Evaluation of the biotechnological potential of the cellulolytic system of Xanthomonas axonopodis pv. citri: a structural, functional and applied approach

• Grant number: 17/14253-9
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: November 01, 2017
• End date: August 06, 2021
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Mário Tyago Murakami
• Grantee: Ricardo Rodrigues de Melo
• 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

Abstract

Due to the heterogeneity and recalcitrance of lignocellulosic biomass its complete and efficient degradation is a challenge to be overcome aiming the economic viability of some bioprocesses, such as the cellulosic ethanol. One of the main factors that contribute for this is the absence of efficient and low cost enzymatic cocktails. Plant pathogens found in Xanthomonadaceae family, particularly, species Xanthomonas axonopodis pv. citri (XAC), that causes citrus canker, affecting several citrus cultivars, and resulting in significant economic losses to the country's agriculture, contains a large repertoire of genes related to the expression of glycoside hydrolases (GHs). The XAC, although it does not perform massive maceration of the plant cell wall, has in its genome more than 100 GHs genes spread into 45 families, comparable in number to other organisms specialized in the lignocellulosic biomass degradation, such as Ruminococcus albus and Clostridium cellulolyticum. Therefore, the aim of this postdoctoral project is to evaluate the GHs still unexplored XAC involved in cellulose degradation (family GHs 3, 5, 8 and 9). According to CAZy database, of the more than 13,971 sequences predicted to be GHs 5, 8 and 9, only 5.6 % of this total has been biochemically characterized, and even less from a structural view. It is understand that the knowledge of only a few members is not enough to reveal all the functional diversity contained in these families, even more if they exhibit a range of activities and modes of action. In this context, we hope to contribute to the greater mechanistic understanding of modes of action found in these families, using a multidisciplinary approach involving biochemical, enzymatic kinetics, immobilization, and structural analysis (X-ray crystallography, DLS, SAXS and Circular Dichroism). In addition, this project aims at one of the main GH families that are present in the cellulose-degrading commercial cocktails, family 3. We intend to evaluate the role of the PA14 domain present in one GH3 obtained from XAC, an enigmatic domain still poorly characterized in the literature to family 3; in addition to investigating the functional (biochemical and glucose tolerant) and structural differences between GH3 belonging to this plant pathogen.

Scientific publications

(10)

(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)

MIYAMOTO, RENAN YUJI; DE MELO, RICARDO RODRIGUES; DE MESQUITA SAMPAIO, ISABELLE LOBO; DE SOUSA, AMANDA SILVA; MORAIS, EDVALDO RODRIGO; SARGO, CINTIA REGINA; ZANPHORLIN, LETICIA MARIA. Paradigm shift in xylose isomerase usage: a novel scenario with distinct applications. CRITICAL REVIEWS IN BIOTECHNOLOGY, OCT 2021. (19/08855-1, 17/14253-9, 18/02865-2)

DE MELO, RICARDO RODRIGUES; DE LIMA, EVANDRO ANTONIO; PERSINOTI, GABRIELA FELIX; VIEIRA, PLINIO SALMAZO; DE SOUSA, AMANDA SILVA; ZANPHORLIN, LETICIA MARIA; DE GIUSEPPE, PRISCILA OLIVEIRA; RULLER, ROBERTO; MURAKAMI, MARIO TYAGO. Identification of a cold-adapted and metal-stimulated beta-1,4-glucanase with potential use in the extraction of bioactive compounds from plants. International Journal of Biological Macromolecules, v. 166, p. 190-199, JAN 1 2021. (15/26982-0, 17/14253-9)

CONTESSOTO, VINICIUS DE GODOI; RAMOS, FELIPE CARDOSO; DE MELO, RICARDO RODRIGUES; DE OLIVEIRA, VINICIUS MARTINS; SCARPASSA, JOSIANE ANIELE; DE SOUSA, AMANDA SILVA; ZANPHORLIN, LETICIA MARIA; SLADE, GABRIEL GOUVEA; PEREIRA LEITE, VITOR BARBANTI; RULLER, ROBERTO. Electrostatic interaction optimization improves catalytic rates and thermotolerance on xylanases. BIOPHYSICAL JOURNAL, v. 120, n. 11, p. 9-pg., 2021-06-01. (16/19766-1, 16/13998-8, 17/14253-9, 18/11614-3, 14/06862-7, 19/22540-3, 17/09662-7)

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)

CONTESSOTO, VINICIUS DE GODOI; RAMOS, FELIPE CARDOSO; DE MELO, RICARDO RODRIGUES; DE OLIVEIRA, VINICIUS MARTINS; SCARPASSA, JOSIANE ANIELE; DE SOUSA, AMANDA SILVA; ZANPHORLIN, LETICIA MARIA; SLADE, GABRIEL GOUVEA; PEREIRA LEITE, VITOR BARBANTI; RULLER, ROBERTO. lectrostatic interaction optimization improves catalytic rates and thermotolerance on xylanase. BIOPHYSICAL JOURNAL, v. 120, n. 11, p. 2172-2180, JUN 1 2021. (17/09662-7, 17/14253-9, 16/19766-1, 19/22540-3, 18/11614-3, 16/13998-8, 14/06862-7)

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)

LEAL MOTTA, MARIA LORENZA; FERREIRA FILHO, JAIRE ALVES; DE MELO, RICARDO RODRIGUES; ZANPHORLIN, LETICIA MARIA; DOS SANTOS, CLELTON APARECIDO; DE SOUZA, ANETE PEREIRA. A novel fungal metal-dependent alpha-l-arabinofuranosidase of family 54 glycoside hydrolase shows expanded substrate specificity. SCIENTIFIC REPORTS, v. 11, n. 1, MAY 26 2021. (17/14253-9, 15/09202-0, 18/19660-4, 16/19775-0, 19/08855-1)

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)

RODRIGUES DE MELO, RICARDO; CARLOS ALNOCH, ROBSON; DE SOUSA, AMANDA SILVA; SATO, HELIA HARUMI; RULLER, ROBERTO; MATEO, CESAR. Cross-Linking with Polyethylenimine Confers Better Functional Characteristics to an Immobilized beta-glucosidase from Exiguobacterium antarcticum B7. CATALYSTS, v. 9, n. 3, MAR 2019. (17/14253-9)

CONTESINI, FABIANO JARES; DAVANCO, MARCELO GOMES; BORIIN, GUSTAVO PAGOTTO; VANEGAS, KATHERINA GARCIA; GONCALVES CIRINO, JOAO PEDRO; DE MELO, RICARDO RODRIGUES; MORTENSEN, UFFE HASBRO; HILDEN, KRISTIINA; CAMPOS, DANIEL ROSSI; CARVALHO, PATRICIA DE OLIVEIRA. Advances in Recombinant Lipases: Production, Engineering, Immobilization and Application in the Pharmaceutical Industry. CATALYSTS, v. 10, n. 9, p. 33-pg., 2020-09-01. (18/13317-6, 17/14253-9)