Elucidation of the mechanistic basis of family GH62 present in organisms specialized in the degradation of plant biomass

• Grant number: 13/24622-0
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: May 01, 2014
• End date: March 31, 2017
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal Investigator: Mário Tyago Murakami
• Grantee: Flavio Henrique Moreira de Souza
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil

Abstract

The bioconversion of lignocellulosic materials, particularly agro-industrial residues for ethanol production has increasingly attracted the attention of researchers as a source of renewable energy that can replace fossil fuels , reducing the environmental impact . Initially, many studies have been focused on the understanding and development of technologies for the bioconversion of cellulose. However, recent studies have shown that the economic sustainability of the use of lignocellulosic residues for biofuel production also depends on the development of technologies for bioconversion of hemicellulose, thus increasing the interest and efforts in this area. However, the hemicellulose has a much more complex structure than cellulose, and hence requires a much larger number of enzymes for hydrolysis. ±-L-arabinofuranosidases appear as essential enzymes to the total hydrolysis of hemicelluloses by increasing the release rate of xylose, the carbohydrate most abundant hemicellulose, and arabinose, the second most abundant carbohydrate found in the hemicellulose . Recent studies have shown that supplementation of commercial enzyme cocktails with ± -L - arabinofuranosidases resulted in greater efficiency in the hydrolysis of plant biomass (Delabona et al ., 2013) , highlighting the importance of these enzymes for biotechnologies related to biomass degradation. Thus, this project aims to study three ±-L-arabinofuranosidases belonging to family GH62 from the bacterium Sorangium cellulosum , which is considered an excellent producer of CAZymes (Carbohydrate-Active Enzymes). So far, there is no structural data available for members of GH62 family and therefore the molecular basis for the mechanism of catalysis and specificity remains obscure. Through a multidisciplinary approach involving biochemistry, enzyme kinetics, biophysics, site-directed mutagenesis and X-ray crystallography, we expect to contribute to the better understanding of this family of enzymes, at functional and structural point of view, and therefore the development of new technologies involving the hydrolysis of hemicellulose, which is of great interest to both academic and industrial domains.

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)

ZANPHORLIN, LETICIA MARIA; BUENO DE MORAIS, MARIANA ABRAHAO; DIOGO, JOSE ALBERTO; DOMINGUES, MARIANE NORONHA; MOREIRA DE SOUZA, FLAVIO HENRIQUE; RULLER, ROBERTO; MURAKAMI, MARIO TYAGO. Structure-guided design combined with evolutionary diversity led to the discovery of the xylose-releasing exo-xylanase activity in the glycoside hydrolase family 43. Biotechnology and Bioengineering, v. 116, n. 4, p. 734-744, APR 2019. (16/19995-0, 13/24622-0, 15/26982-0)

DOMINGUES, MARIANE NORONHA; MOREIRA SOUZA, FLAVIO HENRIQUE; VIEIRA, PLINIO SALMAZO; BUENO DE MORAIS, MARIANA ABRAHAO; ZANPHORLIN, LETICIA MARIA; DOS SANTOS, CAMILA RAMOS; SIQUEIRA PIROLLA, RENAN AUGUSTO; HONORATO, RODRIGO VARGAS; LOPES DE OLIVEIRA, PAULO SERGIO; GOZZO, FABIO CESAR; et al. Structural basis of exo--mannanase activity in the GH2 family. Journal of Biological Chemistry, v. 293, n. 35, p. 13636-13649, AUG 31 2018. (16/06509-0, 15/26982-0, 16/19995-0, 13/24622-0)

DE MORAIS, MARIANA ABRAHAO BUENO; POLO, CARLA CRISTINA; DOMINGUES, MARIANE NORONHA; PERSINOTI, GABRIELA FELIX; PIROLLA, RENAN AUGUSTO SIQUEIRA; DE SOUZA, FLAVIO HENRIQUE MOREIRA; CORREA, JESSICA BATISTA DE LIMA; DOS SANTOS, CAMILA RAMOS; MURAKAMI, MARIO TYAGO. Exploring the Molecular Basis for Substrate Affinity and Structural Stability in Bacterial GH39 beta-Xylosidases. FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY, v. 8, MAY 15 2020. (16/19995-0, 13/24622-0, 15/26982-0)