Produção das enzimas acessórias feruloil esterase e xilanase por fungos filamentosos isolados da região amazônica e sua aplicação na hidrólise do bagaço de canade-açúcar
Braga, Cleiton Márcio Pinto
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Xylanase and feruloyl esterase (FAE) are two enzymes of great importance for plant biomass decomposition. The hemicellulose, one of the biomass constituents, has a great structural variety, so that for its complete deconstruction, several groups of enzymes are required, including mainly xylanases. These enzymes act on the β-1,4 glycosidic bonds between the xylose residues. Xylanase activity can be enhanced in the presence of esterases, such as feruloyl esterase (FAE), since the latter catalyzes the hydrolysis of linkages which make cell wall structure to be more intricated. Such connections are formed mainly by the ferulic acid esterified to sugar residues attached to the carbohydrate main chain. It is noteworthy that the ferulic acid cross-links to hemicellulose, pectin and lignin structures. Both enzymes are of interest in many sectors, for example, during the saccharification for bioethanol production and bleaching stages of pulp and paper industry. Therefore, this study aimed to select and cultivate filamentous fungi from the Amazon rainforest capable of synthesizing FAE and xylanase, and to evaluate the application of these enzymes in the hydrolysis of sugar cane bagasse. Initially, a screening was conducted on agar plates containing ethyl ferulate. Among eleven fungi tested, seven showed a clear halo around the colony, as indicative of FAE production. Then, the fungi were cultivated in liquid nutrient medium supplemented with 1% wheat bran, and, based on productivity values, three strains of Aspergillus oryzae (P21C3, P6B2 and P27C3) were chosen for cultivation in a stirred tank bioreactor. After 24 hours of cultivation of the P21C3 strain in the bioreactor, it was obtained a crude enzyme extract with 74.85 UI.mL-1 and 47.02 UI.L-1 of xylanase and FAE activity, respectively. This extract was used for the hydrolysis of hydrothermal pretreated sugarcane bagasse with supplementation of a commercial cellulolytic enzyme preparation. These hydrolysis reactions resulted in conversions of 51.2% (cellulose to glucose) and 78.1% (hemicellulose to xylose). Furthermore, the addition of the on-site produced enzyme extract increased the glucose release by 35.8% and the xylose release by 53.8%.