Purification and Characterization of an Ethanol-Tolerant β-Glucosidase from Sporidiobolus pararoseus and Its Potential for Hydrolysis of Wine Aroma Precursors

An extracellular ethanol-tolerant β-glucosidase from Sporidiobolus pararoseus was purified to homogeneity and characterized, and its potential use for the enhancement of wine aroma was investigated. The crude enzymatic extract was purified in four steps (concentration, dialysis, ultrafiltration, and chromatography) with a yield of around 40 % for total activity. The purified enzyme (designated Sp-βgl-P) showed a specific activity of approximately 20.0 U/mg, an estimated molecular mass of 63 kDa after sodium dodecyl sulfate polyacrylamide gel electrophoresis, and isoelectric point of 5.0 by isoelectric focusing. Sp-βgl-P has optimal activity at pH 4.0 and at 55 °C. It was stable in a broad pH range at low temperatures and it was tolerant to ethanol and glucose, indicating suitable properties for winemaking. The hydrolysis of glycosidic terpenes was analyzed by adding Sp-βgl-P directly to the wines. The released terpene compounds were evaluated by gas chromatography/mass spectrometry. The enzymatic treatment significantly increased the amount of free terpenes, suggesting that this enzyme could potentially be applicable in wine aroma improvement. © 2013 Springer Science+Business Media New York.

Ethanol addition as a strategy for start-up and acclimation of an AnSBBR for the treatment of landfill leachate

This study evaluated the ethanol addition as a strategy for start-up and acclimation of a pilot scale (1300 L) anaerobic sequencing batch biofilm reactor (AnSBBR) for the treatment of municipal landfill leachate with seasonal biodegradability variations. The treatment was carried out at ambient temperature (23.8 ± 2.1 °C) in the landfill area. In a first attempt, the leachate collected directly from landfill showed to be predominantly recalcitrant to anaerobic treatment and the acclimation was not possible. In a second attempt, adding ethanol to leachate, the reactor was successfully acclimated. After acclimation, without ethanol addition, the CODTotal influent ranged from 4970 to 13040 mg L-1 and the removal efficiencies ranged from 12.1% to 70.7%. A final test was carried out increasing the ammonia and free-ammonia concentration from 2486 mgN L-1 and 184 mgN L-1 to 4519 mgN L-1 and 634 mgN L-1, respectively, with no expressive inhibition verified. The start-up strategy was found to be feasible, providing the acclimation of the biomass in the AnSBBR, and maintaining the biomass active even when the leachate was recalcitrant. © 2013 Elsevier Ltd. All rights reserved.

Ethanol electro-oxidation in an alkaline medium using Pd/C, Au/C and PdAu/C electrocatalysts prepared by electron beam irradiation

Carbon-supported Pd, Au and bimetallic PdAu (Pd:Au 90:10, 50:50 and 30:70 atomic ratios) electrocatalysts were prepared using electron beam irradiation. The obtained materials were characterized by energy dispersive X-ray analysis (EDX), X-ray diffraction (XRD) and transmission electron microscopy (TEM), and their catalytic activities toward ethanol electro-oxidation were evaluated in an alkaline medium using electrochemical techniques, in situ attenuated total reflectance Fourier transformed infrared spectroscopy (ATR-FTIR) analysis and a single alkaline direct ethanol fuel cell (ADEFC). EDX analyses showed that the actual Pd: Au atomic ratios were very similar to the nominal ones. X-ray diffractograms of PdAu/C electrocatalysts evidenced the presence of Pd-rich (fcc) and Au-rich (fcc) phases. TEM analysis showed a homogeneous dispersion of nanoparticles on the carbon support, with an average size in the range of 3-5 nm and broad size distributions. Cyclic voltammetry (CV) and chronoamperometry (CA) experiments revealed the superior ambient activity toward ethanol electro-oxidation of PdAu/C electrocatalysts with Pd: Au ratios of 90:10 and 50:50. In situ ATR-FTIR spectroscopy measurements have shown that the mechanism for ethanol electro-oxidation is dependent on catalyst composition, leading to different reaction products, such as acetaldehyde and acetate, depending on the number of electrons transferred. Experiments on a single ADEFC were conducted between 50 and 900 C, and the best performance of 44 mW cm-2 in 2.0molL-1 ethanol was obtained at 850C for the Pd:Au 90:10 catalysts. This superior performance is most likely associated with enhancement of ethanol adsorption on Pd, oxidation of the intermediates, the presence of gold oxide-hydroxyl species, low mean particle diameters and better distribution of particles on the support. © 2013 Elsevier Ltd. All rights reserved.

Anaerobic digestion of stillage to produce bioenergy in the sugarcane-to-ethanol industry

Stillage is the main wastewater from ethanol production, containing a high chemical oxygen demand in addition to acidic and corrosive characteristics. Though stillage may be used as a soil fertilizer, its land application may be considered problematic due its high polluting potential. Anaerobic digestion represents an effective alternative treatment to reduce the pollution load of stillage. In addition, the methane gas produced within the process may be converted to energy, which can be directly applied to the treatment plant. The objective of this paper was to investigate the energetic potential of anaerobic digestion applied to stillage in the sugarcane ethanol industry. An overall analysis of the results indicates energy recovery capacity (ERC) values for methane ranging from 3.5% to 10%, respectively, for sugarcane juice and molasses. The processes employed to obtain the fermentable broth, as well as the distillation step, represent the main limiting factors to the energetic potential feasibility. Considering financial aspects the annual savings could reach up to US$ 30 million due to anaerobic digestion of stillage in relatively large-scale distilleries (365,000 m3 of ethanol per year). The best scenarios were verified for the association between anaerobic digestion of stillage and combustion of bagasse. In this case, the fossil fuels consumption in distilleries could be fully ceased, such the ERC of methane could reach values ranging from 140% to 890%. © 2013 Taylor & Francis.

Enzymatic transesterification of soybean oil with ethanol using lipases immobilized on highly crystalline PVA microspheres

Polyvinyl alcohol (PVA) microspheres with different degree of crystallinity were used as solid supports for Rhizomucor miehei lipase immobilization, and the enzyme-PVA complexes were used as biocatalysts for the transesterification of soybean oil to fatty acid ethyl esters (FAEE). The amounts of immobilized enzyme on the polymeric supports were similar for both the amorphous microspheres (PVA4) and the high crystalline microspheres (PVA25). However, the enzymatic activity of the immobilized enzymes was depended on the crystallinity degree of the PVA microspheres: enzymes immobilized on the PVA4 microspheres have shown low enzymatic activity (6.13 U mg-1), in comparison with enzymes immobilized on the high crystalline PVA25 microspheres (149.15 U mg-1). A synergistic effect was observed for the enzyme-PVA25 complex during the transesterification reaction of soybean oil to FAEE: transesterification reactions with free enzyme with the equivalent amount of enzyme that were immobilized onto the PVA25 microspheres (5.4 U) have yielded only 20% of FAEE, reactions with the pure highly crystalline microsphere PVA25 have not yielded FAEE, however reactions with the enzyme-PVA25 complexes have yielded 66.3% of FAEE. This synergistic effect of an immobilized enzyme on a polymeric support has not been observed before for transesterification reaction of triacylglycerides into FAEE. Based on ATR-FTIR, 23Na- and 13C-NMR-MAS spectroscopic data and the interaction of the polymeric network intermolecular hydrogen bonds with the lipases residual amino acids a possible explanation for this synergistic effect is provided. © 2013 Elsevier Ltd. All rights reserved.

Influence of ethanol and glycerol concentration over functional and structural properties of zein-oleic acid films

This study was performed in order to determine the effect of the addition of different concentrations of glycerol and ethanol over functional and structural properties of zein-oleic acid films. Films were prepared from zein and oleic acid formulations, containing: 0, 10, 20 and 30% (w/w) of glycerol as plasticizer and 75, 80, 85, 90 and 95% (v/v) of ethanol as zein solvent. Water vapor permeability (WVP) at 4 and 24 C, opacity, water solubility and structural behavior of the film were assessed. The film water barrier properties, WVP and water solubility, were increased when higher ethanol concentration and lower glycerol concentration were used. Furthermore, WVP at 4 C was lower than WVP at 24 C due to the crystalline solid state of oleic acid at lower temperatures. Likewise, opacity, homogeneity and structure of the composite film were improved as ethanol increased and glycerol lowered. © 2013 Elsevier B.V. All rights reserved.

Comportamento fermentativo de linhagens industriais de Saccharomyces cerevisiae em mosto com diferentes concentrações de sacarose e fontes estruturalmente complexas de nitrogênio

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação da tolerância de leveduras a um coquetel de inibidores que simula um hidrolisado de bagaço de cana quando adicionado a um meio sintético

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Seleção de micro-organismos fermentadores de xilose em etanol a partir de diferentes variedades da casca de uvas (Vitis spp)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Genotipagem de leveduras presentes no processo industrial de produção de álcool combustível e estudo do polimorfismo de genes envolvidos no processo fermentativo em Saccharomyces cerevisiae

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Expressão gênica diferencial por cDNA-AFLP em raízes de cana-de-açúcar submetida ao déficit hídrico

Pós-graduação em Agronomia (Genética e Melhoramento de Plantas) - FCAV

Estudo da expressão gênica entre cultivares de cana-de-açúcar contrastantes para a tolerância à seca

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Sugarcane tolerance to ratoon eradication with glyphosate determined by physiological responses

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

The psychobiological model: a new explanation to intensity regulation and (in)tolerance in endurance exercise

The mechanisms underpinning fatigue and exhaustion, and the specific sources of exercise-endurance intensity regulation and (in)tolerance have been investigated for over a century. Although several scientific theories are currently available, over the past five years a new framework called Psychobiological model has been proposed. This model gives greater attention to perceptual and motivational factors than its antecedents, and their respective influence on the conscious process of decision-making and behavioral regulation. In this review we present experimental evidences and summarize the key points of the Psychobiological model to explain intensity regulation and (in)tolerance in endurance exercise. Still, we discuss how the Psychobiological model explains training-induced adaptations related to improvements in performance, experimental manipulations, its predictions, and propose future directions for this investigative area. The Psychobiological model may give a new perspective to the results already published in the literature, helping scientists to better guide their research problems, as well as to analyze and interpret new findings more accurately.

Estudo genômico e transcritômico de isolados selecionados de alto rendimento de dorna de fermentação alcoólica de Saccharomyces cerevisiae

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)