Myotoxic phospholipases A2 isolated from Bothrops brazili snake venom and synthetic peptides derived from their C-terminal region: Cytotoxic effect on microorganism and tumor cells

This paper reports the purification and biochemical/pharmacological characterization of two myotoxic phospholipases A2 (PLA2s) from Bothrops brazili venom, a native snake from Brazil. Both myotoxins (MTX-I and II) were purified by a single chromatographic step on a CM-Sepharose ion-exchange column up to a high purity level, showing Mr ∼ 14,000 for the monomer and 28,000 Da for the dimer. The N-terminal and internal peptide amino acid sequences showed similarity with other myotoxic PLA2s from snake venoms, MTX-I belonging to Asp49 PLA2 class, enzymatically active, and MTX-II to Lys49 PLA2s, catalytically inactive. Treatment of MTX-I with BPB and EDTA reduced drastically its PLA2 and anticoagulant activities, corroborating the importance of residue His48 and Ca2+ ions for the enzymatic catalysis. Both PLA2s induced myotoxic activity and dose-time dependent edema similar to other isolated snake venom toxins from Bothrops and Crotalus genus. The results also demonstrated that MTXs and cationic synthetic peptides derived from their 115-129 C-terminal region displayed cytotoxic activity on human T-cell leukemia (JURKAT) lines and microbicidal effects against Escherichia coli, Candida albicans and Leishmania sp. Thus, these PLA2 proteins and C-terminal synthetic peptides present multifunctional properties that might be of interest in the development of therapeutic strategies against parasites, bacteria and cancer. © 2008 Elsevier Inc. All rights reserved.

Isolamento de fungos produtores de lipases catalisadores de reações de transesterificação para produção de biodiesel

Pós-graduação em Microbiologia - IBILCE

Produção de biodiesel etílico com uso de lipases extracelulares de fungos termofílicos

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Produção de lipases por fungos termofílicos e mesofílicos e uso na produção de biodiesel etílico

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

Análise dos compostos flavorizantes da cana-de-açúcar e otimização da aplicação de extratos ricos em β-glicosidases para liberação de aroma na produção de aguardente de cana

Pós-graduação em Química - IBILCE

Aplicação do fungo termofílico Thermomyces lanuginosus na síntese enzimática de ésteres

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

Oxidação de carboidratos via catálise metálica não enzimática para aplicação em sensores

This work has main aim of is to propose the synthesis and characterization of nanostructured materials for oxidation of carbohydrates such as glucose, with non-enzymatic catalysis. The proposed pathway of synthesis of metal catalysts is the polyol method and techniques of physical characterization proposals for analysis of prepared catalyst pass through diffraction technique of ray-x (DRX), scanning electron microscopy (SEM) and Energy Dispersive Spectroscopy ray-x (EDX). Technical proposals for the electrochemical characterization of the synthesized catalysts are Cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The prospects of this work are compared by the catalytic activity of the sensor designed with non-enzymatic sensors and biosensors also known in the literature

Enzymatic toxins from snake venom: structural characterization and mechanism of catalysis

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

Characterization of glycerol kinase from baker's yeast: Response surface modeling of the enzymatic reaction

The present study describes a methodology of dosage of glycerol kinase (GK) from baker's yeast. The standardization of the activity of the glycerol kinase from baker's yeast was accomplished using the diluted enzymatic preparation containing glycerol phosphate oxidase (GPO) and glycerol kinase. The mixture was incubated at 60 degrees C by 15 min and the reaction was stopped by the SDS solution addition. A first set of experiments was carried out in order to investigate the individual effect of temperature (7), pH and substrate concentration (S), on GK activity and stability. The pH and temperature stability tests showed that the enzyme presented a high stability to pH 6.0-8.0 and the thermal stability were completely maintained up to 50 degrees C during 1 h. The K(m) of the enzyme for glycerol was calculated to be 2 mM and V(max) to be 1.15 U/mL. In addition, modeling and optimization of reaction conditions was attempted by response surface methodology (RSM). Higher activity values will be attained at temperatures between 52 and 56 degrees C, pH around 10.2-10.5 and substrate concentrations from 150 to 170 mM.This low cost method for glycerol kinase dosage in a sequence of reactions is of great importance for many industries, like food, sugar and alcohol. RSM showed to be an adequate approach for modeling the reaction and optimization of reaction conditions to maximize glycerol kinase activity. (C) 2007 Elsevier B.V. All rights reserved.

Effects of Low Molecular Weight Sulfated Galactan Fragments From Botryocladia Occidentalis on the Pharmacological and Enzymatic Activity of Spla2 From Crotalus Durissus Cascavella

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Enzymatic and structural characterization of new PLA2 isoform isolated from white venom of Crotalus durissus ruruima

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

Effect of umbelliferone (7-hydroxycoumarin, 7-HOC) on the enzymatic, edematogenic and necrotic activities of secretory phospholipase A2 (sPLA2) isolated from Crotalus durissus collilineatus venom

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Inhibition of Neurotoxic Secretory Phospholipases A(2) Enzymatic, Edematogenic, and Myotoxic Activities by Harpalycin 2, an Isoflavone Isolated from Harpalyce brasiliana Benth

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)