Mechanisms underlying dioxygen reduction in laccases. Structural and modelling studies focusing on proton transfer

<p>Abstract</p> <p>Background</p> <p>Laccases are enzymes that couple the oxidation of substrates with the reduction of dioxygen to water. They are the simplest members of the multi-copper oxidases and contain at least two types of copper centres; a mononuclear T1 and a trinuclear that includes two T3 and one T2 copper ions. Substrate oxidation takes place at the mononuclear centre whereas reduction of oxygen to water occurs at the trinuclear centre.</p> <p>Results</p> <p>In this study, the CotA laccase from <it>Bacillus subtilis </it>was used as a model to understand the mechanisms taking place at the molecular level, with a focus in the trinuclear centre. The structures of the holo-protein and of the oxidised form of the apo-protein, which has previously been reconstituted <it>in vitro </it>with Cu(I), have been determined. The former has a dioxygen moiety between the T3 coppers, while the latter has a monoatomic oxygen, here interpreted as a hydroxyl ion. The UV/visible spectra of these two forms have been analysed in the crystals and compared with the data obtained in solution. Theoretical calculations on these and other structures of CotA were used to identify groups that may be responsible for channelling the protons that are needed for reduction of dioxygen to water.</p> <p>Conclusions</p> <p>These results present evidence that Glu 498 is the only proton-active group in the vicinity of the trinuclear centre. This strongly suggests that this residue may be responsible for channelling the protons needed for the reduction. These results are compared with other data available for these enzymes, highlighting similarities and differences within laccases and multicopper oxidases.</p

Preparação de extratos de própolis dos Açores e avaliação das suas atividades antimicrobiana e antioxidante

Própolis é uma substância resinosa natural produzida pelas abelhas a partir de material de origem vegetal recolhido na flora circundante, sendo utilizado posteriormente na construção e reparação das suas colmeias [1]. Este produto apresenta uma composição química muito complexa e também muito diversa, bem como diversas atividades biológicas de interesse [2,3]. No presente trabalho estudaram-se cinco amostras de própolis recolhidas em 2010, na ilha Terceira (Açores, Portugal) com o objetivo de avaliar a sua capacidade antioxidante e as suas propriedades antimicrobianas. Para o efeito prepararam-se extratos etanólicos das amostras em estudo. O potencial antioxidante foi avaliado através de uma técnica eletroquímica, a voltametria. A atividade antimicrobiana foi estimada pelo método de incorporação dos extratos em placas com meio de cultura apropriado, utilizando como indicadores de suscetibilidade um painel de leveduras e bactérias de interesse em diferentes áreas. Em relação à atividade antimicrobiana, os extratos foram mais ativos contra leveduras do que contra bactérias, e genericamente mais ativos contra as bactérias Gram-negativas que contra as Gram-positivas. No entanto, os extratos mais ativos contra leveduras não foram os mais eficazes contra bactérias, sugerindo modos de ação distintos sobre procariotas e eucariotas. Os resultados sugerem ainda uma relação entre algumas características macroscópicas das amostras, como por exemplo a cor, o aroma e a dureza, e a sua atividade antifúngica, o que poderá constituir um importante marcador. Quanto ao potencial antioxidante, verificou-se existirem também diferenças entre os cinco extratos estudados, apresentando o extrato 4 maior atividade antioxidante, enquanto que os extratos 1 e 3 apresentam capacidade antioxidante semelhante. A inexistência de uma correlação entre a capacidade antioxidante e os espetros de atividade antimicrobiana revela que os compostos responsáveis por estas bioatividades sejam distintos. Em conclusão, apesar das amostras serem provenientes de uma região relativamente circunscrita, existem diferenças significativas quanto à composição e bioatividades analisadas

Performance of bacterial nanocellulose packaging film functionalised in situ with zinc oxide: Migration onto chicken skin and antimicrobial activity

Supplementary data associated with this article can be found in the online version at doi:10.1016/j.fpsl.2023.101140.Zinc oxide nanoparticles (ZnO) are cost-effective antimicrobial agents with great potential for the active packaging industry. Bacterial NanoCellulose (BNC) features a porous fibre network, with high absorption capacity, flexible and with good mechanical properties, suitable as a carrier of active agents. In this work, BNCZnO films were developed and optimized regarding the particle size and ZnO concentration. The NaOH dropwise addition to BNC membranes immersed in Zn(CH3COO)2-PVOH enabled the production of ZnO nanoparticles with an z-average of 144 nm and a low polydispersity index. High ZnO incorporation (27%mZn/mBNCZnO) was obtained, with uniform distribution all over the BNC membranes. These composites were then characterized and evaluated for Zn migration using food simulants (10%, 20%, and 50% ethanol) with results lower than the limit. Migration into chicken skin, as a real food model, was low at 4 °C but exceeded the migration limit at 10 and 22 °C. Zn migration was also found to be temperature and pH dependent. When applied to chicken skin, BNCZnO was effective against E. coli, Salmonella (0.51.0 log reduction), and Campylobacter spp. (2.0 log reduction), indicating its potential for active packaging applications.The authors appreciate the technical support from CINATE team, especially Susana Teixeira for the assistance in atomic absorption spectroscopy.info:eu-repo/semantics/publishedVersio

Colour profile analysis of Port Wines by different instrumental and visual methods

BACKGROUND: Wine colour is an important quality parameter, being the first sensorial attribute evaluated during wine tasting. The perception of wine colour can be different depending on many factors, including the depth of the sample under observation. The main objectives of the present study were to measure the colour of Port wines, using CIE L*a*b* parameters at different depths with different instrumental techniques (spectrophotometry and colorimetry), and to compare the obtained results with the sensory (visual) perception of colour samples. RESULTS: Representative profiles of Lightness (L*), Hue (H*) and Chroma (C*) at different depths were obtained using Port wine samples from different categories and ages. In general, relatively good correlations between the colorimetric and spectrophotometric methods were obtained for the L* and H* parameters. The results of the sensory tests also showed good correlations between the visually-assessed hue scores and the colorimetric measurements of the H* parameter, particularly at the lower depths tested (up to 4.0 mm). CONCLUSIONS: Overall, the results indicate that the colorimetric method can be used for estimating wine colour parameters, providing useful information about the colour profile of wines at different depthsinfo:eu-repo/semantics/acceptedVersio

Antimicrobial and antioxidant properties of propolis ethanol extracts from Terceira Island (Azores, Portugal)

Publicado em "Planta Medica, vol. 80, nr. 16 (2014) P1L17" (Book of abstracts)Bees produce a resinous mixture named propolis known to have several functions in the beehive, namely structural and sanitary [1,2]. Propolis presents a rich and complex chemical composition, imparted from the several plant materials collected, and several compounds and extracts have been associated with biological activities of high interest [3]. This study focuses on the antimicrobial activity and antioxidant capacity of ethanolic extracts (EE) obtained from propolis samples collected in 2010, in five different areas of the Terceira Island (Azores Archipelago). Antimicrobial activity was evaluated by dilution assay incorporating different EE concentrations in solid media. Bacteria and yeasts of clinical and agrofood relevance were used as indicators of susceptibility. The results showed that Gram-positive bacteria displayed more resistance (best MIC value 200 µg/ml) than the Gram-negative (100 µg/ml). Extracts with higher antibacterial activity (e.g. EE4 and EE5) were generally distinct from those with greater activity against yeast (EE1), but species-specific susceptibilities and extract-specific efficacies against particular microorganisms were also observed, only EE1 was effective against S. epidermidis. A relationship between propolis macroscopic characteristics (colour, hardness and odour) and its antifungal activity was noticed. If confirmed for a broader range of samples, this relationship may constitute an important diagnostic tool. The antioxidant capacity was assessed using cyclic voltammetry [4]. EE4 and EE5 showed the highest antioxidant capacities (39.6 and 23.5 mM eq. gallic acid, respectively), in agreement with results from the antimicrobial assays. Chemical characterization of all EE are underway. Despite being collected in a somewhat confined geographic area propolis samples exhibited diverse antimicrobial activities with different efficiencies and antioxidant capacities

An additional brain endocast of the ictidosaur riograndia guaibensis (Eucynodontia: Probainognathia): Intraspecific variation of endocranial traits

Recently, the morphology and encephalization of the brain endocast of the Triassic non-mammaliaform probainognathian cynodont Riograndia guaibensis were studied. Here, we analyzed the brain endocast of an additional specimen of this species. The new endocast shows well-defined olfactory bulbs and a median sulcus dividing the hemispheres, traits that were not clearly observed in the first studied specimen. Encephalization quotients were also calculated, revealing similar values to other non-mammaliaform cynodonts and lower than those of the first analyzed specimen. The analyzed cranium is slightly larger than the first studied one and may represent an advanced ontogenetic stage. Hence, these differences may be related to the intraspecific variation of this cynodont or alternatively, to the preservation of each specimen.Fil: Kerber, Leonardo. Universidade Federal de Santa Maria; BrasilFil: Ferreira, José Darival. Universidade Federal de Santa Maria; BrasilFil: Fonseca, Pedro Henrique M.. Universidade Federal do Rio Grande do Sul; BrasilFil: Franco, Arymatheia. Universidade Federal de Santa Maria; BrasilFil: Martinelli, Agustín Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; ArgentinaFil: Soares, Marina Bento. Universidade Federal do Rio Grande do Sul; Brasil. Universidade Federal do Rio de Janeiro; BrasilFil: Ribeiro, Ana Maria. Universidade Federal do Rio Grande do Sul; Brasi

Role of active species formed by electrolysis on electrochemical therapy

info:eu-repo/semantics/publishedVersio

Distinctive features of composts of different origin: a thorough examination of the characterization results

The potential of composts produced from different origin residues to be used in environmentally friendly agriculture is addressed in this work. Seven composts obtained from different raw materials and composting methodologies are compared using elemental, thermal and spectroscopic characterization data. Despite the stabilization of the organic matter in all composts being adequate for agricultural applications, they display distinct elemental and structural compositions. Likewise, the fertilisers have very different effects on lettuce growth. Despite the observed differences, some common features were found, namely a mass loss (TGA) of 25.2 g per mol C, association between groups of elements (Fe, Al, Ni, Co, Cr, Cu and S; Mg, Na, K and P, C, Coxi, N and Pb) and correlations between the amount of carbon nanostructures and the characteristic aromaticity parameters. These results suggest that the tuning of the compost features for specific cultures may be possible for sustainable food production.This work was financially supported by the Interreg VA Spain-Portugal Programme (EU) through the project Res2ValHum (0366_RES2VALHUM_1_P). A.C. Silva acknowledges receipt of a PhD grant (UMINHO/BD/40/2016) financed by the Operational Programme Norte 2020 (through the Project “NORTE-08-5369-FSE-000033”). A. Teixeira acknowledges the grant (Res2ValHum 01/2018) to develop experimental work for 11 months on the project. J. Antelo and S. Fiol are also grateful for the financial support provided by Xunta de Galicia—Consellería de Educación e Ordenación Universitaria de Galicia (Consolidation of Competitive Groups of Investigation; GI-1245, ED431C 2018/12 and CRETUS AGRUP2015/02, ref. 2018-PG10)

Propriedades físico-químicas de farinhas de diferentes feijões coloridos (Phaseolus vulgaris) cozidos no vapor da autoclave.

Farinha de feijão é uma excelente matéria prima para a elaboração de produtos sem glúten, pois confere ao produto alto teor proteico, fibras alimentares e é fonte de minerais. No entanto, a farinha de grãos de feijão cru pode apresentar fatores que afetam a sua aceitabilidade na adição em formulações, como textura, solubilidade, e sabor desagradável. Este trabalho avaliou o efeito de pré-tratamentos de grãos de feijões coloridos na melhoria das características tecnológicas e nutricionais das farinhas derivadas.Evento online