Synergistic effect in the catalytic activity of lipase Rhizomucor miehei immobilized on zeolites for the production of biodiesel

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

Antimicrobial activity of an L-amino acid oxidase isolated from Bothrops leucurus snake venom

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Crystal structure of a dimeric Ser49 PLA(2)-like myotoxic component of the Vipera ammodytes meridionalis venomics reveals determinants of myotoxicity and membrane damaging activity

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Partial characterization of protease from a thermophilic fungus, Thermoascus aurantiacus, and its hydrolytic activity on bovine casein

Proteases are one of the most important groups of industrial enzymes, with considerable application in the food industry. The aim of this work was to study a novel protease produced by the thermophilic fungus, Thermoascus aurantiacus, through solid-state fermentation (SSF). The enzyme acted optimally at pH 5.5 and 60 degrees C it was stable up to 60 degrees C for 1 h and in the pH range 3.0-9.5. To elucidate the enzyme's proteolytic activity, its hydrolytic profile on bovine casein, an important protein in the food industry, was studied by enzymatic hydrolysis on skim milk, analyzed by gel electrophoresis (UREA-PAGE), which clearly showed that the protease does not have the same specificity as bovine chymosin. (c) 2006 Elsevier Ltd. All rights reserved.

Colorimetric enzymatic assay of L-malic acid using dehydrogenase from baker's yeast

A colorimetric method has been developed and optimized to measure L-malic acid in samples of fruit juices and wine. This method is based on oxidation of the analyte, catalyzed by malate dehydrogenase (MDH) from dry baker's yeast, and in combination with the reduction of a tetrazolium salt (MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide). In the present study, the method exhibited sensitivity in the range of 500-4000 mu M of L-malic acid in the reaction cuvette, with the lower detection limit of 6.7-10(-2) g/L, the upper limit of 53.6.10(-2) g/L and a maximum standard deviation of only 2.5 % for the analyzed samples. The MDH activity from baker's yeast was also optimized, the enzyme showed a high stability at pH=8.0-9.0 and the activity was maintained completely at temperatures up to 40 degrees C for 1 hour. The results show that the colorimetric method using enzymatic preparations from dry baker's yeast is a simple and low-cost method with possibility of wide application.

The effect of non-surgical periodontal therapy on peroxidase activity in diabetic patients: a case-control pilot study

Objective: To evaluate the effect of periodontal therapy on clinical parameters as well as on total salivary peroxidase (TSP) activity and myeloperoxidase (MPO) activity in the gingival crevicular fluid (GCF) of patients with type 2 diabetes mellitus (DM2) and of systemically healthy individuals.Material and Methods: Twenty DM2 subjects with inadequate metabolic control (test group) and 20 systemically healthy individuals (control group), both groups with chronic periodontitis, were enrolled. Periodontal clinical parameters, namely periodontal probing depth (PD), clinical attachment level (CAL), visible plaque index (VPI), bleeding on probing (BOP), gingival bleeding index (GBI) and presence of suppuration (SUP), as well as TSP activity and GCF MPO activity, were assessed before and 3 months after non-surgical periodontal therapy.Results: At baseline and 3 months post-treatment, the test group presented a higher percentage of sites with VPI and BOP (p < 0.01). MPO activity in the GCF presented lower values (p < 0.05) for the test group at both baseline and the post-treatment period. The periodontal treatment resulted in a significant improvement of most clinical and enzymatic parameters for both groups (p < 0.05).Conclusions: In both groups, the periodontal therapy was effective in improving most clinical parameters and in reducing salivary and GCF enzymatic activity. The diabetic individuals presented lower MPO activity in the GCF.

Quartz crystal microbalance as a tool for kinetic enzymatic assays by variation of pH

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

Geranylation of benzoic acid derivatives by enzymatic extracts from Piper crassinervium (Piperaceae)

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

Catalase activity of different Candida species after exposition to specific antiserum

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

MICROBIAL ACTIVITY of DARK RED LATOSOL SAMPLES STORED AT DIFFERENT TEMPERATURES

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

ISOENZYMATIC POLYMORPHISM and ACTIVITY of PEROXIDASES of COMMON BEAN(Phaseolus vulgaris L.) UNDER SALINE STRESS

Uma das utilizações da técnica de cultura de tecidos para o melhoramento vegetal é a identificação de linhas de células que apresentem tolerância ao estresse salino. Para se estudar os mecanismos bioquímicos envolvidos na expressão genética da tolerância a salinidade, calos oriundos de eixos embrionários de quatro cultivares de feijão (Phaseolus vulgaris L.; cultivares IAC - carioca, IAPAR 14, JALO-EEP 558, BAT - 93), foram cultivados em meio sólido Murashige & Skoog (1962), suplementado com NaCl nas concentrações de 0, 20, 40, 60 e 80 mM. Após 14 dias de incubação, os calos foram coletados e analisados quanto aos padrões isoenzimáticos e de atividade das peroxidases. Os cultivares BAT e IAPAR apresentaram duas zonas de atividade em comum na região anódica e apenas uma zona enzimática específica a cada um deles (migração mais rápida).Possivelmente as duas zonas anódicas intermediárias sejam produtos do mesmo loco enzimático, porém com alelos diferentes, consequentemente diferentes mobilidades eletroforéticas. O cv. JALO apresentou duas zonas anódicas de atividade em comum com os cultivares IAC e IAPAR com uma zona anódica exclusiva de migração mais lenta, a qual apresentou atividade mais intensa de todos os cultivares analisados. Este cultivar revelou ainda uma zona catódica provavelmente dimérica e heterozigota nos indivíduos de todos os tratamentos aplicados. Provavelmente, esta é a mesma zona que ocorre em homozigose com fixação do alelo lento para os indivíduos de todos os tratamentos efetuados nos cultivares BAT e IAPAR. O cv. IAC apresentou duas bandas anódicas em comum com os cv. IAPAR e JALO. Apresentou também a banda anódica mais rápida em comum com o cv. IAPAR e uma banda anódica exclusiva de migração mais lenta. Curiosamente, os indivíduos deste cv. mantidos em meio suplementado com 20 mM de NaCl não apresentaram atividade nas três zonas anódicas mais lentas. Ocorreu no cv. IAC uma única zona de atividade catódica, dimérica e heterozigota para os indivíduos provenientes de todos os tratamentos, composta provavelmente de dois alelos diferentes da zona correspondente ao cv. JALO. Amostras provenientes dos tratamentos 40 e 60 mM de NaCl, desta zona catódica, apresentaram maior atividade enzimática. A análise da atividade da peroxidase no extrato bruto, revelou que os cultivares responderam diferentemente ao aumento da concentração salina no meio de cultura, com aumento pronunciado dessa atividade nos cultivares IAC e JALO.

Effect of broiler breeder age on pancreas enzymes activity and digestive tract weight of embryos and chicks

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

Gastric antiulcer activity of Syngonanthus arthrotrichus SILVEIRA

Syngonanthus arthrotrichus SILVEIRA, popularly known as sempre-vivas mini-saia, is found in mountains of the Espinhaco range in the Brazilian states of Bahia and Minas Gerais. Extracts of this species contain several constituents, including flavonoids which may have antiulcerogenic activity. An ethanolic extract (EEOH), and flavonoid-rich (FRF) and flavonoid-deficient (FDF) fractions obtained from the scapes of S. arthrotrichus were investigated for their ability to prevent ulceration of the gastric mucosa in mice and rats. In the ethanol/HCl-induced ulcer model, lansoprazole (30 mg/kg), EEOH (50, 100, 250 mg/kg) given orally protected the gastric mucosal against injury in mice by 79%, 78%, 73%, and 64% respectively. In the ethanol-induced gastric ulcer model in rats, the lansoprazole (30 mg/kg), FRF and FDF (100 mg/kg) significantly protected the gastric mucosal of rats by 65%, 38% and 25% respectively when compared with the negative control group. In indomethacin/ bethanechol-induced gastric ulcers, cimetidine (100 mg/kg) and the EEOH (100, 250 mg/kg) inhibited gastric ulcer formation by 73%, 55% and 32% respectively. In this exactly model other treatments as cimetidine, FRF and FDF (100 mg/kg) each caused 54%, 36% and 45% inhibition, respectively. In the stress-induced gastric ulcer model, cimetidine (100 mg/kg) and the EEOH (50, 100, 250 mg/kg), inhibited gastric ulcer formation by 63%, 73%, 68% and 69% respectively. In the same model, cimetidine, FRF and FDF (100 mg/kg) significantly protected the gastric mucosal of the mice by 60%, 51% and 47% when compared to the control group. In pylorus-ligated mice, cimetidine (positive control) and FRF significantly decreased gastric acid secretion, increased gastric pH and reduced the acid output when compared to the negative control. FDF had no significant effect on these parameters. The protection provided by FRF probably involved an antisecretory mechanism mediated by flavonoids which were absent in FDF. The amount of adherent mucous in the stomach contents was also evaluated with the treatments carbenoxolone (200 mg/kg), FRF and FDF (100 mg/kg) treatment. Each treatments significantly increased the amount of adherent mucous in the gastric juice (8.67 +/- 1.73, 3.35 +/- 1.59, 2.1 +/- 0.41 mg/g of wet tissue, respectively) compared to the control group, indicating a cytoprotective action on the gastric mucosa. Treatment with FRF plus indomethacin and FDF plus indomethacin reduced the prostaglandin biosyntesis (13.6 +/- 6.5, 27 +/- 5.5 pg/well) by the mucosa, indicating that the cytoprotective action on the gastric mucosa was not related to the level of prostaglandins. Only FDF (38 +/- 17 pg/well) maintained the level of prostaglandins and guaranteed the integrity of the mucosa. The results indicate that the EEOH, FRF and FDF have antisecretory and cytoprotective actions, that may be related to the presence of luteoline in the extract and active fractions.

Dissociation of enzymatic and pharmacological properties of piratoxins-I and -III, two myotoxic phospholipases A(2) from Bothrops pirajai snake venom

Piratoxins (PrTX) I and III are phospholipases A(2) (PLA(2)s) or PLA(2) homologue myotoxins isolated from Bothrops pirajai snake venom, which also induce myonecrosis, bactericidal activity against Escherichia coli, disruption of artificial membranes, and edema. PrTX-III is a catalytically active hemolytic and anticoagulant Asp49 PLA(2), while PrTX-I is a Lys49 PLA, homologue, which is catalytically inactive on artificial substrates, but promotes blockade of neuromuscular transmission. Chemical modifications of His, Lys, Tyr, and Trp residues of PrTX-I and PrTX-III were performed, together with cleavage of the N-terminal octapeptide by CNBr and inhibition by heparin and EDTA. The lethality, bactericidal activity, myotoxicity, neuromuscular effect, edema inducing effect, catalytic and anticoagulant activities, and the liposome-disruptive activity of the modified toxins were evaluated. A complex pattern of functional differences between the modified and native toxins was observed. However, in general, chemical modifications that significantly affected the diverse pharmacological effects of the toxins did not influence catalytic or membrane disrupting activities. Analysis of structural changes by circular dichroism spectroscopy demonstrated significant changes in the secondary structure only in the case of N-terminal octapeptide cleavage. These data indicate that PrTX-I and PrTX-III possess regions other than the catalytic site, which determine their toxic and pharmacological activities. (C) 2001 Academic Press.

Endectocide activity of a new long-action formulation containing 2.25% ivermectin+1.25% abamectin in cattle

The present work aimed to evaluate the endectocide activity of a new injectable long-action formulation, containing ivermectin (IVM) and abamectin (ABA). In each one of the four experiments performed, the following groups were formed: group I: 2.25% IVM (450 mu g/kg) + 1.25% ABA (250 mu g/kg), group II: 3.15% IVM (630 mu g/kg) and group III: control. Eighteen bovine naturally infected by gastrointestinal nematoda were selected for anthelmintic evaluation and necropsied on posttreatment day (PTD) 14 to estimate the total parasitic burden. For the Rhipicephalus (Boophilus) microplus field trial, 30 bovine were selected by means of counts of semi-engorged R. (B.) microplus and the therapeutic and residual efficacy evaluated by tick counts on PTDs 1, 3, 7, 14, 21, 28, 35, 42, 49, 56, 63, 70, 77, 84 and 91. In the stall test, 15 calves were artificially infested with 5000 R. (B.) microplus (Mozzo strain) larvae three times a week and daily collections of all the engorged female ticks detached from each calf were performed until the PTD 80. Forty bovine naturally infected with Dermatobia hominis larvae were selected and the number of larvae was counted by visual and tactile inspection on PTDs 3, 7, 14, 28, 35, 49, 63, 77, 91 and 105. In this trial, a formulation containing 1% doramectin (200 mu g/kg) was also used. IVM + ABA formulation and 3.15% IVM eliminated four of the eight species of nematode identified. The anthelmintic efficacy of the avermectins association against Haemonchus placei, Cooperia spatulata and C. punctata was 89.64%, 98.84% and 97.69%, while 3.15% IVM achieved 30.98%, 84.79% and 75.56%, respectively. The two formulations evaluated showed reduced acaricide action on the PTD I and 3, reaching high efficacy percentages from PTD 14 onward. The IVM + ABA showed efficacy above 95% in the period between PTDs 21 and 49. In the stall test, it observed no difference (P > 0.05) between the two formulations regarding the R. (B.) microplus counts during the entire evaluation period. IVM + ABA reduced the number of ticks from the PTD 1 to 77 (P < 0.05) and 3.15% IVM reduced (P < 0.05) the tick number from PTD 4 up to PTD 80. The three endectocides showed no difference (P > 0.05) regarding the number of D. hominis larvae and prevented this parasite reestablishment until PTD 105. These results indicate that the IVM + ABA association showed higher anthelmintic activity and similar efficacy against arthropods to the formulation containing 3.15% IVM. (C) 2008 Elsevier B.V. All rights reserved.