Influência dos coquetéis enzimáticos produzidos por Trichoderma reesei e Aspergillus niger pelo processo de fermentação sequencial na hidrólise do bagaço de cana-de-açúcar

Detalhes bibliográficos
Autor(a) principal: Florencio, Camila
Data de Publicação: 2016
Tipo de documento: Tese
Idioma: por
eng
Título da fonte: Repositório Institucional da UFSCAR
Texto Completo: https://repositorio.ufscar.br/handle/ufscar/8504
Resumo: Currently, one of the major challenges for second generation ethanol is to reduce the cost of cellulolytic enzymes. Thus, the development of bioprocesses for the enzyme production on-site and strategies to increase the final yield of the enzymatic hydrolysis are required to ensure that biomass conversion to be economically feasible. Therefore, the objective of this work was to study the production and characterization of enzyme cocktails involved in the degradation of plant biomass by filamentous fungi Trichoderma reesei and Aspergillus niger grown in sequential fermentation and evaluate the application of these cocktails in the saccharification process of sugarcane bagasse. Firstly, evaluation and validation of sequential fermentation cultivation methodology to different strains of Trichoderma. Cultivation were made using sugarcane bagasse "in natura" and pretreated by steam explosion, as a carbon source. The result more significantly was observed for T. reesei Rut C30, the endoglucanase production was 4.2-fold higher than the values obtained in conventional submerged fermentation. The enzyme extracts were characterized in terms of optimum pH and temperature and endoglucanase profile. The thermostability was directly influenced by the type of carbon source and type of cultivation method. Subsequently, the proteomic analysis were performed of enzyme cocktails from T. reesei Rut C30 and A. niger A12 produced by submerged and sequential fermentation in the presence of pretreated bagasse. The performance of the enzyme cocktail in saccharification of pretreated bagasse showed that the combination of enzyme cocktails from T. reesei and A. niger produced by sequential fermentation had a yield than 3-fold higher than the enzyme cocktails of submerged fermentation. In order to evaluate the action of the enzyme cocktails produced by T. reesei and A. niger in sugarcane bagasse saccharification, the last step of the work was to study the additives effects during the sugarcane bagasse hydrolysis aiming at reducing non-productive adsorption of enzymes into lignin. The saccharification results in the presence of soybean protein were 2-fold higher than the controls (no additive) to the enzyme cocktails of two fungi studied produced by solid state fermentation, indicating the potential use of soybean protein as an additive to minimize non-productive adsorption of the enzyme into lignin. Overall, this study presents an interesting final contribution in the cellulase production process and the application of the enzyme cocktail in the hydrolysis of sugarcane bagasse.
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spelling Florencio, CamilaFarinas, Cristiane Sanchezhttp://lattes.cnpq.br/9933650905615452Badino Junior, Alberto Collihttp://lattes.cnpq.br/6244428434217018http://lattes.cnpq.br/8317605349425304aec89963-bd66-43eb-900e-f1ff1bb6e19e2017-02-08T12:10:12Z2017-02-08T12:10:12Z2016-04-28FLORENCIO, Camila. Influência dos coquetéis enzimáticos produzidos por Trichoderma reesei e Aspergillus niger pelo processo de fermentação sequencial na hidrólise do bagaço de cana-de-açúcar. 2016. Tese (Doutorado em Biotecnologia) – Universidade Federal de São Carlos, São Carlos, 2016. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8504.https://repositorio.ufscar.br/handle/ufscar/8504Currently, one of the major challenges for second generation ethanol is to reduce the cost of cellulolytic enzymes. Thus, the development of bioprocesses for the enzyme production on-site and strategies to increase the final yield of the enzymatic hydrolysis are required to ensure that biomass conversion to be economically feasible. Therefore, the objective of this work was to study the production and characterization of enzyme cocktails involved in the degradation of plant biomass by filamentous fungi Trichoderma reesei and Aspergillus niger grown in sequential fermentation and evaluate the application of these cocktails in the saccharification process of sugarcane bagasse. Firstly, evaluation and validation of sequential fermentation cultivation methodology to different strains of Trichoderma. Cultivation were made using sugarcane bagasse "in natura" and pretreated by steam explosion, as a carbon source. The result more significantly was observed for T. reesei Rut C30, the endoglucanase production was 4.2-fold higher than the values obtained in conventional submerged fermentation. The enzyme extracts were characterized in terms of optimum pH and temperature and endoglucanase profile. The thermostability was directly influenced by the type of carbon source and type of cultivation method. Subsequently, the proteomic analysis were performed of enzyme cocktails from T. reesei Rut C30 and A. niger A12 produced by submerged and sequential fermentation in the presence of pretreated bagasse. The performance of the enzyme cocktail in saccharification of pretreated bagasse showed that the combination of enzyme cocktails from T. reesei and A. niger produced by sequential fermentation had a yield than 3-fold higher than the enzyme cocktails of submerged fermentation. In order to evaluate the action of the enzyme cocktails produced by T. reesei and A. niger in sugarcane bagasse saccharification, the last step of the work was to study the additives effects during the sugarcane bagasse hydrolysis aiming at reducing non-productive adsorption of enzymes into lignin. The saccharification results in the presence of soybean protein were 2-fold higher than the controls (no additive) to the enzyme cocktails of two fungi studied produced by solid state fermentation, indicating the potential use of soybean protein as an additive to minimize non-productive adsorption of the enzyme into lignin. Overall, this study presents an interesting final contribution in the cellulase production process and the application of the enzyme cocktail in the hydrolysis of sugarcane bagasse.Atualmente, um dos grandes desafios para a produção de etanol de segunda geração consiste em diminuir o custo das enzimas celulolíticas. Assim, o desenvolvimento de bioprocessos para produção das enzimas on-site e estratégias para aumentar o rendimento final da hidrólise enzimática são necessários para assegurar que a conversão de biomassa seja economicamente viável. Para tanto, o objetivo deste trabalho foi estudar a produção e caracterização de coquetéis enzimáticos envolvidos na degradação da biomassa vegetal pelos fungos filamentosos Trichoderma reesei e Aspegillus niger cultivados por fermentação sequencial, bem como avaliar a aplicação dos mesmos no processo de sacarificação do bagaço de cana-de-açúcar. Primeiramente foi realizada a avaliação e validação da metodologia de cultivo de fermentação sequencial para diferentes linhagens de Trichoderma. Os cultivos foram feitos utilizando o bagaço de cana “in natura” e prétratado por explosão a vapor, como fonte de carbono. O melhor resultado foi observado para T. reesei Rut C30, em que a produção de endoglucanase foi 4,2 vezes maior do que os valores obtidos em cultivo convencional de fermentação submersa. Os extratos enzimáticos foram caracterizados em termos de pH e temperatura ótimos e perfil de endoglucanase. A termo-estabilidade foi diretamente influenciada pelo tipo de fonte de carbono e tipo de cultivo. Posteriormente, foram realizadas as análises proteômicas dos coquetéis enzimáticos do T. reesei Rut C30 e A. niger A12 produzidos por fermentação submersa convencional e fermentação sequencial, na presença de bagaço de cana prétratado. A performance dos coquetéis enzimáticos na sacarificação do bagaço de cana pré-tratado mostraram que a combinação dos coquetéis enzimáticos de T. reesei e A. niger produzidos por fermentação sequencial tiveram um rendimento 3 vezes maior do que os coquetéis da fermentação submersa. A fim de explorar melhor a ação dos coquetéis enzimáticos produzidos por T. reesei e A. niger na sacarificação do bagaço, na última etapa do trabalho foi estudo o efeito de aditivos durante a hidrólise do bagaco de cana visando à redução da adsorção improdutiva de enzimas na lignina. Os resultados de sacarificação na presença da proteína de soja foram 2 vezes maiores do que os controles (sem aditivo) para os coquetéis enzimáticos dos dois fungos estudados produzidos por fermentação em estado sólido, indicando o potencial do uso da proteína de soja como aditivo para minimizar a adsorção improdutiva das enzimas na lignina. De modo geral, o presente trabalho conseguiu uma contribuição final interessante no processo de produção das celulases e a aplicação do coquetel enzimático na hidrólise do bagaço de cana.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)porengUniversidade Federal de São CarlosCâmpus São CarlosPrograma de Pós-Graduação em Biotecnologia - PPGBiotecUFSCarTrichoderma reeseiAspergillus nigerEnzimas hemicelulolíticasProcessos fermentativosHidrólise enzimáticaBagaço de cana-de-açúcarHemicellulolític enzymesFermentative processEnzymatic hydrolysisSugarcane bagasseCIENCIAS EXATAS E DA TERRAInfluência dos coquetéis enzimáticos produzidos por Trichoderma reesei e Aspergillus niger pelo processo de fermentação sequencial na hidrólise do bagaço de cana-de-açúcarinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisOnline600600dd9ceb6c-d509-421a-a31e-bcb55bb21e02info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINALTeseCF.pdfTeseCF.pdfapplication/pdf2944475https://repositorio.ufscar.br/bitstream/ufscar/8504/1/TeseCF.pdf0b8b7d83e1195e91eb572351858f7b93MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81957https://repositorio.ufscar.br/bitstream/ufscar/8504/2/license.txtae0398b6f8b235e40ad82cba6c50031dMD52TEXTTeseCF.pdf.txtTeseCF.pdf.txtExtracted texttext/plain359183https://repositorio.ufscar.br/bitstream/ufscar/8504/3/TeseCF.pdf.txt11fcdc09f383279a8751a654f51a8afbMD53THUMBNAILTeseCF.pdf.jpgTeseCF.pdf.jpgIM Thumbnailimage/jpeg10110https://repositorio.ufscar.br/bitstream/ufscar/8504/4/TeseCF.pdf.jpgb3e70f607621f8a1bb21e27c61678db0MD54ufscar/85042024-03-08 01:18:59.262oai:repositorio.ufscar.br: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Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222024-03-08T01:18:59Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false
dc.title.por.fl_str_mv Influência dos coquetéis enzimáticos produzidos por Trichoderma reesei e Aspergillus niger pelo processo de fermentação sequencial na hidrólise do bagaço de cana-de-açúcar
title Influência dos coquetéis enzimáticos produzidos por Trichoderma reesei e Aspergillus niger pelo processo de fermentação sequencial na hidrólise do bagaço de cana-de-açúcar
spellingShingle Influência dos coquetéis enzimáticos produzidos por Trichoderma reesei e Aspergillus niger pelo processo de fermentação sequencial na hidrólise do bagaço de cana-de-açúcar
Florencio, Camila
Trichoderma reesei
Aspergillus niger
Enzimas hemicelulolíticas
Processos fermentativos
Hidrólise enzimática
Bagaço de cana-de-açúcar
Hemicellulolític enzymes
Fermentative process
Enzymatic hydrolysis
Sugarcane bagasse
CIENCIAS EXATAS E DA TERRA
title_short Influência dos coquetéis enzimáticos produzidos por Trichoderma reesei e Aspergillus niger pelo processo de fermentação sequencial na hidrólise do bagaço de cana-de-açúcar
title_full Influência dos coquetéis enzimáticos produzidos por Trichoderma reesei e Aspergillus niger pelo processo de fermentação sequencial na hidrólise do bagaço de cana-de-açúcar
title_fullStr Influência dos coquetéis enzimáticos produzidos por Trichoderma reesei e Aspergillus niger pelo processo de fermentação sequencial na hidrólise do bagaço de cana-de-açúcar
title_full_unstemmed Influência dos coquetéis enzimáticos produzidos por Trichoderma reesei e Aspergillus niger pelo processo de fermentação sequencial na hidrólise do bagaço de cana-de-açúcar
title_sort Influência dos coquetéis enzimáticos produzidos por Trichoderma reesei e Aspergillus niger pelo processo de fermentação sequencial na hidrólise do bagaço de cana-de-açúcar
author Florencio, Camila
author_facet Florencio, Camila
author_role author
dc.contributor.authorlattes.por.fl_str_mv http://lattes.cnpq.br/8317605349425304
dc.contributor.author.fl_str_mv Florencio, Camila
dc.contributor.advisor1.fl_str_mv Farinas, Cristiane Sanchez
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/9933650905615452
dc.contributor.advisor-co1.fl_str_mv Badino Junior, Alberto Colli
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/6244428434217018
dc.contributor.authorID.fl_str_mv aec89963-bd66-43eb-900e-f1ff1bb6e19e
contributor_str_mv Farinas, Cristiane Sanchez
Badino Junior, Alberto Colli
dc.subject.lat.fl_str_mv Trichoderma reesei
Aspergillus niger
topic Trichoderma reesei
Aspergillus niger
Enzimas hemicelulolíticas
Processos fermentativos
Hidrólise enzimática
Bagaço de cana-de-açúcar
Hemicellulolític enzymes
Fermentative process
Enzymatic hydrolysis
Sugarcane bagasse
CIENCIAS EXATAS E DA TERRA
dc.subject.por.fl_str_mv Enzimas hemicelulolíticas
Processos fermentativos
Hidrólise enzimática
Bagaço de cana-de-açúcar
dc.subject.eng.fl_str_mv Hemicellulolític enzymes
Fermentative process
Enzymatic hydrolysis
Sugarcane bagasse
dc.subject.cnpq.fl_str_mv CIENCIAS EXATAS E DA TERRA
description Currently, one of the major challenges for second generation ethanol is to reduce the cost of cellulolytic enzymes. Thus, the development of bioprocesses for the enzyme production on-site and strategies to increase the final yield of the enzymatic hydrolysis are required to ensure that biomass conversion to be economically feasible. Therefore, the objective of this work was to study the production and characterization of enzyme cocktails involved in the degradation of plant biomass by filamentous fungi Trichoderma reesei and Aspergillus niger grown in sequential fermentation and evaluate the application of these cocktails in the saccharification process of sugarcane bagasse. Firstly, evaluation and validation of sequential fermentation cultivation methodology to different strains of Trichoderma. Cultivation were made using sugarcane bagasse "in natura" and pretreated by steam explosion, as a carbon source. The result more significantly was observed for T. reesei Rut C30, the endoglucanase production was 4.2-fold higher than the values obtained in conventional submerged fermentation. The enzyme extracts were characterized in terms of optimum pH and temperature and endoglucanase profile. The thermostability was directly influenced by the type of carbon source and type of cultivation method. Subsequently, the proteomic analysis were performed of enzyme cocktails from T. reesei Rut C30 and A. niger A12 produced by submerged and sequential fermentation in the presence of pretreated bagasse. The performance of the enzyme cocktail in saccharification of pretreated bagasse showed that the combination of enzyme cocktails from T. reesei and A. niger produced by sequential fermentation had a yield than 3-fold higher than the enzyme cocktails of submerged fermentation. In order to evaluate the action of the enzyme cocktails produced by T. reesei and A. niger in sugarcane bagasse saccharification, the last step of the work was to study the additives effects during the sugarcane bagasse hydrolysis aiming at reducing non-productive adsorption of enzymes into lignin. The saccharification results in the presence of soybean protein were 2-fold higher than the controls (no additive) to the enzyme cocktails of two fungi studied produced by solid state fermentation, indicating the potential use of soybean protein as an additive to minimize non-productive adsorption of the enzyme into lignin. Overall, this study presents an interesting final contribution in the cellulase production process and the application of the enzyme cocktail in the hydrolysis of sugarcane bagasse.
publishDate 2016
dc.date.issued.fl_str_mv 2016-04-28
dc.date.accessioned.fl_str_mv 2017-02-08T12:10:12Z
dc.date.available.fl_str_mv 2017-02-08T12:10:12Z
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dc.identifier.citation.fl_str_mv FLORENCIO, Camila. Influência dos coquetéis enzimáticos produzidos por Trichoderma reesei e Aspergillus niger pelo processo de fermentação sequencial na hidrólise do bagaço de cana-de-açúcar. 2016. Tese (Doutorado em Biotecnologia) – Universidade Federal de São Carlos, São Carlos, 2016. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8504.
dc.identifier.uri.fl_str_mv https://repositorio.ufscar.br/handle/ufscar/8504
identifier_str_mv FLORENCIO, Camila. Influência dos coquetéis enzimáticos produzidos por Trichoderma reesei e Aspergillus niger pelo processo de fermentação sequencial na hidrólise do bagaço de cana-de-açúcar. 2016. Tese (Doutorado em Biotecnologia) – Universidade Federal de São Carlos, São Carlos, 2016. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8504.
url https://repositorio.ufscar.br/handle/ufscar/8504
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eng
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eng
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dc.publisher.none.fl_str_mv Universidade Federal de São Carlos
Câmpus São Carlos
dc.publisher.program.fl_str_mv Programa de Pós-Graduação em Biotecnologia - PPGBiotec
dc.publisher.initials.fl_str_mv UFSCar
publisher.none.fl_str_mv Universidade Federal de São Carlos
Câmpus São Carlos
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