47.9% decrement) and 0.675 mg/L (65.4% decrement) when the dosed biosurfactant were 1 and 3 CMC, respectively. The biodegradation kinetics of BaP in contaminated water by the biosurfactant fitted well with the two-compartment kinetic model well (R2 > 0.90). For the bioremediation of BaP contaminated soil, adding 0.1%-0.5% (w/w) biosurfactant biodegraded 39.2%-84.8% of BaP, while the control without biosurfactant was 24.2%. In addition, the application of the biosurfactant significantly improved the properties of the contaminated soil, behaved as the increase in microbial quantity, water holding capacity (WHC) and dehydrogenase (DH) activity of the soil. To sum up, the biosurfactant facilitated the BaP biodegradation and can be effectively used in in-site remediation of polycyclic aromatic hydrocarbons (PAHs) (BaP in this study) contaminated water and soil.


Assuntos
Benzo(a)pireno/análise , Recuperação e Remediação Ambiental/métodos , Poluentes do Solo/análise , Tensoativos/química , Águas Residuárias/química , Poluentes Químicos da Água/análise , Animais , Biodegradação Ambiental , Cinética , Pseudomonas/metabolismo , Solo/química , Solubilidade , Tensoativos/metabolismo , Suínos , Águas Residuárias/microbiologia
4.
PLoS One ; 15(7): e0235508, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32614917

RESUMO

This study examined the influence of bioaugmentation on metal concentrations (aluminum, cadmium, chromium, cobalt, copper, iron, lead, manganese, molybdenum, nickel and zinc) in anaerobically digested sewage sludge. To improve the digestion efficiency, bioaugmentation with a mixture of wild-living Archaea and Bacteria (MAB) from Yellowstone National Park, USA, was used. The total concentration of all metals was higher in the digestate than in the feedstock. During anaerobic digestion, the percent increase in the concentration of most of metals was slightly higher in the bioaugmented runs than in the un-augmented runs, but these differences were not statistically significant. However, the percent increase in cadmium and cobalt concentration was significantly higher in the bioaugmented runs than in the un-augmented runs. At MAB doses of 9 and 13% v/v, cadmium concentration in the digestate was 211 and 308% higher than in the feedstock, respectively, and cobalt concentration was 138 and 165%, respectively. Bioaugmentation increased over 4 times the percentage of Pseudomonas sp. in the biomass that are able to efficiently accumulate metals by both extracellular adsorption and intracellular uptake. Biogas production was not affected by the increased metal concentrations. In conclusion, bioaugmentation increased the concentration of metals in dry sludge, which means that it could potentially have negative effects on the environment.


Assuntos
Metais/metabolismo , Esgotos/química , Adsorção , Anaerobiose , Archaea/genética , Archaea/crescimento & desenvolvimento , Archaea/metabolismo , Bactérias/genética , Bactérias/crescimento & desenvolvimento , Bactérias/metabolismo , Biomassa , Cádmio/análise , Cádmio/metabolismo , Cobalto/análise , Cobalto/metabolismo , Metais/química , Pseudomonas/genética , Pseudomonas/crescimento & desenvolvimento , Pseudomonas/metabolismo , Esgotos/microbiologia , Eliminação de Resíduos Líquidos
5.
PLoS One ; 15(6): e0234865, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32598366

RESUMO

In this study, we selected and characterized different pesticide-tolerant bacteria isolated from a biomixture of a biopurification system that had received continuous applications of a pesticides mixture. The amplicon analysis of biomixture reported that the phyla Proteobacteria, Firmicutes, Bacteroidetes and Actinobacteria were predominant. Six strains grew in the presence of chlorpyrifos and iprodione. Biochemical characterization showed that all isolates were positive for esterase, acid phosphatase, among others, and they were identified as Pseudomonas, Rhodococcus and Achromobacter based on molecular and proteomic analysis. Bacterial growth decreased as both pesticide concentrations increased from 10 to 100 mg L-1 in liquid culture. The Achromobacter sp. strain C1 showed the best chlorpyrifos removal rate of 0.072-0.147 d-1 a half-life of 4.7-9.7 d and a maximum metabolite concentration of 2.10 mg L-1 at 120 h. On the other hand, Pseudomonas sp. strain C9 showed the highest iprodione removal rate of 0.100-0.193 d-1 a half-life of 4-7 d and maximum metabolite concentration of 0.95 mg L-1 at 48 h. The Achromobacter and Pseudomonas strains showed a good potential as chlorpyrifos and iprodione-degrading bacteria.


Assuntos
Achromobacter/metabolismo , Biodegradação Ambiental , Praguicidas/metabolismo , Pseudomonas/metabolismo , Microbiologia do Solo , Achromobacter/isolamento & purificação , Aminoimidazol Carboxamida/análogos & derivados , Aminoimidazol Carboxamida/metabolismo , Aminoimidazol Carboxamida/toxicidade , Clorpirifos/metabolismo , Clorpirifos/toxicidade , Hidantoínas/metabolismo , Hidantoínas/toxicidade , Praguicidas/toxicidade , Pseudomonas/isolamento & purificação , Poluentes do Solo/metabolismo , Poluentes do Solo/toxicidade , Recursos Hídricos
6.
Ecotoxicol Environ Saf ; 201: 110850, 2020 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-32531571

RESUMO

Despite of significant progress in remediation of Cr(VI) or Hg(II) pollution by microorganisms, study on the reduction of both Cr(VI) and Hg(II) by the same microbial strain was not reported so far, which is actually important for bioremediation of contaminated sites with multiple heavy metals. In this study, Pseudomonas umsongensis CY-1 was newly isolated from chromium-contaminated soil and showed remediation potentials for both Cr(VI) and Hg(II) pollution. The highest Cr(VI) (93.9%) and Hg(II) (82.8%) reduction rates were obtained at the initial concentration of 5 mg/L. Comparison between removal by resting cells and heat-treated resting cells demonstrated that P. umsongensis CY-1 removed Cr(VI) and Hg(II) from Tris-HCl buffer (pH 7.0) mainly through reduction instead of adsorption. By comparing the Cr(VI) and Hg(II) reduction rates of different cellular fractions, it was found that Cr(VI) and Hg(II) reductions mainly happened in the cytoplasm of P. umsongensis CY-1, which were further demonstrated by Transmission electron microscopy (TEM) analysis. Furthermore, analysis of X-ray photoelectron spectroscopy demonstrated that the reduction products of Cr(VI) and Hg(II) were mainly in the form of Cr(III) and Hg (0), respectively. The findings in this study will provide a guide for further insights in the bioremediation of contaminated sites with multiple heavy metals.


Assuntos
Cromo/análise , Compostos de Mercúrio/análise , Pseudomonas/metabolismo , Poluentes do Solo/análise , Adsorção , Biodegradação Ambiental , Cromo/metabolismo , Compostos de Mercúrio/metabolismo , Modelos Teóricos , Oxirredução , Filogenia , Pseudomonas/isolamento & purificação , Poluentes do Solo/metabolismo
7.
Appl Environ Microbiol ; 86(16)2020 08 03.
Artigo em Inglês | MEDLINE | ID: mdl-32503910

RESUMO

The enzymatic production of 2,5-furandicarboxylic acid (FDCA) from 5-hydroxymethylfurfural (HMF) has gained interest in recent years, as FDCA is a renewable precursor of poly(ethylene-2,5-furandicarboxylate) (PEF). 5-Hydroxymethylfurfural oxidases (HMFOs) form a flavoenzyme family with genes annotated in a dozen bacterial species but only one enzyme purified and characterized to date (after heterologous expression of a Methylovorus sp. HMFO gene). This oxidase acts on both furfuryl alcohols and aldehydes and, therefore, is able to catalyze the conversion of HMF into FDCA through 2,5-diformylfuran (DFF) and 2,5-formylfurancarboxylic acid (FFCA), with only the need of oxygen as a cosubstrate. To enlarge the repertoire of HMFO enzymes available, genetic databases were screened for putative HMFO genes, followed by heterologous expression in Escherichia coli After unsuccessful trials with other bacterial HMFO genes, HMFOs from two Pseudomonas species were produced as active soluble enzymes, purified, and characterized. The Methylovorus sp. enzyme was also produced and purified in parallel for comparison. Enzyme stability against temperature, pH, and hydrogen peroxide, three key aspects for application, were evaluated (together with optimal conditions for activity), revealing differences between the three HMFOs. Also, the kinetic parameters for HMF, DFF, and FFCA oxidation were determined, the new HMFOs having higher efficiencies for the oxidation of FFCA, which constitutes the bottleneck in the enzymatic route for FDCA production. These results were used to set up the best conditions for FDCA production by each enzyme, attaining a compromise between optimal activity and half-life under different conditions of operation.IMPORTANCE HMFO is the only enzyme described to date that can catalyze by itself the three consecutive oxidation steps to produce FDCA from HMF. Unfortunately, only one HMFO enzyme is currently available for biotechnological application. This availability is enlarged here by the identification, heterologous production, purification, and characterization of two new HMFOs, one from Pseudomonas nitroreducens and one from an unidentified Pseudomonas species. Compared to the previously known Methylovorus HMFO, the new enzyme from P. nitroreducens exhibits better performance for FDCA production in wider pH and temperature ranges, with higher tolerance for the hydrogen peroxide formed, longer half-life during oxidation, and higher yield and total turnover numbers in long-term conversions under optimized conditions. All these features are relevant properties for the industrial production of FDCA. In summary, gene screening and heterologous expression can facilitate the selection and improvement of HMFO enzymes as biocatalysts for the enzymatic synthesis of renewable building blocks in the production of bioplastics.


Assuntos
Proteínas de Bactérias/metabolismo , Ácidos Dicarboxílicos/metabolismo , Furaldeído/análogos & derivados , Furanos/metabolismo , Methylophilaceae/genética , Oxirredutases/metabolismo , Pseudomonas/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Furaldeído/metabolismo , Methylophilaceae/metabolismo , Microrganismos Geneticamente Modificados/genética , Microrganismos Geneticamente Modificados/metabolismo , Pseudomonas/metabolismo
8.
Korean J Parasitol ; 58(2): 173-179, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32418386

RESUMO

Leishmaniasis is a prevalent cause of death and animal morbidity in underdeveloped countries of endemic area. However, there is few vaccine and effective drugs. Antimicrobial peptides are involved in the innate immune response in many organisms and are being developed as novel drugs against parasitic infections. In the present study, we synthesized a 5-amino acid peptide REDLK, which mutated the C-terminus of Pseudomonas exotoxin, to identify its effect on the Leishmania tarentolae. Promastigotes were incubated with different concentration of REDLK peptide, and the viability of parasite was assessed using MTT and Trypan blue dye. Morphologic damage of Leishmania was analyzed by light and electron microscopy. Cellular apoptosis was observed using the annexin V-FITC/PI apoptosis detection kit, mitochondrial membrane potential assay kit and flow cytometry. Our results showed that Leishmania tarentolae was susceptible to REDLK in a dose-dependent manner, disrupt the surface membrane integrity and caused parasite apoptosis. In our study, we demonstrated the leishmanicidal activity of an antimicrobial peptide REDLK from Pseudomonas aeruginosa against Leishmania tarentolae in vitro and present a foundation for further research of anti-leishmanial drugs.


Assuntos
Proteínas de Bactérias/farmacologia , Leishmania/efeitos dos fármacos , Leishmania/crescimento & desenvolvimento , Peptídeos/farmacologia , Pseudomonas/metabolismo , Técnicas In Vitro
9.
Chemosphere ; 256: 126998, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32470727

RESUMO

Efficient degradation of polycyclic aromatic hydrocarbons (PAHs) in a petroleum-contaminated soil was challenging which requires ample PAH-degrading flora and nutrients. In this study, we investigated the effects of 'natural attenuation', 'bioaugmentation', 'compost only (raw materials of compost included pig manure and rice husk mixed at a 1:2 proportion, supplemented with 2.5% charcoal)', and 'compost with bioaugmentation' treatments on degradation of polycyclic aromatic hydrocarbons (PAHs) and microbial community shifts during the remediation of petroleum-contaminated soil. After sixteen weeks of incubation, the removal efficiencies of PAHs were 0.52 ± 0.04%, 6.92 ± 0. 32%, 9.53 ± 0.29%, and 18.2 ± 0.64% in the four treatments, respectively. 'Compost with bioaugmentation' was the most effective for PAH removal among all the treatments. Illumina sequencing analysis suggested that both the 'compost only' and 'compost with bioaugmentation' treatments changed soil microbial community structures and enhanced microbial biodiversity. Some of the microorganisms affiliated with the compost including Azomonas, Luteimonas, Pseudosphingobacterium, and Parapedobacter were able to survive and become dominant in the contaminated soil. The 'bioaugmentation and 'natural attenuation' treatments had no significant effects on soil microbial community structure. Inoculation of the PAH degraders including Bacillus, Pseudomonas, and Acinetobacter directly into the contaminated soil led to lower biodiversity under natural conditions. This result suggested that compost addition increased the α-diversity of both the bacterial and fungal communities in petroleum-contaminated soil, leading to higher PAH degradation efficiency in petroleum-contaminated soil.


Assuntos
Petróleo/metabolismo , Hidrocarbonetos Policíclicos Aromáticos/metabolismo , Microbiologia do Solo , Poluentes do Solo/metabolismo , Animais , Bactérias/metabolismo , Biodegradação Ambiental , Carvão Vegetal/metabolismo , Compostagem , Recuperação e Remediação Ambiental , Microbiota , Hidrocarbonetos Policíclicos Aromáticos/análise , Pseudomonas/metabolismo , Solo/química , Poluentes do Solo/análise , Suínos
10.
Artigo em Inglês | MEDLINE | ID: mdl-32400278

RESUMO

Degradation studies of phenanthrene and naphthalene as a mixture was carried out using a developed bacterial consortium. The isolates used in consortium were identified as Chryseobacterium sp., Sphingobacterium sp., Stenotrophomonas sp., Agromyces sp. and Pseudomonas sp. Limited work is done on genus Agromyces in degradation studies of PAHs. Catechol production was detected using Arnow's assay suggested that the pathway used for degradation is the meta-cleavage pathway. Results showed that Tween 80, as a surfactant, had maximum effect on the growth of isolates during PAH degradation. This suggests that use of Tween 80 as a surfactant enhanced the uptake of PAH by bacterial isolates during degradation. The study further revealed that, bacterial consortium was successfully utilized in the treatment of water contaminated with PAH in a laboratory-scale biofilm bioreactor. The bacterial consortium was able to degrade 99.9% of naphthalene and 92.9% of phenanthrene as a mixture at a high concentration of 2000 mg/L within 6 days. Further scaling up of the biofilm bioreactor can prove beneficial in large scale treatment of PAH contaminated water. This study showed promising results and these bacterial strains can be used as potential tools for bioremediation of PAH in contaminated sites.


Assuntos
Microbiota , Naftalenos/metabolismo , Fenantrenos/metabolismo , Poluentes do Solo/metabolismo , Poluentes Químicos da Água/metabolismo , Biodegradação Ambiental , Biofilmes/crescimento & desenvolvimento , Reatores Biológicos/microbiologia , Catecóis/análise , Testes de Sensibilidade Microbiana , Microbiota/efeitos dos fármacos , Modelos Teóricos , Pseudomonas/isolamento & purificação , Pseudomonas/metabolismo , Sphingobacterium/isolamento & purificação , Sphingobacterium/metabolismo
11.
PLoS One ; 15(4): e0231215, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32267901

RESUMO

Plants interact with a great variety of microorganisms that inhabit the rhizosphere or the epiphytic and endophytic phyllosphere and that play critical roles in plant growth as well as the biocontrol of phytopathogens and insect pests. Avocado fruit damage caused by the thrips species Scirtothrips perseae leads to economic losses of 12-51% in many countries. In this study, a screening of bacteria associated with the rhizosphere or endophytic phyllosphere of avocado roots was performed to identify bacterial isolates with plant growth-promoting activity in vitro assays with Arabidopsis seedlings and to assess the biocontrol activity of the isolates against Scirtothrips perseae. The isolates with beneficial, pathogenic and/or neutral effects on Arabidopsis seedlings were identified. The plant growth-promoting bacteria were clustered in two different groups (G1 and G3B) based on their effects on root architecture and auxin responses, particularly bacteria of the Pseudomonas genus (MRf4-2, MRf4-4 and TRf2-7) and one Serratia sp. (TS3-6). Twenty strains were selected based on their plant growth promotion characteristics to evaluate their potential as thrips biocontrol agents. Analyzing the biocontrol activity of S. perseae, it was identified that Chryseobacterium sp. shows an entomopathogenic effect on avocado thrips survival. Through the metabolic profiling of compounds produced by bacteria with plant growth promotion activity, bioactive cyclodipeptides (CDPs) that could be responsible for the plant growth-promoting activity in Arabidopsis were identified in Pseudomonas, Serratia and Stenotrophomonas. This study unravels the diversity of bacteria from the avocado rhizosphere and highlights the potential of a unique isolate to achieve the biocontrol of S. perseae.


Assuntos
Controle de Insetos/métodos , Persea/crescimento & desenvolvimento , Persea/microbiologia , Controle Biológico de Vetores/métodos , Tisanópteros/microbiologia , Árvores/crescimento & desenvolvimento , Árvores/microbiologia , Animais , Arabidopsis/fisiologia , Técnicas de Cocultura , DNA Bacteriano/genética , Ácidos Indolacéticos/metabolismo , Filogenia , Pseudomonas/metabolismo , Rizosfera , Plântula/metabolismo , Serratia/metabolismo , Stenotrophomonas/metabolismo
12.
Microbes Environ ; 35(2)2020.
Artigo em Inglês | MEDLINE | ID: mdl-32269203

RESUMO

More than 3,000 isolates of fluorescent pseudomonads have been collected from plant roots in Japan and screened for the presence of antibiotic-synthesizing genes. In total, 927 hydrogen cyanide (HCN)-, 47 2,4-diacetylphloroglucinol (PHL)-, 6 pyoluteorin (PLT)-, 14 pyrrolnitrin (PRN)-, and 8 phenazine (PHZ)-producing isolates have been detected. A cluster analysis (≥99% identity) identified 10 operational taxonomic units (OTUs) in antibiotic biosynthesis gene-possessing pseudomonads. OTU HLR (PHL, PLT, and PRN) contained four antibiotics: HCN, PHL, PLT, and PRN, while OTU RZ (PRN and PHZ) contained three: HCN, PRN, and PHZ. OTU H1, H2, H3, H4, H5, H6, and H7 (PHL1-7) contained two antibiotics: HCN and PHL, while OTU H8 (PHL8) contained one: PHL. Isolates belonging to OTU HLR and RZ suppressed damping-off disease in cabbage seedlings caused by Rhizoctonia solani. Effective strains belonging to OTU HLR and RZ were related to Pseudomonas protegens and Pseudomonas chlororaphis, respectively. Antibiotic biosynthesis gene-possessing fluorescent pseudomonads are distributed among different geographical sites in Japan and plant species.


Assuntos
Antibacterianos/biossíntese , Fluorescência , Pseudomonas/classificação , Rizosfera , Agentes de Controle Biológico , Genes Bacterianos , Variação Genética , Japão , Raízes de Plantas/microbiologia , Pseudomonas/metabolismo , Pseudomonas fluorescens/genética , RNA Ribossômico 16S/genética , Microbiologia do Solo
13.
Ecotoxicol Environ Saf ; 194: 110378, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32146194

RESUMO

The primary purpose of this study was to systematically explore the complete metabolic pathway and tolerance mechanism of strain DNB-S1 to dibutyl phthalate (DBP), and the effect of DBP on energy metabolism of DNB-S1. Here, DNB-S1, a strain of Pseudomonas sp. that was highly effective in degrading DBP, was identified, and differentially expressed metabolites and metabolic networks of DBP were studied. The results showed that the differentially expressed metabolites were mainly aromatic compounds and lipid compounds, with only a few toxic intermediate metabolites. It speculated that phthalic acid, salicylic acid, 3-hydroxybenzoate acid, 3-Carboxy-cis, cis-muconate, fumarypyravate were intermediate metabolites of DBP. Their up-regulation indicated that there were two metabolic pathways in the degradation of DBP (protocatechuate pathway and gentisate pathway), which had been verified by peak changes at 290 nm, 320 nm, 330 nm, and 375 nm in the enzymatic method. Also, aspartate, GSH, and other metabolites were up-regulation, indicating that DNB-S1 had a high tolerance to DBP and maintained cell homeostasis, which was also one of the essential reasons to ensure the efficient degradation of DBP. Altogether, this study firstly proposed two pathways to degrade DBP and comprehensively explored the effect of DBP on the metabolic function of DNB-S1, which enriched the study of microbial metabolism of organic pollutants, and which provided a basis for the application of metabolomics.


Assuntos
Dibutilftalato/metabolismo , Poluentes Ambientais/metabolismo , Pseudomonas/metabolismo , Adaptação Fisiológica/efeitos dos fármacos , Biodegradação Ambiental , Dibutilftalato/toxicidade , Metabolismo Energético/efeitos dos fármacos , Poluentes Ambientais/toxicidade , Pseudomonas/efeitos dos fármacos , Pseudomonas/crescimento & desenvolvimento
14.
Ecotoxicol Environ Saf ; 194: 110434, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32155483

RESUMO

A factor that may significantly increase the efficacy of phytoextraction is soil bioaugmentation with specific bacteria, which can alter the composition of rhizospheric and endophytic bacterial communities. The aim of this study was to compare the effect of soil treatment with living (bioaugmentation) and dead (control) cells of the plant growth-promoting metal-resistant endophytic strain Pseudomonas sp. H15 on the bacterial community composition in the rhizo- and endo-sphere of white mustard during enhanced phytoextraction. The bacterial communities in the rhizosphere were dominated (51.7-68.2%) by Proteobacteria, regardless of the soil treatment or sampling point. A temporary increase in the number of sequences belonging to Gammaproteobacteria (up to 37.3%) was only observed 24 h after the soil treatment with living Pseudomonas sp. H15 cells, whereas for the remaining samples, the relative abundance of this class did not exceed 7.1%. The relative abundance of Proteobacteria in the endosphere of the roots, stems, and leaves of white mustard was higher in the control than in bioaugmented plants. The most pronounced dominance of the Gammaproteobacteria sequences was observed in the stems and leaves of the control plants at the first sampling point, which strongly indicates the ability of the plants to rapidly uptake DNA from soil and translocate it to the aboveground parts of the plants. Additionally, the bioaugmentation of the soil caused a diverse shift in the bacterial communities in the rhizo- and endo-sphere of white mustard compared to control. The most distinct differences, which were dependent on the treatment, were observed in the endosphere of plants at the beginning of the experiment and decreased over time. These results indicate that the rhizo- and endo-biome of white mustard reacts to soil bioaugmentation and may influence the efficiency of bacterial-assisted phytoextraction.


Assuntos
Biodegradação Ambiental , Pseudomonas/metabolismo , Sinapis/metabolismo , Microbiologia do Solo , Bactérias/efeitos dos fármacos , Brassica , Desenvolvimento Vegetal , Folhas de Planta/química , Raízes de Plantas/química , Pseudomonas/efeitos dos fármacos , Rizosfera , Solo , Poluentes do Solo/análise
15.
Appl Environ Microbiol ; 86(10)2020 05 05.
Artigo em Inglês | MEDLINE | ID: mdl-32198167

RESUMO

Burkholderia sp. strain SG-MS1 and Pseudomonas sp. strain SG-MS2 have previously been found to mineralize (+)-pinoresinol through a common catabolic pathway. Here, we used comparative genomics, proteomics, protein semipurification, and heterologous expression to identify a flavoprotein from the vanillyl alcohol oxidase/p-cresol methyl hydroxylase (VAO/PCMH) enzyme family in SG-MS2 that carries out the initial hydroxylation of (+)-pinoresinol at the benzylic carbon. The cognate gene is translationally coupled with a downstream cytochrome gene, and the cytochrome is required for activity. The flavoprotein has a unique combination of cofactor binding and cytochrome requirements for the VAO/PCMH family. The heterologously expressed enzyme has a Km of 1.17 µM for (+)-pinoresinol. The enzyme is overexpressed in strain SG-MS2 upon exposure to (+)-pinoresinol, along with 45 other proteins, 22 of which were found to be encoded by genes in an approximately 35.1-kb cluster also containing the flavoprotein and cytochrome genes. Homologs of 18 of these 22 genes, plus the flavoprotein and cytochrome genes, were also found in a 38.7-kb cluster in SG-MS1. The amino acid identities of four of the other proteins within the SG-MS2 cluster suggest they catalyze conversion of hydroxylated pinoresinol to protocatechuate and 2-methoxyhydroquinone. Nine other proteins upregulated in SG-MS2 on exposure to (+)-pinoresinol appear to be homologs of proteins known to comprise the protocatechuate and 2-methoxyhydroquinone catabolic pathways, but only three of the cognate genes lie within the cluster containing the flavoprotein and cytochrome genes.IMPORTANCE (+)-Pinoresinol is an important plant defense compound, a major food lignan for humans and some other animals, and the model compound used to study degradation of the ß-ß' linkages in lignin. We report a gene cluster, in one strain each of Pseudomonas and Burkholderia, that is involved in the oxidative catabolism of (+)-pinoresinol. The flavoprotein component of the α-hydroxylase which heads the pathway belongs to the 4-phenol oxidizing (4PO) subgroup of the vanillyl alcohol oxidase/p-cresol methyl hydroxylase (VAO/PCMH) enzyme family but constitutes a novel combination of cofactor and electron acceptor properties for the family. It is translationally coupled with a cytochrome gene whose product is also required for activity. The work casts new light on the biology of (+)-pinoresinol and its transformation to other bioactive molecules. Potential applications of the findings include new options for deconstructing lignin into useful chemicals and the generation of new phytoestrogenic enterolactones from lignans.


Assuntos
Proteínas de Bactérias/genética , Flavoproteínas/genética , Furanos/metabolismo , Genes Bacterianos/genética , Lignanas/metabolismo , Pseudomonas/genética , Proteínas de Bactérias/metabolismo , Flavoproteínas/metabolismo , Redes e Vias Metabólicas , Família Multigênica , Oxirredução , Pseudomonas/metabolismo
16.
PLoS Pathog ; 16(3): e1008375, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32191776

RESUMO

The extent to which disturbances in the resident microbiota can compromise an animal's health is poorly understood. Hydra is one of the evolutionary oldest animals with naturally occurring tumors. Here, we found a causal relationship between an environmental spirochete (Turneriella spec.) and tumorigenesis in Hydra. Unexpectedly, virulence of this pathogen requires the presence of Pseudomonas spec., a member of Hydra´s beneficial microbiome indicating that dynamic interactions between a resident bacterium and a pathogen cause tumor formation. The observation points to the crucial role of commensal bacteria in maintaining tissue homeostasis and adds support to the view that microbial community interactions are essential for disease. These findings in an organism that shares deep evolutionary connections with all animals have implications for our understanding of cancer.


Assuntos
Carcinogênese , Hydra , Leptospiraceae/metabolismo , Microbiota , Neoplasias , Pseudomonas/metabolismo , Animais , Hydra/metabolismo , Hydra/microbiologia , Neoplasias/metabolismo , Neoplasias/microbiologia
17.
J Appl Microbiol ; 129(3): 575-589, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32147927

RESUMO

OBJECTIVE: The present study was performed to examine the role of pqqE inhabiting rhizobacteria in organic acid production and relationship of the organic acids with phosphate solubilization by the bacteria in vitro as well as in vivo. METHODS AND RESULTS: The pqqE gene was PCR amplified and sequenced in genomic DNA of Pantoea sp. WP-5 and Pseudomonas sp. NN-4. Nucleotide sequence obtained from WP-5 and NN-4 showed maximum sequence similarity (88 and 89%, respectively) with the pqqE gene of Pseudomonas fluorescens strain CMR12a (KM251420). Deduced amino acid sequence from pqqE gene of Pseudomonas sp. NN-4 and Pantoea sp. WP-5 showed 75 and 93% similarity, respectively, with protein pyrroloquinoline quinone. Phosphate solubilization and acid production assay were quantified on spectrophotometer and high-profile liquid chromatograph, respectively, by each bacterial strain. Both strains produced organic acids such as acetic, citric, gluconic, succinic and malic acid and lowered the pH of Pikovskaya broth medium under laboratory conditions. Phosphate solubilization by Pantoea sp. WP-5 was 311 ± 4 and 204 ± 3 µg ml-1 in the culture medium supplemented with glucose and sucrose as carbon source, respectively. Pseudomonas sp. NN-4 solubilized 176 ± 3 and 298 ± 5 µg ml-1 phosphate in Pikovskaya broth medium under similar conditions. In field experiments conducted during two consecutive years, the concentration of acetic acid and gluconic acid was higher in root exudates of plants treated with Pantoea sp. WP-5 at 30% reduced doses of nitrogen (N)- and phosphorus (P)-based chemical fertilizers as compared to non-inoculated plants. Values of chlorophyll contents, crop growth rate, leaf area index, straw yield and P contents were recorded higher in plants inoculated with Pantoea sp. WP-5 and Pseudomonas sp. NN-4 as compared to non-inoculated control. Grain yield was increased by 10-12% due to inoculation with Pantoea sp. WP-5 and Pseudomonas sp. NN-4 over non-inoculated control in the field experiments. CONCLUSIONS: These results lead to the conclusions that the rhizobacteria inhabiting pqqE gene produced organic acids and solubilized the phosphate in vitro. On inoculation to wheat plants in field experiments, these strains produced the organic acids, solubilized the phosphate, and improved the P uptake and productivity of wheat. SIGNIFICANCE AND IMPACT OF THE STUDY: The Pantoea sp. WP-5 and Pseudomonas sp. NN-4 are the potential candidates for inoculation to wheat as phosphate solubilizer even with reduced chemical fertilizer dose. The inoculation of the strains may enhance grain yield and net income of the farmer even with less chemical fertilizer application. This practice will be helpfull inminimizing environmental pollution.


Assuntos
Proteínas de Bactérias/genética , Pantoea/fisiologia , Pseudomonas/fisiologia , Triticum/crescimento & desenvolvimento , Triticum/microbiologia , Ácidos/metabolismo , Proteínas de Bactérias/metabolismo , Grão Comestível/crescimento & desenvolvimento , Grão Comestível/metabolismo , Fertilizantes/análise , Pantoea/genética , Pantoea/metabolismo , Fosfatos/metabolismo , Pseudomonas/genética , Pseudomonas/metabolismo , Solo/química , Microbiologia do Solo , Triticum/metabolismo
18.
Sci Rep ; 10(1): 4502, 2020 03 11.
Artigo em Inglês | MEDLINE | ID: mdl-32161360

RESUMO

Malachite green is a common environmental pollutant that poses a great threat to non-target organisms, including humans. This study reports the characterization of a bacterial strain, Pseudomonas veronii JW3-6, which was isolated from a malachite green enrichment culture. This strain degraded malachite green efficiently in a wide range of temperature and pH levels. Under optimal degradation conditions (32.4 °C, pH 7.1, and inoculum amount of 2.5 × 107 cfu/mL), P. veronii JW3-6 could degrade 93.5% of 50 mg/L malachite green within seven days. Five intermediate products from the degradation of malachite green were identified: leucomalachite green, 4-(dimethylamino) benzophenone, 4-dimethylaminophenol, benzaldehyde, and hydroquinone. We propose a possible degradation pathway based on these findings. The present study is the first to report the degradation of malachite green by P. veronii and the identification of hydroquinone as a metabolite in the degradation pathway.


Assuntos
Biodegradação Ambiental , Redes e Vias Metabólicas , Pseudomonas/metabolismo , Corantes de Rosanilina/metabolismo , Biodiversidade , Microbiologia Ambiental , Cinética , Estrutura Molecular , Filogenia , Pseudomonas/classificação , Pseudomonas/genética , Pseudomonas/isolamento & purificação , RNA Ribossômico 16S , Corantes de Rosanilina/química
19.
Chemosphere ; 250: 126339, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32120155

RESUMO

Di-n-butyl phthalate (DBP), one of the most widely used plasticizers, has been listed as a priority pollutant because of its toxicity to both humans and animals. In this study, Pseudomonas sp. W1, isolated from activated sludge, was capable of degrading 99.88% of DBP (1000 mg L-1) within 8 days. We immobilized the W1 strain using Fe3O4 iron nanoparticles (IONPs) coated with poly-dopamine (PDA), and further evaluated its DBP degradation efficiency. The DBP degradation performance of W1 was improved by immobilization, exhibiting 99.69% of DBP degradation efficiency on the 6th day, which was 25.68% higher than un-immobilized W1. After three cycles of magnetic recycling and utilization, W1-PDA-IONPs retained 99.6% of their original efficiency. W1-PDA-IONPs were then used to degrade DBP in landfill leachate. When the proportion of raw leachate was ≤50%, DBP could be all degraded by W1-PDA-IONPs within 6 days. In 100% landfill leachate, DBP degradation efficiency after 10 days of incubation reached 66.40%. Furthermore, W1-PDA-IONPs cells in a simulated aeration system could be effectively magnetically separated at aeration rates from 60 to 600 mL min-1. These results highlight the potential of W1-PDA-IONPs in the bioremediation of DBP-contaminated waste water.


Assuntos
Dibutilftalato/química , Poluentes Ambientais/química , Biodegradação Ambiental , Fenômenos Magnéticos , Nanopartículas , Plastificantes , Pseudomonas/metabolismo , Esgotos , Águas Residuárias , Poluentes Químicos da Água/química , Poluentes Químicos da Água/metabolismo
20.
Arch Microbiol ; 202(6): 1551-1557, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32125450

RESUMO

The control of pyrimidine nucleotide formation in the bacterium Pseudomonas aurantiaca ATCC 33663 by pyrimidines was studied. The activities of the pyrimidine biosynthetic pathway enzymes were investigated in P. aurantiaca ATCC 33663 cells and from cells of an auxotroph lacking orotate phosphoribosyltransferase activity under selected culture conditions. All activities of the pyrimidine biosynthetic pathway enzymes in ATCC 33663 cells were depressed by uracil addition to the minimal medium when succinate served as the carbon source. In contrast, all pyrimidine biosynthetic pathway enzyme activities in ATCC 33663 cells were depressed by orotic acid supplementation to the minimal medium when glucose served as the carbon source. The orotidine 5'-monophosphate decarboxylase activity in the phosphoribosyltransferase mutant strain increased by more than sixfold in succinate-grown cells and by more than 16-fold in glucose-grown cells after pyrimidine limitation showing possible repression of the decarboxylase by a pyrimidine-related compound. Inhibition by ATP, GTP, UTP and pyrophosphate of the in vitro activity of aspartate transcarbamoylase in ATCC 33663 was observed. The findings demonstrated control at the level of pyrimidine biosynthetic enzyme synthesis and activity for the P. aurantiaca transcarbamoylase. The control of pyrimidine synthesis in P. aurantiaca seemed to differ from what has been observed previously for the regulation of pyrimidine biosynthesis in related Pseudomonas species. This investigation could prove helpful to future work studying pseudomonad taxonomic analysis as well as to those exploring antifungal and antimicrobial agents produced by P. aurantiaca.


Assuntos
Aspartato Carbamoiltransferase/metabolismo , Pseudomonas/metabolismo , Nucleotídeos de Pirimidina/biossíntese , Pirimidinas/metabolismo , Aspartato Carbamoiltransferase/genética , Vias Biossintéticas , Difosfatos , Regulação Bacteriana da Expressão Gênica , Orotato Fosforribosiltransferase/genética , Orotidina-5'-Fosfato Descarboxilase/metabolismo , Pseudomonas/enzimologia , Nucleotídeos de Pirimidina/metabolismo , Ácido Succínico/metabolismo , Uracila/metabolismo
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