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Comparative analysis of the heterologous protein secretion mechanisms in Aspergillus nidulans

Grant number: 14/23051-2
Support Opportunities:Scholarships in Brazil - Master
Effective date (Start): March 01, 2015
Effective date (End): September 30, 2015
Field of knowledge:Biological Sciences - Microbiology - Applied Microbiology
Acordo de Cooperação: Coordination of Improvement of Higher Education Personnel (CAPES)
Principal Investigator:André Ricardo de Lima Damasio
Grantee:Felipe Calzado
Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovações (Brasil). Campinas , SP, Brazil
Associated research grant:12/20549-4 - Secretion of heterologous glycoproteins in Aspergillus: effect of glycosylation pattern in functional parameters of glycosyl hydrolases, AP.BIOEN.JP


As a way to reduce the use of energy from fossil fuels and increase the use of renewable energy, the second-generation ethanol obtained from the conversion of lignocellulosic plant biomass into fermentable sugars has been the focus of much research nowadays as a way to enable its use. Filamentous fungi stand out in the production and secretion of lignocellulolytic homologous and heterologous enzymes, but little is known about the mechanisms of protein secretion used by these organisms. Thought about it, the use of the model organism Aspergillus nidulans becomes an important resource for studies to be genetically well characterized. The objective of this master's project is to perform a comparative analysis of the mechanisms of secretion of different heterologous proteins in A. nidulans obtaining the transcriptome data generated from RNA-seq technique (Next Generation RNA sequencing) and analyze data using bioinformatics tools. To monitor the secretion process, we will adopt four target genes: GH43 arabinase (AbnA), GH7 cellobiohydrolase (CbhI), GH3 beta-glucosidase (BglC) and thermophilic GH5 mananase (1542.1). These genes were previously cloned and transformed into A. nidulans, for AbnA and CbhI was reached high levels of secretion, unlike BglC and 1542.1 in which the secretion levels were minimal. With the obtained results, our main goal is at least to demonstrate how the strain adapts to heterologous expression of different target genes, including a gene of thermophilic trait. (AU)

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Scientific publications
(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)
CALZADO, FELIPE; PRATES, ERICA T.; GONCALVES, THIAGO A.; RUBIO, MARCELO V.; ZUBIETA, MARIANE P.; SQUINA, FABIO M.; SKAF, MUNIR S.; DAMASIO, ANDRE R. L.. Molecular basis of substrate recognition and specificity revealed in family 12 glycoside hydrolases. Biotechnology and Bioengineering, v. 113, n. 12, p. 2577-2586, . (14/06923-6, 12/20549-4, 13/08293-7, 14/23051-2, 13/24988-5)
RUBIO, MARCELO VENTURA; ZUBIETA, MARIANE PALUDETTI; LOURENO FRANCO CAIRO, JOAO PAULO; CALZADO, FELIPE; PAES LEME, ADRIANA FRANCO; SQUINA, FABIO MARCIO; PRADE, ROLF ALEXANDER; DE LIMA DAMASIO, ANDRE RICARDO. Mapping N-linked glycosylation of carbohydrate-active enzymes in the secretome of Aspergillus nidulans grown on lignocellulose. BIOTECHNOLOGY FOR BIOFUELS, v. 9, . (14/15403-6, 14/06923-6, 11/20977-3, 13/24988-5, 12/20549-4, 14/23051-2, 09/54067-3)
CONTESINI, FABIANO JARES; LIBERATO, MARCELO VIZONA; RUBIO, MARCELO VENTURA; CALZADO, FELIPE; ZUBIETA, MARIANE PALUDETTI; RIANO-PACHON, DIEGO MAURICIO; SQUINA, FABIO MARCIO; BRACHT, FABRICIO; SKAF, MUNIR S.; DAMASIO, ANDRE RICARDO. Structural and functional characterization of a highly secreted alpha-L-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1865, n. 12, p. 1758-1769, . (14/15403-6, 12/20549-4, 13/08293-7, 14/04105-4, 14/23051-2, 08/58037-9, 13/24988-5)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
CALZADO, Felipe. Comparative analysis of the heterologous protein secretion in Aspergillus nidulans. 2017. Master's Dissertation - Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia Campinas, SP.

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