69 resultados para Amylases


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Recently’ recognition cnf immobilization ‘technology for the rapid conversion of several substrates into metabolites and repeated reuse of the biocatalysts have drawn the attention of the fermentation scientists and technologists to try these new technologies for the rapid production of pnxkmt and enhancement of the efficiencies of the systems Hence in the present study rice was selected,as a substrate since it is a rich source of starch, available and cultivated throughout the year almost in all part of our country. Rice although known for its use as a staple food in many forms as rice, idli, dosai etc., has not been used in industry extensively. However, it ii; a potential resource for’ the production of alcohol, high protein food anui for sugar and sugar syrups as it is evidenced by the few reports mentioned in the review of literature. Of the several microorganisms available, Bacillus sp, is a known candidate for the production of amylases. Hence in the present study Bacillus sp, was desired for its known efficiencies in starch conversion

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Microbial enzymes are in great demand owing to their importance in several industries such as brewing, baking, leather, laundry detergent, dairy. starch processing and textiles besides pharmaceuticals. About 80% of the enzymes produced through fermentation and sold in the industrial scale are hydrolytic enzymes. Due to recognition of new and new applications, an intensive screening of different kinds of enzymes with novel properties, from various microorganisms, is being pursued all over the world. Bacillus sp are largely known to produce a-amylase, among the different groups of microoganisms, at industrial level. They are known to produce both saccharifying and liquefying a-amylases (Fukumoto 1963; walker and Campbell, 1967a). which are distinguishable by their mechanisms of starch degradation by the fact that the saccharifying asamylases produce an increase in reducing power about twice that of the liquefying enzyme (Fukumoto, 1963; Pazur and Okada, 1966). Under this circumstances, the present study was undertaken, with a View to utilise a fast growing B.coagu1ans isolated from soil, for production of thermostable and alkaline oz-amylase under different fermentation processes

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Seed germination and seedling establishment are critical processes for commercial plantation and depend directly on reserve mobilization as a source of cellular fuels and biosynthetic precursors. In this way, we investigated the coordination among reserve mobilization, metabolite partitioning, and mobilizing enzyme activities in Moringa oleifera Lam (moringa) an oil-seeded species employed in biofuel production. Seeds were germinated under controlled conditions and seedlings were grown hydroponically at a greenhouse. Samples were harvested at 0, 4, 8, 10, 12, 16, and 20 days after imbibition (DAI). The contents of dry mass (DM), neutral lipids (NL), soluble proteins (SP), starch, total soluble sugars (TSS), non-reducing sugars (NRS), and total free amino acids (TFAA) as the activity of isocitrate lyase (ICL), acid proteases, and amylases were determined. The mobilization of storage proteins was initiated during seed germination whereas the mobilization of storage lipids and starch was triggered throughout seedling establishment although all reserves have been depleted until 20 DAI. The partitioning of DM and metabolites to the roots and the shoots was uneven during seedling establishment. Low shoot/root ratio on the basis of DM could be related to the natural occurrence of moringa in drought climates. In the roots, TSS, NRS, and TFAA were accumulated from 12 to 16 DAI and then were consumed until the end of the experiment. In the shoots, TSS and TFAA were consumed in parallel with NRS accumulation from 12 to 20 DAI. The activity of ICL, acid proteases, and amylases was coordinated with the mobilization of lipids, proteins and starch respectively. Thus, we propose that the patterns of reserve mobilization and metabolite partitioning verified in moringa seem distinct from those found to other tree species and may be involved in metabolic strategies to enable environment colonization

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A soil sample was taken from the top 0-20cm at Jaboticabal county, São Paulo State, Brazil, air dried, sieved to 5mm, and placed into pots (2700g per pot). Sewage sludge was air-dried, ground to 2mm, and thoroughly mixed to the top 0-10cm soil of each pot, which were irrigated with distilled water in a total volume equivalent to the last 30years average rainfall in the region. Sorghum was sowed 120days after sewage sludge incorporation and then the irrigation was made according to the plants' requirement. When the plants were about 10 cm high, they were thinned to two per pot. Soil samples (0-10, 10-20, and 20-30 cm depth) were obtained immediately after the incorporation of sewage sludge and at 30, 60, 120, and 170 days after, air dried, sieved to 2 mm and analyzed for organic matter (OM), pH (0,01 mol L-1 CaCl2), extractable P (resin), potassium (K), calcium (Ca), and magnesium (Mg), amylase and cellulase activity. Sewage sludge increased soil OM, pH, extractable phosphorus (P), K. Ca. amylase and cellulase activity, especially at the rate 16 t ha(-1). Organic matter, extractable P, K, Ca, Mg. and amylase activity were higher in the top 0-10cm, while pH was higher in the 20-30cm layer. Amylase activity was not affected by sampling depth. Organic matter, pH, extractable P. K, Ca, and Mg decreased during the experimental period. Amylase activity decreased until sorghum was sowed and increased afterwards. Cellulase activity increased until 90 days after sewage sludge application and then decreased. Sewage sludge used in the experiment should already contain some amylase activity or a substance that was a soil enzyme activator and also a substance that was an inhibitor of soil cellulase inhibitor. Sonic of the plant nutrients contained in sewage sludge, mainly P, did not migrate down the soil column. an indication that sewage sludge should be incorporated into the soil to improve nutrient bioavailability. Sorghum roots increased amylase activity but did not affect cellulase activity.

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O objetivo deste trabalho foi avaliar o efeito da combinação de fitase e do complexo amilase, protease e xilanase, em dietas de milho e soja, formuladas com redução e sem redução dos níveis de energia, cálcio e fósforo sobre o desempenho e a digestibilidade ileal de nutrientes, em frangos de corte. Foi utilizado o delineamento inteiramente casualizado, com esquema fatorial 2x2, com duas dietas-controle: com redução e sem redução dos nutrientes, e duas suplementações das enzimas (sem adição e com adição de fitase e complexo enzimático), com dez repetições de 40 aves. A digesta ileal foi coletada aos 43 dias de idade, para determinação da energia digestível e dos coeficientes de digestibilidade da proteína bruta, matéria seca, cálcio e fósforo. O desempenho apresentou interação em todos os parâmetros analisados. As aves do tratamento com redução dos nutrientes mostraram pior desempenho em relação às aves da dieta sem redução dos nutrientes. Não houve efeito da matéria seca na digestibilidade e na retenção de cálcio. A adição da combinação enzimática melhorou a digestibilidade da proteína e a retenção de fósforo. Houve interação quanto à energia digestível, com efeito apenas nas dietas sem redução dos nutrientes, com maiores valores nas dietas suplementadas.

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

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Branching enzyme catalyzes the formation of alpha-1,6 branch points in either glycogen or starch. We report the 2.3-Angstrom crystal structure of glycogen branching enzyme from Escherichia coli. The enzyme consists of three major domains, an NH2-terminal seven-stranded beta-sandwich domain, a COOH-terminal domain, and a central alpha/beta-barrel domain containing the enzyme active site. While the central domain is similar to that of all the other amylase family enzymes, branching enzyme shares the structure of all three domains only with isoamylase. Oligosaccharide binding was modeled or branching enzyme using the enzyme-oligosaccharide complex structures of various alpha-amylases and cyclodextrin glucanotransferase and residues were implicated in oligosaccharide binding. While most of the oligosaccharides modeled well in the branching enzyme structure, an approximate 50degrees rotation between two of the glucose units was required to avoid steric clashes with Trp(298) of branching enzyme. A similar rotation was observed in the mammalian alpha-amylase structure caused by an equivalent tryptophan residue in this structure. It appears that there are two binding modes for oligosaccharides in these structures depending on the identity and location of this aromatic residue.

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O presente trabalho teve por objetivo analisar resíduos do farelo de mandioca resultantes de processos de hidrólise enzimática para obtenção de etanol; visando o aproveitamento destes como fonte de fibras dietéticas. Foram realizados quatro ensaios enzimáticos utilizando as enzimas amilolíticas, a-amilase e amiloglucosidase, complementadas ou não com celulase e/ou pectinase. Os resíduos foram caracterizados quanto à composição centesimal, pH, acidez, perfil de açúcares e quanto às fibras (FDA, FDN, celulose, hemicelulose, lignina, açúcares neutros). Realizou-se também a análise microscópica dos resíduos. Pelos resultados obtidos na caracterização dos resíduos calculou-se a energia metabolizável aparente (EM). Observou-se que independente do ensaio enzimático todos os resíduos podem ser usados como fonte de fibras insolúveis. Os resíduos resultantes dos ensaios com pectinase apresentaram uma proporção aproximada de 1:1:1 de amido, fibras e açúcares, sendo a glicose o açúcar majoritário, e com energia metabolizável aparente de cerca de 2,6 kcal/g. Já os resíduos, onde não se utilizou a pectinase a proporção foi de 2:1:1 aproximadamente e a energia 3,1 kcal/g. A análise microscópica dos resíduos mostrou a presença de amido não hidrolisado preso às células em todos os ensaios enzimáticos sendo que, nos resíduos dos ensaios com pectinase a quantidade observada foi bem inferior aos demais. Uma possível alternativa para diminuir o valor calórico dos resíduos seria a lavagem com água após a prensagem para extração do hidrolisado para fermentação.

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Glucoamylases have been used with alpha-amylases for the industrial conversion of starch into glucose. However, little is known about the properties of this glycosylated protein retained in the cell wall of Saccharomyces as well as its role in the saccharification and fermentation of amylaceous substrates, notably in high cell density processes. In most of the strains assayed, decreases in biomass formation were followed by increases in glucoamylase secretion (expressed as U/mg(biomass) in 1 ml of culture) when glucose was exchanged for starch as carbon source or the growth temperature was raised from 30 to 35 degrees C. Despite the losses in viability, significant increases in the activity of the wall fraction occurred when cultures of thermotolerant yeasts propagated at 30 degrees C or washed cells resuspended in buffer solution were heated to 60 degrees C for 60-80 min prior to amylolytic assays. Thus, intact cells of thermotolerant yeasts can be used as colloidal biocatalysts in starch degradation processes. (C) 2005 Published by Elsevier Ltd.

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The acute, subchronic and chronic toxicities of 2,4- dichlorophenoxyacetic acid (2,4-D) were studied in rats. Animals were exposed acutely (600 mg/kg), subchronically (200 ppm for 30 d) and chronically (200 ppm for 180 d) to 2,4-D by the oral route. Clinical, laboratory and histopathological methods were used as indicators of toxicity. After acute exposure, the herbicide decreased locomotor activity and induced ataxia, sedation, muscular weakness (mainly of the hind quarters) and gasping for breath; increased aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), alkaline phosphatase (AP), amylase activities and creatinine levels; decreased total protein (TP) and glucose levels; and increased hematocrit values. Subchronic and chronic 2,4-D exposures did not induce overt clinical signs or symptoms of intoxication. However, subchronic herbicide exposure increased AST activity and albumin and hematocrit values, and chronic exposure increased AST, AP and LDH activities, decreased amylase and glucose levels, but did not change hematocrit values. Chromatographic analysis of the serum of chronically exposed rats showed the presence of the herbicide; the amount found (3.76 ± 1.16 mg/ml) suggested the absence of 2,4-D accumulation within the body. Although macroscopic or histopathological lesions were not observed in acutely, subchronically or chronically 2,4-D exposed rats, the laboratory data obtained suggest tissue injuries after dosing, since the results are considered early indicators of primarily hepatic and muscle tissue damage.

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The antimalarial properties of azomethine H represent the basis for its use as a chemotherapeutic agent. This work was carried out in order to verify the biological side effects of azomethine H and to clarify the contribution of reactive oxygen species (ROS) in this process. It was shown that azomethine H increased serum activities of amylase, alanine transaminase (ALT) and the TEARS concentrations, in rats. No changes were observed in glutathione peroxidase and catalase activities. The drug-induced tissue damage might be due to superoxide radicals (O-2(.-)), since Cu-Zn superoxide dismutase activities were increased by azomethine I-I treatment. This study allows tentative conclusions to be drawn regarding which reactive oxygen metabolites play a role in azomethine H activity. We concluded that (O-2(.-)) maybe produced as a mediator of azomethine H action.

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The growth and the extracellular amylase production by Aspergillus ochraceus were studied in a stationary culture medium. Maximum growth rate of this fungus was found after 5 days of incubation at 30° C, but maximum amylase production was obtained after 2 days. The highest amylase production were attained with lactose, maltose, xylose and starch as carbon sources. The extracellular amylase production and mycelial growth were influenced by the concentration of starch. Other carbohydrates supported growth but did not induce amylase synthesis and glucose repressed it, indicating catabolite repression in this microorganism. The presence of both mechanisms of induction and repression suggests that at least these multiple forms of regulation are present in A. ochraceus. Of the nitrogen sources tested, casaminoacids, ammonium nitrate and sodium nitrate stimulated the highest yield of amylase. Optimal amylase production was obtained at pH 5.0, but enzyme activity was found only in the 4.0-6.0 pH range. These results were probably due to the inhibitory effect of NH 4 +-N in the culture medium.

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Neonatal administration of monosodium glutamate (MSG) in rats causes definite neuroendocrine disturbances which lead to alterations in many organ systems. The possibility that MSG could affect tooth and salivary gland physiology was examined in this paper. Male and female pups were injected subcutaneously with MSG (4 mg/g BW) once a day at the 2nd, 4th, 6th, 8th and 10th day after birth. Control animals were injected with saline, following the same schedule. Lower incisor eruption was determined between the 4th and the 10th postnatal days, and the eruption rate was measured between the 43rd and the 67th days of age. Pilocarpine-stimulated salivary flow was measured at 3 months of age; protein and amylase contents were thereby determined. The animals treated with MSG showed significant reductions in the salivary flow (males, -27%; females, -40%) and in the weight of submandibular glands (about -12%). Body weight reduction was only about 7% for males, and did not vary in females. Saliva of MSG-treated rats had increased concentrations of total proteins and amylase activity. The eruption of lower incisors occurred earlier in MSG-treated rats than in the control group, but on the other hand the eruption rate was significantly slowed down. The incisor microhardness was found to be lower than that of control rats. Our results show that neonatal MSG treatment causes well-defined oral disturbances in adulthood in rats, including salivary flow reduction, which coexisted with unaltered protein synthesis, and disturbances of dental mineralization and eruption. These data support the view that some MSG-sensitive hypothalamic nuclei have an important modulatory effect on the factors which determine caries susceptibility.

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Background: Previous experiments have shown that a decoction of Bauhinia forficata leaves reduces the changes in carbohydrate and protein metabolism that occur in rats with streptozotocin-induced diabetes. In the present investigation, the serum activities of enzymes known to be reliable toxicity markers were monitored in normal and streptozotocin-diabetic rats to discover whether the use of B. forficata decoction has toxic effects on liver, muscle or pancreas tissue or on renal microcirculation. Methods: An experimental group of normal and streptozotocin-diabetic rats received an aqueous decoction of fresh B. forficata leaves (150 g/L) by mouth for 33 days while a control group of normal and diabetic rats received water for the same length of time. The serum activity of the toxicity markers lactate dehydrogenase, creatine kinase, amylase, angiotensin-converting enzyme and bilirubin were assayed before receiving B. forficata decoction and on day 19 and 33 of treatment. Results: The toxicity markers in normal and diabetic rats were not altered by the diabetes itself nor by treatment with decoction. Whether or not they received B. forficata decoction the normal rats showed a significant increase in serum amylase activity during the experimental period while there was a tendency for the diabetic rats, both treated and untreated with decoction, to have lower serum amylase activities than the normal rats. Conclusions: Administration of an aqueous decoction of B. forficata is a potential treatment for diabetes and does not produce toxic effects measurable with the enzyme markers used in our study. © 2004 Pepato et al; licensee BioMed Central Ltd.

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The salivary activity in pups of spontaneously hypertensive rats (SHR) and Wistar (W) rats treated with atenolol during pregnancy, and lactation was evaluated. Atenolol's anti-hypertensive effect on the SHR rats was noticed from the beginning of treatment. Atenolol-treated SHR and Wistar rat pups showed a decrease in salivary gland weight, salivary flow, and protein concentration, with no alteration in salivary amylase activity. Atenolol's effect on salivary glands can interfere with oral health maintenance. Copyright © Informa Healthcare USA, Inc.