Development of techniques for immobilization, purification, and characterization of Trametes hirsuta laccases aiming at biotechnological applications

• Grant number: 22/06275-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: September 01, 2022
• End date: August 31, 2024
• Field of knowledge: Biological Sciences - Biochemistry - Biochemistry of Microorganisms
• Principal Investigator: Maria de Lourdes Teixeira de Moraes Polizeli
• Grantee: Guilherme Guimarães Ortolan
• Host Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 14/50884-5 - INCT 2014: National Institute of Science and Technology of Bioethanol, AP.TEM

Abstract

Mushrooms belonging to the Basidiomycota phylum produce an arsenal of enzymes responsible for degrading organic matter and other synthetic molecules. Some of the enzymes with the highest catalytic efficiency and, therefore, greater interest from researchers' point of view in the biotechnology sector are laccases. These phenol oxidases catalyze the oxidation reaction of aromatic or phenolic derivatives. In addition, they have low substrate specificity, which allows their application in different areas of technology and industry. For example, laccases are essential enzymes in the degradation of lignin, one of the bottlenecks for obtaining second-generation bioethanol. The immobilization process is one option to increase an enzyme's efficiency and catalytic stability. In this sense, in a study currently developed by the proponent, the measurement of several enzyme activities belonging to the lignocellulolytic complex of the fungus Trametes hirsute was carried out, and the laccases presented the best performance among the others. Regarding the positive results for the possible application of this enzyme, this study proposes the innovative objective to optimize the laccase production by T. hirsuta through factorial design methodologies, purify and immobilize these enzymes in ionic adsorption supports, and characterize the free, immobilized and purified enzyme. Furthermore, the laccases will be integrated into enzymatic cocktails already developed in the Laboratory of Microbiology and Cell Biology, LMBC, to improve their hydrolysis performance in different biomass substrates. This proposal is bound to the thematic project "INCT Bioethanol," coordinated by Professor Marcos Buckeridge, Ph.D., approved by FAPESP/CNPq.(AU)