Influência dos íons metálicos na atividade e estabilidade de (hemi)celulases e no processo de sacarificação da biomassa
Autor(a) principal: | |
---|---|
Data de Publicação: | 2015 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFSCAR |
Texto Completo: | https://repositorio.ufscar.br/handle/ufscar/7616 |
Resumo: | The enzymes involved in the conversion process of lignocellulosic biomass into second generation (2G) ethanol add a high cost to the process. Studies reported in the literature indicate that some metal ions, easily found in nature, improve the activity and stability of enzymes and the biomass saccharification process when added to the enzymatic cocktail. In this context, this work aimed to evaluate the strategy of addition of metal ions in the crude enzymatic extract in order to increase the enzyme activity and stability and to improve the saccharification process. For the production of (hemi)cellulase enzymatic complex step, a selected strain of Aspergillus niger was cultivated under solid-state fermentation (SSF), submerged fermentation (SmF) and sequential fermentation (FS). Enzyme production and thermostability were evaluated also considering the effects of pretreatment (using steam-explosion and liquid-hot-water) of the sugarcane bagasse used as carbon source and inducer. For endoglucanase and xylanase enzymes, cultivation under SSF favored production when using steam-exploded and liquid-hot-water pretreated bagasse (both washed). Removal of inhibitors from the pretreated biomass employed in the cultivation media was necessary, because the presence of phenolic compounds restricted fungal growth. The findings revealed that endoglucanase and β-glucosidase produced under SSF were less prone to thermal deactivation. From these results, enzyme complex produced by SSF using the hydrothermal bagasse was selected to evaluate the influence of the divalent metal ions Ca, Co, Cu, Fe, Mg, Mn, Ni, Zn in two concentrations (2 and 10 mM) on enzymatic activity and stability. Most of the ions influenced differently the enzymatic activity and stability, at both concentrations. The most favorable influence was obtained by the Mn+2 ion, increasing by 57% the endoglucanase activity and maintaining the enzyme stable for 72 hours, besides it also showed no negative effects on β-glucosidase and xylanase. The Mn+2 ion was then selected for the saccharification of sugarcane bagasse submitted to different pretreatments. The addition of Mn2+ ion (10 mM) in the saccharification process using enzymatic extracts produced "in house" proved to be very effective by increasing the release of glucose up to 120% when using the acid pretreated bagasse (Bác). The increase was 55% for the washed exploded bagasse (BEXL) and 70% for the liquid-hot-water pretreated bagasse (BHT). Therefore, it was possible to validate that the addition of metal ions is able to positively influence the activity and stability of enzymes, and also the process of enzymatic saccharification of lignocellulose biomass. |
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Vasconcellos, Vanessa Molina deFarinas, Cristiane Sanchezhttp://lattes.cnpq.br/9933650905615452Giordano, Raquel de Lima Camargohttp://lattes.cnpq.br/9695542424889786http://lattes.cnpq.br/90507297570662727cd6bae4-9dde-4277-bc5a-baba500b2e332016-10-04T17:34:24Z2016-10-04T17:34:24Z2015-02-26VASCONCELLOS, Vanessa Molina de. Influência dos íons metálicos na atividade e estabilidade de (hemi)celulases e no processo de sacarificação da biomassa. 2015. Dissertação (Mestrado em Engenharia Química) – Universidade Federal de São Carlos, São Carlos, 2015. Disponível em: https://repositorio.ufscar.br/handle/ufscar/7616.https://repositorio.ufscar.br/handle/ufscar/7616The enzymes involved in the conversion process of lignocellulosic biomass into second generation (2G) ethanol add a high cost to the process. Studies reported in the literature indicate that some metal ions, easily found in nature, improve the activity and stability of enzymes and the biomass saccharification process when added to the enzymatic cocktail. In this context, this work aimed to evaluate the strategy of addition of metal ions in the crude enzymatic extract in order to increase the enzyme activity and stability and to improve the saccharification process. For the production of (hemi)cellulase enzymatic complex step, a selected strain of Aspergillus niger was cultivated under solid-state fermentation (SSF), submerged fermentation (SmF) and sequential fermentation (FS). Enzyme production and thermostability were evaluated also considering the effects of pretreatment (using steam-explosion and liquid-hot-water) of the sugarcane bagasse used as carbon source and inducer. For endoglucanase and xylanase enzymes, cultivation under SSF favored production when using steam-exploded and liquid-hot-water pretreated bagasse (both washed). Removal of inhibitors from the pretreated biomass employed in the cultivation media was necessary, because the presence of phenolic compounds restricted fungal growth. The findings revealed that endoglucanase and β-glucosidase produced under SSF were less prone to thermal deactivation. From these results, enzyme complex produced by SSF using the hydrothermal bagasse was selected to evaluate the influence of the divalent metal ions Ca, Co, Cu, Fe, Mg, Mn, Ni, Zn in two concentrations (2 and 10 mM) on enzymatic activity and stability. Most of the ions influenced differently the enzymatic activity and stability, at both concentrations. The most favorable influence was obtained by the Mn+2 ion, increasing by 57% the endoglucanase activity and maintaining the enzyme stable for 72 hours, besides it also showed no negative effects on β-glucosidase and xylanase. The Mn+2 ion was then selected for the saccharification of sugarcane bagasse submitted to different pretreatments. The addition of Mn2+ ion (10 mM) in the saccharification process using enzymatic extracts produced "in house" proved to be very effective by increasing the release of glucose up to 120% when using the acid pretreated bagasse (Bác). The increase was 55% for the washed exploded bagasse (BEXL) and 70% for the liquid-hot-water pretreated bagasse (BHT). Therefore, it was possible to validate that the addition of metal ions is able to positively influence the activity and stability of enzymes, and also the process of enzymatic saccharification of lignocellulose biomass.As enzimas envolvidas no processo de degradação da biomassa lignocelulósica para a produção do etanol de segunda geração (2G) agregam um alto custo ao processo. Estudos já reportados na literatura indicam que alguns íons metálicos, facilmente encontrados na natureza, ao serem adicionados ao coquetel enzimático atuam na melhoria da atividade e estabilidade das enzimas e no processo de sacarificação da biomassa. Nesse contexto, este trabalho teve como objetivo avaliar a estratégia de adição de íons metálicos ao extrato enzimático bruto com o intuito de aumentar a atividade e estabilidade enzimática e a melhoria na conversão do processo de sacarificação. Para a etapa de seleção do complexo (hemi)celulásico foi utilizada uma cepa selecionada de Aspergillus niger cultivada em fermentação em estado sólido (FES), fermentação submersa (FSm) e fermentação sequencial (FS). A produção de enzimas e a termoestabilidade foram avaliadas considerando os efeitos do pré-tratamento (explosão a vapor e hidrotérmico) do bagaço de cana-de-açúcar utilizado como fonte de carbono e indutor. A produção de endoglucanase e xilanase foi favorecida pelo cultivo em FES a partir do bagaço explodido e hidrotérmico (submetidos ao processo de lavagem). A remoção de inibidores da biomassa prétratada mostrou-se necessária, pois a presença de compostos fenólicos restringiu o crescimento fúngico em FES. Os resultados revelaram que endoglucanase e β-glicosidase produzidas sob FES foram menos propensas à desativação térmica. A partir desses resultados, foi selecionado o complexo enzimático produzido por FES utilizando o bagaço hidrotérmico lavado, para avaliação da influência dos íons metálicos bivalentes Ca, Co, Cu, Fe, Mg, Mn, Ni, Zn em duas concentrações (2 e 10 mM) na atividade e estabilidade enzimática. A maioria dos íons metálicos utilizados influenciou, de forma distinta, a atividade e estabilidade enzimática em ambas as concentrações para as enzimas estudadas. O íon Mn2+ se destacou, propiciando um aumento de 57% na atividade de endoglucanase e mantendo a enzima estável por 72 horas, além disso não apresentou efeito negativo para β-glicosidase e xilanase. Assim, o íon Mn2+ foi selecionado para o estudo da sacarificação dos bagaços de cana submetidos a diferentes pré-tratamentos. A adição do íon Mn2+ (10 mM) no processo de sacarificação utilizando extratos enzimáticos produzidos “in house” mostrou-se bastante eficaz no aumento da liberação de glicose, com ganhos percentuais de até 120% para o bagaço pré-tratado em meio ácido (Bác). Para o bagaço explodido lavado (BEXL) o aumento foi de 55 % e para o hidrotérmico (BHT) foi de 70%. Desse modo, foi possível validar que a adição de íons metálicos é capaz de influenciar positivamente tanto a atividade e estabilidade, como o processo de sacarificação enzimática da biomassa lignocelulósica.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)porUniversidade Federal de São CarlosCâmpus São CarlosPrograma de Pós-Graduação em Engenharia Química - PPGEQUFSCarAspergillus nigerÍons metálicosCompostos fenólicosMetal ionsPhenolic compoundsENGENHARIAS::ENGENHARIA QUIMICAInfluência dos íons metálicos na atividade e estabilidade de (hemi)celulases e no processo de sacarificação da biomassainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisOnline600600dd9ceb6c-d509-421a-a31e-bcb55bb21e02info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINALDissVMV.pdfDissVMV.pdfapplication/pdf2967521https://repositorio.ufscar.br/bitstream/ufscar/7616/1/DissVMV.pdf9bebee517d7d20c1b36810aa5e5744dfMD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81957https://repositorio.ufscar.br/bitstream/ufscar/7616/2/license.txtae0398b6f8b235e40ad82cba6c50031dMD52TEXTDissVMV.pdf.txtDissVMV.pdf.txtExtracted texttext/plain161935https://repositorio.ufscar.br/bitstream/ufscar/7616/3/DissVMV.pdf.txtbc027228a58ec91fbf42319c3f40c16aMD53THUMBNAILDissVMV.pdf.jpgDissVMV.pdf.jpgIM Thumbnailimage/jpeg5875https://repositorio.ufscar.br/bitstream/ufscar/7616/4/DissVMV.pdf.jpg1f78d0b0fd30a2dba90e4cdb331070a3MD54ufscar/76162023-09-18 18:31:48.725oai:repositorio.ufscar.br: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Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:31:48Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false |
dc.title.por.fl_str_mv |
Influência dos íons metálicos na atividade e estabilidade de (hemi)celulases e no processo de sacarificação da biomassa |
title |
Influência dos íons metálicos na atividade e estabilidade de (hemi)celulases e no processo de sacarificação da biomassa |
spellingShingle |
Influência dos íons metálicos na atividade e estabilidade de (hemi)celulases e no processo de sacarificação da biomassa Vasconcellos, Vanessa Molina de Aspergillus niger Íons metálicos Compostos fenólicos Metal ions Phenolic compounds ENGENHARIAS::ENGENHARIA QUIMICA |
title_short |
Influência dos íons metálicos na atividade e estabilidade de (hemi)celulases e no processo de sacarificação da biomassa |
title_full |
Influência dos íons metálicos na atividade e estabilidade de (hemi)celulases e no processo de sacarificação da biomassa |
title_fullStr |
Influência dos íons metálicos na atividade e estabilidade de (hemi)celulases e no processo de sacarificação da biomassa |
title_full_unstemmed |
Influência dos íons metálicos na atividade e estabilidade de (hemi)celulases e no processo de sacarificação da biomassa |
title_sort |
Influência dos íons metálicos na atividade e estabilidade de (hemi)celulases e no processo de sacarificação da biomassa |
author |
Vasconcellos, Vanessa Molina de |
author_facet |
Vasconcellos, Vanessa Molina de |
author_role |
author |
dc.contributor.authorlattes.por.fl_str_mv |
http://lattes.cnpq.br/9050729757066272 |
dc.contributor.author.fl_str_mv |
Vasconcellos, Vanessa Molina de |
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 |
Giordano, Raquel de Lima Camargo |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://lattes.cnpq.br/9695542424889786 |
dc.contributor.authorID.fl_str_mv |
7cd6bae4-9dde-4277-bc5a-baba500b2e33 |
contributor_str_mv |
Farinas, Cristiane Sanchez Giordano, Raquel de Lima Camargo |
dc.subject.por.fl_str_mv |
Aspergillus niger Íons metálicos Compostos fenólicos |
topic |
Aspergillus niger Íons metálicos Compostos fenólicos Metal ions Phenolic compounds ENGENHARIAS::ENGENHARIA QUIMICA |
dc.subject.eng.fl_str_mv |
Metal ions Phenolic compounds |
dc.subject.cnpq.fl_str_mv |
ENGENHARIAS::ENGENHARIA QUIMICA |
description |
The enzymes involved in the conversion process of lignocellulosic biomass into second generation (2G) ethanol add a high cost to the process. Studies reported in the literature indicate that some metal ions, easily found in nature, improve the activity and stability of enzymes and the biomass saccharification process when added to the enzymatic cocktail. In this context, this work aimed to evaluate the strategy of addition of metal ions in the crude enzymatic extract in order to increase the enzyme activity and stability and to improve the saccharification process. For the production of (hemi)cellulase enzymatic complex step, a selected strain of Aspergillus niger was cultivated under solid-state fermentation (SSF), submerged fermentation (SmF) and sequential fermentation (FS). Enzyme production and thermostability were evaluated also considering the effects of pretreatment (using steam-explosion and liquid-hot-water) of the sugarcane bagasse used as carbon source and inducer. For endoglucanase and xylanase enzymes, cultivation under SSF favored production when using steam-exploded and liquid-hot-water pretreated bagasse (both washed). Removal of inhibitors from the pretreated biomass employed in the cultivation media was necessary, because the presence of phenolic compounds restricted fungal growth. The findings revealed that endoglucanase and β-glucosidase produced under SSF were less prone to thermal deactivation. From these results, enzyme complex produced by SSF using the hydrothermal bagasse was selected to evaluate the influence of the divalent metal ions Ca, Co, Cu, Fe, Mg, Mn, Ni, Zn in two concentrations (2 and 10 mM) on enzymatic activity and stability. Most of the ions influenced differently the enzymatic activity and stability, at both concentrations. The most favorable influence was obtained by the Mn+2 ion, increasing by 57% the endoglucanase activity and maintaining the enzyme stable for 72 hours, besides it also showed no negative effects on β-glucosidase and xylanase. The Mn+2 ion was then selected for the saccharification of sugarcane bagasse submitted to different pretreatments. The addition of Mn2+ ion (10 mM) in the saccharification process using enzymatic extracts produced "in house" proved to be very effective by increasing the release of glucose up to 120% when using the acid pretreated bagasse (Bác). The increase was 55% for the washed exploded bagasse (BEXL) and 70% for the liquid-hot-water pretreated bagasse (BHT). Therefore, it was possible to validate that the addition of metal ions is able to positively influence the activity and stability of enzymes, and also the process of enzymatic saccharification of lignocellulose biomass. |
publishDate |
2015 |
dc.date.issued.fl_str_mv |
2015-02-26 |
dc.date.accessioned.fl_str_mv |
2016-10-04T17:34:24Z |
dc.date.available.fl_str_mv |
2016-10-04T17:34:24Z |
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info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
VASCONCELLOS, Vanessa Molina de. Influência dos íons metálicos na atividade e estabilidade de (hemi)celulases e no processo de sacarificação da biomassa. 2015. Dissertação (Mestrado em Engenharia Química) – Universidade Federal de São Carlos, São Carlos, 2015. Disponível em: https://repositorio.ufscar.br/handle/ufscar/7616. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/7616 |
identifier_str_mv |
VASCONCELLOS, Vanessa Molina de. Influência dos íons metálicos na atividade e estabilidade de (hemi)celulases e no processo de sacarificação da biomassa. 2015. Dissertação (Mestrado em Engenharia Química) – Universidade Federal de São Carlos, São Carlos, 2015. Disponível em: https://repositorio.ufscar.br/handle/ufscar/7616. |
url |
https://repositorio.ufscar.br/handle/ufscar/7616 |
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por |
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por |
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openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de São Carlos Câmpus São Carlos |
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Programa de Pós-Graduação em Engenharia Química - PPGEQ |
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UFSCar |
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Universidade Federal de São Carlos Câmpus São Carlos |
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