Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 5.698
Filtrar
1.
Metab Eng ; 72: 325-336, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35513297

RESUMO

Ligand-induced system plays an important role for microbial engineering due to its tunable gene expression control over timings and levels. An oleic acid (OA)-induced system was recently constructed based on protein FadR, a transcriptional regulator involved in fatty acids metabolism, for metabolic control in Escherichia coli. In this study, we constructed a synthetic FadR-based OA-induced systems in Halomonas bluephagenesis by hybridizing the porin promoter core region and FadR-binding operator (fadO). The dynamic control range was optimized over 150-fold, and expression leakage was significantly reduced by tuning FadR expression and positioning fadO, forming a series of OA-induced systems with various expression strengths, respectively. Additionally, ligand orthogonality and cross-species portability were also studied and showed highly linear correlation among Halomonas spp., Escherichia coli and Pseudomonas spp. Finally, OA-induced systems with medium- and small-dynamic control ranges were employed to dynamically control the expression levels of morphology associated gene minCD, and monomer precursor 4-hydroxybutyrate-CoA (4HB-CoA) synthesis pathway for polyhydroxyalkanoates (PHA), respectively, in the presence of oleic acid as an inducer. As a result, over 10 g/L of poly-3-hydroxybutyrate (PHB) accumulated by elongated cell sizes, and 6 g/L of P(3HB-co-9.57 mol% 4HB) were obtained by controlling the dose and induction time of oleic acid only. This study provides a systematic approach for ligand-induced system engineering, and demonstrates an alternative genetic tool for dynamic control of industrial biotechnology.


Assuntos
Halomonas , Poli-Hidroxialcanoatos , Coenzima A/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Halomonas/genética , Halomonas/metabolismo , Ligantes , Engenharia Metabólica , Ácido Oleico/metabolismo , Poliésteres/metabolismo , Poli-Hidroxialcanoatos/genética , Pseudomonas/genética , Pseudomonas/metabolismo
2.
Sci Total Environ ; 838(Pt 2): 156076, 2022 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-35597344

RESUMO

Biochar and ryegrass have been used in the remediation of polycyclic aromatic hydrocarbons (PAHs)-contaminated soils; however, the effects of different biochar application levels on the dissipation of PAHs, bacterial communities, and PAH-ring hydroxylating dioxygenase (PAH-RHDα) genes in rhizosphere soil remain unclear. In this study, enzyme activity tests, real-time quantitative polymerase chain reaction (PCR), and high-throughput sequencing were performed to investigate the effects of different proportions of rape straw biochar (1%, 2%, and 4% (w/w)) on the degradation of PAHs, as well as the associated changes in the soil bacterial community and PAH-RHDα gene expression. The results revealed that biochar enhanced the rhizoremediation of PAH-contaminated soil and that 2% biochar-treated rhizosphere soil was the most effective in removing PAHs. Furthermore, urease activity, abundance and activity of total bacteria, and PAH-degrading bacteria were enhanced in soil that was amended with biochar and ryegrass. Additionally, the activity of 16S rDNA and PAH-RHDα gram-negative (GN) genes increased with increasing biochar dosage and had a positive correlation with the removal of PAHs. Biochar changed the rhizosphere soil bacterial composition and α-diversity, and promoted the growth of Pseudomonas and Zeaxanthinibacter. In addition, the relative abundance of Pseudomonas was positively correlated with PAH removal. These findings imply that rape straw biochar can enhance the rhizoremediation of PAH-contaminated soil by changing soil bacterial communities and stimulating the expression of PAH-RHDα GN genes. The 2% of rape straw biochar combined with ryegrass would be an effective method to remediate the PAH-contaminated soil.


Assuntos
Lolium , Hidrocarbonetos Policíclicos Aromáticos , Poluentes do Solo , Bactérias/genética , Bactérias/metabolismo , Biodegradação Ambiental , Carvão Vegetal , Expressão Gênica , Lolium/metabolismo , Hidrocarbonetos Policíclicos Aromáticos/análise , Pseudomonas/metabolismo , Solo , Microbiologia do Solo , Poluentes do Solo/análise
3.
NPJ Biofilms Microbiomes ; 8(1): 32, 2022 04 28.
Artigo em Inglês | MEDLINE | ID: mdl-35484166

RESUMO

Bioprospecting of microorganisms suitable for bioremediation of fuel or oil spills is often carried out in contaminated environments such as gas stations or polluted coastal areas. Using next-generation sequencing (NGS) we analyzed the microbiota thriving below the lids of the fuel deposits of diesel and gasoline cars. The microbiome colonizing the tank lids differed from the diversity found in other hydrocarbon-polluted environments, with Proteobacteria being the dominant phylum and without clear differences between gasoline or diesel-fueled vehicles. We observed differential growth when samples were inoculated in cultures with gasoline or diesel as the main carbon source, as well as an increase in the relative abundance of the genus Pseudomonas in diesel. A collection of culturable strains was established, mostly Pseudomonas, Stenotrophomonas, Staphylococcus, and Bacillus genera. Strains belonging to Bacillus, Pseudomonas, Achromobacter, and Isoptericola genera showed a clear diesel degradation pattern when analyzed by GC-MS, suggesting their potential use for bioremediation and a possible new species of Isoptericola was further characterized as hydrocarbon degrader.


Assuntos
Automóveis , Gasolina , Bactérias/genética , Biodegradação Ambiental , Hidrocarbonetos/metabolismo , Pseudomonas/genética , Pseudomonas/metabolismo
4.
Environ Microbiol Rep ; 14(3): 464-474, 2022 06.
Artigo em Inglês | MEDLINE | ID: mdl-35388632

RESUMO

Assistive eco-physiological traits are necessary for microbes to adapt and colonize at polluted niches, enabling efficient clean-up. To demarcate species distinctiveness and eco-physiological traits of aromatic compounds metabolizing Pseudomonas sp. CSV86T (earlier identified as Pseudomonas putida), an Indian isolate from a petrol station soil, comparative genome mining, taxono-genomic, and physiological analyses were performed. A 6.79 Mbp genome (62.72 G + C mol%) of CSV86T encodes 6798 CDS and 238 unique genes. Naphthalene metabolism and Co-Zn-Cd resistance gene clusters were part of distinct genomic islands. Abundance of transporters (aromatics, organic acids, amino acids, and metals) and mobile elements (integrases, transposases, conjugative proteins) differentiated CSV86T from its closest relatives. Enhanced siderophore production for Fe-uptake during aromatic metabolism, indole acetic acid production, and fusaric acid resistance wasvalidated by genomic attributes. Full-length 16S-rRNA phylogeny revealed Pseudomonas japonica WLT as a closest relative of CSV86T . However, lower genomic indices (<97% gyrB-rpoB-rpoD homology, <90% ANI, <50% DNA-DNA relatedness) and taxonomic differences (assimilation of organic acids, amino acids, fatty acids composition) substantially differentiated CSV86T from its closest relatives, indicating it to be a novel species as Pseudomonas bharatica. Preferential metabolism of aromatics with advantageous eco-physiological traits renders CSV86T an ideal candidate for bioremediation and host for metabolic engineering.


Assuntos
Pseudomonas putida , Pseudomonas , Aminoácidos/metabolismo , Técnicas de Tipagem Bacteriana , DNA Bacteriano/genética , DNA Bacteriano/metabolismo , Ácidos Graxos/química , Genes Bacterianos , Genômica , Filogenia , Pseudomonas/genética , Pseudomonas/metabolismo , Pseudomonas putida/genética , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
5.
Bioresour Technol ; 354: 127162, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35429594

RESUMO

Performance and molecular changes of an aerobic denitrifying phosphorus accumulating bacteria Pseudomonas psychrophila HA-2 have been investigated under different temperatures and ZnO nanoparticles (NPs) exposures. Strain HA-2 removed 95.7% of total nitrogen (TN) and 24.6% of phosphorus at 10 °C, which was attributed to the joint up-regulation of intracellular energy metabolism and ribosome. Moreover, with the increase of ZnO NPs from 0 to 100 mg/L, TN and phosphurs removal efficiencies decreased from 95.7% to 44.5% and 24.6% to 6.8% at 10 °C, respectively, whereas phosphorus removal rate increased from 10.5% to 24.5% at 20 °C. Further transcriptomics and proteomics revealed that significant down-regulation of purine and amino acid metabolisms was the main reason for the inhibitory effect at 10 °C, while the up-regulation of antioxidant pathways and functional genes expressions was responsible for the promoted phosphorus accumulation at 20 °C. This study provides a potential solution for improving biological nutrients removal processes in winter months.


Assuntos
Nanopartículas , Óxido de Zinco , Bactérias Aeróbias/metabolismo , Reatores Biológicos/microbiologia , Desnitrificação , Nitrogênio/metabolismo , Fósforo/metabolismo , Pseudomonas/metabolismo , Temperatura , Óxido de Zinco/farmacologia
6.
Arch Microbiol ; 204(5): 247, 2022 Apr 09.
Artigo em Inglês | MEDLINE | ID: mdl-35397008

RESUMO

Genomes of three strains-phenazine producers-Pseudomonas chlororaphis subsp. aurantiaca (B-162 (wild type), mutant strain B-162/255, and its derivative B-162/17) were sequenced and compared. Comparison of a wild-type strain and B-162/255 mutant genomes revealed 32 mutations. 19 new mutations were detected in the genome of B-162/17. Further bioinformatics analysis allowed us to predict mutant protein functions and secondary structures of five gene products, mutations which might potentially influence phenazine synthesis and secretion in Pseudomonas bacteria. These genes encode phenylalanine hydroxylase transcriptional activator PhhR, type I secretion system permease/ATPase, transcriptional regulator MvaT, GacA response regulator, and histidine kinase. Amino acid substitutions were found in domains of studied proteins. One deletion in an intergenic region could affect a potential transcription factor binding site that participates in the regulation of gene that encodes ABC transporter.


Assuntos
Fenazinas , Pseudomonas , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Fenazinas/metabolismo , Pseudomonas/genética , Pseudomonas/metabolismo
7.
ACS Synth Biol ; 11(4): 1650-1657, 2022 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-35389616

RESUMO

Pseudomonas is a large genus that inhabits diverse environments due to its distinct metabolic versatility. Its applications range from environmental to industrial biotechnology. Molecular tools that allow precise and efficient genetic manipulation are required to understand and harness its full potential. Here, we report the development of a highly efficient adenine base editing system, i.e., dxABE-PS, for Pseudomonas species. The system allows A:T → G:C transition with up to 100% efficiency along a broad target spectrum because we use xCas9 3.7, which recognizes NG PAM. To enhance the dxABE-PS utility, we develop a prediction workflow for protein dysfunction using ABE, namely, DABE-CSP (dysfunction via ABE through CRISPOR-SIFT prediction). We applied DABE-CSP to inactivate several genes in Pseudomonas putida KT2440 to accumulate a nylon precursor, i.e., muconic acid from catechol with 100% yield. Moreover, we expanded the ABE to non-model Pseudomonas species by developing an nxABE system for P. chengduensisDY56-96, isolated from sediment samples from the seamount area in the West Pacific Ocean. Taken together, the establishment of the ABE systems along with DABE-CSP will fast-track research on Pseudomonas species.


Assuntos
Sistemas CRISPR-Cas , Edição de Genes , Adenina/metabolismo , Pseudomonas/genética , Pseudomonas/metabolismo , Fluxo de Trabalho
8.
Int J Mol Sci ; 23(8)2022 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-35457206

RESUMO

Bacterial biofilm represents a multicellular community embedded within an extracellular matrix attached to a surface. This lifestyle confers to bacterial cells protection against hostile environments, such as antibiotic treatment and host immune response in case of infections. The Pseudomonas genus is characterised by species producing strong biofilms difficult to be eradicated and by an extraordinary metabolic versatility which may support energy and carbon/nitrogen assimilation under multiple environmental conditions. Nutrient availability can be perceived by a Pseudomonas biofilm which, in turn, readapts its metabolism to finally tune its own formation and dispersion. A growing number of papers is now focusing on the mechanism of nutrient perception as a possible strategy to weaken the biofilm barrier by environmental cues. One of the most important nutrients is amino acid L-arginine, a crucial metabolite sustaining bacterial growth both as a carbon and a nitrogen source. Under low-oxygen conditions, L-arginine may also serve for ATP production, thus allowing bacteria to survive in anaerobic environments. L-arginine has been associated with biofilms, virulence, and antibiotic resistance. L-arginine is also a key precursor of regulatory molecules such as polyamines, whose involvement in biofilm homeostasis is reported. Given the biomedical and biotechnological relevance of biofilm control, the state of the art on the effects mediated by the L-arginine nutrient on biofilm modulation is presented, with a special focus on the Pseudomonas biofilm. Possible biotechnological and biomedical applications are also discussed.


Assuntos
GMP Cíclico , Pseudomonas aeruginosa , Arginina/metabolismo , Arginina/farmacologia , Proteínas de Bactérias/metabolismo , Biofilmes , Carbono/metabolismo , Carbono/farmacologia , GMP Cíclico/metabolismo , Nitrogênio/metabolismo , Nitrogênio/farmacologia , Nutrientes , Pseudomonas/metabolismo , Pseudomonas aeruginosa/fisiologia
9.
Arch Microbiol ; 204(5): 272, 2022 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-35445985

RESUMO

A psychrotolerant bacterial strain of Pseudomonas sp. (P. palleroniana GBPI_508), isolated from the Indian Himalayan region, is studied for analyzing its potential for degrading bisphenol A (BPA). Response surface methodology using Box-Behnken design was used to statistically optimize the environmental factors during BPA degradation and the maximum degradation (97%) was obtained at optimum conditions of mineral salt media pH 9, experimental temperature 25 °C, an inoculum volume of 10% (v/v), and agitation speed 130 rpm at the BPA concentration 270 mg L-1. The Monod model was used for understanding bacterial degradation kinetics, and 37.5 mg-1 half saturation coefficient (KS) and 0.989 regression coefficient (R2) were obtained. Besides, the utmost specific growth rate µmax was witnessed as 0.080 h-1 with the GBPI_508 during BPA degradation. Metabolic intermediates detected in this study by GC-MS were identified as valeric acid, propionic acid, diglycolic acid, and phenol. The psychrotolerant bacterial strain of Pseudomonas sp. (P. palleroniana GBPI_508), isolated from the Indian Himalayan region has shown good potential for remediation of BPA at variable conditions.


Assuntos
Compostos Benzidrílicos , Microbiologia do Solo , Compostos Benzidrílicos/metabolismo , Biodegradação Ambiental , Fenóis , Pseudomonas/metabolismo
10.
Bioprocess Biosyst Eng ; 45(6): 1057-1064, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35412074

RESUMO

Pseudomonas taetrolens has previously been shown to convert cellobiose to cellobionic acid (CBA), which can potentially be used in cosmetics, food, and pharmaceutical industries. The cellobiose-oxidizing activity of the P. taetrolens strain, which expressed the homologous quinoprotein glucose dehydrogenase (GDH), was increased by approximately 50.8% compared to the original strain. Whole-cell biocatalyst (WCB) of the genetically modified P. taetrolens strain [pDSK-GDH] was prepared simply by fermentation and washing processes. Reaction conditions for the proper use of WCB, such as reaction temperature, cell density to be added, and cell harvest time for preparing WCB, were investigated. The highest CBA productivity (18.2 g/L/h) was achieved when WCB prepared in the late-exponential phase of cell culture was used at 35 °C with cell density of 10 at OD600nm. Under these conditions, 200 g/L of cellobiose was all converted to CBA in 11 h, and the WCB of P. taetrolens [pDSK-GDH] maintained the maximum catalytic activity during at least six cycles without a significant decline in the productivity. Our results suggest that the manufacture of WCB based on genetically engineered P. taetrolens and its optimized use could be further developed as an economically viable option for the large-scale production of CBA.


Assuntos
Celobiose , Dissacarídeos , Pseudomonas/genética , Pseudomonas/metabolismo
11.
Appl Biochem Biotechnol ; 194(7): 2901-2918, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35294734

RESUMO

Thiourea is used in agriculture and industry as a metal scavenger, synthetic intermediate, and nitrification inhibitor. However, in wastewater, it can inhibit the nitrification process and induce the collapse of the nitrification system. In such a case, ammonia-oxidizing bacteria (AOB) lose their ability to remove ammonia. We investigated the nitrification system of a 60,000-t/d municipal sewage treatment plant in Nanjing, which collapsed after receiving 5-15 ppm (5-15 mg/L) thiourea. Ammonia nitrogen removal quickly recovered to more than 95% after inoculation with 10 t high-efficiency nitrification sludge, which was collected from a kitchen waste treatment plant. A heterotrophic nitrification strain was isolated from the inoculated sludge and identified as wild Pseudomonas by 16S rDNA sequencing and named "BT1." Based on thiourea tolerance tests, BT1 can tolerate a thiourea content of more than 500 ppm. For comparison, the in situ process was imitated by the simulation system, and the wastewater shocked by 10 ppm thiourea could still meet the emission standard after adding 1% (V/V) BT1. High-throughput sequencing analysis was applied to study microbial succession during thiourea shock loading. The results showed that Hydrogenophaga and Thiobacillus grew with the growth of BT1. Pseudomonas BT1 was used for a 6,000-t/d printed circuit board (PCB) wastewater treatment system, the nitrification system returned to normal in 15 days, and the degradation rate stabilized at more than 95%.


Assuntos
Amônia , Esgotos , Reatores Biológicos , Desnitrificação , Nitrificação , Nitrogênio/metabolismo , Pseudomonas/metabolismo , Esgotos/microbiologia , Tioureia , Águas Residuárias
12.
Appl Biochem Biotechnol ; 194(6): 2747-2761, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35267121

RESUMO

Endosulfan remains as a lipophilic insecticide that causes serious medical problems because of biological stability and toxicity also found in air, water, soil sediments, and foodstuffs. Henceforward, the present study reveals a novel bacterial species isolated from pesticide-contaminated soil for enhanced endosulfan degradation. Next, isolated bacterial species was characterized with biochemical assays and 16S rRNA sequencing technique. Subsequently, the optimal conditions for endosulfan biodegradation such as pH, concentration of endosulfan, and bacterial growth were estimated with non-sulfur medium (NSM). Sequentially, the amount of endosulfan and compound degradation were analyzed through thin-layer chromatography and gas chromatography/mass spectrometry. Overall, the obtained results revealed the endosulfan acting as primary carbon source for bacterial growth. From the GC-MS analysis, the metabolic products released during endosulfan degradation by Pseudomonas sp. MSCAS BT01 were compared with standard GC-MS spectra. The highest (98%) endosulfan degradation was obtained at pH 7.0. The complete endosulfan degradation was achieved at 14th day of incubation and the less toxic endosulfan diol produced was observed via GC-MS. To conclude, the pesticide-contaminated isolate Pseudomonas sp. MSCAS BT01 emerged as a promising bioremediation tool and effectively employed to degrade endosulfan from contaminated soils, sediments, and wastewaters in the days yet to come.


Assuntos
Inseticidas , Praguicidas , Poluentes do Solo , Bactérias/metabolismo , Biodegradação Ambiental , Endossulfano/química , Endossulfano/metabolismo , Inseticidas/metabolismo , Pseudomonas/metabolismo , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/metabolismo , Solo , Microbiologia do Solo , Poluentes do Solo/metabolismo
13.
J Hazard Mater ; 433: 128778, 2022 07 05.
Artigo em Inglês | MEDLINE | ID: mdl-35358812

RESUMO

The oxidation of aqueous arsenite (As(III)) by As(III)-oxidizing bacteria is known to attenuate the mobilization and toxicity of arsenic, and is regarded as potential method for As(III)-pollution remediation. However, during the interactions between As(III)-oxidizing bacteria and different As(III)-adsorbed soil Fe-minerals, the oxidation and partitioning of solid-phase As(III), as well as the controlling mechanisms, remain unclear. In this study, we therefore incubated three As(III)-adsorbed Fe-minerals with a typical As(III)-oxidizing bacteria (Pseudomonas sp. HN-1) at different pH conditions. After microbial oxidation, the percentage of arsenate (As(V)) was significantly higher at pH 7 (15-94%) and 9 (12-89%) than at pH 4 (6-50%) in all Fe-minerals. Incubation of As(III)-oxidizing bacteria promoted As-immobilization under acidic-conditions but As-mobilization under alkaline-conditions. Arsenic-X-ray adsorption spectroscopy results showed that solid-phase As(V) fraction in goethite, hematite and magnetite was 27-64%, 5-12% and 50-91%, respectively. Compared with the corner-sharing As(III)-adsorption complexes formed on magnetite, the edge-sharing complexes on hematite were significantly more stable towards microbial-oxidation. Additionally, the strong adhesion between strain HN-1 and hematite probably limit bacterial-activity and mobility, thereby inhibiting microbial As(III)-oxidation. Our findings elucidate the controlling mechanisms of microbial As(III)-oxidation in different As(III)-adsorbed Fe-minerals and demonstrate strain HN-1 is an excellent candidate for As(III)-remediation in soils containing goethite and magnetite.


Assuntos
Arsênio , Ferro , Arsênio/metabolismo , Arsenitos , Bactérias/metabolismo , Compostos Férricos/química , Óxido Ferroso-Férrico , Concentração de Íons de Hidrogênio , Ferro/metabolismo , Minerais/química , Oxirredução , Pseudomonas/metabolismo , Solo/química
14.
Sci Rep ; 12(1): 2123, 2022 02 08.
Artigo em Inglês | MEDLINE | ID: mdl-35136142

RESUMO

Lactic acid is an important platform chemical used for the production of various compounds including polylactic acid (PLA). Optically pure L- and D-lactic acids are required to obtain high quality PLA. To advance the development and selection of microbial strains for improved production of lactic acid enantiomers, a high-throughput screening, dynamic pathway control, or real-time monitoring are often applied. Inducible gene expression systems and their application in the genetically encoded biosensors contribute to the development of these techniques and are important devices for the advancement of lactic acid biotechnology. Here, we identify and characterize eleven lactate-inducible systems from Escherichia coli, Cupriavidus necator, and Pseudomonas spp. The specificity and dynamics of these systems in response to L- and D-lactate, or structurally similar compounds are investigated. We demonstrate that the inducible systems EcLldR/PlldP and CnGntR/PH16_RS19190 respond only to the L-lactate, exhibiting approximately 19- and 24-fold induction, respectively. Despite neither of the examined bacteria possess the D-lactate-specific inducible system, the PaPdhR/PlldP and PfPdhR/PlldP are induced approximately 37- and 366-fold, respectively, by D-lactate and can be used for developing biosensor with improved specificity. The findings of this study provide an insight into understanding of L- and D-lactate-inducible systems that can be employed as sensing and tuneable devices in synthetic biology.


Assuntos
Cupriavidus necator/metabolismo , Escherichia coli/metabolismo , Ácido Láctico/biossíntese , Família Multigênica , Pseudomonas/metabolismo , Técnicas Biossensoriais , Cupriavidus necator/genética , Escherichia coli/genética , Pseudomonas/genética , Biologia Sintética
15.
Microbiol Spectr ; 10(1): e0034521, 2022 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-35196785

RESUMO

Polyphosphate (polyP) accumulation is an important trait of microorganisms. Implication of polyP accumulating bacteria (PAB) in enhanced biological phosphate removal, heavy metal sequestration, and dissolution of dental enamel is well studied. Phosphorous (P) accumulated within microbial biomass also regulates labile P in soil; however, abundance and diversity of the PAB in soil is still unexplored. Present study investigated the genetic and functional diversity of PAB in rhizosphere soil. Here, we report the abundance of Pseudomonas spp. as high PAB in soil, suggesting their contribution to global P cycling. Additional subset analysis of functional genes i.e., polyphosphate kinase (ppk) and exopolyphosphatase (ppx) in all PAB, indicates their significance in bacterial growth and metabolism. Distribution of functional genes in phylogenetic tree represent a more biologically realistic discrimination for the two genes. Distribution of ppx gene disclosed its phylogenetic conservation at species level, however, clustering of ppk gene of similar species in different clades illustrated its environmental condition mediated modifications. Selected PAB showed tolerance to abiotic stress and strong correlation with plant growth promotary (PGP) traits viz. phosphate solubilization, auxin and siderophore production. Interaction of PAB with A. thaliana enhanced the growth and phosphate status of the plant under salinity stress, suggestive of their importance in P cycling and stress alleviation. IMPORTANCE Study discovered the abundance of Pseudomonas genera as a high phosphate accumulator in soil. The presence of functional genes (polyphosphate kinase [ppk] and exopolyphosphatase [ppx]) in all PAB depicts their importance in polyphosphate metabolism in bacteria. Genetic and functional diversity reveals conservation of the ppx gene at species level. Furthermore, we found a positive correlation between PAB and plant growth promotary traits, stress tolerance, and salinity stress alleviation in A. thaliana.


Assuntos
Arabidopsis/crescimento & desenvolvimento , Polifosfatos/metabolismo , Pseudomonas/genética , Pseudomonas/metabolismo , Microbiologia do Solo , Hidrolases Anidrido Ácido/genética , Hidrolases Anidrido Ácido/metabolismo , Arabidopsis/microbiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Variação Genética , Ácidos Indolacéticos/metabolismo , Fósforo/metabolismo , Fosfotransferases (Aceptor do Grupo Fosfato)/genética , Fosfotransferases (Aceptor do Grupo Fosfato)/metabolismo , Filogenia , Pseudomonas/classificação , Pseudomonas/enzimologia , Rizosfera , Sideróforos/biossíntese , Solo/química
16.
Microbiol Spectr ; 10(1): e0103321, 2022 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-35196816

RESUMO

Bdellovibrio bacteriovorus is a predatory, Gram-negative bacteria that feeds on many pathogenic bacteria and has been investigated as a possible solution for mitigating biofilms in different fields. The application depends on more fundamental ecological studies into the dynamics between Bdellovibrio and their prey. To do so requires an accurate, reliable, and, preferably rapid, way of enumerating the cells. Flow cytometry (FCM) is potentially a rapid, accurate, and inexpensive tool for this, but it has yet to be validated in the enumeration of Bdellovibrio. In this study, we developed a protocol to measure the number of Bdellovibrio in samples of various densities using FCM and compared the results with those of other methods: optical density (OD), PFU assay (PFU), and quantitative PCR (qPCR). We observed a strong correlation between values obtained using FCM and PFU (ρ = 0.923) and FCM and qPCR (ρ = 0.987). Compared to optical density there was a much weaker correlation (ρ = 0.784), which was to be expected given the well-documented uncertainty in converting optical density (OD) to cell numbers. The FCM protocol was further validated by demonstrating its ability to distinguish and count mixed populations of Bdellovibrio and the prey Pseudomonas. Thus, the accuracy of FCM as well as its speed and reproducibility make it a suitable alternative for measuring Bdellovibrio cell numbers, especially where many samples are required to capture the dynamics of predator-prey interactions. IMPORTANCE The rise of antibiotic resistance and the unwanted growth of bacteria is a universally growing problem. Predatory bacteria can be used as a biological alternative to antibiotics because they grow by feeding on other bacteria. To apply this effectively requires further study and a deeper understanding of the forces that drive a prey population to elimination. Initially, such studies require more reliable methods to count these cells. Flow cytometry (FCM) is potentially a rapid, accurate, and inexpensive tool for this, but it has yet to be validated for predatory bacteria. This study develops a protocol to count the predatory bacteria Bdellovibrio bacteriovorus and its Pseudomonas prey using FCM and compare the results with those of other methods, demonstrating its ability for studies into B. bacteriovorus predation dynamics. This could lead to the use of B. bacteriovorus for killing bacterial biofilms in fields, such as drinking water and agriculture.


Assuntos
Bdellovibrio bacteriovorus/fisiologia , Citometria de Fluxo/métodos , Pseudomonas/metabolismo , Biofilmes
17.
J Agric Food Chem ; 70(7): 2136-2145, 2022 Feb 23.
Artigo em Inglês | MEDLINE | ID: mdl-35147028

RESUMO

Microbial bioremediation is a very potent and eco-friendly approach to alleviate pesticide pollution in agricultural ecosystems, and hydrolase is an effective element for contaminant degradation. In the present study, a novel Mn2+-dependent esterase, PchA, that efficiently hydrolyzes carbamate pesticides with aromatic structures was identified from Pseudomonas sp. PS21. The hydrolytic activity was confirmed to be related closely to the core catalytic domain, which consists of six residues. The crucial residues indirectly stabilized the position of carbaryl via chelating Mn2+ according to the binding model clarified by molecular simulations, and the additional hydrophobic interactions between carbaryl with several hydrophobic residues also stabilized the binding conformation. The residue Glu398, by serving as the general base, might activate a water molecule and facilitate PchA catalysis. This work offers valuable insights into the binding interaction and hydrolytic mechanism of carbaryl with the hydrolase PchA and will be crucial to designing strategies leading to the protein variants that are capable of degrading related contaminants.


Assuntos
Carbaril , Pseudomonas , Biodegradação Ambiental , Carbaril/metabolismo , Ecossistema , Esterases/metabolismo , Pseudomonas/genética , Pseudomonas/metabolismo
18.
Biodegradation ; 33(2): 169-180, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-35103887

RESUMO

The present study is aimed to develop a microbial system for efficient naphthalene bioremediation. A phytotoxicity study was carried out to check the naphthalene detoxification efficiency of Pseudomonas sp. strain SA3 in mung bean (Vigna radiata). For this, administration of the degraded product (supernatant) of 500 mg L-1 naphthalene by Pseudomonas sp. strain SA3 was studied on V. radiata till 168 h. The growth parameters of mung bean seedlings exposed to treated naphthalene solution were statistically similar to distilled water but a twofold decrease when exposed to untreated naphthalene solution. Further, through the soil microcosm study, the naphthalene degradation by pure colonies of Pseudomonas sp. strain SA3 was 6.8% higher as compared to when the natural microflora was mixed with Pseudomonas sp. strain SA3. Further naphthalene degradation by a microcosm model revealed that with an increased concentration of glucose, the carbon dioxide trap rate decreases.


Assuntos
Pseudomonas , Vigna , Biodegradação Ambiental , Naftalenos/metabolismo , Pseudomonas/metabolismo , Solo , Microbiologia do Solo
19.
Sci Rep ; 12(1): 2666, 2022 02 17.
Artigo em Inglês | MEDLINE | ID: mdl-35177693

RESUMO

The continuing reports of plastic pollution in various ecosystems highlight the threat posed by the ever-increasing consumption of synthetic polymers. Therefore, Pseudomonas capeferrum TDA1, a strain recently isolated from a plastic dump site, was examined further regarding its ability to degrade polyurethane (PU) compounds. The previously reported degradation pathway for 2,4-toluene diamine, a precursor and degradation intermediate of PU, could be confirmed by RNA-seq in this organism. In addition, different cell fractions of cells grown on a PU oligomer were tested for extracellular hydrolytic activity using a standard assay. Strikingly, purified outer membrane vesicles (OMV) of P. capeferrum TDA1 grown on a PU oligomer showed higher esterase activity than cell pellets. Hydrolases in the OMV fraction possibly involved in extracellular PU degradation were identified by mass spectrometry. On this basis, we propose a model for extracellular degradation of polyester-based PUs by P. capeferrum TDA1 involving the role of OMVs in synthetic polymer degradation.


Assuntos
Fenilenodiaminas/metabolismo , Poliuretanos/metabolismo , Pseudomonas/metabolismo , Biodegradação Ambiental
20.
Genes (Basel) ; 13(2)2022 02 18.
Artigo em Inglês | MEDLINE | ID: mdl-35205417

RESUMO

CbrAB is a two-component system, unique to bacteria of the family Pseudomonaceae, capable of integrating signals and involved in a multitude of physiological processes that allow bacterial adaptation to a wide variety of varying environmental conditions. This regulatory system provides a great metabolic versatility that results in excellent adaptability and metabolic optimization. The two-component system (TCS) CbrA-CbrB is on top of a hierarchical regulatory cascade and interacts with other regulatory systems at different levels, resulting in a robust output. Among the regulatory systems found at the same or lower levels of CbrAB are the NtrBC nitrogen availability adaptation system, the Crc/Hfq carbon catabolite repression cascade in Pseudomonas, or interactions with the GacSA TCS or alternative sigma ECF factor, such as SigX. The interplay between regulatory mechanisms controls a number of physiological processes that intervene in important aspects of bacterial adaptation and survival. These include the hierarchy in the use of carbon sources, virulence or resistance to antibiotics, stress response or definition of the bacterial lifestyle. The multiple actions of the CbrAB TCS result in an important competitive advantage.


Assuntos
Proteínas de Bactérias , Regulação Bacteriana da Expressão Gênica , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Carbono/metabolismo , Nitrogênio/metabolismo , Pseudomonas/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...