Protein Hydrolysis by Immobilized and Stabilized Trypsin

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Isolation, solubility and in vitro hydrolysis of chickpea vicilin-like protein

The chickpea vicilin-like globulin was isolated and chromatographed on Sepharose CL-6B and Sephacryl S-300. The native globulin with a molecular weight of 140 kDa was resolved in Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in seven polypeptide bands in the range of 12.4-67 kDa. The solubility profile of the protein in water and NaCl solutions was typical of a legume globulin. The purified vicilin-like globulin, native and heated, was hydrolyzed by pepsin, trypsin and chymotrypsin. The hydrolysis patterns indicated that the native vicilin-like protein was only partially degraded by the enzymes in comparison with casein. Heating increased its susceptibility to hydrolysis relative to the native form, for all the enzymes. However, the results obtained by the pH-drop method revealed that the in vitro digestibility of the vicilin-like protein was not altered by heating, while 11 S-like and total globulins suffered a small increase, indicating that the structural characteristics of storage globulins may be important factors limiting the protein digestion. (c) 2007 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.

Recombinant expression of glycerol-3-phosphate dehydrogenase using the Pichia pastoris system

In the present study, the GPD2 gene from Saccharomyces cerevisiae, which codifies for the enzyme glycerol-3-phosphate dehydrogenase (GPDH), was cloned from the pPICZ-alpha expression vector and used with the purpose of inducing the extracellular expression of the glycerol-3-phosphate dehydrogenase under the control of the methanol-regulated AOX promoter. The presence of the GPD2 insert was confirmed by PCR analysis. Pichia pastoris X-33 (Mut(+)) was transformed with linearized plasmids by electroporation and transformants were selected on YPDS plates containing 100 mu g/mL of zeocin. Several clones were selected and the functionality of this enzyme obtained in a culture medium was assayed. Among the mutants tested, one exhibited 3.1 x 10(-2) U/mg of maximal activity. Maximal enzyme activity was achieved at 6 days of growth. Medium composition and pre-induction osmotic stress influenced protein production. Pre-induction osmotic stress (culturing cells in medium with either 0.35 M sodium chloride or 1.0 M sorbitol for 4h prior to induction) led to an increase in cell growth with sorbitol and resulted in a significant increase in GPDH productivity with sodium chloride in 24h of induction approximately fivefold greater than under standard conditions (without pre-induction). (C) 2010 Elsevier B.V. All rights reserved.

Synthesis, ex Vivo and in Vitro Hydrolysis Study of an Indoline Derivative Designed as an Anti-Inflammatory with Reduced Gastric Ulceration Properties

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Phosphate forms in plant and their internal buffering in five soybean cultivars

Diferenças inter e intra-específicas na habilidade de suportar períodos de estresse nutricional podem dever-se à capacidade de armazenar e liberar íons dos vacúolos, e, ou, à intensidade de retranslocação de nutrientes em tais condições. Neste trabalho, pretendeu-se avaliar diferenças varietais quanto ao tamanho do pool não-metabólico de Pi; velocidade de liberação do Pi previamente armazenado (VLPi), quando o P citoplasmático cai a um valor limite; capacidade de transportar Pi de regiões menos ativas para aquelas mais ativas metabolicamente e definir compartimentos que são preferencialmente fontes e os que são preferencialmente drenos para o Pi, em condições de absorção limitada de P. Avaliaram-se a produção de matéria seca e os teores internos de Pi, orgânico (Po) e total solúvel em ácido (Pts), de diferentes órgãos de plantas dos cultivares de soja (Glycine max L. Merrill) Santa Rosa, Uberaba, IAC8, Doko e UFV1, submetidos a oito dias de omissão do elemento. A VLPi foi estimada como tangente às equações obtidas para Pi como função do perído de omissão no ponto médio do período de omissão em que houve maior decréscimo em Pi (zero a quatro dias de omissão de P), t = dois dias, considerando-se que -deltaPi/deltat expressa a velocidade de liberação de Pi. A capacidade interna de tamponamento de Pi (CTIPi) foi calculada como o inverso da VLPi. O cultivar Santa Rosa apresentou maior capacidade de armazenar Pi, quando o suprimento externo foi alto, liberando-o mais intensamente sob condições de baixo suprimento de P que os cultivares IAC8 e UFV1. O cultivar Uberaba mostrou-se superior ao Doko em sua habilidade de armazenar e utilizar o Pi. Folhas superiores mostraram ser o principal dreno para o Pi armazenado em folhas medianas e inferiores, seguidas por raízes e caules. Raízes comportaram-se como fontes ou drenos para o Pi. Raízes e folhas superiores apresentaram maiores (VLPi) e menores valores de CTIPi que folhas medianas e folhas inferiores, sendo o caule o compartimento com menor VLPi e maior CTIPi. Dentre as variedades, as diferenças foram pequenas, destacando-se a maior VLPi e menor CTIPi do cultivar Santa Rosa. O cultivar Doko apresentou a menor VLPi e maior CTIPi, enquanto Uberaba, IAC8 e UFV1 ocuparam posição intermediária quanto a essas características.

Effect of lentil tannins on albumin hydrolysis by trypsin

Os taninos da casca da semente de lentilha foram extraídos e purificados, levados à interação com albumina isolada de lentilha e com caseína; e estudados por turbidimetria. As interações da albumina e caseína com taninos purificados, a várias relações tanino-proteína, mostraram ser independente e dependente do pH, respectivamente. Hidrólise in vitro com tripsina das proteínas sem taninos indicou que o aquecimento a 99°C/15 min reduzia a susceptibilidade da albumina e aumentava a da caseína à tripsina. A influência de diferentes relações tanino:proteína (1:40; 1:20; 1:5; 1:2,5) na hidrólise mostrou maior inibição para caseína que para albumina de lentilha, independente de aquecimento. Após aquecimento ambas proteínas foram mais hidrolizadas para qualquer das relações tanino proteínas estudadas. A eletroforese em gel de poliacrilamida-dodecilsulfato de sódio do transcurso da hidrólise da interação tanino-albumina nativa mostra a dependência da relação tanino:proteína.

Thermal analysis as a screening technique for the characterization of babassu flour and its solid fractions after acid and enzymatic hydrolysis

The babassu (Orbignya phalerata) is a native tree found in northern Brazil. Extracts of the babassu coconut have been widely used in industry. Babassu flour has about 60% starch, thus, besides nourishment it can be used as an alternative biofuel source. However, the properties of this starch lack of study and understanding. The main purpose of this study was to investigate the thermal behavior of raw babassu flour and its solid hydrolyzed fraction. The analyses were carried out using SHIMADZU DSC and TG thermic analyzers. The results demonstrated a reduction in thermal stability of the solid hydrolyzed fraction compared to raw matter. The kinetic parameters were investigated using non-isothermal methods and the parameters obtained for its decomposition process were an E(a) of 166.86 kJ mol(-1) and a frequency factor (beta) of 6.283 x 1014 min(-1); this was determined to be a first order reaction (n = 1). (C) 2011 Elsevier B.V. All rights reserved.

Optimization of acid hydrolysis from the hemicellulosic fraction of Eucalyptus grandis residue using response surface methodology

Biotechnological conversion of biomass into fuels and chemicals requires hydrolysis of the polysaccharide fraction into monomeric sugars. Hydrolysis can be performed enzymatically and with dilute or concentrate mineral acids. The present study used dilute sulfuric acid as a catalyst for hydrolysis of Eucalyptus grandis residue. The purpose of this paper was to optimize the hydrolysis process in a 1.41 pilot-scale reactor and investigate the effects of the acid concentration, temperature and residue/acid solution ratio on the hemicellulose removal and consequently on the production of sugars (xylose, glucose and arabinose) as well as on the formation of by-products (furfural, 5-hydroxymethylfurfural and acetic acid). This study was based on a model composition corresponding to a 2 3 orthogonal factorial design and employed the response surface methodology (RSM) to optimize the hydrolysis conditions, aiming to attain maximum xylose extraction from hemicellulose of residue. The considered optimum conditions were: H2SO4 concentration of 0.65%, temperature of 157 degrees C and residue/acid solution ratio of 1/8.6 with a reaction time of 20 min. Under these conditions, 79.6% of the total xylose was removed and the hydrolysate contained 1.65 g/l glucose, 13.65 g/l xylose, 1.55 g/l arabinose, 3.10 g/l acetic acid, 1.23 g/l furfural and 0.20 g/l 5-hydroxymethylfurfural. (c) 2006 Published by Elsevier Ltd.

Evaluation of the kinetics of xylose formation from dilute sulfuric acid hydrolysis of forest residues of Eucalyptus grandis

Dilute acid hydrolysis studies were performed on forest residues of Eucalyptus grandis, in a cylindrical reactor of stainless steel. The kinetics of this hydrolysis reaction was investigated employing 0.65% sulfuric acid, a residue/acid solution ratio of 1/9 (w/w), temperatures of 130, 140, 150, and 160 degrees C, and reaction times in the range 20-100 min. The results showed that, under the optimized conditions of acid hydrolysis employed in this study, the variables temperature and reaction time had a strong influence on hemicellulose removal and a small influence on the degree of lignin and cellulose removal. The highest xylose extraction yield was 87.6% attained at 160 degrees C, after 70 min reaction time, simultaneously with the formation of decomposition products, namely 2.8% acetic acid, 0.6% furfural, and 0.06% 5-hydroxymethylfurfural. A similar xylose extraction yield (82.8%) was observed at 150 degrees C after 100 min, with the formation of 3.2% acetic acid, 1.0% furfural, and 0.07% 5-hydroxymethylfurfural. The kinetic parameters determined at 130, 140, 150, and 160 degrees C for degradation of xylan present in the hemicellulose of the eucalyptus forest residue during the formation of xylose were the first-order reaction rate constants (k) for each temperature, 1.22 x 10(-4), 2.12 x 10(-4), 5.43 x 10(-4), and 9.05 x 10(-4) s(-1), respectively, and an activation energy (E-a) of 101.3 kJ mol(-1).

Growth of Calcium Phosphate Using Chemically Treated Titanium Oxide Nanotubes

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

PREPARATION and CHARACTERIZATION of CELLULOSE/HYDROUS NIOBIUM PHOSPHATE HYBRID

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

Quantitative microscopy characterization of hydrous niobium phosphate into bleached cellulose

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

Crystallization kinetics in phosphate sodium-based glass studied by DSC technique

In this work, differential scanning calorimetry (DSC) was used to study the mechanism of crystallization of 5OP(2)O(5-)27.8Na(2)O-16ZnO-6.2Al(2)O(3) glass. DSC measurements were performed on bulk and powdered glasses with different particle size. The curve for bulk glass shows one crystallization peak while powdered glasses presented two distinct crystallization peaks. Based on DSC studies, the activation energies obtained were 336 +/- 6 and 213 +/- 3 U mol(-1), associated with first and second crystallization peaks, respectively. Analyzing the DSC parameters as a function of particle size, the Avrami n parameter suggests that the peak at low temperature may be associated with surface crystallization while the peak at high temperature is associated with bulk crystallization. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Effect of PbS impurity on crystallization mechanism of phosphate glasses studied by differential scanning calorimetry

In this work, differential scanning calorimetry (DSC) was used to study effect of PbS impurity on crystallization mechanism of phosphate glasses. Bulk glasses presented one crystallization peak while powdered glasses presented two distinct crystallization peaks. For both undoped and doped glasses were determined the activation energies for the crystallization and the Avrami n parameters. The activation energies for undoped phosphate glass were 336 +/- 6 and 213 +/- 3 kJ mol(-1), respectively, associated with first and second crystallization peaks. For doped glass, the obtained energies were 373 +/- 9 and 286 +/- 7 kJ mol(-1). The calculated Avrami parameters, based on first crystallization peaks, for undoped and doped glasses were 2.25 +/- 0.01 and 1.75 +/- 0.02, respectively. These values suggest that the first DSC peak, in both glasses, may be associated with surface crystallization. (C) 2002 Elsevier B.V. B.V. All rights reserved.

Evidence of distinct phase transformations in lithium phosphate glass Li2O-P2O5

The purpose of this work is to study the Li2O-P2O5 glass using the differential scanning calorimetry (DSC) and x-ray diffraction (XRD) techniques to understand the crystallization process in this glass matrix. To study the glass by DSC, screened samples with different particle sizes to resolve the crystallization peaks were used. Both crystallization peaks were attributed to Li6P6O18 and LiPO3 phases. This evidence was corroborated by XRD analysis on glasses annealed at different temperatures in order to crystallize these phases.