Resumo: |
Cellulases are enzymes that degrade plant cell wall. These enzymes are produced by various microorganisms, which filaments fungi are the main producers. Pathogenic microorganisms produced enzymes available to act the degradation of plant cell wall, allowing their invasion in the plant. In addition with a possible involvement in the pathogenic mechanism of Moniliophthora perniciosa, cellulases are enzymes that have wide application in the food and biofuels. In this context, this study aimed to produce, characterize, apply and determine the tree-dimensional of glycosidic enzymes. The results showed that the best conditions for its production, in liquid fermentation, were using 7 g L-1 yeast extract in 19 days of incubation. The pH optimum and temperature for maximum cellulase activity were 7.2 and 47°C, respectively. Studies of the effects of salts on enzyme activity showed that the potassium chloride and potassium bromide causes increase in enzyme activity and the presence of 100 mM. Kinetic studies showed a Km and Vmax of 0.03732 mg / mL substrate CMC and 27.47 μmoles / mg / min, respectively. The sequence of the cellulase of M. perniciosa was obtained from the database of Genome / Proteome M. Pernicious with 2193 base pairs and 731 amino acids, and using it to build the three-dimensional model for comparative modeling. After molecular dynamics simulations of the model presented an early stable simulation between 2.00 and 2.36 nanoseconds. The complex cellulolytic obtained from M. pot, was efficient in the hydrolysis of bagasse-cane sugar and reducing the viscosity of cashew apple juice. |
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