Aspergillus nidulans as a model for heterologous expression of cellulases and hemicellulases

• Grant number: 11/02169-7
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: August 01, 2011
• End date: October 31, 2013
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Principal Investigator: Fábio Márcio Squina
• Grantee: André Ricardo de Lima Damasio
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil
• Associated research grant: 08/58037-9 - Library generation for biomass-conversion enzymes from soil metagenome, AP.BIOEN.JP

Abstract

Placed within the field of renewable energy, the enzymatic pre-treatment of biomass (the enzymatic degradation of plant wall polysaccharide) is becoming a more attractive alternative than the use of chemicals pre-treatment or mechanical processes. Furthermore, the biodegradation of cellulose by cellulases and cellulosomes produced by numerous microorganisms, represents the largest carbon flow from carbon fixed for atmospheric CO2. Likewise, this process is very important for various processes for agro-industrial waste that can be used for bioenergy to replace fossil fuels. Enzymatic hydrolysis of cellulose involves the synergistic action of at least three enzymes, endoglucanase, exoglucanase or cellobiohydrolase, and ²-glucosidase. One of the groups most abundant polysaccharide in nature, hemicellulose are classified according to the sugar present in its molecule, and the basic structure is composed by: xylan, galacto-glucomannan, ±-1,5-L-arabinan and arabinogalactan. Xylan is a polymer of xylose, but in nature, is usually associated with other sugars forming glucuronoxylan, glucuroarabinoxylan, glucomannan, arabinogalactans, and galactoglucomannan. In this context, hemicellulases are a diverse group of enzymes that hydrolyze hemicelluloses. These enzymes have many biotechnological applications and their link structure/ function, is a subject of intense research. The attraction of filamentous fungi as expression machinery is based on your natural ability to secrete large amounts of protein (mainly hydrolytic enzymes) to the medium. Compared with other systems available for expression of heterologous proteins in liquid cultures, filamentous fungi show great performance, high yield and an inexpensive option with respect to nutritional requirements. The main focus of this proposal is to study and consolidate Aspergillus nidulans (strain A773) as a model for rapid and stable heterologous expression and secretion of cellulases and hemicellulases. Our main objective is to develop a collection of recombinant strains which overexpress the different enzyme activities involved in the degradation of plant cell wall polysaccharides. In addition to study the functional properties of these enzymes, with broad biotechnological applications that include the production of bioethanol, in this study we intend to develop strains that over-express two or more synergistic cellulolytic and hemicellulolycic activities.

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)

DOS REIS ALMEIDA, FAUSTO BRUNO; PIGOSSO, LAURINE LACERDA; DE LIMA DAMASIO, ANDRE RICARDO; MONTEIRO, VALDIRENE NEVES; DE ALMEIDA SOARES, CELIA MARIA; SILVA, ROBERTO NASCIMENTO; ROQUE-BARREIRA, MARIA CRISTINA. alpha-(1,4)-Amylase, but not alpha- and beta-(1,3)-glucanases, may be responsible for the impaired growth and morphogenesis of Paracoccidioides brasiliensis induced by N-glycosylation inhibition. YEAST, v. 31, n. 1, p. 1-11, JAN 2014. (11/02169-7, 06/60642-2, 09/51197-3)

DAMASIO, ANDRE R. L.; PINTO BRAGA, CLEITON MARCIO; BRENELLI, LIVIA B.; CITADINI, ANA PAULA; MANDELLI, FERNANDA; COTA, JUNIO; DE ALMEIDA, RODRIGO FERREIRA; SALVADOR, VICTOR HUGO; ALVAREDO PAIXAO, DOUGLAS ANTONIO; SEGATO, FERNANDO; et al. Biomass-to-bio-products application of feruloyl esterase from Aspergillus clavatus. Applied Microbiology and Biotechnology, v. 97, n. 15, p. 6759-6767, AUG 2013. (11/02169-7, 09/17956-4, 08/58037-9)

COTA, JUNIO; OLIVEIRA, LEANDRO C.; DAMASIO, ANDRE R. L.; CITADINI, ANA P.; HOFFMAM, ZAIRA B.; ALVAREZ, THABATA M.; CODIMA, CARLA A.; LEITE, VITOR B. P.; PASTORE, GLAUCIA; DE OLIVEIRA-NETO, MARIO; et al. Assembling a xylanase-lichenase chimera through all-atom molecular dynamics simulations. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1834, n. 8, p. 1492-1500, AUG 2013. (11/13242-7, 11/02169-7, 08/58037-9)

DE LUCAS, ROSYMAR COUTINHO; DE OLIVEIRA, TASSIO BRITO; LIMA, MATHEUS SANITA; PASIN, THIAGO MACHADO; DE ALMEIDA SCARCELLA, ANA SILVIA; COSTA RIBEIRO, LILIANE FRAGA; CARVALHO, CAIO; DE LIMA DAMASIO, ANDRE RICARDO; BUCKERIDGE, MARCOS SILVEIRA; PRADE, ROLF ALEXANDER; et al. The profile secretion of Aspergillus clavatus: Different pre-treatments of sugarcane bagasse distinctly induces holocellulases for the lignocellulosic biomass conversion into sugar. RENEWABLE ENERGY, v. 165, n. 1, p. 748-757, MAR 2021. (17/09000-4, 11/02169-7)

DE LIMA DAMASIO, ANDRE RICARDO; RUBIO, MARCELO VENTURA; GONCALVES, THIAGO AUGUSTO; PERSINOTI, GABRIELA FELIX; SEGATO, FERNANDO; PRADE, ROLF ALEXANDER; CONTESINI, FABIANO JARES; DE SOUZA, AMANDA PEREIRA; BUCKERIDGE, MARCOS SILVEIRA; SQUINA, FABIO MARCIO. Xyloglucan breakdown by endo-xyloglucanase family 74 from Aspergillus fumigatus. Applied Microbiology and Biotechnology, v. 101, n. 7, p. 2893-2903, APR 2017. (11/02169-7, 08/58037-9, 12/20549-4, 14/18714-2)

DE LUCAS, ROSYMAR COUTINHO; DE OLIVEIRA, TASSIO BRITO; LIMA, MATHEUS SANITA; PASIN, THIAGO MACHADO; DE ALMEIDA SCARCELLA, ANA SILVIA; COSTA RIBEIRO, LILIANE FRAGA; CARVALHO, CAIO; DE LIMA DAMASIO, ANDRE RICARDO; BUCKERIDGE, MARCOS SILVEIRA; PRADE, ROLF ALEXANDER; et al. The profile secretion of Aspergillus clavatus: Different pre-treatments of sugarcane bagasse distinctly induces holocellulases for the lignocellulosic biomass conversion into sugar. RENEWABLE ENERGY, v. 165, p. 10-pg., 2021-03-01. (17/09000-4, 11/02169-7)

GONCALVES, T. A.; DAMASIO, A. R. L.; SEGATO, F.; ALVAREZ, T. M.; BRAGATTO, J.; BRENELLI, L. B.; CITADINI, A. P. S.; MURAKAMI, M. T.; RULLER, R.; PAES LEME, A. F.; et al. Functional characterization and synergic action of fungal xylanase and arabinofuranosidase for production of xylooligosaccharides. Bioresource Technology, v. 119, p. 293-299, SEP 2012. (11/02169-7, 10/18198-3, 10/11469-1)