Analysis of structural and functional diversity of GH43 enzymes from Xanthomonas axonopodis pv. citri: biological implications and potential biotechnological applications

• Grant number: 16/19995-0
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Effective date (Start): August 01, 2017
• Effective date (End): June 14, 2020
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal Investigator: Mário Tyago Murakami
• Grantee: Mariana Abrahão Bueno de Morais
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil
• Associated research grant: 15/26982-0 - Exploring novel strategies for depolymerization of plant cell-wall polysaccharides: from structure, function and rational design of glycosyl hydrolases to biological implications and potential biotechnological applications, AP.TEM
• Associated scholarship(s): 18/22138-8 - Unveiling the calcium role on GH43 beta-xylosidases / alpha-L-arabinofuranosidases catalysis by QM/MM molecular dynamics methods, BE.EP.PD

Abstract

Plant cell wall degradative enzymes have attracted great interest of scientists due to their many biological roles and industrial applications such as in biofuels, paper, textiles, food and beverage production processes. However, the depolymerization of plant cell wall by enzymatic routes is neither a fully consolidated process nor economically viable, being necessary the development of new strategies and biochemically more efficient processes. Phytopathogenic organisms, such as fungi and bacteria, have a large arsenal of glycoside hydrolase (GH), enzymes which act on plant cell wall polysaccharides, favoring the infectious process, providing nutrients and mediating plant-pathogen interactions. Therefore, the elucidation of the molecular mechanisms underlying these enzymes is very important not only because of their biotechnological appeal, but also to provide a better understanding of their role in pathogen development as well as in biological events associated with the plant-pathogen interaction. Xanthomonas axonopodis pv. citri (Xac), causative agent of citrus canker, has a large number of GHs, including enzymes from the GH43 family, which are represented by multiple genes, suggesting the existence of functional divergence and distinct biological roles. Besides that, GH43 family is notoriously ubiquitous in plant biomass maceration specialized organisms, acting on the degradation of key polysaccharides that constitute the hemicellulose such as xylan and arabinans. The GH43 family is defined as polyspecific due to its large functional diversity, so this family was latter divided into 37 subfamilies. According to this new classification, several subfamilies remain completely unknown such as the subfamily 9, which is present in Xac's genome. Thus, this project aims to provide a mechanistic understanding of Xac GH43 enzymes using structural, biochemical and cellular approaches that will allow us to infer the roles of these enzymes for the pathogen (either in virulence, signaling, plant-pathogen interaction or nutrition). On the light of these results, we will also be able to estimate the potential biotechnological applications for these enzymes, considering that the GH43 family is still poorly explored and has strong correlation with plant cell wall maceration.

Scientific publications

(12)

(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)

DE SOUSA, AMANDA S.; DE MELO, RICARDO R.; MIYAMOTO, RENAN Y.; MORAIS, MARIANA A. B.; ANDRADE, LILIANE P.; MILAN, NATALIA; DE AVILA, MAYARA C.; DE SOUZA, CLAUDIA M.; ADAO, REGINA C.; SCARPASSA, JOSIANE A.; et al. A rationally identified marine GH1 beta-glucosidase has distinguishing functional features for simultaneous saccharification and fermentation. BIOFUELS BIOPRODUCTS & BIOREFINING-BIOFPR, AUG 2020. (16/19995-0, 17/14253-9, 18/02865-2, 15/26982-0, 16/06509-0, 19/08855-1)

CABRAL, LUCELIA; PERSINOTI, GABRIELA F.; PAIXAO, DOUGLAS A. A.; MARTINS, MARCELE P.; MORAIS, MARIANA A. B.; CHINAGLIA, MARIANA; DOMINGUES, MARIANE N.; SFORCA, MAURICIO L.; PIROLLA, RENAN A. S.; GENEROSO, WESLEY C.; et al. ut microbiome of the largest living rodent harbors unprecedented enzymatic systems to degrade plant polysaccharide. NATURE COMMUNICATIONS, v. 13, n. 1, FEB 2 2022. (15/26982-0, 16/19995-0)

VIEIRA, PLINIO S.; BONFIM, ISABELA M.; ARAUJO, EVANDRO A.; MELO, RICARDO R.; LIMA, AUGUSTO R.; FESSEL, MELISSA R.; PAIXAO, DOUGLAS A. A.; PERSINOTI, GABRIELA F.; ROCCO, SILVANA A.; LIMA, TATIANI B.; et al. Xyloglucan processing machinery in Xanthomonas pathogens and its role in the transcriptional activation of virulence factors. NATURE COMMUNICATIONS, v. 12, n. 1, JUN 30 2021. (17/14253-9, 15/26982-0, 16/06509-0, 15/13684-0, 18/03724-3, 19/13936-0, 17/00203-0, 16/19995-0)

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)

SANTOS, CLELTON A.; MORAIS, MARIANA A. B.; TERRETT, OLIVER M.; LYCZAKOWSKI, JAN J.; ZANPHORLIN, LETICIA M.; FERREIRA, JAIRE A.; TONOLI, CELISA C. C.; MURAKAMI, MARIO T.; DUPREE, PAUL; SOUZA, ANETE P.. An engineered GH1 ss-glucosidase displays enhanced glucose tolerance and increased sugar release from lignocellulosic materials. SCIENTIFIC REPORTS, v. 9, MAR 20 2019. (16/19995-0, 15/26982-0, 17/17390-7, 15/09202-0)

MORAIS, MARIANA ABRAHAO BUENO; PAIVA, JOICE HELENA; MURAKAMI, MARIO TYAGO. Molecular plasticity of CBM3 ancillary domain leads to conformational changes in the cellulose binding interface. Biochemical and Biophysical Research Communications, v. 645, p. 8-pg., 2023-01-19. (08/10555-1, 16/19995-0, 15/26982-0)

MANDELLI, FERNANDA; DE MORAIS, MARIANA ABRAHAO BUENO; DE LIMA, EVANDRO ANTONIO; OLIVEIRA, LEANE; PERSINOTI, GABRIELA FELIX; MURAKAMI, MARIO TYAGO. Spatially remote motifs cooperatively affect substrate preference of a ruminal GH26-type endo-?-1,4-mannanase. Journal of Biological Chemistry, v. 295, n. 15, p. 5012-5021, APR 10 2020. (16/19995-0, 15/26982-0)

MARIUTTI, RICARDO B.; HERNANDEZ-GONZALEZ, JORGE E.; NASCIMENTO, ANDREY F. Z.; DE MORAIS, MARIANA A. B.; MURAKAMI, MARIO T.; CARARETO, CLAUDIA M. A.; ARNI, RAGHUVIR K.. A single P115Q mutation modulates specificity in the Corynebacterium pseudotuberculosis arginine repressor. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, v. 1864, n. 7, JUL 2020. (18/07977-3, 15/13765-0, 16/19995-0, 15/18868-2, 18/10736-8, 16/24587-9)

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)

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)

MORAIS, MARIANA A. B.; COINES, JOAN; DOMINGUES, MARIANE N.; PIROLLA, RENAN A. S.; TONOLI, CELISA C. C.; SANTOS, CAMILA R.; CORREA, JESSICA B. L.; GOZZO, FABIO C.; ROVIRA, CARME; MURAKAMI, MARIO T.. Two distinct catalytic pathways for GH43 xylanolytic enzymes unveiled by X-ray and QM/MM simulations. NATURE COMMUNICATIONS, v. 12, n. 1, JAN 14 2021. (18/22138-8, 16/19995-0, 15/26982-0)

DE SOUSA, AMANDA S.; DE MELO, RICARDO R.; MIYAMOTO, RENAN Y.; MORAIS, MARIANA A. B.; ANDRADE, LILIANE P.; MILAN, NATALIA; DE AVILA, MAYARA C.; DE SOUZA, CLAUDIA M.; ADAO, REGINA C.; SCARPASSA, JOSIANE A.; et al. A rationally identified marine GH1 beta-glucosidase has distinguishing functional features for simultaneous saccharification and fermentation. BIOFUELS BIOPRODUCTS & BIOREFINING-BIOFPR, v. 14, n. 6, p. 17-pg., 2020-08-20. (19/08855-1, 17/14253-9, 18/02865-2, 15/26982-0, 16/06509-0, 16/19995-0)