Thermodynamic analysis of ferrous ion binding to Escherichia coli ferritin EcFtnA

Iron oxidation in the bacterial ferritin EcFtnA from Escherichia coli shows marked differences from its homologue human H-chain ferritin (HuHF). While the amino acid residues that constitute the dinuclear center in these proteins are highly conserved, EcFtnA has a third iron-binding site (C site) in close proximity to the dinuclear center that is seemingly responsible for these differences. Here, we describe the first thermodynamic study of Fe2+ binding to EcFtnA and its variants to determine the location of the primary ferrous ion-binding sites on the protein and to better understand the role of the third C site in iron binding. Isothermal titration calorimetric analyses of the wild-type protein reveal the presence of two main classes of binding sites in the pH range of 6.5-7.5, ascribed to Fe2+ binding, first at the A and then the B sites. Site-directed mutagenesis of ligands in the A, B, or C sites affects the apparent Fe2+-binding stoichiometries at the unaltered sites. The data imply some degree of inter- and intrasubunit negative cooperative interaction between sites. Unlike HuHF where only the A site initially binds Fe2+, both A and B sites in EcFtnA bind Fe2+, implying a role for the C site in influencing the binding of Fe2+ at the B site of the di-iron center of EcFtnA. The ITC equations describing a binding model for three classes of independent binding sites are reported here for the first time.

Photocatalytic oxidation of humic substances and lignins in aqueous solutions

Tässä tutkimuksessa tarkastellaan kahden yleisen, veden ympäristökuormitusta aiheuttavan kemikaaliryhmän, ligniinin ja humusaineiden, fotokatalyyttistahapetusta (photocatalytic oxidation, PCO) vesiliuoksessa. Fotokatalyyttina käytettiin titaanidioksidia, jota säteilytettiin ultraviolettivalolla. Työssä selvitettiin useiden eri olosuhdeparametrien vaikutusta fotokatalyysiin. Tutkittavia parametreja olivat mm. kontaminanttien alkukonsentraatio, pH, vetyperoksidilisäys, rauta-ionien lisäys, fotokatalyysimenetelmä, fotokatalyytin pintakonsentraatioja titaanidioksidin määrä lasisissa mikropartikkeleissa. Ultraviolettivalon lähteinä käytettiin sekä keinovaloa että auringonvaloa. Katalyytin kantoaineena käytettiin huokoisia lasisia mikropartikkeleita, joiden pintaan kiinnittynyt titaanidioksidi pystyi hyvin vähentämään kontaminanttien määrää vedessä. Fotokatalyysin tehokkuus kasvoi humusaine- ja ligniinikonsentraatioiden kasvaessa. Korkeimmat hapetustehokkuudet kumallakin kontaminantilla saavutettiin neutraaleissa jalievästi emäksisissä olosuhteissa huolimatta siitä, että paras adsorboituminen tapahtui happamissa olosuhteissa. Tämän perusteella voidaan olettaa, että humusaineiden ja ligniinin hapetus tapahtuu pääosin radikaalimekanismilla. Vetyperoksidin lisääminen humusaineliuokseen lisäsi hapettumisnopeutta, vaikka näennäinen hapetustehokkuus ei muuttunut. Tämän perusteella vetyperoksidi hapetti myös humusaineita referenssinäytteessä. Ligniinin fotokatalyyttinen hapettuminen parani vetyperoksidilisäyksellä happamissa olosuhteissa johtuen lisääntyneestä OH-radikaalien muodostumisesta. Ligniini ei hapettunut vetyperoksidilla, jos fotokatalyyttiä ei¿ollut läsnä. Rauta-ionit eivät lisänneet humushappojen fotokatalyyttistähapettumista, mutta Fe2+-ionien lisäys aina konsentraatioon 0.05 mM johti ligniinin hapettumistehokkuuden voimakkaaseen kasvuun. Rauta-ionikonsentraation kasvattaminen edelleen johti ligniinin hapetustehokkuuden alenemiseen.

Um fotômetro multi-led microcontrolado, portátil e de baixo custo

A microcontrolled, portable and inexpensive photometer is described. It uses six light-emitting diodes (LEDs) as radiation sources and a phototransistor as detector, as well as a microcontroller (PIC - Programmable Controller of Interruption). This device provided total autonomy to the proposed photometer, which was successfully applied to determination of Fe2+ in ferrous syrups and of seven clinical biochemical parameters. As the components are cheap (~U$30.00) and easy to find, the proposed photometer is an economical alternative for routine chemical analyses in small laboratories, for research and teaching. Being portable and microcontrolled, it allows doing field chemical analyses.

Extraction optimization of antioxidant polysaccharides from leaves of Gynura bicolor (Roxb. & Willd.) DC

Orthogonal design was employed to study the effect of extraction time, temperature and liquid-to-solid ratio on the production of antioxidant polysaccharides from leaves of Gynura bicolor (PLG). Analysis of variance was performed on the data obtained. The most relevant variable was extraction time. A liquid-solid ratio of 30:1 (v/w), a temperature of 80 °C and an extraction time of 3 h were found to be optimal for PLG. The optimal extraction yield of 4.9% was obtained through additional verification test. Hydroxyl radical-scavenging activity, reducing power and ferrous ion chelating ability of PLG were determined. PLG possess concentration-dependent antioxidant potency and IC50 of PLG was 4.67, 0.24 and 4.31 mg/mL for hydroxyl radical-scavenging and ferric ion chelating abilities as well as reducing power, respectively. The results suggest that G. bicolor polysaccharides could be potential source of natural antioxidant and be contributor to the health benefits of G. bicolor.

Antimalarial drugs based on artemisinin: DFT calculations on the principal reactions

Theoretical calculations have been carried out on the interactions of several endoperoxides which are potential antimalarials, including the clinically useful artemisinin, with two possible sources of iron in the parasite, namely the hexa-aquo ferrous ion [Fe(H2O)(6)](2+) and haeme. DFT calculations show that the reactions of all endoperoxides considered, with both sources of iron, initially generate a Fe-O bond followed by cleavage of the O-O bond to oxygen radical species. Subsequently, they can be transformed into carbon-centred radicals of greater stability. However, with [Fe(H2O)(6)](2+) as the iron source, the oxygen-centred radical species are more likely to react further akin to Fenton's reagent, whereby iron salts encourage hydrogen peroxide to act as an oxidizing agent, and that solvent plays a major role. In contrast, when reacting with haeme, the oxygen-centred radicals interconvert to more stable carbon-centred radicals, which can then alkylate haeme. Subsequent cleavage of the Fe-O bond leads to stable and inactive antimalarial products. These results indicate that the reactivity of the endoperoxides as antimalarials is greater with iron hexahydrates for radical-mediated damage as opposed to haeme, which leads to unreactive species. Since only nanomolar quantities of hydrated metal ions could catalyse the reactions leading to damage to the parasites, this could be an alternative or competitive reaction responsible for the antimalarial activity. (c) 2005 Elsevier B.V. All rights reserved.

Determinação do íon ferro em preparações farmacêuticas através da cromatografia planar e espectrofotometria

Os autores estudaram o comportamento cromatográfico de preparações farmacêuticas comerciais contendo o íon Fe (II). Utilizando celulose microcristalina/Propanol: ácido clorídrico 4 N: ácido acético concentrado: ácido nítrico concentrado: clorofórmio (40: 5: 5: 10: 10), como sistema cromatográfico e alizarina como reagente de detecção, Fe (II), Mn (II), Mg (II), Cu (II), Zn (II) e Ca (II) foram separados e identificados pela Cromatografia Planar. O Fe (II) foi determinado pela reação com a ortofenantrolina, resultando em solução adequada para quantificação colorimétrica.

Immobilized cells of Acidithiobacillus ferrooxidans in PVC strands and sultite removal in a pilot-scale bioreactor

The biooxidation of ferrous ion into ferric ion by Acidithiobacillus ferrooxidans can be potentially used for the removal of H2S from industrial gases. In this work, Fe3+ ions were obtained through the oxidation of Fe2+ using the LR strain of At. ferrooxidans immobilized in PVC stands in a pilot-scale bioreactor, while H2S was removed in an absorption tower equipped with Rasching rings. At. ferrooxidans LR strain cells were immobilized by inoculating the bacterium in a Fe2+-mineral medium and percolating it through the support. After complete Fe2+ oxidation, which took around 90 h, the reactor was washed several times with sulfuric acid (pH 1.7) before a new cycle was started. Four additional cycles using fresh Fe2+ mineral medium were then run. During these colonization cycles, the time required for complete iron oxidation decreased, dropping to about 60 h in the last cycle. The batch experiments in the H2S gas removal trials resulted in a gas removal rate of about 98-99% under the operational conditions employed. In the continuous experiments with the bioreactor coupled to the gas absorption column, a gas removal efficiency of almost 100% was reached after 500 min. Precipitate containing mainly sulfur formed during the experimental trial was identified by EDX. (c) 2005 Elsevier B.V. All rights reserved.

Solar photodegradation of dichloroacetic acid and 2,4-dichlorophenol using an enhanced photo-Fenton process

The photo-Fenton process using potassium ferrioxalate as a mediator was investigated for the photodegradation of dichloracetic acid (DCA) and 2,4-dichlorophenol (DCP) in aqueous medium using solar light as source of irradiation. The influence of the solution depth, the light intensity and the effect of stirring the solution during irradiation process were evaluated using DCA as a model compound. A negligible influence of stirring the solution was observed when the concentration of ferrioxalate (FeOx) was 0.8 mM and solution depth was 4.5 or 14 cm. The optimum FeOx concentration determined for solution depths between 4.5 and 14 cm was 0.8 mM considering total organic carbon (TOC) removal during DCA irradiation. The high efficiency of the photo-Fenton process was demonstrated on summer days, when only 10 min of exposition (around noon) were sufficient to completely destroy the organic carbon of a 1.0 mM DCA solution in the presence of 0.8 mM FeOx and 6.0 mM H2O2 using a solution depth of 4.5 cm. It was observed that the photodegradation efficiency increases linearly with the solar light intensity up to values around 15 Wm-2 but this linear relationship does not hold above this value showing a square root dependence. The photodegradation of a solution of DCP/FeOx showed a lower TOC removal rate than that observed for DCA/FeOx, achieving ∼90% after 35 min irradiation under 19 Wm-2, while under this light intensity, the same TOC removal of DCA/FeOx was achieved in only 10 min irradiation. © 2002 Elsevier Science Ltd. All rights reserved.

TL, EPR and optical absorption studies on natural alexandrite compared to natural chrysoberyl

The TL, optical absorption (OA) and EPR properties of natural Brazilian alexandrite and chrysoberyl have been investigated. The TL measurements for natural alexandrite show five peaks between 100 and 450°C, with their emission spectrum having 370 and/or 570 nm components. The intensity of the 320°C TL peak was found to be enhanced with pre-annealing treatment, more prominently above 600°C. The OA and EPR measurements showed that this kind of heat treatment induces the Fe2→ Fe3+ conversion in the natural sample. Chrysoberyl samples exhibited the TL peaks at the same temperatures as alexandrite samples, but the glow curves were more than 200 times less intense than alexandrite ones.

Evaluation of the combined solar TiO2/photo-Fenton process using multivariate analysis

The effect of combining the photocatalytic processes using TiO 2 and the photo-Fenton reaction with Fe3+ or ferrioxalate as a source of Fe2+ was investigated in the degradation of 4-chlorophenol (4CP) and dichloroacetic acid (DCA) using solar irradiation. Multivariate analysis was used to evaluate the role of three variables: iron, H2O2 and TiO2 concentrations. The results show that TiO2 plays a minor role when compared to iron and H2O2 in the solar degradation of 4CP and DCA in the studied conditions. However, its presence can improve TOC removal when H2O2 is totally consumed. Iron and peroxide play major roles, especially when Fe(NO3)3 used in the degradation of 4CP. No significant synergistic effect was observed by the addition of TiO 2 in this process. On the other hand, synergistic effects were observed between FeOx and TiO2 and between H 2O2 and TiO2 in the degradation of DCA. © IWA Publishing 2004.

Development and application of a highly selective biomimetic sensor for detection of captopril, an important ally in hypertension control

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

Baccharis dracunculifolia, the main source of green propolis, exhibits potent antioxidant activity and prevents oxidative mitochondrial damage

Baccharis dracunculifolia DC (Asteraceae) is the main botanical source used by honeybees to produce Brazilian green propolis whose hepatoprotective properties have been already described. In this work we investigated the protective effects of the glycolic extract of B. dracunculifolia (GEBd) against oxidative stress in isolated rat liver mitochondria (RLM). The GEBd was prepared by fractionated percolation using propylene glycol as solvent. The total phenols and flavonoids, which are substances with recognized antioxidant action, were quantified in GEBd and the phytochemical analysis was carried out by HPLC. GEBd exhibited significant scavenger activity towards DPPH radicals and superoxide anions in a concentration-dependent manner, and also a Fe 2+ chelating activity. GEBd decreased the basal H 2O 2 generation and the Fe 2+- or t-BuOOH-induced ROS production in isolated mitochondria. Lipid oxidation of mitochondrial membranes, protein thiol groups and GSH oxidation were also prevented by GEBd. This shows that B. dracunculifolia exhibit potent antioxidant activity protecting liver mitochondria against oxidative damage and such action probably contribute to the antioxidant and hepatoprotective effects of green propolis. © 2011 Elsevier Ltd.

Estudo da oxidação de covelita (CuS) e molibdenita (MoS2) sintéticas por Acidithiobacillus ferrooxidans

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Oxidação da calcopirita (CuFe S'IND.2') por Acidithiobacillus ferooxidans em presença de cisteína e de Acidithiobacillus thiooxidans

Pós-graduação em Biotecnologia - IQ

Análise proteômica diferencial de Acidithiobacillus ferrooxidans em resposta aos sulfetos minerais calcopirita e bornita e efeito de uma proteína recombinante de A. ferrooxidans LR contendo cisteína na oxidação bacteriana da calcopirita

Pós-graduação em Biotecnologia - IQ