Enzymatic hydrolysis of botryosphaeran and laminarin by beta-1,3-glucanases produced by Botryosphaeria rhodina and Trichoderma harzianum Rifai

Botryosphaeran, a (1 -> 3; 1 -> 6)-beta-D-glucan produced by Botryosphaeria rhodina, and laminarin were hydrolysed by two fungal beta-glucanases predominantly of the 1,3-type produced by B. rhodina and Trichoderma harzianum Rifai grown on botryosphaeran as sole carbon source. Both beta-glucanase preparations presented different modes of attack on botryosphaeran and laminarin. Laminarin was hydrolysed to the extent of similar to 50% in 1 hand 100% within 24 h, and its hydrolysis products were mainly glucose and gentiobiose, and lesser amounts of laminaribiose and oligosaccharides of DP 3-4 during the early stages of hydrolysis, while botryosphaeran 'yielded mainly glucose and gentiobiose with some trisaccharide, but no laminaribiose or tetrasaccharide when hydrolysed by the T. harzianum enzyme. By contrast, B. rhodina beta-1,3-glucanases produced predominantly glucose during all stages of botryosphaeran hydrolysis. Some physicochemical properties of the 1,3- and 1,6-beta-glucanases, and beta-glucosidases contained in the two fungal P-glucanase preparations are also described for the first time. (c) 2006 Elsevier Ltd. All rights reserved.

Ionizing radiation induced degradation of poly (2-methoxy-5-(2 '-ethyl-hexyloxy)-1,4-phenylene vinylene) in solution

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Evaluation of the beta-glucanolytic enzyme complex of Trichoderma harzianum Rifai for the production of gluco-oligosaccharide fragments by enzymatic hydrolysis of 1,3;1,6-beta-D-glucans

Botryosphaeran, a new exopolysaccharide from the endophytic fungus Botryosphaeria rhodina MAMB-05, and algal laminarin were hydrolyzed by partially-fractionated enzymes of the beta-glucanolytic complex from Trichoderma harzianum Rifai. beta-Glucanase fractions (F-I and F-II) separated by gel permeation chromatography presented different modes of attack on botryosphaeran and laminarin. Botryosphaeran was hydrolyzed to the extent of 66% (F-I) and 98% (F-II) within 30 min, and its main hydrolysis products were gluco-oligosaccharides of DP >= 4, with lesser amounts of glucose, di- and tri-saccharides. The action of enzyme fractions I and II on laminarin resulted in 15% conversion to glucose, while the percentage of saccharification was radically different (70% for F-I and 25% for F-II). The different product arrays within the polysaccharide hydrolysates can be explained by the difference in the enzymes' specificities within each enzyme fraction, and the molecular structures of the polysaccharides and their complexity.

Gelatinization properties of native starches and their Naegeli dextrins

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

Refolding and purification of bothropstoxin-1, a Lys49 - Phospholipase A(2) homologue, expressed as inclusion bodies in Escherichia coli

Hydrolysis of phospholipids by Group II phospholipase A(2) enzymes involves a nucleophilic attack on the sn-2 ester bond by the His48 residue and stabilization of the reaction intermediate by a Ca2+ ion cofactor bound to the Asp49 residue in the protein active site region, Bothropstoxin-I (BthTX-I) is a PLA, variant present in the venom of the snake Bothrops jararacussu which shows a Asp49 to Lys substitution and which lacks hydrolytic activity yet damages artificial membranes by a noncatalytic Ca2+-independent mechanism. In order to better characterize this unusual mechanism of membrane damage, we have established an expression system for BthTX-I in Escherichia coli. The DNA-coding sequence for BthTX-I was subcloned into the vector pET11-d, and the BthTX-I was expressed as inclusion bodies in E, coli BL21(DE3). The native BthTX-I contains seven disulfide bonds, and a straightforward protocol has been developed to refold the recombinant protein at high protein concentration in the presence of surfactants using a size-exclusion chromatography matrix. After refolding, recovery yields of 2.5% (corresponding to 4-5 mg of refolded recombinant BthTX-I per liter of bacterial culture) were routinely obtained. After refolding, identical fluorescent and circular dichroism spectra were obtained for the recombinant BthTX-I compared to those of the native protein. Furthermore, the native and refolded recombinant protein demonstrated identical membrane-damaging properties as evaluated by measuring the release of an entrapped fluorescent marker from liposomes, (C) 2001 Academic Press.

Surface degradation of glass ceramics after exposure to acidulated phosphate fluoride

Objective: This study evaluated the surface degradation effect of acidulated phosphate fluoride (APF) gel exposure on the glassy matrix ceramics as a function of time. Material and methods: Disc-shaped ceramic specimens (N = 120, 10/per ceramic material) were prepared in stainless steel molds (inner diameter: 5 mm, height: 2 mm) using 6 dental ceramics: 3 indicated for ceramic-fused-to-metal (Vita Omega 900, Carmen and Vita Titankeramik), 2 for all-ceramic (Vitadur Alpha and Finesse (R) Low Fusing) and 1 for both types of restorations (IPS d. SIGN). The specimens were wet ground finished, ultrasonically cleaned and auto-glazed. All specimens were subjected to calculation of percentage of mass loss, surface roughness analysis and topographical description by scanning electron microscopy (SEM) before (0 min) and after exposure to 1.23 % APF gel for 4 min and 60 min representing short-and long-term etching effect, respectively. The data were analyzed using two-way ANOVA with repeated measures and Tukey` s test (alpha=0.05). Results: Significant effect of the type of the ceramics (p=0.0000, p=0.0031) and exposure time (p=0.0000) was observed in both surface roughness and percentage of mass loss values, respectively. The interaction factor between both parameters was also significant for both parameters (p=0.0904, p=0.0258). Both 4 min (0.44 +/- 0.1-0.81 +/- 0.2 mu m) and 60 min (0.66 +/- 0.1 - 1.04 +/- 0.3 mu m) APF gel exposure created significantly more surface roughness for all groups when compared to the control groups (0.33 +/- 0.2-0.68 +/- 0.2 mu m) (p<0.05). There were no significant differences in percentage of mass loss between the ceramics at 4 min (p>0.05) but at 60 min exposure, IPS d. SIGN showed the highest percentage of mass loss (0.1151 +/- 0.11). The mean surface roughness for Vita Titankeramik (0.84 +/- 0.2 mu m) and Finesse (R) Low Fusing (0.74.+/- 0.2 mu m) was significantly higher than those of the other ceramics (0.59 +/- 0.1 mu m - 0.49 +/- 0.1 mu m) and Vita Titankeramik (p<0.05) regardless of the exposure time. A positive correlation was found between surface roughness and percentage of mass loss for all ceramic materials [(r=0.518 (Vitadur Alpha), r=0.405 (Vita Omega 900), r=0.580 (Carmen), r=0.687 (IPS d. SIGN), r=0.442 (Finesse (R) Low Fusing), r=0.572 (Vita Titankeramik), Pearson's correlation coefficient)]. The qualitative SEM analysis showed evidence of corrosive attack on all of ceramics at varying degrees. Conclusions: The ceramics indicated for either metal-ceramic or all-ceramic restorations were all vulnerable to surface texture changes and mass loss after short-term and long-term APF gel exposure.

FACTORS THAT AFFECT THE ENZYMATIC DEGRADATION OF NATURAL STARCH GRANULES - EFFECT OF THE SIZE OF THE GRANULES

In this study the relationship between the enzymatic susceptibility and the size of the com and cassava starch granules was studied. The starch granules were separated by size and classified according to their average diameter in: a) larger than 16 mum; b) between 15 and 10 mum and c) smaller than 10 mum. The starch granules of various sizes were hydrolyzed by bacterial alpha-amylase and fungal amyloglucosidase. The results showed a relationship between the enzymatic susceptibility and the size of the starch granules; smaller size of the starch granules resulted in a higher percentage of hydrolysis. A basic difference in the mode of action of enzymes on small and large granules was observed. Enzymatic attack on the large granules was characterized by considerable surface corrosion, mainly at the radial axis. For small granules, the enzymatic action occurred on the surface of the granules and was characterized by an erosion with solubilization of the granules. Chemical and physical analysis of the starches suggested that hydrolysis should occur mainly at the amorphous areas of the granules.

An eco-friendly protocol for synthesis of thiourea derivatives: 1-benzoyl-3-benzylguanidine and 1-benzoyl-3-benzyl-O-ethylisourea. A possible non-purely thermal microwave assisted reaction

1-Benzoyl-3-benzylguanidine and 1-benzoyl-3-benzyl-O-ethylisourea were synthesized in good yields (68 and 76%, respectively) from 1-benzoyl-3-benzylthiourea and benzoyl-ethylthiocarbamate in dry media conditions using KF-Al2O3 under microwave irradiation. Strong nucleophilic amines promoted the sulfur elimination by attack on the thiocarbonyl group in both thiourea and thiocarbamates to afford guanidines and isourea, respectively. Transesterification products were obtained from p-TsOH catalyzed reaction of thiocarbamate with alcohols under MW-solvent-free conditions. Very important non-purely thermal MW specific effects were evidenced and attributed to stabilization by coulombic interactions between materials and waves. (c) 2005 Elsevier Ltd. All rights reserved.

Épocas de semeadura e manejos de corte de milheto cultivado no outono-inverno e efeitos na cultura da soja em sucessão sob plantio direto

Pós-graduação em Agronomia (Agricultura) - FCA

Em busca do Império: a trajetória intelectual e política de Eduardo Prado

Pós-graduação em História - FCHS

Blendas de PVC/PCL foto/termo e biotratadas com fungos de solo (Phanerochaete chrysosporium e Aspergillus fumigatus)

Pós-graduação em Ciências Biológicas (Microbiologia Aplicada) - IBRC

Estudo termoanalítico e eletroquímico das reações em estado sólido na interfase metálica dos sistemas Pt-Rh/Hg, empregando a liga de composição Pt-Rh 80:20% (m/m)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo das reações no estado sólido entre a liga de composição 'PT'-'RH'85:15%(m/m) e 'HG'

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Resistência de clones de seringueira ao ataque de Calacarus heveae feres (Acari, Eriphyidae): aspectos biológicos e ecológicos da acarofauna e análises fisiológicas de plantas infestadas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Produção de beta-glucanases por Trichoderma harzianum Rifai para obtenção gluco-oligossacarídeos a partir de botriosferana

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