Global warming is an environmental problem that has generated worldwide concern. This phenomenon results from the emission of greenhouse gases (CO2, for example) into the atmosphere by human activities, leaving it up to new technologies to find renewable and clean energy sources. One of the most promising solution is the development of lignocellulosic biofuels, a field of study that has been growing in recent decades. Considering that lignocellulosic biomass is the largest reserve of carbohydrates available, and that Brazil has a large territory with climatic conditions favorable to the production of biomass, besides being a major producer of agro-industrial waste, we become one of the countries with the greatest potential in this industry. The biggest obstacle to the viability of this process is related to the enzymatic hydrolysis step, considering the high cost of enzymatic cocktails and the monopoly of foreign companies. An efficient enzymatic cocktail must contain a mix of cellulases, hemicellulases and ligninases capable of acting synergistically and producing a high yield of fermentable sugars at the end of the hydrolysis. Most of the enzymes used for the production of enzymatic cocktails are secreted by filamentous fungi, and one of the best-described species is Trichoderma reesei, an efficient producer of enzymes. However, the secretion of certain types of enzymes by Trichoderma sp. is relatively low and mutagenesis-based approaches are often applied to improve these deficiencies. Although biotechnology offers important tools to reach these objectives, the production of enzymatic cocktails is still a challenge to be overcome. To this purpose, the identification of enzymes capable of hydrolyzing cellulose and hemicellulose becomes essential, leading to a cheaper process and contributing to the viability of the production of lignocellulosic biofuels. This project aims to identify different enzymes produced by a Trichoderma sp. fungus, identified as being highly efficient in the hydrolysis of lignocellulosic materials. This fungus was found to be a better producer of xylanases, b-glucosidases and endoglucanases, as well as more efficient in the hydrolysis of lignocellulosic materials, in comparison to the reference species Trichoderma reesei. To this end, it is necessary to achieve the following:- Standardization of the cultivation of the selected fungus and the control T. reesei on two different carbon sources (pre-treated straw and pre-treated energy-cane).- Identification of the enzymes secreted by both species under the two proposed conditions by mass spectrometry.- Genetic engineering of Trichoderma reesei to insert identified genes from the studied species in order to validate the relationship between these genes and the observed high performance of the studied fungus during hydrolysis.
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