Busca avançada
Ano de início
Entree

Structure and function of enzymes and auxiliary proteins from Trichoderma, active in cell-wall hydrolysis

Processo: 08/56255-9
Linha de fomento:Auxílio à Pesquisa - Programa BIOEN - Temático
Vigência: 01 de novembro de 2009 - 30 de setembro de 2014
Área do conhecimento:Ciências Biológicas - Biofísica - Biofísica Molecular
Convênio/Acordo: CNPq - Pronex
Pesquisador responsável:Igor Polikarpov
Beneficiário:Igor Polikarpov
Instituição-sede: Instituto de Física de São Carlos (IFSC). Universidade de São Paulo (USP). São Carlos , SP, Brasil
Pesquisadores principais:
Munir Salomao Skaf ; Valtencir Zucolotto
Auxílios(s) vinculado(s):10/18849-4 - Purificação e caracterização de hidrolases de glicosídeos, produzidas pelo fungo filamentoso Trichoderma harzianum, com perspectiva de aplicabilidade dessas enzimas em coquetéis para hidrólise da biomassa e produção de bioetanol, AV.EXT
Bolsa(s) vinculada(s):13/13766-1 - Estrutura e função de enzimas e proteínas auxiliares de Trichoderma, atuantes na hidrólise da parede celular, BP.TT
12/17298-0 - Apoio técnico ao grupo de biotecnologia molecular do Instituto de Física de São Carlos, IFSC/USP, BP.TT
12/08402-8 - Expressão, purificação, cristalização e ensaios funcionais da enzima beta-galactosidase da arqueobactéria haloterrigena turkmenica, BP.IC
+ mais bolsas vinculadas 11/19100-0 - Estudos funcionais e estruturais da classe de enzimas endoglucanases GHF45 visando suas aplicações para hidrólise de biomassa, BP.DD
11/20505-4 - Duas classes importantes de glicosil hidrolases: estudos funcionais e análise estrutural, BP.PD
11/22670-2 - Estrutura e função de enzimas e proteínas auxiliares de Trichoderma, atuantes na hidrólise da parede celular, BP.TT
11/05712-3 - Estudos estruturais e funcionais das enzimas beta-galactosidases de Xanthomonas campestris, BP.DD
10/20681-4 - Projeto e desenvolvimento de um reator enzimático para a produção de etanol a partir de bagaço de cana-de-açúcar, BP.PD
10/18773-8 - Clonagem molecular, expressão, purificação e caracterização estrutural da endoglucanase de Trichoderma harzianum visando o desenvolvimento de coquetéis enzimáticos para a produção de etanol lignocelulósico, BP.DD
10/17569-8 - Estrutura e função de enzimas e proteínas auxiliares de Trichoderma, ativas na hidrólise da parede celular, BP.TT
10/16542-9 - Estudos de enzimas celulolíticas com potencial na transformação enzimática da biomassa de cana-de-açúcar utilizando espalhamento de raios-X a baixo ângulo, BP.PD
10/11135-6 - Aspectos estruturais e morfológicos da degradação da parede celular e seus biopolímeros, BP.PD
10/08680-2 - Aspectos moleculares da degradação de biomassa lignocelulósica: dinâmica de enzimas e nanoarquitetura de paredes celulares de plantas, BP.DR
09/13185-3 - Análise da estrutura genômica de regiões do genoma do fungo Trichoderma harzianum contendo genes envolvidos na produção de enzimas com atividade celulolítica, BP.MS
09/11536-3 - Estudos bioquímicos e estruturais dos sistemas celulolíticos de Trichoderma reesei e Phanerochaete chrysosporium aplicados à melhoria da eficiência de degradação de biomassa, BP.PD
09/14107-6 - Dinâmica Molecular de Hidrolases para Sacarificação de Celulose e Proteínas Correlatas, BP.DR
09/08233-9 - Estudos moleculares, estruturais e funcionais da Cel12A de Gloeophyllum trabeum, uma endo-1,4-²-glucanase da família 12 de hidrolases de glicosídeos, BP.DD - menos bolsas vinculadas
Assunto(s):Biologia molecular  Fungos mitospóricos  Trichoderma  Enzimas  Bioenergia 
Publicação FAPESP:https://media.fapesp.br/bv/uploads/publicacoes/pasta_bioen_jun2012_70.pdf

Resumo

Lignocellulosic biomass, such as sugarcane bagasse, holds a promise of environmentally friendly bioenergy production in Brazil. However, enzymatic hydrolysis, currently considered a method of choice in biomass saccharification, is hampered by considerable cell-wall recalcitrance. To make this technology sustainable and cost effective, our comprehension of cellulose enzymatic hydrolysis should be significantly improved. Here we propose to conduct systematic structure-functional studies of Trichoderma cellulases and auxiliary proteins active in cell-wall degradation using a combination of X-ray protein crystallography, biophysical and biochemical studies, molecular dynamics simulations, statistical coupling analysis aligned with the site-directed mutagenesis and enzymatic assays aiming to obtain in-depth comprehension of cellulose hydrolysis. We plan to contribute toward structural analysis of Trichoderma reesei endoglucanases by solving a crystal structure of endoglucanase II (Cel5A), main enzymatically active, but structurally uncharacterized endoglucanase of this important industrial fungus. Moreover, we will contribute toward our knowledge of Trichoderma cellulases molecular organization by solving X-ray structures of main Trichoderma harzianum endo- and exoglucanases (primarily focusing on Cel7A and Cel5A) and by comparing them with the correspondent T. reesei enzymes. We also aim to structurally characterize swollenins, non-hydrolytic proteins, shown to enhance cellulose hydrolysis catalyzed by celulases, and to study thermodynamically its interactions with cellulose. In addition, we will construct chimeric enzymes by fusing of swollenin with the cellulases and will study enzymatic properties of such chimeras. Furthermore, we will conduct systematic molecular dynamics studies of the cellulases and swollenin, and investigate their flexibility by hydrogen deuterium exchange followed by massspectrometry. Finally, we will use all these acquired knowledge to modify the proteins using site-directed mutagenesis aiming to better comprehend molecular basis of their function and to produce enzymes and their mixtures with enhanced hydrolytic properties. (AU)

Matéria(s) publicada(s) na Agência FAPESP sobre o auxílio:
Matéria(s) publicada(s) em Outras Mídias (0 total):
Mais itensMenos itens
VEICULO: TITULO (DATA)
VEICULO: TITULO (DATA)

Publicações científicas (27)
(Referências obtidas automaticamente do Web of Science e do SciELO, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores)
BERNARDINELLI, OIGRES DANIEL; LIMA, MARISA APARECIDA; REZENDE, CAMILA ALVES; POLIKARPOV, IGOR; DEAZEVEDO, EDUARDO RIBEIRO. Quantitative C-13 MultiCP solid-state NMR as a tool for evaluation of cellulose crystallinity index measured directly inside sugarcane biomass. BIOTECHNOLOGY FOR BIOFUELS, v. 8, . (10/52362-5, 09/18354-8, 10/08370-3, 10/11135-6, 08/56255-9)
ROSSETO, FLAVIO RODOLFO; MANZINE, LIVIA REGINA; NETO, MARIO DE OLIVEIRA; POLIKARPOV, IGOR. Biophysical and biochemical studies of a major endoglucanase secreted by Xanthomonas campestris pv. campestris.. Enzyme and Microbial Technology, v. 91, p. 1-7, . (09/54035-4, 10/16542-9, 10/08370-3, 08/56255-9)
LIBERATO, MARCELO V.; SILVEIRA, RODRIGO L.; PRATES, ERICA T.; DE ARAUJO, EVANDRO A.; PELLEGRINI, VANESSA O. A.; CAMILO, CESAR M.; KADOWAKI, MARCO A.; NETO, MARIO DE O.; POPOV, ALEXANDER; SKAF, MUNIR S.; et al. Molecular characterization of a family 5 glycoside hydrolase suggests an induced-fit enzymatic mechanism. SCIENTIFIC REPORTS, v. 6, . (10/18773-8, 13/08293-7, 13/15582-5, 09/52840-7, 14/10448-1, 08/56255-9)
TOMAZINI JUNIOR, ATILIO; DOLCE, LUCIANO GRACIANI; DE OLIVEIRA NETO, MARIO; POLIKARPOV, IGOR. Xanthomonas campestris expansin-like X domain is a structurally disordered beta-sheet macromolecule capable of synergistically enhancing enzymatic efficiency of cellulose hydrolysis. Biotechnology Letters, v. 37, n. 12, p. 2419-2426, . (07/08706-9, 09/05349-6, 08/56255-9, 10/52362-5)
LIMA, LEONARDO H. F.; SERPA, VIVIANE I.; ROSSETO, FLAVIO R.; SARTORI, GERALDO RODRIGUES; DE OLIVEIRA NETO, MARIO; MARTINEZ, LEANDRO; POLIKARPOV, IGOR. Small-angle X-ray scattering and structural modeling of full-length: cellobiohydrolase I from Trichoderma harzianum. Cellulose, v. 20, n. 4, p. 1573-1585, . (07/08706-9, 09/52840-7, 10/16947-9, 08/05637-9, 08/56255-9)
DE ARAUJO, EVANDRO ARES; TOMAZINI, JR., ATILIO; SEIKI KADOWAKI, MARCO ANTONIO; MURAKAMI, MARIO TYAGO; POLIKARPOV, IGOR. Crystallization and preliminary X-ray diffraction analysis of a new xyloglucanase from Xanthomonas campestris pv. campestris. ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, v. 69, n. 6, p. 676-678, . (09/05349-6, 10/52362-5, 07/08706-9, 08/56255-9)
BERNARDES, AMANDA; TEXTOR, LARISSA C.; SANTOS, JADEMILSON C.; CUADRADO, NAZARET HIDALGO; KOSTETSKY, EDUARD YA.; ROIG, MANUEL G.; BAVRO, VASSILIY N.; MUNIZ, JOAO R. C.; SHNYROV, VALERY L.; POLIKARPOV, IGOR. Crystal structure analysis of peroxidase from the palm tree Chamaerops excelsa. Biochimie, v. 111, p. 58-69, . (12/22802-9, 09/05349-6, 08/56255-9, 10/52362-5)
DOS SANTOS, JADEMILSON CELESTINO; BERNARDES, AMANDA; GIAMPIETRO, LETIZIA; AMMAZZALORSO, ALESSANDRA; DE FILIPPIS, BARBARA; AMOROSO, ROSA; POLIKARPOV, IGOR. Different binding and recognition modes of GL479, a dual agonist of Peroxisome Proliferator-Activated Receptor alpha/gamma. Journal of Structural Biology, v. 191, n. 3, p. 332-340, . (07/58443-4, 08/56255-9, 09/14333-6)
GODOY, ANDRE SCHUTZER; CAMILO, CESAR MOISES; KADOWAKI, MARCO ANTONIO; MUNIZ, HELOISA DOS S.; SANTO, MELISSA ESPIRITO; MURAKAMI, MARIO TYAGO; NASCIMENTO, ALESSANDRO S.; POLIKARPOV, IGOR. Crystal structure of beta 1 -> 6-galactosidase from Bifidobacterium bifidum S17: trimeric architecture, molecular determinants of the enzymatic activity and its inhibition by alpha-galactose. FEBS Journal, v. 283, n. 22, p. 4097-4112, . (09/52840-7, 14/06565-2, 11/05712-3, 09/05328-9, 11/20505-4, 08/56255-9)
TSUCHIDA, JEFFERSON ESQUINA; REZENDE, CAMILA ALVES; DE OLIVEIRA-SILVA, RODRIGO; LIMA, MARISA APARECIDA; D'EURYDICE, MARCEL NOGUEIRA; POLIKARPOV, IGOR; BONAGAMBA, TITO JOSE. Nuclear magnetic resonance investigation of water accessibility in cellulose of pretreated sugarcane bagasse. BIOTECHNOLOGY FOR BIOFUELS, v. 7, . (12/22119-7, 09/18354-8, 10/08370-3, 10/11135-6, 09/54880-6, 08/56255-9)
MIOTTO, LIS SCHWARTZ; DE REZENDE, CAMILA ALVES; BERNARDES, AMANDA; SERPA, VIVIANE ISABEL; TSANG, ADRIAN; POLIKARPOV, IGOR. The Characterization of the Endoglucanase Cel12A from Gloeophyllum trabeum Reveals an Enzyme Highly Active on beta-Glucan. PLoS One, v. 9, n. 9, . (10/52362-5, 12/22802-9, 09/52840-7, 08/56255-9, 09/08233-9)
FLORINDO, RENATA N.; SOUZA, VALQUIRIA P.; MUTTI, HEMILY S.; CAMILO, CESAR; MANZINE, LIVIA REGINA; MARANA, SANDRO R.; POLIKARPOV, IGOR; NASCIMENTO, ALESSANDRO S.. Structural insights into beta-glucosidase transglycosylation based on biochemical, structural and computational analysis of two GH1 enzymes from Trichoderma harzianum. NEW BIOTECHNOLOGY, v. 40, n. B, p. 218-227, . (08/56255-9, 14/06565-2, 14/19439-5)
PRATES, ERICA T.; STANKOVIC, IVANA; SILVEIRA, RODRIGO L.; LIBERATO, MARCELO V.; HENRIQUE-SILVA, FLAVIO; PEREIRA, JR., NEI; POLIKARPOV, IGOR; SKAF, MUNIR S.. X-ray Structure and Molecular Dynamics Simulations of Endoglucanase 3 from Trichoderma harzianum: Structural Organization and Substrate Recognition by Endoglucanases That Lack Cellulose Binding Module. PLoS One, v. 8, n. 3, . (10/08680-2, 09/54035-4, 08/56255-9)
LIMA, MARISA A.; OLIVEIRA-NETO, MARIO; KADOWAKI, MARCO ANTONIO S.; ROSSETO, FLAVIO R.; PRATES, ERICA T.; SQUINA, FABIO M.; LEME, ADRIANA F. P.; SKAF, MUNIR S.; POLIKARPOV, IGOR. Aspergillus niger beta-Glucosidase Has a Cellulase-like Tadpole Molecular Shape INSIGHTS INTO GLYCOSIDE HYDROLASE FAMILY 3 (GH3) beta-GLUCOSIDASE STRUCTURE AND FUNCTION. Journal of Biological Chemistry, v. 288, n. 46, p. 32991-33005, . (10/16542-9, 09/54035-4, 08/56255-9)
ROSSETO, FLAVIO R.; PUHL, ANA C.; ANDRADE, MAXUEL O.; POLIKARPOV, IGOR. Crystallization and preliminary diffraction analysis of the catalytic domain of major extracellular endoglucanase from Xanthomonas campestris pv. campestris. ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, v. 69, n. 2, p. 137-140, . (07/08706-9, 09/05349-6, 08/56255-9, 10/52362-5)
PELLEGRINI, VANESSA O. A.; SERPA, VIVIANE ISABEL; GODOY, ANDRE S.; CAMILO, CESAR M.; BERNARDES, AMANDA; REZENDE, CAMILA A.; PEREIRA JUNIOR, NEI; CAIRO, JOAO PAULO L. FRANCO; SQUINA, FABIO M.; POLIKARPOV, IGOR. Recombinant Trichoderma harzianum endoglucanase I (Cel7B) is a highly acidic and promiscuous carbohydrate-active enzyme. Applied Microbiology and Biotechnology, v. 99, n. 22, p. 9591-9604, . (10/18773-8, 11/20977-3, 09/52840-7, 11/05712-3, 09/05328-9, 08/56255-9)
DOS REIS, CAIO VINICIUS; BERNARDES, AMANDA; POLIKARPOV, IGOR. Expression, purification, crystallization and preliminary X-ray diffraction analysis of Bifidobacterium adolescentis xylose isomerase. ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, v. 69, n. 5, p. 588-591, . (08/56255-9, 09/52840-7)
SKAF, MUNIR SALOMAO; POLIKARPOV, IGOR; STANKOVIC, IVANA M.. A linker of the proline-threonine repeating motif sequence is bimodal. Journal of Molecular Modeling, v. 26, n. 7, . (08/56255-9, 10/08680-2, 09/54035-4)
GODOY, ANDRE S.; DE LIMA, MARIANA Z. T.; CAMILO, CESAR M.; POLIKARPOV, IGOR. Crystal structure of a putative exo-beta-1,3-galactanase from Bifidobacterium bifidum S17. ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, v. 72, n. 4, p. 288-293, . (09/52840-7, 11/05712-3, 09/05328-9, 11/20505-4, 08/56255-9)
LIMA, MARISA A.; GOMEZ, LEONARDO D.; STEELE-KING, CLARE G.; SIMISTER, RACHAEL; BERNARDINELLI, OIGRES D.; CARVALHO, MARCELO A.; REZENDE, CAMILA A.; LABATE, CARLOS A.; DEAZEVEDO, EDUARDO R.; MCQUEEN-MASON, SIMON J.; et al. Evaluating the composition and processing potential of novel sources of Brazilian biomass for sustainable biorenewables production. BIOTECHNOLOGY FOR BIOFUELS, v. 7, . (10/52362-5, 09/18354-8, 10/08370-3, 10/11135-6, 08/56255-9)
NASCIMENTO, ALESSANDRO S.; MUNIZ, JOAO RENATO C.; APARICIO, RICARDO; GOLUBEV, ALEXANDER M.; POLIKARPOV, IGOR. Insights into the structure and function of fungal beta-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme. FEBS Journal, v. 281, n. 18, SI, p. 4165-4178, . (08/56255-9, 09/52840-7)
LIMA, MARISA A.; LAVORENTE, GABRIELA B.; DA SILVA, HANA K. P.; BRAGATTO, JULIANO; REZENDE, CAMILA A.; BERNARDINELLI, OIGRES D.; DEAZEVEDO, EDUARDO R.; GOMEZ, LEONARDO D.; MCQUEEN-MASON, SIMON J.; LABATE, CARLOS A.; et al. Effects of pretreatment on morphology, chemical composition and enzymatic digestibility of eucalyptus bark: a potentially valuable source of fermentable sugars for biofuel production - part 1. BIOTECHNOLOGY FOR BIOFUELS, v. 6, . (10/52362-5, 09/18354-8, 10/08370-3, 10/11135-6, 08/56255-9)
BROGNARO, HEVILA; ALMEIDA, VITOR MEDEIROS; DE ARAUJO, EVANDRO ARES; PIYADOV, VASILY; MORIM SANTOS, MARIA AUXILIADORA; MARANA, SANDRO ROBERTO; POLIKARPOV, IGOR. Biochemical Characterization and Low-Resolution SAXS Molecular Envelope of GH1 beta-Glycosidase from Saccharophagus degradans. MOLECULAR BIOTECHNOLOGY, v. 58, n. 12, p. 777-788, . (08/56255-9)
CAMILO, CESAR M.; POLIKARPOV, IGOR. High-throughput cloning, expression and purification of glycoside hydrolases using Ligation-Independent Cloning (LIC). Protein Expression and Purification, v. 99, p. 35-42, . (09/11536-3, 10/08680-2, 09/54035-4, 08/56255-9)
MELLO, BRUNO LUAN; POLIKARPOV, IGOR. Family 1 carbohydrate binding-modules enhance saccharification rates. AMB EXPRESS, v. 4, . (10/52362-5, 09/05349-6, 08/56255-9)
FLORINDO, RENATA N.; SOUZA, VALQUIRIA P.; MANZINE, LIVIA R.; CAMILO, CESAR M.; MARANA, SANDRO R.; POLIKARPOV, IGOR; NASCIMENTO, ALESSANDRO S.. Structural and biochemical characterization of a GH3 beta-glucosidase from the probiotic bacteria Bifidobacterium adolescentis. Biochimie, v. 148, p. 107-115, . (08/56255-9, 14/06565-2, 14/19439-5)

Por favor, reporte erros na lista de publicações científicas escrevendo para: cdi@fapesp.br.