Studies of the structural and functional behavior of enzymes evolutionarily specialized in the degradation of plant biomass with potential biotechnological applications

• Grant number: 13/13309-0
• Support Opportunities: Regular Research Grants
• Start date: October 01, 2013
• End date: March 31, 2016
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Mário Tyago Murakami
• Grantee: Mário Tyago Murakami
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil
• Associated researchers: Camila Ramos dos Santos
• Associated research grant(s): 14/07135-1 - Multi-user equipament approved in grant 2013/13309-0: Beckman Coulter capillary electrophoresis systems with LIF detection system, AP.EMU

Abstract

The glycosyl hydrolases and feruloyl esterases have a range of industrial applications, and more recently, they have attracted great interest due to its importance for the use of lignocellulosic biomass, the largest source of renewable resource on earth, to produce biofuels. However, the high costs involving enzymatic routes have limited the economic production of cellulosic ethanol, mainly due to lower enzymatic performance, inhibition by sub(products) and reduced stability of enzymes under typical conditions in the industry. Thus, the search for new enzymes evolutionarily sculpted for the degradation of plant biomass and in-depth knowledge of the molecular basis of their modes of action, selectivity and stability represent an essential step for the development of more robust and efficient biocatalysts. In this project, we have targeted novel enzymes with great potential for the "digestion" of lignocellulosic material derived from microbial sources related to the bovine rumen and pathogenicity in plants. The data generated will assist the development of new biotechnological applications and the rational design of enzymes in order to tune extremophilicity (pH and temperature), action modes and catalytic performance. Furthermore, these results will be of paramount importance for a better understanding of the biology of the superfamily of glycoside hydrolases (GHs), with more than 132 families described, participating in numerous biological events such as plant development, cell wall remodeling , cellular homeostasis, among others. (AU)

Scientific publications

(14)

(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)

DIOGO, JOSE ALBERTO; ZANPHORLIN, LETICIA MARIA; SATO, HELIA HARUMI; MURAKAMI, MARIO TYAGO; RULLER, ROBERTO. Molecular cloning, overexpression, purification and crystallographic analysis of a GH43-xylosidase from Bacillus licheniformis. ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, v. 71, n. 8, p. 962-965, AUG 2015. (13/13309-0, 13/10443-7, 14/07135-1)

ALVAREZ, THABATA M.; PAIVA, JOICE H.; RUIZ, DIEGO M.; CAIRO, JOAO PAULO L. F.; PEREIRA, ISABELA O.; PAIXAO, DOUGLAS A. A.; DE ALMEIDA, RODRIGO F.; TONOLI, CELISA C. C.; RULLER, ROBERTO; SANTOS, CAMILA R.; et al. Structure and Function of a Novel Cellulase 5 from Sugarcane Soil Metagenome. PLoS One, v. 8, n. 12, DEC 17 2013. (13/13309-0, 11/20977-3, 10/11469-1, 09/08312-6, 08/58037-9)

FURTADO, GILVAN P.; SANTOS, CAMILA R.; CORDEIRO, ROSA L.; RIBEIRO, LUCAS F.; DE MORAES, LUIZ A. B.; DAMASIO, ANDR R. L.; POLIZELI, MARIA DE LOURDES T. M.; LOURENZONI, MARCOS R.; MURAKAMI, MARIO T.; WARD, RICHARD J.. Enhanced xyloglucan-specific endo-beta-1,4-glucanase efficiency in an engineered CBM44-XegA chimera. Applied Microbiology and Biotechnology, v. 99, n. 12, p. 5095-5107, JUN 2015. (10/18850-2, 12/20549-4, 13/18910-3, 13/13309-0, 14/07135-1, 10/07133-8)

DOS SANTOS, CAMILA RAMOS; CORDEIRO, ROSA LORIZOLLA; WONG, DOMINIC W. S.; MURAKAMI, MARIO TYAGO. Structural Basis for Xyloglucan Specificity and alpha-D-Xylp(1 -> 6)-D-Glcp Recognition at the-1 Subsite within the GH5 Family. BIOCHEMISTRY, v. 54, n. 10, p. 1930-1942, MAR 17 2015. (13/13309-0, 10/51890-8)

DE GIUSEPPE, PRISCILA OLIVEIRA; CAMPOS BRASIL SOUZA, TATIANA DE ARRUDA; MOREIRA SOUZA, FLAVIO HENRIQUE; ZANPHORLIN, LETICIA MARIA; MACHADO, CARLA BOTELHO; WARD, RICHARD JOHN; JORGE, JOAO ATILIO; MELO FURRIEL, ROSA DOS PRAZERES; MURAKAMI, MARIO TYAGO. Structural basis for glucose tolerance in GH1 beta-glucosidases. ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, v. 70, p. 9-pg., 2014-06-01. (09/08312-6, 13/13309-0)

DE GIUSEPPE, PRISCILA OLIVEIRA; CAMPOS BRASIL SOUZA, TATIANA DE ARRUDA; MOREIRA SOUZA, FLAVIO HENRIQUE; ZANPHORLIN, LETICIA MARIA; MACHADO, CARLA BOTELHO; WARD, RICHARD JOHN; JORGE, JOAO ATILIO; MELO FURRIEL, ROSA DOS PRAZERES; MURAKAMI, MARIO TYAGO. Structural basis for glucose tolerance in GH1 beta-glucosidases. ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, v. 70, n. 6, p. 1631-1639, JUN 2014. (13/13309-0, 09/08312-6)

DOS SANTOS, CAMILA RAMOS; DE GIUSEPPE, PRISCILA OLIVEIRA; MOREIRA DE SOUZA, FLAVIO HENRIQUE; ZANPHORLIN, LETICIA MARIA; DOMINGUES, MARIANE NORONHA; SIQUEIRA PIROLLA, RENAN AUGUSTO; HONORATO, RODRIGO VARGAS; COSTA TONOLI, CELISA CALDANA; BUENO DE MORAIS, MARIANA ABRAHAO; DE MATOS MARTINS, VANESA PEIXOTO; et al. The mechanism by which a distinguishing arabinofuranosidase can cope with internal di-substitutions in arabinoxylans. BIOTECHNOLOGY FOR BIOFUELS, v. 11, AUG 11 2018. (13/13309-0, 14/17264-3, 15/26982-0, 14/07135-1)

CRESPIM, ELAINE; ZANPHORLIN, LETICIA M.; DE SOUZA, FLAVIO H. M.; DIOGO, JOSE A.; GAZOLLA, ALEX C.; MACHADO, CARLA B.; FIGUEIREDO, FERNANDA; SOUSA, AMANDA S.; NOBREGA, FELIPE; PELLIZARI, VIVIAN H.; et al. A novel cold-adapted and glucose-tolerant GH1 beta-glucosidase from Exiguobacterium antarcticum B7. International Journal of Biological Macromolecules, v. 82, p. 375-380, JAN 2016. (13/13309-0, 14/07135-1)

SANTOS, CLELTON A.; ZANPHORLIN, LET-CIA M.; CRUCELLO, ALINE; TONOLI, CELISA C. C.; RULLER, ROBERTO; HORTA, MARIA A. C.; MURAKAMI, MARIO T.; DE SOUZA, ANETE PEREIRA. Crystal structure and biochemical characterization of the recombinant ThBgl, a GH1 beta-glucosidase overexpressed in Trichoderma harzianum under biomass degradation conditions. BIOTECHNOLOGY FOR BIOFUELS, v. 9, MAR 22 2016. (14/09720-9, 08/52197-4, 14/18856-1, 13/13309-0)

ZANPHORLIN, LETICIA MARIA; DE GIUSEPPE, PRISCILA OLIVEIRA; HONORATO, RODRIGO VARGAS; COSTA TONOLI, CELISA CALDANA; FATTORI, JULIANA; CRESPIM, ELAINE; LOPES DE OLIVEIRA, PAULO SERGIO; RULLER, ROBERTO; MURAKAMI, MARIO TYAGO. Oligomerization as a strategy for cold adaptation: Structure and dynamics of the GH1 beta-glucosidase from Exiguobacterium antarcticum B7. SCIENTIFIC REPORTS, v. 6, MAR 31 2016. (13/13309-0, 14/07135-1)

RUIZ, DIEGO M.; TUROWSKI, VALERIA R.; MURAKAMI, MARIO T.. Effects of the linker region on the structure and function of modular GH5 cellulases. SCIENTIFIC REPORTS, v. 6, JUN 23 2016. (13/13309-0, 13/01583-0)

SANTOS, CAMILA RAMOS; HOFFMAM, ZAIRA BRUNA; DE MATOS MARTINS, VANESA PEIXOTO; ZANPHORLIN, LETICIA MARIA; DE PAULA ASSIS, LEANDRO HENRIQUE; HONORATO, RODRIGO VARGAS; LOPES DE OLIVEIRA, PAULO SERGIO; RULLER, ROBERTO; MURAKAMI, MARIO TYAGO. Molecular Mechanisms Associated with Xylan Degradation by Xanthomonas Plant Pathogens. Journal of Biological Chemistry, v. 289, n. 46, p. 32186-32200, NOV 14 2014. (13/13309-0, 10/51890-8, 14/07135-1)

SANTOS, CAMILA R.; POLO, CARLA C.; COSTA, MARIA C. M. F.; NASCIMENTO, ANDREY F. Z.; MEZA, ANDREIA N.; COTA, JUNIO; HOFFMAM, ZAIRA B.; HONORATO, RODRIGO V.; OLIVEIRA, PAULO S. L.; GOLDMAN, GUSTAVO H.; et al. Mechanistic Strategies for Catalysis Adopted by Evolutionary Distinct Family 43 Arabinanases. Journal of Biological Chemistry, v. 289, n. 11, p. 7362-7373, MAR 14 2014. (13/13309-0, 10/51890-8)

DIOGO, JOSE ALBERTO; ZANPHORLIN, LETICIA MARIA; SATO, HELIA HARUMI; MURAKAMI, MARIO TYAGO; RULLER, ROBERTO. Molecular cloning, overexpression, purification and crystallographic analysis of a GH43-xylosidase from Bacillus licheniformis. ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, v. 71, p. 4-pg., 2015-08-01. (13/10443-7, 13/13309-0, 14/07135-1)