Characterization of calcium-dependent signaling pathway in the secretion and cellulase expression by Trichoderma reesei (Hypocrea jecorina

• Grant number: 16/23233-9
• Support Opportunities: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): March 01, 2017
• Effective date (End): May 21, 2019
• Field of knowledge: Biological Sciences - Microbiology - Biology and Physiology of Microorganisms
• Acordo de Cooperação: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Roberto do Nascimento Silva
• Grantee: Renato Graciano de Paula
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

Trichoderma reesei is a saprophytic fungus involved with polysaccharides cell wall depolymerization. This fungus has an impressive capacity in produce and secretes cellulases, being the most important industrial source of these enzymes, which has been used for bioethanol production. The degradation of the lignocellulosic wall component is a complex process which requires a number of regulatory proteins to make it more efficient. This project aims to contribute to the understanding of the molecular mechanisms involved in the process of deconstruction of plant biomass by T. reesei through the characterization of calcium transporters related to calcium-dependent intracellular signaling pathway, which are differentially expressed by cellulose, sophorose and sugarcane bagasse induction. Initially, nine calcium transporters involved in intracellular signaling calcium, identified previously by our group through in silico analysis of RNA-Seq data were selected according to the level of gene expression in QM4914 during cultivation cellulose and sophorose and QM6a in sugarcane bagasse. Our study is focused on functional genomics for characterization of T. reesei carriers calcium and its role in the regulation of transcription and secretion of cellulases by: 1) Construction of mutant strains for each gene, 2) Determination of the cellulolytic activity and gene expression of cellulases and signaling pathway genes in the mutants obtained, 3 ) phenotypic characterization of mutant strains for calcium transporters 4) Assay of tolerance to calcium using different concentrations of calcium, 5) Intracellular calcium measurement, 6) Construction retransformantes strains for target genes, 7) Determination of the intracellular location of calcium carriers by fluorescence microscopy, 8) Purification and characterization of extracellular vesicles of T. reesei in cellulose, sophorose and sugarcane bagasse induction and 9) Proteomic analysis of proteic content of extracellular vesicles of T. reesei. The results obtained from the functional characterization of these transporters will permit the construction of a model relating their role in calcium-dependent signaling pathway as well as understanding the involvement of the calcium signaling pathway in regulating the expression of cellulase genes by the fungus T. reesei. (AU)

Scientific publications

(7)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

LOPES, DOUGLAS CHRISTIAN BORGES; CARRARO, CLAUDIA BATISTA; SILVA, ROBERTO NASCIMENTO; DE PAULA, RENATO GRACIANO. Molecular Characterization of Xyloglucanase cel74a from Trichoderma reesei. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v. 22, n. 9, MAY 2021. (16/23233-9, 16/09708-4)

CAMPOS ANTONIETO, AMANDA CRISTINA; VIEIRA NOGUEIRA, KAROLINE MARIA; DE PAULA, RENATO GRACIANO; NORA, LUFSA CZAMANSKI; ANZOLINI CASSIANO, MURILO HENRIQUE; GUAZZARONI, MARIA-EUGENIA; ALMEIDA, FAUSTO; DA SILVA, THIAGO APARECIDO; RIES, LAURE NICOLAS ANNICK; DE ASSIS, LEANDRO JOSE; et al. A Novel Cys2His2 Zinc Finger Homolog of AZF1 Modulates Holocellulase Expression in Trichoderma reesei. MSYSTEMS, v. 4, n. 4, JUL-AUG 2019. (16/23233-9, 16/03322-7, 15/04309-1, 17/04217-5, 16/03763-3, 18/03766-8, 15/09553-8, 12/22921-8)

RIBEIRO, MARCELA SURIANI; DE PAULA, RENATO GRACIANO; VOLTAN, ALINE RAQUEL; DE CASTRO, RAPHAELA GEORG; CARRARO, CLAUDIA BATISTA; DE ASSIS, LEANDRO JOSE; STEINDORFF, ANDREI STECCA; GOLDMAN, GUSTAVO HENRIQUE; SILVA, ROBERTO NASCIMENTO; ULHOA, CIRANO JOSE; et al. Endo-beta-1,3-glucanase (GH16 Family) from Trichoderma harzianum Participates in Cell Wall Biogenesis but Is Not Essential for Antagonism Against Plant Pathogens. BIOMOLECULES, v. 9, n. 12, DEC 2019. (16/23233-9, 17/04206-3)

DE PAULA, RENATO GRACIANO; CAMPOS ANTONIETO, AMANDA CRISTINA; CARRARO, CLAUDIA BATISTA; BORGES LOPES, DOUGLAS CHRISTIAN; PERSINOTI, GABRIELA FELIX; AGUIAR PERES, NALU TEIXEIRA; MARTINEZ-ROSSI, NILCE MARIA; SILVA-ROCHA, RAFAEL; SILVA, ROBERTO NASCIMENTO. The Duality of the MAPK Signaling Pathway in the Control of Metabolic Processes and Cellulase Production in Trichoderma reesei. SCIENTIFIC REPORTS, v. 8, OCT 8 2018. (17/04206-3, 16/23233-9, 14/23653-2)

DE PAULA, RENATO GRACIANO; CAMPOS ANTONIETO, AMANDA CRISTINA; COSTA RIBEIRO, LILIANE FRAGA; SRIVASTAVA, NEHA; O'DONOVAN, ANTHONIA; MISHRA, P. K.; GUPTA, VIJAI K.; SILVA, ROBERTO N.. Engineered microbial host selection for value-added bioproducts from lignocellulose. BIOTECHNOLOGY ADVANCES, v. 37, n. 6, NOV 1 2019. (16/23233-9, 17/04206-3)

DE PAULA, RENATO GRACIANO; CAMPOS ANTONIETO, AMANDA CRISTINA; VIEIRA NOGUEIRA, KAROLINE MARIA; COSTA RIBEIRO, LILIANE FRAGA; ROCHA, MARINA CAMPOS; MALAVAZI, IRAN; ALMEIDA, FAUSTO; SILVA, ROBERTO NASCIMENTO. Extracellular vesicles carry cellulases in the industrial fungus Trichoderma reesei. BIOTECHNOLOGY FOR BIOFUELS, v. 12, JUN 15 2019. (17/04206-3, 16/23233-9)

DE PAULA, RENATO GRACIANO; CAMPOS ANTONIETO, AMANDA CRISTINA; COSTA RIBEIRO, LILIANE FRAGA; CARRARO, CLAUDIA BATISTA; VIEIRA NOGUEIRA, KAROLINE MARIA; BORGES LOPES, DOUGLAS CHRISTIAN; SILVA, ALINNE COSTA; ZERBINI, MARIANA TAISE; PEDERSOLI, WELLINGTON RAMOS; COSTA, MARIANA DO NASCIMENTO; et al. New Genomic Approaches to Enhance Biomass Degradation by the Industrial Fungus Trichoderma reesei. INTERNATIONAL JOURNAL OF GENOMICS, 2018. (16/23233-9, 17/04206-3, 16/20358-5)