Speciation of lead in seawater and river water by using Saccharomyces cerevisiae immobilized in agarose gel as a binding agent in the diffusive gradients in thin films technique

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Use of Saccharomyces cerevisiae immobilized in agarose gel as a binding agent for diffusive gradients in thin films

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Speciation analysis of inorganic arsenic in river water by Amberlite IRA 910 resin immobilized in a polyacrylamide gel as a selective binding agent for As(v) in diffusive gradient thin film technique

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Determination of labile barium in petroleum-produced formation water using paper-based DGT samplers

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Paper-based diffusive gradients in thin films technique coupled to energy dispersive X-ray fluorescence spectrometry for the determination of labile Mn, Co, Ni, Cu, Zn and Pb in river water

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Immobilization of lipases and assay in continuous fixed bed reactor

Lipases are versatile enzymes regarding the range of reactions they catalyse and substrates on which they act. They are as well important as catalyst in organic synthesis. Their immobilization on appropriate supports confer them greater stability besides the possibility of operating in continuous reactors. In order to explore these abilities, the reactions involving hydrolysis of p-nitrophenyl acetate (PNPA) and transesterification of PNPA with n-butanol were chosen. Lipases from two different sources were assayed, namely: microbial (Candida rugosa, CRL, Sigma Type VII) and pancreatic (PPL, Sigma, Type 11). Two immobilization methods were also used, namely: 1) adsorption, using as support the following silica derivatives (150-300μm e 450μ): phenyl, epoxy, amino and without derivation, and 2) covalent binding, using glutaraldehyde as binding agent and silica amino as support. This later method led to better results. Hydrolytic activity was 6.1 U/gsupport for CRL and 0.97U/gsupport for PPL, and of transesterification, 2,8U/gsupport for CRL and 1,9U/gsupport for PPL. Stability of the immobilized enzyme as a function of temperature was evaluated for CRL at 40°C and 50°C and for PPL at 32°C and 40°C. The assays were initially carried out batchwise, both for soluble and immobilized enzymes, aiming to the obtention of parameters for the continues reactor. Lipases immobilized by covalent binding were used in the assays of operacional stability in continuos reactors. For PPL in aqueous medium, at 32°C, and CRL in organic medium at 40°C, both operating continuously, no significant loss of activity was detected along the analysis period of 17 days. In the case of CRL in aqueous medium at 40°C there was a loss of activity around 40% after 18 days. For PPL in organic medium at 40°C the loss was 33% after 20 days. Compairing both sources with each other, very different results were obtained. Higher activitiy was found for CRL, both for hydrolysis and for transesterification reactions, with higher stability in organic medium. PPL showed lower activity as well as higher stability in aqueous medium. The immobilization method by covalent binding showed to be the most appropriate. Immobilized lipases are therefore relatively stable both in aqueous and organic medium.

Microstructure of ceramics TiO2 obtained by protein consolidation

Porous ceramics can be produced by adding starch (corn, potato) and protein (animal or vegetable) to raw material as pore forming element. In this study, titanium dioxide ceramics were formed by vegetable protein consolidation. Soybean was chosen as the binding agent and pore forming. The samples, which were produced in cylindrical shape, had the following processing: material mixture, gelling, drying, pre-sintering and sintering. Heated platinum microscopy were performed by using suspensions with different compositions in order to verify protein gelling capacity and better know the temperature in which this process occurs. The samples were characterized by apparent porosity and roughness measurement. Besides, imaging by light microscopy was also performed in order to determine the sample morphology and porosity. © (2012) Trans Tech Publications, Switzerland.

Biodisponibilidade de metais na baía da Ilha Grande, RJ: avaliação pelas técnicas de bivalves transplantados (Nodipecten nodosus) e difusão de filmes finos por gradiente de concentração

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Determinação de Cd, Cu, Ni, Pb e Zn em águas marinhas oriundas da Bacia Potiguar utilizando a técnica de difusão em filmes finos por gradiente de concentração (DGT)

The DGT technique has been used to determine and better understand the dynamics and bioavailability of metals in the sea near to marine outfalls. In this work, the DGT was assessed for the determination of Cd, Pb, Cu, Zn and Ni in marine water samples from Potiguar Basin in its conventional aspect (binding agent, Chelex-100 and diffusive agent: Diffusive polyacrylamide gel (D) and restrictive (R)) and determination of Pb and Cd through its alternative aspect (binding agent: Saccharomyces cerevisiae immobilized in agarose gels and diffusive agent: 3MM Chromatography Paper). The deployment curves for long periods of immersion in the sample, showed a good linear correlation for the conventional aspect to Zn (D: R²=0.9586, R: R²=0.9444), Ni (D: R²=0.9789, R: R²=0.9286) and Cu (D: R²=0.764, R: R²=0.8143), and alternative to Pb (R²=0.9228) and Cd (R²=0.9673). The results of the organic and inorganic composition analysis in conventional aspect, showed that for every element there is no significant fraction of organic compounds in the sample. In addition, considering the alternative aspect, the comparison of obtained and expected masses suggests that some of the Pb is not labile and that the main retention mechanism of Pb for S. Cerevisiae occurs through ion exchange

Avaliação da membrana P81 como agente ligante da técnica de difusão em filmes finos por gradiente de concentração (DGT) para determinação de mercúrio em sistemas aquáticos

Worldwide environmental degradation is an undesirable byproduct resulting from the increasing demand for natural resources. Water sources are suffering intense contamination since they usually receive a huge amount of domestic and industrial effluents - which are mostly wasted without proper treatment - inserting a large number of pollutants in the environment, heavy metals included. Mercury holds great toxicological importance because, under some physicochemical conditions in a water environment, Hg (II) ion turns into methylated compounds stemming from this element, such as methylmercury CH3Hg, which is highly toxic for the aquatic community in which bioaccumulation occurs. Nowadays passive sampling techniques are being developed to enable the analytical procedures which are applied in environmental monitoring. Diffusive gradients in thin-films technique (DGT) has been proven an interesting tool for the determination of labile metal species due to its in situ application. The DGT technique consists of a piston-like device on which the following series of agents is disposed: a binding agent (conventionally Chelex 100 resin), a diffusive agent, usually a polyacrylamide gel, and a membrane filter. Nevertheless, the agents conventinally used for this technique don't usually show satisfactory results in mercury sampling. The main goal of this study was to evaluate the phosphate-treated cellulose membrane (Whatman P 81), an alternative material, as binding agent in the DGT to determine labile mercury fractions in aquatic systems. In this context, we conducted a study of the behavior of this material in relation with system variables, pH and ionic strength. Afterwards we performed immersions of the DGT devices in real and enriched samples and in situ aiming the determination of mercury

Determinação de vanádio em águas usando a técnica de difusão em filmes finos por gradientes de concentração

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Crystallographic and pre-steady-state kinetics studies on binding of NADH to wild-type and isoniazid-resistant enoyl-ACP(CoA) reductase enzymes from Mycobacterium tuberculosis

An understanding of isoniazid (INH) drug resistance mechanism in Mycobacterium tuberculosis should provide significant insight for the development of newer anti-tubercular agents able to control INH-resistant tuberculosis (TB). The inhA-encoded 2-trans enoyl-acyl carrier protein reductase enzyme (InhA) has been shown through biochemical and genetic studies to be the primary target for INH. In agreement with these results, mutations in the inhA structural gene have been found in INH-resistant clinical isolates of M. tuberculosis, the causative agent of TB. In addition, the InhA mutants were shown to have higher dissociation constant values for NADH and lower values for the apparent first-order rate constant for INH inactivation as compared to wild-type InhA. Here, in trying to identify structural changes between wild-type and INH-resistant InhA enzymes, we have solved the crystal structures of wild-type and of S94A, I47T and I21V InhA proteins in complex with NADH to resolutions of, respectively, 2.3 angstrom, 2.2 angstrom, 2.0 angstrom, and 1.9 angstrom. The more prominent structural differences are located in, and appear to indirectly affect, the dinucleotide binding loop structure. Moreover, studies on pre-steady-state kinetics of NADH binding have been carried out. The results showed that the limiting rate constant values for NADH dissociation from the InhA-NADH binary complexes (k(off)) were eleven, five, and tenfold higher for, respectively, I21V, I47T and S94A INH-resistant mutants of InhA as compared to INH-sensitive wildtype InhA. Accordingly, these results are proposed to be able to account for the reduction in affinity for NADH for the INH-resistant InhA enzymes. (c) 2006 Elsevier Ltd. All rights reserved.

Incorporation of dithiooxamide as a complexing agent into cellulose for the removal and pre-concentration of Cu(II) and Cd(II) ions from natural water samples

The present study describes the incorporation of a complexing agent, dithiooxamide, into microcrystalline cellulose for use in the pre-concentration of Cu(II) and Cd(II) ions from aqueous samples. The FTIR spectrum of the adsorbent exhibited an absorption band in the region of 800 cm-1, which confirmed the binding of the silylating agent to the matrix. Elemental analysis indicated the amount of 0.150 mmol g-1 of the complexing agent. The adsorption data were fit to the modified Langmuir equation, and the maximum amount of metal species extracted from the solution, Ns, was determined to be 0.058 and 0.072 mmol g-1 for Cu(II) and Cd(II), respectively. The covering fraction φ, which was 0.39 and 0.48 for Cu(II) and Cd(II), respectively, was used to estimate a 1:2 (metal:ligand) ratio in the formed complex, and a binding model was proposed based on this information. The adsorbent was applied in the pre-concentration of natural water samples and exhibited an enrichment factor of approximately 50-fold for the species studied, which enabled its use in the analysis of trace metals in aqueous samples. The system was validated by the analysis of certified standard (1643e), and the adsorbent was stable for more than 20 cycles, thus enabling its safe reutilization. © 2012 Elsevier B.V. All rights reserved.