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Identification of enzymes with potential for biotechnological applications in Trichoderma erinaceum

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Author(s):
Michelle Alexandrino de Assis
Total Authors: 1
Document type: Master's Dissertation
Press: Campinas, SP.
Institution: Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia
Defense date:
Examining board members:
Gonçalo Amarante Guimarães Pereira; André Ricardo de Lima Damásio; Roberto Ruller
Advisor: Gonçalo Amarante Guimarães Pereira
Abstract

Energy generation through the combustion of fossil fuels dominated the world scene in the 20th century. However, such anthropic activity had detrimental effects on the environment, in particular through the emission of gases responsible for global warming which is a cause of great concern these days. As a consequence, it is left to the new technologies to try to find renewable energy sources and one of the most promising alternatives is the development of lignocellulosic biofuels. One of the major obstacles to the industrial-scale viability of this process is related to the enzymatic hydrolysis of the lignocellulosic material, taking into account the high cost of enzymatic cocktails and the monopoly of foreign companies. Most of the enzymes used in the production of enzymatic cocktails are secreted by filamentous fungi and Trichoderma reesei is the most utilized in enzymatic formulations used by bio refineries. In addition to improving well-described fungi, bioprospecting studies seeking to identify new fungal strains producers of enzymes are important for an increase in efficiency and national technologies. The present work used a strain of Trichoderma erinaceum, which was isolated by the company GranBio Celere and had as main objectives: 1) comparative tests of enzymatic activities, hydrolysis and exoproteomics in relation to reference strains (T. reesei WT and T. reesei RUT C30), 2) study of relation between enzymatic secretion of the fungus in question with substrate complexity and time of cultivation, 3) the selection of enzymes that were promising and distinct from those currently described, intending a possible aid in the improvement of the available enzymatic cocktails. The comparison assays demonstrated a greater specific enzymatic activity of T. erinaceum culture supernatant for xylanase, beta-xylosidase, alpha-L-arabinofuranosidase, alpha-galactosidase and beta-glycosidase groups, and when comparing the enzyme identifications, 133 proteins identified in T. erinaceum were exclusive. An adaptation of the enzymatic secretion of T. erinaceum was also performed according to the complexity of the carbon source (glucose, Avicel ®, Straw or Integral Cane Integral pretreated) and culture time (96h, 120h and 144h), in addition to identification of two enzymes related to the phenotype evidenced with a similarity of less than 65% in comparison to those found in T. reesei being these transformed into two different expression systems (AU)

FAPESP's process: 16/02792-0 - Identification of enzymes with potential for biotechnological applications in a Trichoderma sp fungus
Grantee:Michelle Alexandrino de Assis
Support Opportunities: Scholarships in Brazil - Master