Imobilização de enzimas em suportes cromatográficos: uma ferramenta na busca por substâncias bioativas

The development and characterization of bioreactors or IMER (immobilized enzyme reactors) as research tools are important in the scope of medicinal chemistry and constitute an alternative for the rational development of drugs. This approach does not require highly purified enzymes or a great amount of protein, but increase the enzymatic stability against heat, organic solvents and pH, without too much loss of catalyst activity. Immobilized enzyme reactors (IMER) can be used for the accomplishment of high efficiency screening on-line and, thus inhibitors can be quickly identified. Here, we emphasize the development of IMER by use of different methods of immobilization and chromatographic supports. Their applications, in different areas of research, are also fully discussed.

Biodiesel: visão crítica do status atual e perspectivas na academia e na indústria

This article presents a bibliographic review of research carried out on different alternative processes for biodiesel production. The supercritical and subcritical (non catalytic) reaction conditions, the use of solid basic, solid acid and other heterogeneous catalysts, including the use of immobilized enzymes and whole-cell catalysts are also critically compared with the traditional homogeneous alkaline or acid catalysts that are common on industrial applications. Advantages and limitations of all these processes for the transference from the laboratory to the industry are discussed. A correlation of the chemical composition with the quality parameters of the produced biodiesel is done with aim to stablish adequate procedures for the right selection of the raw-material. Castor bean oil is used as an example of inappropriate oil in order to produce a B100 that fulfill all the international physico-chemical quality standards. In this article are presented research results to adequate the values of viscosity, density and iodine number of the castor and soybean biodiesel to the international standard limits by means blending these both biodiesels at the right ratio.

Fotodegradación heterogénea de bisfenol A en agua con dióxido de titanio

Bisphenol A (BPA) is a monomer used in epoxy resin and polycarbonate manufacture. This molecule is considered as an endocrine disruptor that causes different diseases. The human exposition to this non biodegrable substance is increasing in the time; in particular, water is contaminated by industrial remainder flow. In this article heterogeneous photo degradation of a solution of BPA in water solution using a catalytic photo reactor with UV light and titanium dioxide (TiO2) was evaluated. High performance liquid chromatography (HPLC) was used to analyze the photo degradation of BPA solutions. The influence of titanium dioxide amount, BPA concentration, reaction temperature and the catalyst state like suspension and immobilized were also determinated. The highest elimination of BPA was 83.2%, in 240 min, beginning with 0.05 mM of BPA and 100 mg/L of TiO2 in suspension.

Imobilização da lacase em micropartículas de quitosana obtidas por spray drying e usadas na construção de biossensores

Biosensors based on laccase immobilized on microparticles of chitosan crosslinked with tripolyphosphate (biosensor I) and glyoxal (biosensor II) obtained by spray drying for the determinations of rutin in pharmaceutical formulations were developed. Under optimized operational conditions (pH 4.0, frequency of 30 Hz, pulse amplitude of 40 mV and scan increment of 2.0 mV) two analytical curves were obtained for both biosensors showing a detection limit of 6.2x10-8 mol L-1 for biosensor (I) and 2.0x10-8 mol L-1 for biosensor (II). The recovery of rutin from pharmaceutical sample ranged from 90.7 to 105.0% and the lifetime of these biosensors were 4 months (at least 400 determinations).

On-line preconcentration system using a microcolumn packed with Alizarin Red S-modified alumina for zinc determination by flame atomic absorption spectrometry

A simple and sensitive on-line flow injection system for determination of zinc with FAAS has been described. The method is based on the separation and preconcentration of zinc on a microcolumn of immobilized Alizarin Red S on alumina. The adsorbed analyte is then eluted with 250 µL of nitric acid (1 mol L-1) and is transported to flame atomic absorption spectrometer for quantification. The effect of pH, sample and eluent flow rates and presence of various cations and anions on the retention of zinc was investigated. The sorption of zinc was quantitative in the pH range of 5.5-8.5. For a sample volume of 25 mL an enrichment factor of 144 and a detection limit (3S) of 0.2 µg L-1 was obtained. The precision (RSD, n=7) was 3.0% at the 20 µg L-1 level. The developed system was successfully applied to the determination of zinc in water samples, hair, urine and saliva.

Cromatografia de afinidade por íons metálicos imobilizados (IMAC) de biomoléculas: aspectos fundamentais e aplicações tecnológicas

Immobilized Metal Ion Affinity Cromatography - IMAC - is a group-specific based adsorption applied to the purification and structure-function studies of proteins and nucleic acids. The adsorption is based on coordination between a metal ion chelated on the surface of a solid matrix and electron donor groups at the surface of the biomolecule. IMAC is a highly selective, low cost, and easily scaled-up technique being used in research and commercial operations. A separation process can be designed for a specific molecule by just selecting an appropriate metal ion, chelating agent, and operational conditions such as pH, ionic strength, and buffer type.

Estudo da oxidação dos sulfetos sintéticos molibdenita (MoS2) e covelita (CuS) por Acidithiobacillus ferrooxidans via respirometria celular

This paper analyses the oxidation of covellite and molybdenite by Acidithiobacillus ferrooxidans strain LR using respirometric experiments. The results showed that both sulfides were oxidized by A. ferrooxidans, however, the covellite oxidation was much higher than molybdenite. Regarding the kinetic oxidation, the findings revealed that just molybdenite oxidation followed the classical Michaelis-Menten kinetic. It is probably associated with the pathway which these sulfides react to chemistry-bacterial attack, what is influenced by its electronic structures. Besides, experiments conducted in the presence of Fe3+ did not indicate alterations in molybdenite oxidation. Thus, ferric ions seem not to be essential to the sulfide oxidations.

Immobilization of laccase on hybrid layered double hydroxide

Crystals of Mg/Al layered double hydroxide were synthesized by alkaline precipitation and treated in an aqueous solution of glutamic acid. The glutamate ions were not intercalated into the interlayer space, but were detected in the material by Fourier transform infrared spectroscopy, suggesting that only the external surfaces of crystals were modified with glutamate ions. The resulting hybrid material was tested as a support for immobilization of the enzyme laccase (Myceliophthora thermophila). The immobilized enzyme preparation was characterized by electronic paramagnetic resonance spectroscopy and by assays of catalytic activity. The activity of the immobilized laccase was 97% of the activity in the free enzyme. Layered double hydroxide is a suitable support for use in remediation of soil studies.

Bioacetilação de álcoois catalisada por Saccharum officinarum

Lipase-catalysed esterifications of alcohols using immobilized enzyme system from sugar cane (Saccharum officinarum) as biocatalyst afforded the corresponding esters in considerable yields (68-93%). Under optimized conditions, the material was utilized for reactions of acetylation with several advantage. It also investigated the possibility of reuse of immobilized enzymes of S. officinarum as biocatalyst under optimal reaction conditions.

Síntese enzimática de butirato de isoamila empregando lipases microbianas comerciais

Isoamyl butyrate production was investigated using free and immobilized lipases by esterification of butyric acid with isoamyl alcohol in a solvent-free system and in an organic media. Among the enzymes studied, Lipozyme TL IM was found to be the most active catalyst in n-hexane as a solvent. The effects of different solvents and the amount of water added on conversion rates were studied. A maximum conversion yield of 80% in n-hexano at 48 h was obtained under the following conditions: 3 g L-1 of Lipozyme TL IM, 30 ºC, 180 rpm of agitation, isoamyl alcohol to butyric acid molar ratio of 1:1 and acid substrate concentration of 0.06 M.

Alternativa potencial para aproveitamento do glicerol gerado na produção de biodiesel: síntese enzimática de monolaurina por esterificação

Esterification reactions of glycerol with lauric acid in solvent free system were carried out using lipases from several sources. All lipases were immobilized on polysiloxane-polyvinyl alcohol particles by covalent binding with high activity recovered. Among the tested enzymes, the Candida antarctica lipase allowed to attain the highest molar conversion (76%), giving similar proportions of monolaurin, dilaurin and low amount of trilaurin. To further improve the process, the Response Surface Methodology (RSM) was used and optima temperature and molar ratio glycerol to lauric acid were found to be 45 ºC and 5:1, respectively. Under these conditions, 31.35% of monolaurin concentrations were attained and this result was in close agreement with the statistical model prediction.

Development of a flotation method for preconcentration-separation of trace amounts of some metal ions in plant tissues prior to their FAAS determinations

An efficient flotation method based on the combination of flame atomic absorption spectrometry (FAAS) and separation and preconcentration step for determination of Cr3+, Cu 2+, Co2+, Ni2+, Zn2+, Cd 2+, Fe3+ and Pb2+ ions in various real samples by the possibility of applying bis(2-hydroxyacetophenone)-1,4-butanediimine (BHABDI) as a new collector was studied. The influence of pH, amount of BHABDI as collector, sample matrix, type and amount of eluting agent, type and amount of surfactant as floating agent, ionic strength and air flow rates i.e. variables affecting the efficiency of the extraction system was evaluated. It is ascertained that metal ions such as iron can be separated simultaneously from matrix in the presence of 0.012 mM ligand, 0.025% (w/v) of CTAB to a test sample of 750 mL at pH 6.5. These ions can be eluted quantitatively with 6 mL of 1.0 mol L-1 HNO3 in methanol which lead to the enrichment factor of 125. The detection limits for analyte ions were in the range of 1.3-2.4 ng mL-1. The method has been successfully applied for determination of trace amounts of ions in various real samples.

Adsorption of glyphosate in a forest soil: a study using Mössbauer and FT-IR spectroscopy

We studied the adsorption of glyphosate (GPS) onto soil mineral particles, using FT-IR and Mössbauer spectroscopy. From IR measurements for samples collected under native vegetation of a forest reserve, bands at 1632 and 1407 cm-1 could be attributed to the interaction between the carboxylic group of GPS and structural Al3+ and Fe3+ on the surface of mineral particles; bands at 1075 and 1000 cm-1 were observed only for cultivated soil. Mössbauer spectra for these soils were definitely fitted using a broad central doublet in addition to the magnetic component. This multiple quadrupolar component may be attributed to all non-magnetic Fe3+ contributions, including that of the GPS/Fe3+ complex.

Análise cristalográfica da solução sólida com estrutura tipo Tungstênio Bronze de niobato de potássio e estrôncio dopado com ferro

Solid solution of iron doped potassium strontium niobate with KSr2(FeNb4)O15-Δ stoichiometry was prepared by high efficiency ball milling method. Structural characterization was carried out by X-ray diffraction. Crystalline structure was analyzed by the Rietveld refinements using the FullProf software. The results showed a tetragonal system with the tetragonal tungsten bronze structure - TTB (a = 12.4631 (2) Ã… and c = 3.9322 (6) Ã…, V = 610.78 (2) ų). In this work, the sites occupancy by the K+, Sr2+ and Fe3+ cations on the TTB structure were determined. NbO6 polihedra distortion and its correlation with the theoretical polarization are discussed.

Separation and preconcentration trace amounts of gold by using modified organo nanoclay closite 15A

The application of organo nanoclay 5-(4-dimethylamino-benzylidene) rhodanine-immobilized as a new, easily prepared, and stable solid sorbent for preconcentration trace amounts of Au(III) ions in aqueous solution is presented. The sorption of Au(III) ions was quantitative in the pH range of 2-4, and quantitative desorption occurred instantaneously with 10.0 mL of a mixture containing 0.5 mol L-1 Na2S2O3 and KSCN. Various parameters, such as the effect of pH, breakthrough volume, extraction time, and interference of a large number of anions and cations have been studied. The proposed method has been applied for determination of trace amount of gold in water samples.