Influência da morfologia e do cisalhamento na produção de enzimas celulolíticas por Aspergillus niger

Detalhes bibliográficos
Autor(a) principal: Buffo, Mariane Molina
Data de Publicação: 2020
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFSCAR
Texto Completo: https://repositorio.ufscar.br/handle/ufscar/12802
Resumo: Issues related to sustainability and development of renewable and less polluting processes are increasingly in evidence, due to the various environmental crises and a greater concern with the scarcity of non-renewable resources, such as oil. Therefore, a bio-based world economy is emerging in response to the global and environmental challenges. An example of a bio-based product are cellulolytic enzymes. This enzymes are produced mainly by filamentous fungi Trichoderma and Aspergillus. Such enzymes are important in several industrial sectors, how for example in the production of second generation ethanol, another important product of biological basis, renewable and environmentally friendly since it uses agro-industrial waste in its manufacture. However, one of the bottlenecks in the production of second generation ethanol, which makes it still uncompetitive compared to first generation ethanol and petrol oil, is the high cost of cellulolytic enzymes. The present thesis studied two important variables related to the production of cellulolytic enzymes, cell morphology and shear conditions, in an attempt to improve the production of cellulite enzymes by A.niger in bioreactors. The shear suffered by A.niger was quantified through the fragmentation suffered by the microorganism’s pellets in different bioreactors, impellers and operating conditions. Through image analysis and measurements of the average diameter of the pellets over time it was possible to propose a kinetics for the pellet fragmentation. The fragmentation of the pellets was mainly dependent on the type of bioreactor and the agitation speed. The airlift bioreactor was less harmful to the microorganism and higher agitation speeds took to more abrupt fragmentation. The shear kinetic parameters were directly related to the production of cellulolytic enzymes, with lower shear conditions favoring the production of endoglucanase and more intense conditions producing β-glucosidase. Regarding with the morphology, a technique of concomitant change of pH and spore concentration in the inoculum was successfully used to obtain different morphologies of A. niger, since larger pellets to almost completely dispersed morphology. Such morphological change was quantified through a proposed morphological parameter, which considered the frequency of each morphological class and a form factor that represents a characteristic dimension of each morphological form. The measures were possible through image treatment of the filamentous fungi, the images were obtained under a microscope and stereoscope using specific software to be treats. Inoculums with morphologies of pellets, clumps and dispersed were then tested in the production of cellulolytic enzymes, and a greater production of endoglucanase was obtained with inoculum in the pelletized form, value approximately three times higher when compared to dispersed morphology, for shaker results. In oposite a greater production of β-glucosidase was obtained with inoculum in the dispersed form, this value is four times higher in the flask assays and 160% higher in the bioreactors.Such results show the importance of the shear and morphology variables in the production of cellulolytic enzymes by A. niger and how such influences may be different, depending on the metabolite under study.
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spelling Buffo, Mariane MolinaBadino Júnior, Alberto Collihttp://lattes.cnpq.br/6244428434217018Farinas, Cristiane Sanchezhttp://lattes.cnpq.br/9933650905615452http://lattes.cnpq.br/4067015829295547b558d5a9-e4e2-4619-a964-f6e2de29b7cb2020-05-23T00:58:56Z2020-05-23T00:58:56Z2020-03-26BUFFO, Mariane Molina. Influência da morfologia e do cisalhamento na produção de enzimas celulolíticas por Aspergillus niger. 2020. Tese (Doutorado em Engenharia Química) – Universidade Federal de São Carlos, São Carlos, 2020. Disponível em: https://repositorio.ufscar.br/handle/ufscar/12802.https://repositorio.ufscar.br/handle/ufscar/12802Issues related to sustainability and development of renewable and less polluting processes are increasingly in evidence, due to the various environmental crises and a greater concern with the scarcity of non-renewable resources, such as oil. Therefore, a bio-based world economy is emerging in response to the global and environmental challenges. An example of a bio-based product are cellulolytic enzymes. This enzymes are produced mainly by filamentous fungi Trichoderma and Aspergillus. Such enzymes are important in several industrial sectors, how for example in the production of second generation ethanol, another important product of biological basis, renewable and environmentally friendly since it uses agro-industrial waste in its manufacture. However, one of the bottlenecks in the production of second generation ethanol, which makes it still uncompetitive compared to first generation ethanol and petrol oil, is the high cost of cellulolytic enzymes. The present thesis studied two important variables related to the production of cellulolytic enzymes, cell morphology and shear conditions, in an attempt to improve the production of cellulite enzymes by A.niger in bioreactors. The shear suffered by A.niger was quantified through the fragmentation suffered by the microorganism’s pellets in different bioreactors, impellers and operating conditions. Through image analysis and measurements of the average diameter of the pellets over time it was possible to propose a kinetics for the pellet fragmentation. The fragmentation of the pellets was mainly dependent on the type of bioreactor and the agitation speed. The airlift bioreactor was less harmful to the microorganism and higher agitation speeds took to more abrupt fragmentation. The shear kinetic parameters were directly related to the production of cellulolytic enzymes, with lower shear conditions favoring the production of endoglucanase and more intense conditions producing β-glucosidase. Regarding with the morphology, a technique of concomitant change of pH and spore concentration in the inoculum was successfully used to obtain different morphologies of A. niger, since larger pellets to almost completely dispersed morphology. Such morphological change was quantified through a proposed morphological parameter, which considered the frequency of each morphological class and a form factor that represents a characteristic dimension of each morphological form. The measures were possible through image treatment of the filamentous fungi, the images were obtained under a microscope and stereoscope using specific software to be treats. Inoculums with morphologies of pellets, clumps and dispersed were then tested in the production of cellulolytic enzymes, and a greater production of endoglucanase was obtained with inoculum in the pelletized form, value approximately three times higher when compared to dispersed morphology, for shaker results. In oposite a greater production of β-glucosidase was obtained with inoculum in the dispersed form, this value is four times higher in the flask assays and 160% higher in the bioreactors.Such results show the importance of the shear and morphology variables in the production of cellulolytic enzymes by A. niger and how such influences may be different, depending on the metabolite under study.Temas relacionados à sustentabilidade e ao desenvolvimento de processos renováveis e menos poluentes estão cada vez mais em evidência, devido às diversas crises ambientais e uma maior preocupação com a escassez de recursos não renováveis, como o petróleo. Assim, uma economia mundial de base biológica está emergindo em resposta a tais desafios globais e ambientais. Um exemplo de produto de base biológica são as enzimas celulolíticas produzidas principalmente por fungos filamentosos do gênero Trichoderma e Aspergillus. Tais enzimas são importantes em diversos setores industriais e um deles é na produção de etanol de segunda geração, outro importante produto de base biológica, renovável e ambientalmente correto, uma vez que utiliza de resíduos agroindústrias na sua fabricação. Porém, um dos gargalos da produção de etanol de segunda geração que o torna ainda não competitivo frente ao etanol de primeira geração e à gasolina é o alto custo das enzimas celulolíticas. Assim, a presente tese buscou estudar duas importantes variáveis relacionadas com a produção de enzimas celulolíticas, a morfologia celular e as condições de cisalhamento, na tentativa de melhorar a produção dessas enzimas por A.niger em biorreatores. O cisalhamento sofrido pelo A. niger foi quantificado através da fragmentação sofrida por pellets do microrganismo em diferentes biorreatores, impelidores e condições de operação. Através de análises de imagens e medidas do diâmetro médio dos pellets ao longo do tempo foi possível propor uma cinética de fragmentação dos pellets. A fragmentação dos pellets foi dependente principalmente do tipo de biorreator e da velocidade de agitação. O biorreator airlift foi menos danoso ao microrganismo e maiores velocidades de agitação levaram a uma fragmentação mais abrupta. Os parâmetros cinéticos de cisalhamento também tiveram relação direta com a produção de enzimas celulolíticas, sendo que menores condições de cisalhamento favoreceram a produção de endoglucanase e condições mais intensas a produção de β-glicosidase. Com relação à morfologia, utilizou-se com sucesso técnica de mudança concomitante do pH e da concentração de esporos do inóculo para se obter diferentes morfologias de A. niger, desde pellets maiores à morfologia quase que totalmente dispersa. Tal mudança morfológica pôde ser quantificada através de um parâmetro morfológico proposto, que considera a frequência de cada classe morfológica e um fator de forma que representa uma dimensão característica de cada forma morfológica. Tais medidas foram possíveis através do tratamento das imagens dos fungos filamentosos obtidas em microscópio e estereoscópio a partir de software específico. Inóculos com morfologias de pellets, clumps e dispersa foram então testados na produção de enzimas celulolíticas, sendo que uma maior produção de endoglucanase foi obtida com inóculo na forma pelletizada, valor aproximadamente três vezes maior quando comparada a morfologia dispersa, para os resultados em shaker. Já uma maior produção de β-glicosidase foi obtida com inóculo na forma dispersa, valor esse 4 vezes maior nos ensaios em frascos e 160% maior em ensaios em biorreator. Tais resultados mostram a importância das variáveis cisalhamento e morfologia na produção de enzimas celulolíticas por A. niger e como tais influências podem ser diferentes dependendo do metabólito em estudo.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)CAPES: Código de Financiamento 001porUniversidade Federal de São CarlosCâmpus São CarlosPrograma de Pós-Graduação em Engenharia Química - PPGEQUFSCarAttribution-NonCommercial-NoDerivs 3.0 Brazilhttp://creativecommons.org/licenses/by-nc-nd/3.0/br/info:eu-repo/semantics/openAccessBiorreatoresImpelidoresCisalhamentoFungo filamentosoMorfologiaEnzimasBioreactorsImpellersShearFilamentous fungiMorphologyEnzymesAspergillus nigerENGENHARIAS::ENGENHARIA QUIMICA::PROCESSOS INDUSTRIAIS DE ENGENHARIA QUIMICAInfluência da morfologia e do cisalhamento na produção de enzimas celulolíticas por Aspergillus nigerInfluence of morphology and shearing in the production of cellulolytic enzymes by Aspergillus nigerinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis60029d76c9b-aac3-4e11-bdb2-5988dc628937reponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINALTese_Mariane_Molina_Buffo_PPGEQ-Ufscar.pdfTese_Mariane_Molina_Buffo_PPGEQ-Ufscar.pdfTese de doutorado versão finalapplication/pdf5032191https://repositorio.ufscar.br/bitstream/ufscar/12802/1/Tese_Mariane_Molina_Buffo_PPGEQ-Ufscar.pdf7a3cab621289e9d4f3a62449eba7776eMD51Documentos_Mariane_declaracao_orientador.pdfDocumentos_Mariane_declaracao_orientador.pdfCarta_comprovante_orientadorapplication/pdf133407https://repositorio.ufscar.br/bitstream/ufscar/12802/3/Documentos_Mariane_declaracao_orientador.pdfa956f4ab727bc0158bcd9a7a71fc703bMD53CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8811https://repositorio.ufscar.br/bitstream/ufscar/12802/4/license_rdfe39d27027a6cc9cb039ad269a5db8e34MD54TEXTTese_Mariane_Molina_Buffo_PPGEQ-Ufscar.pdf.txtTese_Mariane_Molina_Buffo_PPGEQ-Ufscar.pdf.txtExtracted texttext/plain331900https://repositorio.ufscar.br/bitstream/ufscar/12802/5/Tese_Mariane_Molina_Buffo_PPGEQ-Ufscar.pdf.txtf43c5e13ab6d5142137e6803cf01b4c5MD55Documentos_Mariane_declaracao_orientador.pdf.txtDocumentos_Mariane_declaracao_orientador.pdf.txtExtracted texttext/plain915https://repositorio.ufscar.br/bitstream/ufscar/12802/7/Documentos_Mariane_declaracao_orientador.pdf.txt5043ed21adfa2705b61fc28dec8ff30bMD57THUMBNAILTese_Mariane_Molina_Buffo_PPGEQ-Ufscar.pdf.jpgTese_Mariane_Molina_Buffo_PPGEQ-Ufscar.pdf.jpgIM Thumbnailimage/jpeg6758https://repositorio.ufscar.br/bitstream/ufscar/12802/6/Tese_Mariane_Molina_Buffo_PPGEQ-Ufscar.pdf.jpg041856d06dcdb43199511472be0a0272MD56Documentos_Mariane_declaracao_orientador.pdf.jpgDocumentos_Mariane_declaracao_orientador.pdf.jpgIM Thumbnailimage/jpeg6958https://repositorio.ufscar.br/bitstream/ufscar/12802/8/Documentos_Mariane_declaracao_orientador.pdf.jpg878373e59c2cd2835b00d5102c32680bMD58ufscar/128022023-09-18 18:31:55.098oai:repositorio.ufscar.br:ufscar/12802Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:31:55Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false
dc.title.por.fl_str_mv Influência da morfologia e do cisalhamento na produção de enzimas celulolíticas por Aspergillus niger
dc.title.alternative.eng.fl_str_mv Influence of morphology and shearing in the production of cellulolytic enzymes by Aspergillus niger
title Influência da morfologia e do cisalhamento na produção de enzimas celulolíticas por Aspergillus niger
spellingShingle Influência da morfologia e do cisalhamento na produção de enzimas celulolíticas por Aspergillus niger
Buffo, Mariane Molina
Biorreatores
Impelidores
Cisalhamento
Fungo filamentoso
Morfologia
Enzimas
Bioreactors
Impellers
Shear
Filamentous fungi
Morphology
Enzymes
Aspergillus niger
ENGENHARIAS::ENGENHARIA QUIMICA::PROCESSOS INDUSTRIAIS DE ENGENHARIA QUIMICA
title_short Influência da morfologia e do cisalhamento na produção de enzimas celulolíticas por Aspergillus niger
title_full Influência da morfologia e do cisalhamento na produção de enzimas celulolíticas por Aspergillus niger
title_fullStr Influência da morfologia e do cisalhamento na produção de enzimas celulolíticas por Aspergillus niger
title_full_unstemmed Influência da morfologia e do cisalhamento na produção de enzimas celulolíticas por Aspergillus niger
title_sort Influência da morfologia e do cisalhamento na produção de enzimas celulolíticas por Aspergillus niger
author Buffo, Mariane Molina
author_facet Buffo, Mariane Molina
author_role author
dc.contributor.authorlattes.por.fl_str_mv http://lattes.cnpq.br/4067015829295547
dc.contributor.author.fl_str_mv Buffo, Mariane Molina
dc.contributor.advisor1.fl_str_mv Badino Júnior, Alberto Colli
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/6244428434217018
dc.contributor.advisor-co1.fl_str_mv Farinas, Cristiane Sanchez
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/9933650905615452
dc.contributor.authorID.fl_str_mv b558d5a9-e4e2-4619-a964-f6e2de29b7cb
contributor_str_mv Badino Júnior, Alberto Colli
Farinas, Cristiane Sanchez
dc.subject.por.fl_str_mv Biorreatores
Impelidores
Cisalhamento
Fungo filamentoso
Morfologia
Enzimas
topic Biorreatores
Impelidores
Cisalhamento
Fungo filamentoso
Morfologia
Enzimas
Bioreactors
Impellers
Shear
Filamentous fungi
Morphology
Enzymes
Aspergillus niger
ENGENHARIAS::ENGENHARIA QUIMICA::PROCESSOS INDUSTRIAIS DE ENGENHARIA QUIMICA
dc.subject.eng.fl_str_mv Bioreactors
Impellers
Shear
Filamentous fungi
Morphology
Enzymes
Aspergillus niger
dc.subject.cnpq.fl_str_mv ENGENHARIAS::ENGENHARIA QUIMICA::PROCESSOS INDUSTRIAIS DE ENGENHARIA QUIMICA
description Issues related to sustainability and development of renewable and less polluting processes are increasingly in evidence, due to the various environmental crises and a greater concern with the scarcity of non-renewable resources, such as oil. Therefore, a bio-based world economy is emerging in response to the global and environmental challenges. An example of a bio-based product are cellulolytic enzymes. This enzymes are produced mainly by filamentous fungi Trichoderma and Aspergillus. Such enzymes are important in several industrial sectors, how for example in the production of second generation ethanol, another important product of biological basis, renewable and environmentally friendly since it uses agro-industrial waste in its manufacture. However, one of the bottlenecks in the production of second generation ethanol, which makes it still uncompetitive compared to first generation ethanol and petrol oil, is the high cost of cellulolytic enzymes. The present thesis studied two important variables related to the production of cellulolytic enzymes, cell morphology and shear conditions, in an attempt to improve the production of cellulite enzymes by A.niger in bioreactors. The shear suffered by A.niger was quantified through the fragmentation suffered by the microorganism’s pellets in different bioreactors, impellers and operating conditions. Through image analysis and measurements of the average diameter of the pellets over time it was possible to propose a kinetics for the pellet fragmentation. The fragmentation of the pellets was mainly dependent on the type of bioreactor and the agitation speed. The airlift bioreactor was less harmful to the microorganism and higher agitation speeds took to more abrupt fragmentation. The shear kinetic parameters were directly related to the production of cellulolytic enzymes, with lower shear conditions favoring the production of endoglucanase and more intense conditions producing β-glucosidase. Regarding with the morphology, a technique of concomitant change of pH and spore concentration in the inoculum was successfully used to obtain different morphologies of A. niger, since larger pellets to almost completely dispersed morphology. Such morphological change was quantified through a proposed morphological parameter, which considered the frequency of each morphological class and a form factor that represents a characteristic dimension of each morphological form. The measures were possible through image treatment of the filamentous fungi, the images were obtained under a microscope and stereoscope using specific software to be treats. Inoculums with morphologies of pellets, clumps and dispersed were then tested in the production of cellulolytic enzymes, and a greater production of endoglucanase was obtained with inoculum in the pelletized form, value approximately three times higher when compared to dispersed morphology, for shaker results. In oposite a greater production of β-glucosidase was obtained with inoculum in the dispersed form, this value is four times higher in the flask assays and 160% higher in the bioreactors.Such results show the importance of the shear and morphology variables in the production of cellulolytic enzymes by A. niger and how such influences may be different, depending on the metabolite under study.
publishDate 2020
dc.date.accessioned.fl_str_mv 2020-05-23T00:58:56Z
dc.date.available.fl_str_mv 2020-05-23T00:58:56Z
dc.date.issued.fl_str_mv 2020-03-26
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dc.identifier.citation.fl_str_mv BUFFO, Mariane Molina. Influência da morfologia e do cisalhamento na produção de enzimas celulolíticas por Aspergillus niger. 2020. Tese (Doutorado em Engenharia Química) – Universidade Federal de São Carlos, São Carlos, 2020. Disponível em: https://repositorio.ufscar.br/handle/ufscar/12802.
dc.identifier.uri.fl_str_mv https://repositorio.ufscar.br/handle/ufscar/12802
identifier_str_mv BUFFO, Mariane Molina. Influência da morfologia e do cisalhamento na produção de enzimas celulolíticas por Aspergillus niger. 2020. Tese (Doutorado em Engenharia Química) – Universidade Federal de São Carlos, São Carlos, 2020. Disponível em: https://repositorio.ufscar.br/handle/ufscar/12802.
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rights_invalid_str_mv Attribution-NonCommercial-NoDerivs 3.0 Brazil
http://creativecommons.org/licenses/by-nc-nd/3.0/br/
eu_rights_str_mv openAccess
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Câmpus São Carlos
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