Elucidation of cellular signaling pathways mediated by protein kinase A (PKA), MAPK and calcium during expression and secretion of cellulases by the fungus Trichoderma reesei

Abstract

Brazil is one of the countries with the greater diversity in terms of lignocellulosic biomass. The lignocellulolytic enzymes production by filamentous fungi has been the constant subject of study, mainly because such enzymes are of crucial importance in the processes of generation biofuels from plant biomass. However, no micro-organism considered to be highly effective for the hydrolysis of biomass was yet achieved. This fact is due to the lack of a solid knowledge in regulating of the production of these enzymes. One of the main mechanisms of regulation of protein synthesis occurs by activation of phosphorylation cascades that lead to the activation of transcription factors. Thus, the understanding the signaling pathways activated in certain conditions can provide knowledge enabling the engineering the microorganism to becomes a better producer of lignocellulolytic enzymes, even in conditions with high concentrations of glucose in catabolic repression. Thus, the objectives of this project are: (1) Elucidation of the cell signaling pathways mediated by protein kinase A (PKA), MAPK and calcium, (2) phosphoproteomic identification of potential targets for phosphorylation by MAPK, PKA. (3) study of functional genomics of genes that are part of the signaling pathways by PKA, MAPK and calcium transporter identified in T. reesei. (4) to purify and characterize the secretory vesicles of T. reesei induced by sugarcane bagasse. (5) determine and identify the protein content of extracellular vesicles of T. reesei. At the end of the project we intend to contribute to the greater understanding of the biology of this important industrial fungus, producing mutant strains to increase the production of hollocelulases and lower the cost of production of enzyme cocktails that still constitute a large proportion of biotechnological products obtained from T. reesei, mainly in the production of bioethanol. (AU)