Comparison of Botryosphaeran production by the ascomyceteous fungus Botryosphaeria sp., grown on different carbohydrate carbon sources, and their partial structural features

The influence of glucose concentration and other carbohydrates (monosaccharides: fructose, galactose, mannose; polyols: mannitol and sorbitol; disaccharides: lactose, sucrose and commercial sucrose; and industrial sugarcane molasses) were compared as sole carbon sources for the production of Botryosphaeran, an exopolysaccharide (EPS) produced by Botryosphaeria sp. The optimum glucose concentration for EPS production was 50 g 1(-1). With the exception of mannitol, the fungus produced EPS on all carbon sources studied, with highest yields occurring with sucrose followed by glucose. All EPS showed exclusively glucose after acid hydrolysis and monosaccharide analysis. FTIR spectroscopy demonstrated the presence of beta-anomers indicating that all the EPS produced by Botryosphaeria sp. on the different carbon sources were essentially of the beta-D-glucan type.

Botryosphaeran, a new substrate for the production of beta-1,3-glucanases by Botryosphaeria rhodina and Trichoderma harzianum Rifai

Botryosphaeria rhodina and Trichoderma harzianum Rifai were grown on botryosphaeran (an exopolysaccharide (EPS) of the beta-1,3; 1,6-D-Glucan type produced by B. rhodina) as sole carbon source with the objective of producing beta-glucanases of the beta-type. Conditions for beta-1,3-glucanase production by T harzianum were examined by a statistical response surface method, and showed maximal enzyme production at 5 days growth in media containing 1.5 g/1 of EPS. Good agreement was obtained between the experimental values of beta-1, 3-glucanase activity and the corresponding values predicted by the mathernatical model. The crude beta-1,3-glucanase preparations were active towards a number of different beta-1,3-glucans and beta-glucosides. The mycelium of B. rhodina also proved to be a good substrate for beta-1,3-glucanase production by both fungal species. (c) 2005 Published by Elsevier Ltd.

Thermal and Rheological Properties of a Family of Botryosphaerans Produced by Botryosphaeria rhodina MAMB-05

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

Proteomic Characterization of the Hyaluronidase (EC 3.2.1.35) from the Venom of the Social Wasp Polybia paulista

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

Shikimate Kinase (EC 2.7.1.71) from Mycobacterium tuberculosis: Kinetics and Structural Dynamics of a Potential Molecular Target for Drug Development

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

ATIVIDADE DE PEROXIDASES (EC 1.11.1.7) E TEOR DE PROLINA em FEIJOEIRO Phaseolus vulgaris L. CULTIVADO em CONDIÇÕES DE SALINIDADE

Alterações nos níveis de prolina e na atividade da peroxidase foram estudadas em folhas e raízes de feijoeiro em função da resposta à salinidade do meio. A atividade da peroxidase (E.C. 1.11.1.7) e o teor de prolina das folhas foi maior do que nas raízes. Os resultados mostraram que a atividade da peroxidase diminuiu, enquanto os teores de prolina aumentaram gradualmente nas plantas cultivadas em meio salino. Ocorreu diminuição do teor de potássio e aumento de sódio nos órgãos analisados, com o aumento da concentração salina.

ATIVIDADE DE PEROXIDASES (EC 1.11.1.7) E TEOR DE PROLINA NO EMBRIÃO E COTILÉDONES DE FEIJOEIRO Phaseolus vulgaris L. em CONDIÇÕES DE SALINIDADE

Os efeitos da salinidade sobre a atividade de peroxidases foram acompanhadas no embrião e em cotilédones de feijoeiro Phaseolus vulgaris L. cv. Carioca, observando-se em ambos, aumento na atividade das enzimas. Com relação ao teor de prolina, ocorreu decréscimo constante no embrião de feijoeiro cultivado em condições salinas, entretanto, nos cotilédones, verificou-se aumento progressivo em condições de presença ou ausência de salinidade.

Purification and properties of acid phosphatase (EC 3.1.3.2) secreted by strain 74A of the mould Neurospora crassa

Both P-i-repressible acid phosphatases, IIb (mycelial) and IIc (extracellular), synthesized by Neurospora crassa and purified to apparent homogeneity by 7.5% PAGE, are monomers, are inhibited by 2 mM ZnCl2 and are nonspecifically stimulated by salts. However, the IIc form is activated by p-nitrophenylphosphate (in a negative cooperativity effect with a K-0.5 of 2.5 mM) whereas form IIb shows Michaelis kinetics, with a K-m of 0.5 mM. Thus, since both enzymatic forms may be expressed by the same gene (pho-3), it is possible that post-translational modifications lead to the excretion of an enzymatic form with altered Michaelis kinetics compared with the enzymatic form retained by the mycelium.

THE PHO-2A MUTANT OF NEUROSPORA-CRASSA WHICH IS DEFICIENT IN PI-REPRESSIBLE ALKALINE-PHOSPHATASE (EC 3.1.3.1) IS ALSO DEFECTIVE IN PI-REPRESSIBLE ACID-PHOSPHATASE (EC 3.1.3.2)

1. The mycelial Pi-repressible acid phosphatase presented p-nitrophenylphosphatase activity with negative cooperativity and Michaelian behavior when synthesized by the wild-type and pho-2A mutant strains of Neurospora crassa, respectively.2. The major acid phosphatase present in cell extracts of the pho-2A mutant of N. crassa grown in low Pi medium is more thermolabile (t1/2 = 4 min at 54-degrees-C, pH 5.4) than that of the wild strain (stable for at least 80 min at 54-degrees-C, pH 5.4).3. The pho-2A mutant of N. crassa secreted a more thermolabile acid phosphatase (t1/2 = 30 min at 50-degrees-C, pH 5.4) than the wild strain (t1/2 of at least 80 min at 50-degrees-C, pH 5.4).4. The pho-2A mutant of N. crassa synthesized a more thermolabile acid phosphatase (t1/2 = 37 min at 54-degrees-C, pH 5.4) than the wild strain in high Pi medium (t1/2 = 14 min al 54-degrees-C, pH 5.4).5. The pleiotropic nature of the pho-2 locus and its possible involvement in the mechanism of phosphatase secretion by N. crassa are proposed.

Secretion of metalloendopeptidase 24.15 (EC 3.4.24.15)

The metalloendopeptidase EP24.15 (EC3.4.24.15) is a neuropeptide-metabolizing enzyme present in neural and endocrine tissues, presumably functioning extracellularly, Because the majority of the EP24.15 activity is identified in the soluble fraction of cellular homogenates, suggesting that the enzyme is primarily an intracellular protein, we addressed the issue of how EP24.15 arrives in the extracellular environment, We utilized a model system of neuroendocrine secretion, the AtT20 cell, According to both enzymatic activity and immunologic assays, EP24.15 was synthesized in and released from AtT20 cells. Under basal conditions and after stimulation by corticotropin-releasing hormone or the calcium ionophore A23187, EP24.15 activity accumulated in the culture medium. This secretion was not attributable to cell damage, as judged by the absence of release of cytosolic enzyme markers and the ability to exclude trypan blue dye. Pulse-chase analysis and subcellular fractionation of AtT20 cell extracts suggested that the mechanism of EP24.15 secretion is not solely via classical secretory pathways, Additionally, drugs which disrupt the classical secretory pathway, such as Brefeldin A and nocodazole, blocked A23187-stimulated EP24.15 release yet had no effect on basal EP24.15 release, suggesting differences in the basal and stimulated pathways of secretion for EP24.15. In summary, EP24.15 appears to be secreted from AtT20 pituitary cells into the extracellular milieu, where the enzyme can participate in the physiologic metabolism of neuropeptides.

Sulfonation and anticoagulant activity of botryosphaeran from Botryosphaeria rhodina MAMB-05 grown on fructose

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

Kinetic properties of de-repressible alkaline phosphatase (EC 3.1.3.1) from Neurospora crassa

De-repressible alkaline phosphatase from N. crassa shows inhibition by PNP-P and a hyperbolic mixed-type inhibition by Pi. Both increasing concentrations of Pi and decreases in assay pH abolished inhibition by the substrate. Also, Pi promoted polymerization of the enzyme molecule, whose effect may account for the inhibitory behaviour shown by the enzyme in the presence of low Pi concentrations. © 1991 Rapid Communications of Oxford Ltd.

SISTEMAS QUIMILUMINESCENTES COM PEROXIDASE (EC:1.11.1.7) E SUAS APLICACOES EM ANALISES CLINICAS

The enzyme horseradish peroxidase HRP (EC:1.11.1.7), has both acid and basic isoenzymes, catalyses a wide range of reactions (acting as an oxiredutase or an oxidase) and is thought capable of one- or two-electrons oxidations depending on the substrate employed. Today, the methodology for these assay can be chemiluminescent reactions and enhanced chemiluminescent. The enhanced chemiluminescent assay with system HRP, luminol, peroxide and an enhancer has provided the basis for a convenient and sensitive assay for peroxidase and peroxidase conjugates, DNA probe and blotting assay. It is particularly more advantageous than the others, because is very rapid, more sensitive (attomoles), easy to do and technically simple, and is relatively specific for HRP (reduces the effect of the interference).