Estudos estruturais e funcionais de duas β-glicosidases de Trichoderma harzianum
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFSCAR |
Texto Completo: | https://repositorio.ufscar.br/handle/ufscar/7042 |
Resumo: | The depletion of fossil fuels, the growing energy demand and the great need to reduce carbon emissions gradually increased the demand for new sources of renewable and clean energy. Bioethanol is obtained from the fermentation of lignocellulosic biomass, for example sugar cane, and can be considered a viable alternative. For this biomass can be used it is necessary to degrade the constituent molecules of the cell wall to fermentable sugars through the enzymes called cellulases. Among them there are three classes with different functions: the cellobiohydrolases, endoglucanases and β-glucosidases. In order to contribute to the viability and deployment of ethanol production technologies, purification of two β- glucosidases of the fungus Trichoderma harzianum was performed, named in this work as ThBgl1 e ThBgl2, as well as their biophysical and biochemical characterization. Clones in study were obtained by a cloning platform, expression and purification of recombinant proteins in high performance and expressed in bacteria Escherichia coli. Through the functional characterization of proteins under study it was found that they had the optimum pH in the range of 5.5 and optimum temperature around 40ºC. Results of enzyme kinetics showed that ThBgl1 has higher preference (specificity) by cellobiose, while in ThBgl2 cellobiose is a substrate having low specificity. In ThBgl2 specificity is higher by fucosideos. Furthermore, it was observed the occurrence of transglycosylation catalyzed by β-glucosidases in study and the action of some inhibitors on its enzymatic activity. The three-dimensional analysis of these proteins revealed the presence of the barrel tim typical of family 1 of glycoside hydrolases. |
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Mutti, Hêmily SanchesPolikarpov, Igorhttp://lattes.cnpq.br/9669532724764871http://lattes.cnpq.br/2114370378373874c9778d41-2c04-4732-a28e-a7cc7aff31942016-08-17T18:39:52Z2015-02-092016-08-17T18:39:52Z2014-08-29MUTTI, Hêmily Sanches. Estudos estruturais e funcionais de duas β-glicosidases de Trichoderma harzianum. 2014. 110 f. Dissertação (Mestrado em Multidisciplinar) - Universidade Federal de São Carlos, São Carlos, 2014.https://repositorio.ufscar.br/handle/ufscar/7042The depletion of fossil fuels, the growing energy demand and the great need to reduce carbon emissions gradually increased the demand for new sources of renewable and clean energy. Bioethanol is obtained from the fermentation of lignocellulosic biomass, for example sugar cane, and can be considered a viable alternative. For this biomass can be used it is necessary to degrade the constituent molecules of the cell wall to fermentable sugars through the enzymes called cellulases. Among them there are three classes with different functions: the cellobiohydrolases, endoglucanases and β-glucosidases. In order to contribute to the viability and deployment of ethanol production technologies, purification of two β- glucosidases of the fungus Trichoderma harzianum was performed, named in this work as ThBgl1 e ThBgl2, as well as their biophysical and biochemical characterization. Clones in study were obtained by a cloning platform, expression and purification of recombinant proteins in high performance and expressed in bacteria Escherichia coli. Through the functional characterization of proteins under study it was found that they had the optimum pH in the range of 5.5 and optimum temperature around 40ºC. Results of enzyme kinetics showed that ThBgl1 has higher preference (specificity) by cellobiose, while in ThBgl2 cellobiose is a substrate having low specificity. In ThBgl2 specificity is higher by fucosideos. Furthermore, it was observed the occurrence of transglycosylation catalyzed by β-glucosidases in study and the action of some inhibitors on its enzymatic activity. The three-dimensional analysis of these proteins revealed the presence of the barrel tim typical of family 1 of glycoside hydrolases.O esgotamento das fontes de combustíveis fósseis, a crescente demanda energética e a grande necessidade de redução da emissão de carbono fizeram com que aumentasse gradativamente a procura por novas fontes de energia renováveis e limpas. O bioetanol obtido da fermentação de biomassas lignocelulósicas, como o bagaço de cana-de-açúcar, pode ser considerado uma alternativa viável. Para que essa biomassa possa ser utilizada, faz-se necessário a degradação das moléculas constituintes da parede celular a açúcares fermentáveis por meio das enzimas chamadas celulases. Dentre elas existem três classes com diferentes funções: as celobiohidrolases, as endoglucanases e as β-glicosidades. Com o objetivo de contribuir para a viabilização e implantação de tecnologias de produção de etanol, foi realizada a purificação de duas β-glicosidases do fungo Trichoderma harzianum, denominadas nesse trabalho como ThBgl1 e ThBgl2, bem como sua caracterização bioquímica e biofísica. Os clones em estudo foram obtidos por meio de uma plataforma de clonagem, expressão e purificação de proteínas recombinantes em alto desempenho e expressos na bactéria Escherichia coli. Através da caracterização funcional das proteínas em estudo, verificou-se que elas apresentaram o pH ótimo na faixa de 5,5 e temperatura ótima em torno de 40ºC. Os resultados de cinética enzimática mostraram que ThBgl1 tem maior preferência (especificidade) por celobiose, enquanto na ThBgl2 a celobiose é um substrato com baixa especificidade. Na ThBgl2 a especificidade é maior por fucosídeos. Além disso, constatou-se a ocorrência de transglicosilação catalisada pelas β-glicosidases em estudo e a ação de alguns inibidores em sua atividade enzimática. A análise tridimensional dessas proteínas revelou a presença de um barril tim típico das hidrolases de glicosídeos da família 1.Financiadora de Estudos e Projetosapplication/pdfporUniversidade Federal de São CarlosPrograma de Pós-Graduação em Biotecnologia - PPGBiotecUFSCarBRBiotecnologiaCinética enzimáticaTrichoderma harzianumEnzyme kineticsOUTROSEstudos estruturais e funcionais de duas β-glicosidases de Trichoderma harzianuminfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis-1-106f7e56b-278e-4aaa-9ad7-e73f28005c67info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINAL6489.pdfapplication/pdf6160107https://repositorio.ufscar.br/bitstream/ufscar/7042/1/6489.pdf81c1fddd8e696b53fe5fd98c9174ba24MD51TEXT6489.pdf.txt6489.pdf.txtExtracted texttext/plain0https://repositorio.ufscar.br/bitstream/ufscar/7042/2/6489.pdf.txtd41d8cd98f00b204e9800998ecf8427eMD52THUMBNAIL6489.pdf.jpg6489.pdf.jpgIM Thumbnailimage/jpeg5913https://repositorio.ufscar.br/bitstream/ufscar/7042/3/6489.pdf.jpgab2937122fede3b51b7a1441f7712231MD53ufscar/70422023-09-18 18:31:26.981oai:repositorio.ufscar.br:ufscar/7042Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:31:26Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false |
dc.title.por.fl_str_mv |
Estudos estruturais e funcionais de duas β-glicosidases de Trichoderma harzianum |
title |
Estudos estruturais e funcionais de duas β-glicosidases de Trichoderma harzianum |
spellingShingle |
Estudos estruturais e funcionais de duas β-glicosidases de Trichoderma harzianum Mutti, Hêmily Sanches Biotecnologia Cinética enzimática Trichoderma harzianum Enzyme kinetics OUTROS |
title_short |
Estudos estruturais e funcionais de duas β-glicosidases de Trichoderma harzianum |
title_full |
Estudos estruturais e funcionais de duas β-glicosidases de Trichoderma harzianum |
title_fullStr |
Estudos estruturais e funcionais de duas β-glicosidases de Trichoderma harzianum |
title_full_unstemmed |
Estudos estruturais e funcionais de duas β-glicosidases de Trichoderma harzianum |
title_sort |
Estudos estruturais e funcionais de duas β-glicosidases de Trichoderma harzianum |
author |
Mutti, Hêmily Sanches |
author_facet |
Mutti, Hêmily Sanches |
author_role |
author |
dc.contributor.authorlattes.por.fl_str_mv |
http://lattes.cnpq.br/2114370378373874 |
dc.contributor.author.fl_str_mv |
Mutti, Hêmily Sanches |
dc.contributor.advisor1.fl_str_mv |
Polikarpov, Igor |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/9669532724764871 |
dc.contributor.authorID.fl_str_mv |
c9778d41-2c04-4732-a28e-a7cc7aff3194 |
contributor_str_mv |
Polikarpov, Igor |
dc.subject.por.fl_str_mv |
Biotecnologia Cinética enzimática |
topic |
Biotecnologia Cinética enzimática Trichoderma harzianum Enzyme kinetics OUTROS |
dc.subject.lat.fl_str_mv |
Trichoderma harzianum |
dc.subject.eng.fl_str_mv |
Enzyme kinetics |
dc.subject.cnpq.fl_str_mv |
OUTROS |
description |
The depletion of fossil fuels, the growing energy demand and the great need to reduce carbon emissions gradually increased the demand for new sources of renewable and clean energy. Bioethanol is obtained from the fermentation of lignocellulosic biomass, for example sugar cane, and can be considered a viable alternative. For this biomass can be used it is necessary to degrade the constituent molecules of the cell wall to fermentable sugars through the enzymes called cellulases. Among them there are three classes with different functions: the cellobiohydrolases, endoglucanases and β-glucosidases. In order to contribute to the viability and deployment of ethanol production technologies, purification of two β- glucosidases of the fungus Trichoderma harzianum was performed, named in this work as ThBgl1 e ThBgl2, as well as their biophysical and biochemical characterization. Clones in study were obtained by a cloning platform, expression and purification of recombinant proteins in high performance and expressed in bacteria Escherichia coli. Through the functional characterization of proteins under study it was found that they had the optimum pH in the range of 5.5 and optimum temperature around 40ºC. Results of enzyme kinetics showed that ThBgl1 has higher preference (specificity) by cellobiose, while in ThBgl2 cellobiose is a substrate having low specificity. In ThBgl2 specificity is higher by fucosideos. Furthermore, it was observed the occurrence of transglycosylation catalyzed by β-glucosidases in study and the action of some inhibitors on its enzymatic activity. The three-dimensional analysis of these proteins revealed the presence of the barrel tim typical of family 1 of glycoside hydrolases. |
publishDate |
2014 |
dc.date.issued.fl_str_mv |
2014-08-29 |
dc.date.available.fl_str_mv |
2015-02-09 2016-08-17T18:39:52Z |
dc.date.accessioned.fl_str_mv |
2016-08-17T18:39:52Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
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masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
MUTTI, Hêmily Sanches. Estudos estruturais e funcionais de duas β-glicosidases de Trichoderma harzianum. 2014. 110 f. Dissertação (Mestrado em Multidisciplinar) - Universidade Federal de São Carlos, São Carlos, 2014. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/7042 |
identifier_str_mv |
MUTTI, Hêmily Sanches. Estudos estruturais e funcionais de duas β-glicosidases de Trichoderma harzianum. 2014. 110 f. Dissertação (Mestrado em Multidisciplinar) - Universidade Federal de São Carlos, São Carlos, 2014. |
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https://repositorio.ufscar.br/handle/ufscar/7042 |
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info:eu-repo/semantics/openAccess |
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openAccess |
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Universidade Federal de São Carlos |
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Programa de Pós-Graduação em Biotecnologia - PPGBiotec |
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UFSCar |
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BR |
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Universidade Federal de São Carlos |
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