Eucalyptus ESTs involved in the production of 9-cis epoxycarotenoid dioxygenase, a regulatory enzyme of abscisic acid production

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

Production of saccharogenic and dextrinogenic amylases by Rhizomucor pusillus A 13.36

A newly-isolated thermophilic strain of the zygomycete fungus Rhizomucor pusillus 13.36 produced highly active dextrinogenic and saccharogenic enzymes. Cassava pulp was a good alternative substrate for amylase production. Dextrinogenic and saccharogenic amylases exhibited optimum activities at a pH of 4.0-4.5 and 5.0 respectively and at a temperature of 75°C. The enzymes were highly thermostable, with no detectable loss of saccharogenic or dextrinogenic activity after 1 h and 6 h at 60°C, respectively. The saccharogenic activity was inhibited by Ca2+ while the dextrinogenic was indifferent to this ion. Both activities were inhibited by Fe2+ and Cu2+ Hydrolysis of soluble starch by the crude enzyme yielded 66% glucose, 19.5% maltose, 7.7% maltotriose and 6.6% oligosaccharides. Copyright © 2005, The Microbiological Society of Korea.

Effect of prostaglandins on the production of H2O2 and cytokines that modulate the fungicidal activity of human monocytes against Paracoccidioides brasiliensis

Human monocytes lack fungicidal activity against high virulent strains of Paracoccidioides brasiliensis, the etiological agent of paracoccidioidomycosis, even after IFN-γ activation. However, monocytes treated with indomethacin (INDO) or INDO plus IFN-γ effectively killed this fungus, suggesting an inhibitory role of prostaglandins in this process. Thus, the purpose of this work was to test if this regulatory effect of prostaglandin was associated with alterations on H2O2 production and/or on modulatory cytokines levels, such as TNF-α, IL-10, and IL-6. Peripheral blood monocytes obtained from 10 healthy donors were incubated for 18 hours in the presence or absence of IFN-γ, INDO, or IFN-γ plus INDO, and further challenged with a high virulent strain of P. brasiliensis (Pb18) for 4 hours. Then, the monocytes cultures were evaluated for H2O2 release and fungicidal activity calculated by counting the colony forming units after plating. Moreover, on supernatants of the same cultures, TNF-α, IL-10, IL-6, and PGE2 concentrations were evaluated by ELISA. Monocytes treated with INDO or INDO plus IFN-γ presented higher fungicidal activity associated with the release of higher levels of H2O2 and TNF-α, but lesser levels of PGE2, when compared to nontreated cells. However, the levels of IL-10 and IL-6 were similar between treated and nontreated cells. The results suggest that human monocytes when challenged with high virulent strains of P. brasiliensis produce prostaglandins that inhibit the fungicidal activity of these cells by reducing H2O2 and TNF-α levels.

Production of monoclonal antibodies against Streptococcus mutans antigens

Several studies have been conducted in the last decades aiming to obtain an anti-canies vaccine, however some studies have demonstrated cross reactivity between Streptococcus mutans surface antigens and the human cardiac tissue. In this work, the reactivity of five anti-Streptococcus mutans monoclonal antibodies (MoAb) (24A, 56G, G8, E8 and F6) was tested against oral streptococci, cardiac antigens and skeletal and cardiac myosins, aiming to evaluate the specificity of these MoAb. The hybrid producers of immunoglobulins of the IgG 2b class were cloned by limit dilution and expanded in vivo. MoAb were tested by ELISA. The hybrid 24A reacted with S. mutans CCT 1910, S. salivarius CCT 0365 and S. pyogenes T23. No reactivity difference was observed among the tested species. Cross reactivity with heart and cardiac myosin was not confirmed and only reaction with myosin of skeletal muscle was observed (p = 0.0381). The hybrid 56G reacted with all the tested microorganisms and there was statistically significant difference between S. mutans and S. pyogenes T23 (p < 0.001). This hybrid also reacted with myosin of skeletal muscle (p = 0.0095). C8, E8 and F6 presented low reactivity against oral streptococci strains and no reactivity against cardiac antigens. The data of this study showed that the 24A and 56G anti-S. mutans MoAb presented reactivity with S. pyogenes and S. salivarius, reinforcing the occurrence of common antigens between these species. The tested MoAb presented low cross-reactivity with myosin of skeletal muscle, but anti-heart activity could not be confirmed.

Production of four cultivars of iceberg lettuce, under protected cultivation, with and without black polyethylene ground cover

The aim of the work was to evaluate the performance of four cultivars of iceberg lettuce, in a greenhouse, with and without ground cover. The experimental design adopted was that of subdivided parcels, where parcels were provided with and without ground cover consisting of black polyethylene and the subparcels consisted of the cultivars. There were three repetitions per treatment and six central plants of each parcel were evaluated. Drip irrigation was utilized. The cultivars studied were: 'Laurel', 'AF-389', 'Mayara' and 'Tainá'. The seeds were planted in Styrofoam trays with 288 cells containing the substrate Plantmax Hortaliças® A. The trays were kept in a greenhouse until transplanting, 25 days after seeding. The parameters examined were: fresh weight of the aerial part per plant, longitudinal (DL) and transverse (DT) diameters of the head, height of the plant and mean number of leaves per plant. Only among the cultivars there were significant differences in transverse and longitudinal diameters of the head and height of the plant. No significant interactions were detected between ground cover and cultivars. With regard to longitudinal and transverse diameters, 'AF-839' (DL = 27.11 cm; DT = 24.53 cm) showed smaller diameters than those of 'Laurel' (DL = 31.13 cm; DT = 30.55 cm), but did not differ from the other cultivars. In regard to height of the plants, 'Laurel' (20.66 cm) had the greatest value and 'AF-839' (16.02 cm) the lowest. The cultivars did not differ with respect to mean number of leaves and fresh weight of the aerial part of the plant. It is concluded that there were no significant differences in the cultivation of the iceberg lettuce with and without ground cover. The cultivar 'Laurel' stood out with respect to longitudinal and transverse diameters and height of the plant, only differing from the 'AF-839' with regard to the diameters.

Production of cellulolytic and hemicellulolytic enzymes from Aureobasidium pulluans on solid state fermentation

This article investigates a strain of the yeast Aureobasidium pullulans for cellulase and hemicellulase production in solid state fermentation. Among the substrates analyzed, the wheat bran culture presented the highest enzymatic production (1.05 U/mL endoglucanase, 1.3 U/mL β-glucosidase, and 5.0 U/mL xylanase). Avicelase activity was not detected. The optimum pH and temperature for xylanase, endoglucanase and β-glucosidase were 5.0 and 50, 4.5 and 60, 4.0 and 75°C, respectively. These enzymes remained stable between a wide range of pH. The β-glucosidase was the most thermostable enzyme, remaining 100% active when incubated at 75°C for 1 h. © 2007 Humana Press Inc.

The effect of pH on horseradish peroxidase-catalyzed oxidation of melatonin: Production of N1-acetyl-N2-formyl-5- methoxykynuramine versus radical-mediated degradation

There is a growing body of evidence that melatonin and its oxidation product, N1-acetyl-N2-formyl-5-methoxykynuramine (AFMK), have anti-inflammatory properties. From a nutritional point of view, the discovery of melatonin in plant tissues emphasizes the importance of its relationship with plant peroxidases. Here we found that the pH of the reaction mixture has a profound influence in the reaction rate and products distribution when melatonin is oxidized by the plant enzyme horseradish peroxidase. At pH 5.5, 1 mm of melatonin was almost completely oxidized within 2 min, whereas only about 3% was consumed at pH 7.4. However, the relative yield of AFMK was higher in physiological pH. Radical-mediated oxidation products, including 2-hydroxymelatonin, a dimer of 2-hydroxymelatonin and O-demethylated dimer of melatonin account for the fast consumption of melatonin at pH 5.5. The higher production of AFMK at pH 7.4 was explained by the involvement of compound III of peroxidases as evidenced by spectral studies. On the other hand, the fast oxidative degradation at pH 5.5 was explained by the classic peroxidase cycle. © 2007 The Authors.

Bio-safety technology in production of bovine herpesvirus type 5 (BoHV-5) using an alternative serum-free medium

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

Application of a serum/protein-free medium for the growth of mammalian cell lines and high level production of classical, variant and very virulent strain of infectious bursal disease virus (IBDV)

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

Effect of plastic film mulch on the production of butterhead lettuce cultivars under protected cultivation

The study was performed in a greenhouse with the aim of evaluating the production of butterhead lettuce cultivars with and without plastic film mulch in the summer. The cultivars studied were 'Elisa', 'Lídia', 'Herman' and 'Luísa'. The experimental design was a split plot, in which the main plot was plastic film mulch (with or without black polyethylene film) and the subplots were the cultivars, with three repetitions. The seedlings were grown in Styrofoam trays of 288 cells and transplanted 25 days after seeding in a space of 25 × 25 cm. Harvests were carried out 45 days after transplanting and the following variables were determined: fresh and dry weights of the aerial part, number of leaves per plant, plant height and the longitudinal and transverse diameters of the plant. Significant differences were found only among the cultivars for the characteristics studied, with no cover × cultivar interaction. Fresh weight of the aerial part of 'Lídia' (270.9 g) was greater than that of 'Herman' (142.5 g), but did not differ from those of 'Luísa' (229.8 g) or 'Elisa' (204.8). The dry weight of the aerial part of 'Lídia' (12.4 g) differed statistically only with respect to that of 'Herman' (7.9 g). The cultivars 'Luísa' and 'Elisa' had an aerial part dry weight of 11.4 and 11.2 g, respectively. The number of leaves per plant of the cultivars 'Luísa' (45.1) and 'Elisa' (43.0) was higher than for 'Herman' (24.05), but did not differ from that of the cultivar 'Lídia' (39.5). The mean transverse and longitudinal diameters, as well as the mean height of the plants, showed that 'Lídia', 'Elisa' and 'Luísa' had the greatest values. The cultivars 'Lídia', 'Elisa' and 'Luísa' were the most productive and plastic film mulch had no effect on lettuce production under protected cultivation.

Search for resonant pair production of neutral long-lived particles decaying to bb̄ in pp̄ collisions at s=1.96TeV

We report on a first search for resonant pair production of neutral long-lived particles (NLLP) which each decay to a bb̄ pair, using 3.6fb-1 of data recorded with the D0 detector at the Fermilab Tevatron collider. We search for pairs of displaced vertices in the tracking detector at radii in the range 1.6-20cm from the beam axis. No significant excess is observed above background, and upper limits are set on the production rate in a hidden-valley benchmark model for a range of Higgs boson masses and NLLP masses and lifetimes. © 2009 The American Physical Society.

Influence of medium composition in the production of ethanol by Zymomonas mobilis

The production of ethanol using Zymomonas mobilis had been reported to be three to four times larger than with Saccharomyces cereviseae. The influence of pH, temperature and composition of the means of fermentation are parameters that can direct the metabolism for the production of ethanol. The objective of this study was to evaluate the production of ethanol by Zymomonas mobilis CCT 4494, by variations of the initial pH, temperature and concentrations KCl, K 2SO4, MgSO4, CaCl2 and sucrose, by a factorial experimental design of type 27-2, according to the model proposed by Box et al. (1978). For this, the broth of sugar cane was used as sole carbon source, because it is cheap and easily accessible in the region of São José do Rio Preto, São Paulo State. According to the experimental design, the bacteria Zymomonas mobilis CCT 4494 has adapted in the fermentation mean containing high concentrations of sucrose, and supported the change of pH and temperature of fermentation. The highest amount of ethanol produced was 8.89 mg mL-1. This is not similar to the levels of secondary metabolites produced by Zymomonas mobilis CCT 4494.

Influence of hydric stress in the production of toxic principles in Nerium oleander L.

Many plants utilized in the urban center shows substances considered toxic whose production could be influenced by some factors, like hydric stress, including ornamental Nerium oleander L., widely used in gardens in various parts of the world, which presents production of cardioative glucosides, considered toxic. This study had the objective to evaluate the effect of field capacity in the biomass and the level of cardioative glucosides in seedlings of Nerium oleander. The experiment was carried at UNIDERP, in Campo Grande City, Mato Grosso do Sul State, Brazil, at the University for Development of State and Pantanal Region, using the experimental delineation in randomized blocks. There were 4 treatments (25%; 50%; 75% and 100% of the field capacity), 5 replications and 4 plants by parcel, totalling 80 plants. The evaluations were realized 60 days after the seedlings were planted. The quantitative analysis of the cardioative glucosides was realized by gravimetric test, after selective extraction of the glucosides. Were conclude that increase of the quantity of water in the soil raised the biomass production until 75% of the field capacity and increased the level of cardioative glucosides, showing that water management is very important and should be provided only the necessary to development of the plant.

Production of pectate lyase by penicillium viridicatum RFC3 in solid-state and submerged fermentation

Pectate lyase (PL) was produced by the filamentous fungus Penicillium viridicatum RFC3 in solid-state cultures of a mixture of orange bagasse and wheat bran (1 : 1 w/w), or orange bagasse, wheat bran and sugarcane bagasse (1 : 1 : 0.5 w/w), and in a submerged liquid culture with orange bagasse and wheat bran (3%) as the carbon source. PL production was highest (1,500U mL -1 or 300Ug -1 of substrate) in solid-state fermentation (SSF) on wheat bran and orange bagasse at 96 hours. PL production in submerged fermentation (SmF) was influenced by the initial pH of the medium. With the initial pH adjusted to 4.5, 5.0, and 5.5, the peak activity was observed after 72, 48, and 24 hours of fermentation, respectively, when the pH of the medium reached the value 5.0. PL from SSF and SmF were loaded on Sephadex-G75 columns and six activity peaks were obtained from crude enzyme fromSSF and designated PL I, II, III, IV, V, andVI, while five peaks were obtained fromcrude enzyme fromSmF and labeled PL I', II', III', IV', and VII'. Crude enzyme and fraction III from each fermentative process were tested further. The optimum pH for crude PL from either process was 5.5, while that for PL III was 8.0. The maximum activity of enzymes from SSF was observed at 35°C, but crude enzyme was more thermotolerant than PL III, maintaining its maximum activity up to 45°C. Crude enzyme from SmF and PL III' showed thermophilic profiles of activity, with maximum activity at 60 and 55°C, respectively. In the absence of substrate, the crude enzyme from SSF was stable over the pH range 3.0-10.0 and PL III was most stable in the pH range 4.0-7.0. Crude enzyme from SmF retained 70%-80% of its maximum activity in the acid-neutral pH range (4.0-7.0), but PIII showed high stability at alkaline pH (7.5-9.5). PL from SSF was more thermolabile than that from SmF. The latter maintained 60% of its initial activity after 1 h at 55°C. The differing behavior of the enzymes with respect to pH and temperature suggests that they are different isozymes. Copyright © 2010 Viviani Ferreira et al.

Agaricus blazei as a Substrate for the Production of β-1,3-Glucanase by Trichoderma harzianum Rifai

Extracellular β-1,3-glucanase was produced by Trichoderma harzianum Rifai cultivated in the Agaricus blazei (Agaricus brasiliensis) extract as a substrate in submerged fermentation. A 22-central composite factorial design was developed using the time of culture (x1/day) and Agaricus blazei extract concentration (x2/(g/L)) as variables, and the results were analyzed using response surface methodology (RSM). The results showed that the Agaricus blazei extract concentration was the most important variable in the production of β-1,3-glucanase, and the maximum β-1,3-glucanase activity (0.77 U/mL) was obtained in one day of cultivation. The β-glucan present in the cell wall of Agaricus blazei mushroom proved to be a good substrate for inducing the production of specific β-1,3-glucanase by Trichoderma harzianum Rifai.