Studies of the mode of action of two lytic polysaccharide monooxygenases from insect (order: isoptera): molecular structure, bioinorganic chemistry and biotechnological applications

• Grant number: 17/17275-3
• Support Opportunities: Regular Research Grants
• Start date: December 01, 2017
• End date: May 31, 2020
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal Investigator: Wanius José Garcia da Silva
• Grantee: Wanius José Garcia da Silva
• Host Institution: Centro de Ciências Naturais e Humanas (CCNH). Universidade Federal do ABC (UFABC). Ministério da Educação (Brasil). Santo André , SP, Brazil

Abstract

The lytic polysaccharides monooxygenases (LPMOs) constitute a new class of metalloenzymes copper-dependent involved in the biodegradation of recalcitrant polysaccharides, such as cellulose, via oxidative mechanisms. These metalloenzymes have been identified and characterized only in the last ten years and have attracted the interest of researchers due to its great potential of biotechnological applications, mainly in the field of conversion of structural polysaccharides in bioenergy and bioproducts. LPMOs can increase the efficiency of a commercial lignocellulolytic cocktail by up to 40%. In this context, these metalloenzymes have become a promising key component with the potential to finally to make enzymatic biomass conversion economically attractive. In this research project, we propose to study the mode of action of two LPMOs from insect (order isoptera). The two recombinant metalloenzymes will be produced in E. coli and purified by chromatographic methods. The enzymatic activities of the purified enzymes will be studied varying the substrates, physicochemical conditions and nature of the metal ions. The thermodynamics of interaction with metallic ions will be studied by spectroscopic methods and isothermal titration calorimetry. The structures of the enzymes will be studied by spectroscopic methods, small-angle X-ray scattering and protein crystallography. The results obtained from this project will allow a better understanding of the mode of action of LPMOs from insect and the role of metal ions in the catalytic mechanism of these enzymes. In addition, such metalloenzymes may be employed as supplements in commercial cellulolytic cocktails. (AU)

Scientific publications

(17)

(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)

FERNANDES, DIOGENES G. DA S.; ANDRADE, VIVIANE BRITO; LUCENA, LETICIA NEVES; AMBROSIO, FELIPE NOGUEIRA; DE SOUZA, ANDRE L. M.; BATISTA, BRUNO L.; ROLIM, WALLACE ROSADO; SEABRA, AMEDEA BAROZZI; LOMBELLO, CHRISTIANE B.; DA SILVA, FERNANDA DIAS; et al. ytotoxicity and Antimicrobial Properties of Photosynthesized Silver Chloride Nanoparticles Using Plant Extract from Stryphnodendron adstringens (Martius) Covill. JOURNAL OF CLUSTER SCIENCE, v. 33, n. 2, FEB 2021. (18/08194-2, 17/17275-3)

FRANCO CAIRO, JOAO PAULO L.; CANNELLA, DAVID; OLIVEIRA, LEANDRO C.; GONCALVES, THIAGO A.; RUBIO, MARCELO V.; TERRASAN, CESAR R. F.; TRAMONTINA, ROBSON; MOFATTO, LUCIANA S.; CARAZZOLLE, MARCELO F.; GARCIA, WANIUS; et al. On the roles of AA15 lytic polysaccharide monooxygenases derived from the termite Coptotermes gestroi. Journal of Inorganic Biochemistry, v. 216, MAR 2021. (17/17275-3, 16/09950-0, 15/50590-4, 17/11952-3, 15/50612-8, 17/22669-0, 17/16089-1)

ARAUJO, JUSCEMACIA N.; TOFANELLO, ARYANE; SATO, JULIANA A. P.; CRUZ, LUCIANO S.; NANTES-CARDOSO, ISELI L.; FERREIRA, FABIO F.; BATISTA, BRUNO L.; GARCIA, WANIUS. Rapid Synthesis via Green Route of Plasmonic Protein-Coated Silver/Silver Chloride Nanoparticles with Controlled Contents of Metallic Silver and Application for Dye Remediation. Journal of Inorganic and Organometallic Polymers and Materials, v. 28, n. 6, p. 2812-2818, NOV 2018. (17/17275-3, 17/16976-8, 14/03682-8, 15/02897-3)

TOFANELLO, ARYANE; ARAUJO, JUSCEMACIA N.; NANTES-CARDOSO, ISELI L.; FERREIRA, FABIO F.; SOUZA, JOSE A.; LIM, DAE-WOON; KITAGAWA, HIROSHI; GARCIA, WANIUS. Ultrafast fabrication of thermally stable protein-coated silver iodide nanoparticles for solid-state superionic conductors. COLLOIDS AND SURFACES B-BIOINTERFACES, v. 176, p. 47-54, APR 1 2019. (17/16976-8, 17/17275-3)

DE LIMA, MARIANA Z. T.; DE ALMEIDA, LEONARDO R.; MERA, ALAIN M.; BERNARDES, AMANDA; GARCIA, WANIUS; MUNIZ, JOAO R. C.. Crystal Structure of a Sucrose-6-phosphate Hydrolase from Lactobacillus gasseri with Potential Applications in Fructan Production and the Food Industry. Journal of Agricultural and Food Chemistry, v. 69, n. 35, p. 10223-10234, SEP 8 2021. (16/09152-6, 14/50897-0, 17/17275-3, 17/16291-5)

DA SILVA, VIVIAM M.; CABRAL, ALINE D.; SPERANCA, MARCIA A.; SQUINA, FABIO M.; MUNIZ, JOAO RENATO C.; MARTIN, LYDIE; NICOLET, YVAIN; GARCIA, WANIUS. High-resolution structure of a modular hyperthermostable endo-beta-1,4-mannanase from Thermotoga petrophila: The ancillary immunoglobulin-like module is a thermostabilizing domain. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1868, n. 8, AUG 2020. (13/26096-4, 16/15836-5, 17/17275-3, 17/16291-5, 14/02065-5, 16/14514-4, 14/50897-0, 15/50590-4)

BOMEDIANO CAMILLO, LIVIA DE MORAES; FERREIRA, GRAZIELE CRISTINA; ALVES DURAN, ADRIANA FELICIANO; SANTOS DA SILVA, FLAVIA RIBEIRO; GARCIA, WANIUS; SCOTT, ANA LIGIA; SASAKI, SERGIO DAISHI. Structural modelling and thermostability of a serine protease inhibitor belonging to the Kunitz-BPTI family from the Rhipicephalus microplus tick. Biochimie, v. 181, p. 226-233, FEB 2021. (11/07001-7, 17/19077-4, 18/11874-5, 17/17275-3)

DOS SANTOS, OTAVIO AUGUSTO LEITAO; GONCALVES, THIAGO AUGUSTO; SODRE, VICTORIA; VILELA, NATHALIA; TOMAZETTO, GEIZECLER; SQUINA, FABIO M.; GARCIA, WANIUS. Recombinant expression, purification and characterization of an active bacterial feruloyl-CoA synthase with potential for application in vanillin production. Protein Expression and Purification, v. 197, p. 7-pg., 2022-05-11. (17/16089-1, 15/50590-4, 18/18101-1, 17/08166-6, 20/15595-3, 20/05784-3, 17/17275-3)

SANTOS, VALDENICE F.; ARAUJO, ANA C. J.; FREITAS, PRISCILLA R.; SILVA, ANA L. P.; SANTOS, ANA L. E.; DA ROCHA, BRUNO ANDERSON MATIAS; SILVA, ROMERIO R. S.; ALMEIDA, DNANE V.; GARCIA, WANIUS; COUTINHO, HENRIQUE D. M.; et al. Enhanced antibacterial activity of the gentamicin against multidrug-resistant strains when complexed with Canavalia ensiformis lectin. Microbial Pathogenesis, v. 152, MAR 2021. (17/17275-3, 18/13500-5)

DE ALCANTARA, NAYRA RODRIGUES; DE OLIVEIRA, FABIO MUNIZ; GARCIA, WANIUS; LEITAO DOS SANTOS, OTAVIO AUGUSTO; JUNQUEIRA-KIPNIS, ANA PAULA; KIPNIS, ANDRE. Dps protein is related to resistance of Mycobacterium abscessus subsp. massiliense against stressful conditions. Applied Microbiology and Biotechnology, v. 104, n. 11, APR 2020. (17/17275-3)

GARCIA, FELIPE BAENA; CABRAL, ALINE DINIZ; FUHLENDORF, MAX MARIO; DA CRUZ, GEOMAR FEITOSA; DOS SANTOS, JULIETE VITORINO; FERREIRA, GRAZIELE CRISTINA; CARNEIRO DE REZENDE, BERNARD ROBIN; SANTANA, CARLA MOREIRA; PUZER, LUCIANO; SASAKI, SERGIO DAISHI; et al. Functional and structural characterization of an ecotin-like serine protease inhibitor from Trypanosoma cruzi. International Journal of Biological Macromolecules, v. 151, p. 459-466, MAY 15 2020. (13/26096-4, 17/17275-3, 16/14514-4)

MERA, ALAIN; THEODORO DE LIMA, MARIANA ZULIANI; BERNARDES, AMANDA; GARCIA, WANIUS; CARVALHO MUNIZ, JOAO RENATO. Low-resolution structure, oligomerization and its role on the enzymatic activity of a sucrose-6-phosphate hydrolase from Bacillus licheniformis. Amino Acids, v. 51, n. 4, p. 599-610, APR 2019. (17/17275-3, 17/16291-5)

GONCALVES, THIAGO AUGUSTO; SODRE, VICTORIA; DA SILVA, STEPHANIE NEMESIO; VILELA, NATHALIA; TOMAZETTO, GEIZECLER; ARAUJO, JUSCEMACIA NASCIMENTO; MUNIZ, JOAO RENATO C.; FILL, TAICIA PACHECO; DAMASIO, ANDRE; GARCIA, WANIUS; et al. Applying biochemical and structural characterization of hydroxycinnamate catabolic enzymes from soil metagenome for lignin valorization strategies. Applied Microbiology and Biotechnology, v. 106, n. 7, p. 14-pg., 2022-03-30. (17/16976-8, 20/05784-3, 18/18101-1, 17/17275-3, 17/22669-0, 15/50612-8, 15/50590-4, 15/23279-6)

CRUZ, GEOMAR F.; TOFANELLO, ARYANE; ARAUJO, JUSCEMACIA N.; NANTES-CARDOSO, ISELI L.; FERREIRA, FABIO F.; GARCIA, WANIUS. Fast One-Pot Photosynthesis of Plasmonic Protein-Coated Silver/Silver Bromide Nanoparticles with Efficient Photocatalytic Performance. Journal of Inorganic and Organometallic Polymers and Materials, v. 28, n. 5, p. 2056-2062, SEP 2018. (15/02897-3, 17/16976-8, 17/17275-3, 15/17688-0)

SODRE, VICTORIA; ARAUJO, JUSCEMACIA NASCIMENTO; GONCALVES, THIAGO AUGUSTO; VILELA, NATHALIA; KIMUS BRAZ, ANTONIO SERGIO; FRANCO, TELMA TEIXEIRA; NETO, MARIO DE OLIVEIRA; DE LIMA DAMASIO, ANDRE RICARDO; GARCIA, WANIUS; SQUINA, FABIO MARCIO. An alkaline active feruloyl-CoA synthetase from soil metagenome as a potential key enzyme for lignin valorization strategies. PLoS One, v. 14, n. 2, FEB 25 2019. (17/05901-7, 17/22669-0, 17/16976-8, 15/50612-8, 17/16089-1, 15/50590-4, 17/08166-6, 18/18101-1, 17/17275-3)

LIBERATO, MARCELO VIZONA; CAMPOS, BRUNA MEDEIA; TOMAZETTO, GEIZECLER; CROUCH, LUCY ISOBEL; GARCIA, WANIUS; DE MATTOS ZERI, ANA CAROLINA; BOLAM, DAVID NICHOL; SQUINA, FABIO MARCIO.

Unique properties of a Dictyostelium discoideum carbohydrate-binding module expand our understanding of CBM-ligand interactions

. Journal of Biological Chemistry, v. 298, n. 5, p. 10-pg., 2022-05-02. (15/50590-4, 13/06336-0, 15/23279-6, 20/15595-3, 14/04105-4, 20/05784-3, 15/14009-5, 17/17275-3)

FERNANDES, DIOGENES G. DA S.; ANDRADE, VIVIANE BRITO; LUCENA, LETICIA NEVES; AMBROSIO, FELIPE NOGUEIRA; DE SOUZA, ANDRE L. M.; BATISTA, BRUNO L.; ROLIM, WALLACE ROSADO; SEABRA, AMEDEA BAROZZI; LOMBELLO, CHRISTIANE B.; DA SILVA, FERNANDA DIAS; et al. Cytotoxicity and Antimicrobial Properties of Photosynthesized Silver Chloride Nanoparticles Using Plant Extract from Stryphnodendron adstringens (Martius) Coville. JOURNAL OF CLUSTER SCIENCE, v. 33, n. 2, p. 9-pg., 2021-02-05. (18/08194-2, 17/17275-3)