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.

Revista de la CEPAL no.88

Vigencia de los aportes de Celso Furtado al estructuralismo / Ricardo Bielschowsky. -- Obsolescencia de la protección a los inversores extranjeros después de la crisis argentina / Michael Mortimore y Leonardo Stanley. -- Una aproximación al enfoque de derechos en las estrategias y políticas de desarrollo / Víctor Abramovich. -- ¿Pueden los países de América Latina y el Caribe emular el modelo irlandés para atraer inversión extranjera directa? / Ruth Ríos-Morales y David O ’Donovan. -- El lento retorno de las políticas industriales en América Latina y el Caribe / Wilson Peres. -- Un modelo de bajo crecimiento: la informalidad como restricción structural / Mario Cimoli, Annalisa Primi y Maurizio Pugno. -- El mercado de trabajo argentino en la globalización financier / Mario Damill y Roberto Frenkel. -- Precariedad social en México y Argentina: tendencias, expresiones y trayectorias nacionales / María Cristina Bayón. -- Pacto Fiscal en Guatemala: lecciones de una negociación / Juan Alberto Fuentes K. y Maynor Cabrera. -- Cambio de la estructura productiva en Chile, 1986-1996: producción e interdependencia industrial / José Miguel Albala-Bertrand. -- Orientaciones para los colaboradores de la Revista de la CEPAL. -- La Revista de la CEPAL en Internet. -- Publicaciones recientes de la CEPAL.

CEPAL Review no.88

Celso Furtado’s contributions to structuralism and their relevance today / Ricardo Bielschowsky. -- Has investor protection been rendered obsolete by the Argentine crisis? / Michael Mortimore and Leonardo Stanley. -- The rights-based approach in development policies and strategies / Victor Abramovich. -- Can the Latin American and Caribbean countries emulate the Irish model of FDI attraction? / Ruth Rios-Morales and David O’Donovan. -- The slow comeback of industrial policies in Latin America and the Caribbean / Wilson Peres. -- A low-growth model: informality as a structural constraint / Mario Cimoli, Annalisa Primi and Maurizio Pugno. -- The Argentine labour market in a financially globalized world / Mario Damill and Roberto Frenkel. -- Social precarity in Mexico and Argentina: trends, manifestations and national trajectories / María Cristina Bayón. -- The Fiscal Covenant in Guatemala: lessons learned from the negotiations / Juan Alberto Fuentes K. and Maynor Cabrera. -- Changes in Chile’s production structure, 1986-1996: output and industrial interdependence / José Miguel Albala-Bertrand. -- Guidelines for contributors to the CEPAL Review. -- CEPAL Review on the Internet. -- Recent ECLAC publications.

Identification of folding intermediates of streblin, the most stable serine protease: biophysical analysis

Streblin, a serine proteinase from plant Streblus asper, has been used to investigate the conformational changes induced by pH, temperature, and chaotropes. The near/far UV circular dichroism activities under fluorescence emission spectroscopy and 8-aniline-1-naphthalene sulfonate (ANS) binding have been carried out to understand the unfolding of the protein in the presence of denaturants. Spectroscopic studies reveal that streblin belongs to the alpha+beta class of proteins and exhibits stability towards chemical denaturants, guanidine hydrochloride (GuHCl). The pH-induced transition of this protein is noncooperative for transition phases between pH 0.5 and 2.5 (midpoint, 1.5) and pH 2.5 and 10.0 (midpoint, 6.5). At pH 1.0 or lower, the protein unfolds to form acid-unfolded state, and for pH 7.5 and above, protein turns into an alkaline denatured state characterized by the absence of ANS binding. At pH 2.0 (1M GuHCl), streblin exists in a partially unfolded state with characteristics of amolten globule state. The protein is found to exhibit strong and predominant ANS binding. In total, six different intermediate states has been identified to show protein folding pathways.

Selfing rate estimation in sugarcane under unfavorable natural conditions of crossing by using microsatellite markers

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

Production and Characterization of a Milk-clotting Protease Produced in Submerged Fermentation by the Thermophilic Fungus Thermomucor indicae-seudaticae N31

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Seedling Development and Evaluation of Genetic Stability of Cryopreserved Dendrobium Hybrid Mature Seeds

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Molecular and Kinetic Characterization of Two Extracellular Xylanases Isolated from Leucoagaricus gongylophorus

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

The effect of biomass immobilization support material and bed porosity on hydrogen production in an upflow anaerobic packed-bed bioreactor

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

Investigation, expression, and molecular modeling of ORF2, a metagenomic lipolytic enzyme

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

Production and Characterization of β-glucosidase Obtained by the Solid-State Cultivation of the Thermophilic Fungus Thermomucor indicae-seudaticae N31

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

Cocaine effects on mouse incentive-learning and human addiction are linked to alpha 2 subunit-containing GABA(A) receptors

Because GABA(A) receptors containing alpha 2 subunits are highly represented in areas of the brain, such as nucleus accumbens (NAcc), frontal cortex, and amygdala, regions intimately involved in signaling motivation and reward, we hypothesized that manipulations of this receptor subtype would influence processing of rewards. Voltage-clamp recordings from NAcc medium spiny neurons of mice with alpha 2 gene deletion showed reduced synaptic GABA(A) receptor-mediated responses. Behaviorally, the deletion abolished cocaine`s ability to potentiate behaviors conditioned to rewards (conditioned reinforcement), and to support behavioral sensitization. In mice with a point mutation in the benzodiazepine binding pocket of alpha 2-GABA(A) receptors (alpha 2H101R), GABAergic neurotransmission in medium spiny neurons was identical to that of WT (i.e., the mutation was silent), but importantly, receptor function was now facilitated by the atypical benzodiazepine Ro 15-4513 (ethyl 8-amido-5,6-dihydro-5-methyl-6-oxo-4H-imidazo [1,5-a] [1,4] benzodiazepine-3-carboxylate). In alpha 2H101R, but not WT mice, Ro 15-4513 administered directly into the NAcc-stimulated locomotor activity, and when given systemically and repeatedly, induced behavioral sensitization. These data indicate that activation of alpha 2-GABA(A) receptors (most likely in NAcc) is both necessary and sufficient for behavioral sensitization. Consistent with a role of these receptors in addiction, we found specific markers and haplotypes of the GABRA2 gene to be associated with human cocaine addiction.

Effects of Gastric Bypass Surgery in Patients With Type 2 Diabetes and Only Mild Obesity

OBJECTIVE-Roux-en-Y gastric bypass (RYGB) ameliorates type 2 diabetes in severely obese patients through mechanisms beyond just weight loss, and it may benefit less obese diabetic patients. We determined the long-term impact of RYGB on patients with diabetes and only class 1 obesity. RESEARCH DESIGN AND METHODS-Sixty-six consecutively selected diabetic patients with BMI 30-35 kg/m(2) underwent RYGB in a tertiary-care hospital and were prospectively studied for up to 6 years (median 5 years [range 1-6]), with 100% follow-up. Main outcome measures were safety and the percentage of patients experiencing diabetes remission (HbA(1c) <6.5% without diabetes medication). RESULTS-Participants had severe, longstanding diabetes, with disease duration 12.5 +/- 7.4 years and HbA(1c) 9.7 +/- 1.5%, despite insulin and/or oral diabetes medication usage in everyone. For up to 6 years following RYGB, durable diabetes remission occurred in 88% of cases, with glycemic improvement in 11%. Mean HbA(1c) fell from 9.7 +/- 1.5 to 5.9 +/- 0.1% (P < 0.001), despite diabetes medication cessation in the majority. Weight loss failed to correlate with several measures of improved glucose homeostasis, consistent with weight-independent antidiabetes mechanisms of RYGB. C-peptide responses to glucose increased substantially, suggesting improved beta-cell function. There was no mortality, major surgical morbidity, or excessive weight loss. Hypertension and dyslipidemia also improved, yielding 50-84% reductions in predicted 10-year cardiovascular disease risks of fatal and nonfatal coronary heart disease and stroke. CONCLUSIONS-This is the largest, longest-term study examining RYGB for diabetic patients without severe obesity. RYGB safely and effectively ameliorated diabetes and associated comorbidities, reducing cardiovascular risk, in patients with a BMI of only 30-35 kg/m(2).

Effect of Feed Strategy on Methane Production and Performance of an AnSBBR Treating Effluent from Biodiesel Production

The aim of this work was to investigate the effect of different feeding times (2, 4 and 6 h) and applied volumetric organic loads (4.5, 6.0 and 7.5 gCOD L-1 day(-1)) on the performance of an anaerobic sequencing batch biofilm reactor (AnSBBR) treating effluent from biodiesel production. Polyurethane foam cubes were used as inert support in the reactor, and mixing was accomplished by recirculating the liquid phase. The effect of feeding time on reactor performance showed to be more pronounced at higher values of applied volumetric organic loads (AVOLs). Highest organic material removal efficiencies achieved at AVOL of 4.5 gCOD L-1 day(-1) were 87 % at 4-h feeding against 84 % at 2-h and 6-h feeding. At AVOL of 6.0 gCOD L-1 day(-1), highest organic material removal efficiencies achieved with 4-h and 6-h feeding were 84 %, against 71 % at 2-h feeding. At AVOL of 7.5 gCOD L-1 day(-1), organic material removal efficiency achieved with 4-h feeding was 77 %. Hence, longer feeding times favored minimization of total volatile acids concentration during the cycle as well as in the effluent, guaranteeing process stability and safety.

Effect of Substrate Concentration on Dark Fermentation Hydrogen Production Using an Anaerobic Fluidized Bed Reactor

The effect of substrate (glucose) concentration on the stability and yield of a continuous fermentative process that produces hydrogen was studied. Four anaerobic fluidized bed reactors (AFBRs) were operated with a hydraulic retention time (HRT) from 1 to 8 h and an influent glucose concentration from 2 to 25 gL(-1). The reactors were inoculated with thermally pre-treated anaerobic sludge and operated at a temperature of 30 degrees C with an influent pH around 5.5 and an effluent pH of about 3.5. The AFBRs with a HRT of 2 h and a feed strength of 2, 4, and 10 gL(-1) showed satisfactory H-2 production performance, but the reactor fed with 25 gL(-1) of glucose did not. The highest hydrogen yield value was obtained in the reactor with a glucose concentration of 2 gL(-1) when it was operated at a HRT of 2 h. The maximum hydrogen production rate value was achieved in the reactor with a HRT of 1 h and a feed strength of 10 gL(-1). The AFBRs operated with glucose concentrations of 2 and 4 gL(-1) produced greater amounts of acetic and butyric acids, while AFBRs with higher glucose concentrations produced a greater amount of solvents.