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Library generation for biomass-conversion enzymes from soil metagenome


The gradual shift from petroleum to renewable biomass resources is generally seen as an important contribution to the development of a sustainable industrial society and the effective management of green house emissions. Lignocellulosic materiais are an abundant and low-cost source of stored energy in the biosphere. Thus, biomass conversion into feedstock sugars has moved towards the forefront of the biofuel industry. However, the saccharification of plant biomass is a complicated and lengthy process, mainly due to the inherent recalcitrance and the complex heterogeneity of the polymers comprising plant cell walls. Lignocellulosic biomass must go through an intensive pretreatment step, after which enzymes are used to break down the polysaccharides biomass into simple sugar suitable for fermentation and ethanol production. Aiming at the entire exploitation of the plant cell wall polysaccharides, as an environmentally renewable energy source, an extensive repertoire of hydrolytic enzymes would play a major role for biofuel production. The objective of this proposal is the generation of a toolkit of Iignocellulolytic enzymes with a wide range of biotechnological applications, including their use as players for the development of strategies for second generation ethanol production. The prospection of these enzymes will be done from soil metagenome, which contemplates a pioneering strategy towards the prospection of biomass conversion enzymes from microorganisms not conventionally cultivable. Additionally, this study may intensively contribute to the development of the field of bioenergy by improving techniques in rapid detection of hydrolysis using capillary electrophoresis and implementing heterologous gene expression in filamentous fungi. (AU)

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Scientific publications (27)
(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)
ALVAREZ, THABATA M.; GOLDBECK, ROSANA; DOS SANTOS, CAMILA RAMOS; PAIXAO, DOUGLAS A. A.; GONCALVES, THIAGO A.; FRANCO CAIRO, JOAO PAULO L.; ALMEIDA, RODRIGO FERREIRA; PEREIRA, ISABELA DE OLIVEIRA; JACKSON, GEORGE; COTA, JUNIO; et al. Development and Biotechnological Application of a Novel Endoxylanase Family GH10 Identified from Sugarcane Soil Metagenome. PLoS One, v. 8, n. 7, . (11/20977-3, 10/11469-1, 08/58037-9)
DIOGO, JOSE A.; HOFFMAM, ZAIRA B.; ZANPHORLIN, LETICIA M.; COTA, JUNIO; MACHADO, CARLA B.; WOLF, LUCIA D.; SQUINA, FABIO; DAMASIO, ANDRE R. L.; MURAKAMI, MARIO T.; RULLER, ROBERTO. Development of a chimeric hemicellulase to enhance the xylose production and thermotolerance. Enzyme and Microbial Technology, v. 69, p. 31-37, . (13/10443-7, 08/58037-9, 11/14200-6)
HOFFMAM, ZAIRA B.; OLIVEIRA, LEANDRO C.; COTA, JUNIO; ALVAREZ, THABATA M.; DIOGO, JOS A.; NETO, MARIO DE OLIVEIRA; CITADINI, ANA PAULA S.; LEITE, VITOR B. P.; SQUINA, FABIO M.; MURAKAMI, MARIO T.; et al. Characterization of a Hexameric Exo-Acting GH51 alpha-l-Arabinofuranosidase from the Mesophilic Bacillus subtilis. MOLECULAR BIOTECHNOLOGY, v. 55, n. 3, p. 260-267, . (10/51890-8, 08/58037-9, 11/13242-7, 11/17658-3, 11/14200-6)
TRAMONTINA, ROBSON; FRANCO CAIRO, JOAO PAULO L.; LIBERATO, MARCELO V.; MANDELLI, FERNANDA; SOUSA, AMANDA; SANTOS, SAMANTHA; RABELO, SARITA CANDIDA; CAMPOS, BRUNA; IENCZAK, JACIANE; RULLER, ROBERTO; et al. The Coptotermes gestroi aldo-keto reductase: a multipurpose enzyme for biorefinery applications. BIOTECHNOLOGY FOR BIOFUELS, v. 10, . (12/20549-4, 14/20576-7, 14/04105-4, 08/58037-9, 13/06336-0)
CAMPOS, BRUNA MEDEIA; LIBERATO, MARCELO VIZONA; ALVAREZ, THABATA MARIA; ZANPHORLIN, LETICIA MARIA; EMATSU, GABRIELA CRISTINA; BARUD, HERNANE; POLIKARPOV, IGOR; RULLER, ROBERTO; GILBERT, HARRY J.; DE MATTOS ZERI, ANA CAROLINA; et al. A Novel Carbohydrate-binding Module from Sugar Cane Soil Metagenome Featuring Unique Structural and Carbohydrate Affinity Properties. Journal of Biological Chemistry, v. 291, n. 45, p. 23734-23743, . (13/06336-0, 10/11469-1, 14/04105-4, 08/58037-9)
HAHN SCHNEIDER, WILLIAN DANIEL; GONCALVES, THIAGO AUGUSTO; UCHIMA, CRISTIANE AKEMI; COUGER, MATTHEW BRIAN; PRADE, ROLF; SQUINA, FABIO MARCIO; PINHEIRO DILLON, ALDO JOSE; CAMASSOLA, MARLI. Penicillium echinulatum secretome analysis reveals the fungi potential for degradation of lignocellulosic biomass. BIOTECHNOLOGY FOR BIOFUELS, v. 9, . (14/06923-6, 12/19040-0, 08/58037-9)
GOLDBECK, ROSANA; DAMASIO, ANDRE R. L.; GONCALVES, THIAGO A.; MACHADO, CARLA B.; PAIXAO, DOUGLAS A. A.; WOLF, LUCIA D.; MANDELLI, FERNANDA; ROCHA, GEORGE J. M.; RULLER, ROBERTO; SQUINA, FABIO M.. Development of hemicellulolytic enzyme mixtures for plant biomass deconstruction on target biotechnological applications. Applied Microbiology and Biotechnology, v. 98, n. 20, p. 8513-8525, . (13/18910-3, 12/18859-5, 08/58037-9)
SEGATO, FERNANDO; DAMASIO, ANDRE R. L.; DE LUCAS, ROSYMAR C.; SQUINA, FABIO M.; PRADE, ROLF A.. Genomics Review of Holocellulose Deconstruction by Aspergilli. MICROBIOLOGY AND MOLECULAR BIOLOGY REVIEWS, v. 78, n. 4, p. 588-613, . (13/18910-3, 12/20549-4, 08/58037-9, 10/18198-3)
ALVAREZ, THABATA MARIA; LIBERATO, MARCELO VIZONA; FRANCO CAIRO, JOO PAULO L.; PAIXO, DOUGLAS A. A.; CAMPOS, BRUNA M.; FERREIRA, MARCEL R.; ALMEIDA, RODRIGO F.; PEREIRA, ISABELA O.; BERNARDES, AMANDA; EMATSU, GABRIELA C. G.; et al. A Novel Member of GH16 Family Derived from Sugarcane Soil Metagenome. Applied Biochemistry and Biotechnology, v. 177, n. 2, p. 304-317, . (10/11469-1, 08/58037-9)
PIMENTEL, AGNES C.; EMATSU, GABRIELA C. G.; LIBERATO, MARCELO V.; PAIXAO, DOUGLAS A. A.; FRANCO CAIRO, JOAO PAULO L.; MANDELLI, FERNANDA; TRAMONTINA, ROBSON; GANDIN, CESAR A.; DE OLIVEIRA NETO, MARIO; SQUINA, FABIO M.; et al. Biochemical and biophysical properties of a metagenome-derived GH5 endoglucanase displaying an unconventional domain architecture. International Journal of Biological Macromolecules, v. 99, p. 384-393, . (16/01926-2, 14/06923-6, 10/11469-1, 14/12861-3, 14/04105-4, 08/58037-9)
MANDELLI, F.; BRENELLI, L. B.; ALMEIDA, R. F.; GOLDBECK, R.; WOLF, L. D.; HOFFMAM, Z. B.; RULLER, R.; ROCHA, G. J. M.; MERCADANTE, A. Z.; SQUINA, F. M.. Simultaneous production of xylooligosaccharides and antioxidant compounds from sugarcane bagasse via enzymatic hydrolysis. INDUSTRIAL CROPS AND PRODUCTS, v. 52, p. 770-775, . (13/03061-0, 12/18859-5, 08/58037-9, 11/14200-6)
DAMASIO, ANDRE R. L.; RUBIO, MARCELO V.; OLIVEIRA, LEANDRO C.; SEGATO, FERNANDO; DIAS, BRUNO A.; CITADINI, ANA P.; PAIXAO, DOUGLAS A.; SQUINA, FABIO M.. Understanding the Function of Conserved Variations in the Catalytic Loops of Fungal Glycoside Hydrolase Family 12. Biotechnology and Bioengineering, v. 111, n. 8, p. 1494-1505, . (11/13242-7, 08/58037-9, 12/12859-3)
FRANCO CAIRO, JOAO PAULO L.; LEONARDO, FLAVIA C.; ALVAREZ, THABATA M.; RIBEIRO, DANIELA A.; BUECHLI, FERNANDA; COSTA-LEONARDO, ANA M.; CARAZZOLLE, MARCELO F.; COSTA, FERNANDO F.; PAES LEME, ADRIANA F.; PEREIRA, GONCALO A. G.; et al. Functional characterization and target discovery of glycoside hydrolases from the digestome of the lower termite Coptotermes gestroi. BIOTECHNOLOGY FOR BIOFUELS, v. 4, . (10/11469-1, 08/58037-9)
DAMASIO, ANDRE R. L.; PINTO BRAGA, CLEITON MARCIO; BRENELLI, LIVIA B.; CITADINI, ANA PAULA; MANDELLI, FERNANDA; COTA, JUNIO; DE ALMEIDA, RODRIGO FERREIRA; SALVADOR, VICTOR HUGO; ALVAREDO PAIXAO, DOUGLAS ANTONIO; SEGATO, FERNANDO; et al. Biomass-to-bio-products application of feruloyl esterase from Aspergillus clavatus. Applied Microbiology and Biotechnology, v. 97, n. 15, p. 6759-6767, . (09/17956-4, 11/02169-7, 08/58037-9)
COTA, JUNIO; OLIVEIRA, LEANDRO C.; DAMASIO, ANDRE R. L.; CITADINI, ANA P.; HOFFMAM, ZAIRA B.; ALVAREZ, THABATA M.; CODIMA, CARLA A.; LEITE, VITOR B. P.; PASTORE, GLAUCIA; DE OLIVEIRA-NETO, MARIO; et al. Assembling a xylanase-lichenase chimera through all-atom molecular dynamics simulations. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1834, n. 8, p. 1492-1500, . (11/02169-7, 08/58037-9, 11/13242-7)
HAHN SCHNEIDER, WILLIAN DANIEL; GONCALVES, THIAGO AUGUSTO; UCHIMA, CRISTIANE AKEMI; DOS REIS, LAISA; FONTANA, ROSELEI CLAUDETE; SQUINA, FABIO MARCIO; PINHEIRO DILLON, ALDO JOSE; CAMASSOLA, MARLI. Comparison of the production of enzymes to cell wall hydrolysis using different carbon sources by Penicillium echinulaturn strains and its hydrolysis potential for lignocelullosic biomass. Process Biochemistry, v. 66, p. 162-170, . (08/58037-9)
CONTESINI, FABIANO JARES; LIBERATO, MARCELO VIZONA; RUBIO, MARCELO VENTURA; CALZADO, FELIPE; ZUBIETA, MARIANE PALUDETTI; RIANO-PACHON, DIEGO MAURICIO; SQUINA, FABIO MARCIO; BRACHT, FABRICIO; SKAF, MUNIR S.; DAMASIO, ANDRE RICARDO. Structural and functional characterization of a highly secreted alpha-L-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1865, n. 12, p. 1758-1769, . (14/15403-6, 12/20549-4, 13/08293-7, 14/04105-4, 14/23051-2, 08/58037-9, 13/24988-5)
FRANCO CAIRO, JOAO P. L.; CARAZZOLLE, MARCELO F.; LEONARDO, FLAVIA C.; MOFATTO, LUCIANA S.; BRENELLI, LIVIA B.; GONCALVES, THIAGO A.; UCHIMA, CRISTIANE A.; DOMINGUES, ROMANIA R.; ALVAREZ, THABATA M.; TRAMONTINA, ROBSON; et al. Expanding the Knowledge on Lignocellulolytic and Redox Enzymes of Worker and Soldier Castes from the Lower Termite Coptotermes gestroi. FRONTIERS IN MICROBIOLOGY, v. 7, . (15/06971-3, 14/10351-8, 12/19040-0, 11/20977-3, 10/11469-1, 14/20576-7, 08/58037-9, 13/03061-0, 14/50371-8)
DE LIMA DAMASIO, ANDRE RICARDO; RUBIO, MARCELO VENTURA; GONCALVES, THIAGO AUGUSTO; PERSINOTI, GABRIELA FELIX; SEGATO, FERNANDO; PRADE, ROLF ALEXANDER; CONTESINI, FABIANO JARES; DE SOUZA, AMANDA PEREIRA; BUCKERIDGE, MARCOS SILVEIRA; SQUINA, FABIO MARCIO. Xyloglucan breakdown by endo-xyloglucanase family 74 from Aspergillus fumigatus. Applied Microbiology and Biotechnology, v. 101, n. 7, p. 2893-2903, . (11/02169-7, 08/58037-9, 12/20549-4, 14/18714-2)
COLUSSI, FRANCIELI; DA SILVA, VIVIAM M.; MILLER, IAN; COTA, JUNIO; DE OLIVEIRA, LEANDRO C.; DE OLIVEIRA NETO, MARIO; SQUINA, FABIO M.; GARCIA, WANIUS. Oligomeric state and structural stability of two hyperthermophilic beta-glucosidases from Thermotoga petrophila. Amino Acids, v. 47, n. 5, p. 937-948, . (12/21054-9, 12/03503-0, 08/58037-9, 11/13242-7)
COTA, JUNIO; CORREA, THAMY L. R.; DAMASIO, ANDRE R. L.; DIOGO, JOSE A.; HOFFMAM, ZAIRA B.; GARCIA, WANIUS; OLIVEIRA, LEANDRO C.; PRADE, ROLF A.; SQUINA, FABIO M.. Comparative analysis of three hyperthermophilic GH1 and GH3 family members with industrial potential. NEW BIOTECHNOLOGY, v. 32, n. 1, p. 13-20, . (11/13242-7, 12/21054-9, 08/58037-9, 10/18198-3)
BRAGATTO, J.; SEGATO, F.; SQUINA, F. M.. Production of xylooligosaccharides (XOS) from delignified sugarcane bagasse by peroxide-HAc process using recombinant xylanase from Bacillus subtilis. INDUSTRIAL CROPS AND PRODUCTS, v. 51, p. 123-129, . (08/58037-9)
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, . (09/08312-6, 11/20977-3, 10/11469-1, 13/13309-0, 08/58037-9)
SQUINA, FABIO MARCIO; PRADE, ROLF ALEXANDER; WANG, HONGLIANG; MURAKAMI, MARIO TYAGO. Expression, purification, crystallization and preliminary crystallographic analysis of an endo-1,5-alpha-L-arabinanase from hyperthermophilic Thermotoga petrophila. ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, v. 65, n. 9, p. 902-905, . (08/58037-9)
DE OLIVEIRA, LEANDRO C.; DA SILVA, VIVIAM M.; COLUSSI, FRANCIELI; CABRAL, ALINE D.; DE OLIVEIRA NETO, MARIO; SQUINA, FABIO M.; GARCIA, WANIUS. Conformational Changes in a Hyperthermostable Glycoside Hydrolase: Enzymatic Activity Is a Consequence of the Loop Dynamics and Protonation Balance. PLoS One, v. 10, n. 2, . (12/21054-9, 12/03503-0, 08/58037-9, 11/13242-7)
FRANCO CAIRO, JOAO PAULO L.; OLIVEIRA, LEANDRO C.; UCHIMA, CRISTIANE A.; ALVAREZ, THABATA M.; CITADINI, ANA PAULA DA S.; COTA, JUNIO; LEONARDO, FLAVIA COSTA; COSTA-LEONARDO, ANA M.; CARAZZOLLE, MARCELO F.; COSTA, FERNANDO F.; et al. Deciphering the synergism of endogenous glycoside hydrolase families 1 and 9 from Coptotermes gestroi. Insect Biochemistry and Molecular Biology, v. 43, n. 10, p. 970-981, . (12/19040-0, 11/08609-9, 11/20977-3, 10/11469-1, 08/58037-9, 11/13242-7)
MANDELLI, F.; FRANCO CAIRO, J. P. L.; CITADINI, A. P. S.; BUECHLI, F.; ALVAREZ, T. M.; OLIVEIRA, R. J.; LEITE, V. B. P.; PAES LEME, A. F.; MERCADANTE, A. Z.; SQUINA, F. M.. The characterization of a thermostable and cambialistic superoxide dismutase from Thermus filiformis. Letters in Applied Microbiology, v. 57, n. 1, p. 40-46, . (11/17658-3, 08/58037-9)

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Filed patent(s) as a result of this research project

COMPOSIÇÃO DE ENZIMAS LIGNOCELULOLÍTICAS, MÉTODO DE CONVERSÃO ENZIMÁTICA E VETOR DE EXPRESSÃO DE UMA SUPERÓXIDO DISMUTASE BR1020150172567 - Centro Nacional de Pesquisa em Energia e Materiais (CNPEM) ; Universidade Estadual de Campinas (UNICAMP) ; Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP) . Falsarella Carazzole; Fernanda Mandelli; Jose Geraldo Da Cruz Pradella; Thabata Maria Alvarez; Flávia Costa Leonardo; Fernando Ferreira Costa; Ana Maria Costa Leonardo; Robson Tramontina; Livia Beatriz Brenelli De Paiva; Thiago Augusto Gonçalves; Diogo Robl - July 2015, 20

COMPOSITION OF LIGNOCELLULOLYTIC ENZYMES, ENZYMATIC CONVERSION METHOD AND SUPEROXIDE DISMUTASE EXPRESSION VECTOR PCT/BR2016/000069 - Centro Nacional de Pesquisa em Energia e Materiais (CNPEM) ; Universidade Estadual de Campinas (UNICAMP) ; Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP) . Gonçalo Amarante Guimarães Pereira; João Paulo Lourenço Cairo; Fábio Marcio Squina; Marcelo Falsarella Carazzole; Fernanda Mandelli; José Geraldo da Cruz Pradella; Thabata Maria Alvarez; Flávia Costa Leonardo; Fernando Ferreira Costa; Ana Maria Costa Leonardo; Robson Tramontina; Livia Beatriz Brenelli de Paiva; Thiago Augusto Gonçalves; Diogo Robl - July 2016, 18