RESUMO
The increase of industrial discharges is the first cause of the contamination of water bodies. The bacterial survival strategies contribute to the equilibrium restoration of ecosystems being useful tools for the development of innovative environmental biotechnologies. The aim of this work was to study the Cu(II) and Cd(II) biosensing, removal and recovery, mediated by whole cells, exopolymeric substances (EPS) and biosurfactants of the indigenous and non-pathogenic Pseudomonas veronii 2E to be applied in the development of wastewater biotreatments. An electrochemical biosensor was developed using P. veronii 2E biosorption mechanism mediated by the cell surface associated to bound exopolymeric substances. A Carbon Paste Electrode modified with P. veronii 2E (CPEM) was built using mineral oil, pre-washed graphite power and 24 h-dried cells. For Cd(II) quantification the CPEM was immersed in Cd(II) (1-25 µM), detected by Square Wave Voltammetry. A similar procedure was used for 1-50 µM Cu(II). Regarding Cd(II), removal mediated by immobilized EPS was tested in a 50 ml bioreactor with 0.13 mM Cd(II), pH 7.5. A 54% metal retention by EPS was achieved after 7 h of continuous operation, while a 40% was removed by a control resin. In addition, surfactants produced by P. veronii 2E were studied for recovery of Cd(II) adsorbed on diatomite, obtaining a 36% desorption efficiency at pH 6.5. Cu(II) adsorption from a 1 mM solution was tested using P. veronii 2E purified soluble EPS in 50 mL- batch reactors (pH = 5.5, 32°C). An 80% of the initial Cu(II) was retained using 1.04 g immobilized EPS. Focusing on metal recovery, Cu nanoparticles (NPs) biosynthesis by P. veronii 2E was carried out in Cu(II)-PYG Broth at 25°C for 5 days. Extracellular CuNPs were characterized by UV-Vis spectral analysis while both extracellular and intracellular NPs were analyzed by SEM and TEM techniques. Responses of P. veronii 2E and its products as biosurfactants, bound and soluble EPS allowed Cu(II) and Cd(II) removal, recovery and biosensing resulting in a multiple and versatile tool for sustainable wastewater biotreatments.
RESUMO
Pseudomonas veronii 2E, an autochthonous bacterium isolated from sediments associated to a high-polluted watershed, produces a complex matrix of exopolymers with carbohydrates as main components. In this work, four polysaccharides were isolated from the extracellular material. The major acidic polysaccharide named EPO2, was purified and its structure was elucidated using Matrix-assisted laser desorption/ionization and Electrospray ionization mass spectrometry, Infrared spectroscopy, Nuclear magnetic resonance spectroscopy and chemical treatments. This heteropolysaccharide consists in an α(1-4) glucan substituted with N-Acetylglucosamine residues and with a branching α-D-GlcpA-(1-3)-L-Fucp disaccharide. The biosorption capacity of EPO2 and of the whole exopolysaccharide to Pb(II), Zn(II), Cu(II) and Fe(II) was evaluated. EPO2 showed a remarkable sorption capacity for Fe(II) with an efficiency of 70% and for Zn(II) 39%. When the whole exopolysaccharide fraction was tested it showed a significantly lower metal sorption ability than purified EPO2 suggesting the involvement of the distinct acidic branching disaccharide in this interaction.
Assuntos
Cobre/química , Ferro/química , Chumbo/química , Polissacarídeos Bacterianos/química , Pseudomonas/metabolismo , Zinco/química , Adsorção , Matriz Extracelular de Substâncias Poliméricas/química , Espectroscopia de Ressonância Magnética , Polissacarídeos Bacterianos/isolamento & purificação , Solubilidade , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
The assessment of water quality is critical to implement preventive and emergency interventions aimed to limit/avoid environmental contamination and human exposure to toxic compounds. While established high-resolution techniques allow quantitative and qualitative determination of contaminants, their widespread application is not feasible due to cost, time, and need for trained personnel. In this context, the development of easy-to-implement approaches for preliminary detection of contaminants is of the utmost importance. Herein, a portable self-powered microbial electrochemical sensor enabling online monitoring of Cr(VI) is reported. The biosensor employs a bio-inspired redox mediating system to allow extracellular electron transfer between a bacterial isolate from chromium-contaminated environments and the electrode, providing a clear response to Cr(VI) presence. The biosensor shows good linearity (R2 = 0.983) and a limit of detection of 2.4 mg L-1 Cr(VI), with a sensitivity of 0.31 ± 0.02 µA cm-2 mgCr(VI)-1 L. The presented microbial bioanode architecture enhanced biosensor performance thanks to the improved "electrical wiring" between biological entities and the abiotic electrode surface. This approach could be easily implemented in engineered electrode surfaces, such as paper-based multi-anodes that maximize bacterial colonization, further improving biosensor response. Graphical abstract.
Assuntos
Técnicas Biossensoriais/métodos , Cromo/análise , Poluentes Ambientais/análise , Pseudomonas/metabolismo , Fontes de Energia Bioelétrica , Eletrodos , Transporte de Elétrons , Humanos , Modelos Moleculares , OxirreduçãoRESUMO
Siderophores are low-molecular weight ligands secreted by bacteria as a survival strategy in Fe(III)-lacking environments. They bind not only Fe(III), but Co(II), Zn(II), Mn(II), Ni(II), Ga(III) as a detoxification alternative. The synthesis, purification and characterization of siderophores produced by Pseudomonas veronii 2E were evaluated to be applied in future environmental technologies. Optimal production was obtained in Fe(III)-free M9-succinate at 25 °C, 40 h and pH 6.9. Siderophores were chemically characterized as hydroxamate and catechol mixed-type. Spectroscopic analysis indicated their belonging to the pyoverdine family, behaving as ligand to Cd(II), Zn(II), Cu(II), Ni(II) and Cr(III), which promoted siderophoregenesis during growth. Siderophore-Cd(II) complexation was studied by electrochemical monitored titration revealing one family of moderate-strength binding sites. Mass spectral analysis evidenced the secretion of a variety of molecules (molecular mass ca.1200 u). Non pathogenic Pseudomonas veronii 2E siderophores represent a safe alternative for the concrete application of environmental technologies and clinical procedures.
Assuntos
Pseudomonas/metabolismo , Sideróforos/biossíntese , Sideróforos/química , Sideróforos/isolamento & purificação , Compostos Férricos , Concentração de Íons de Hidrogênio , Quelantes de Ferro , Ligantes , Espectrometria de Massas , Metais/farmacologia , Oligopeptídeos/química , Pseudomonas/efeitos dos fármacos , Pseudomonas/crescimento & desenvolvimentoRESUMO
Polyvalent gold nanoparticle oligonucleotide conjugates are subject of intense research. Even though 2nm diameter AuNPs have been previously modified with DNA, little is known about their structure and electrochemical behavior. In this work, we examine the influence of different surface modification strategies on the interplay between the meso-organization and the molecular recognition properties of a 27-mer DNA strand. This DNA strand is functionalized with different sulfur-containing moieties and immobilized on 2nm gold nanoparticles confined on a nanoporous alumina, working the whole system as an electrode array. Surface coverages were determined by EXAFS and the performance as recognition elements for impedance-based sensors is evaluated. Our results prove that low DNA coverages on the confined nanoparticles prompt to a more sensitive response, showing the relevance in avoiding the DNA strand overcrowding. The system was able to determine a concentration as low as 100pM of the complementary strand, thus introducing the foundations for the construction of label-free genosensors at the nanometer scale.
Assuntos
Técnicas Biossensoriais/métodos , DNA/análise , Ouro/química , Ácidos Nucleicos Imobilizados/química , Nanopartículas Metálicas/química , Óxido de Alumínio/química , Técnicas Eletroquímicas/métodos , Eletrodos , Nanoestruturas/química , Hibridização de Ácido Nucleico/métodos , PorosidadeRESUMO
A controlled architecture of nanoelectrodes, of a similar size to small molecule-binding aptamers, is synthesized inside nanoporous alumina. Gold nanoparticles with a controlled size (about 2 nm) are electrogenerated in the alumina cavities, showing a fast electron transfer process toward ferrocyanide. These uncapped nanoparticles are easily modified with a thiol-containing aptamer for label-free detection of adenosine monophosphate by electrochemical impedance spectroscopy. Our results show that the use of a limited electrical conducting surface inside an insulating environment can be very sensitive to conformational changes, introducing a new approach to the detection of small molecules, exemplified here by the direct and selective detection of adenosine monophosphate at the nanomolar scale.
Assuntos
Aptâmeros de Nucleotídeos/química , Ouro/química , Nanopartículas Metálicas/química , Nanotecnologia/métodos , Monofosfato de Adenosina/análise , Óxido de Alumínio/química , Impedância Elétrica , PorosidadeRESUMO
Adsorption properties of bacterial biomass were tested for Cd removal from liquid effluents. Experimental conditions (pH, time, cellular mass, volume, metal concentration) were studied to develop an efficient biosorption process with free or immobilised cells of Pseudomonas veronii 2E. Surface fixation was chosen to immobilise cells on inert surfaces including teflon membranes, silicone rubber and polyurethane foam. Biosorption experiments were carried out at 32 degrees C and controlled pH; maximal Cd(II) retention was observed at pH 7.5. The isotherm followed the Langmuir model (K(d)=0.17 mM and q(max)=0.48 mmol/g cell dry weight). Small changes in the surface negative charge of cells were observed by electrophoretic mobility experiments in presence of Cd(II). In addition, biosorption of 40% Cu(II) (pH 5 and 6.2) and 50% Zn(II) and 50% Cd(II) (pH 7.5) was observed from mixtures of Cu(II), Zn(II) and Cd(II) 0.5mM each.