Produção de celulases por Penicillium sp. FSDE15 e hidrólise enzimática do sabugo de milho

Detalhes bibliográficos
Autor(a) principal: Santos, Felipe Augusto
Data de Publicação: 2021
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UFPB
Texto Completo: https://repositorio.ufpb.br/jspui/handle/123456789/21200
Resumo: Given the world scenario, where more and more waste is generated by the various sectors of industry and agriculture, special attention has been given to the reuse of these materials, such as corncob and straw and wheat bran. One of the alternatives is the production of products with high added value, such as enzymes, using these residues as substrates. Among the various enzymes, cellulases stand out (CMCases, FPases and -glucosidases), capable of converting part of the lignocellulosic biomass into fermentable sugars, which can be converted into cellulosic or second-generation ethanol. Therefore, this study aimed to produce cellulase enzymes through the filamentous fungus Penicillium sp. FSDE15, to carry out the enzymatic hydrolysis of the pretreated corncob aiming at the release of fermentable sugars for the production of second-generation ethanol. Initially, the pretreatment of the corncob was carried out. Three types of pre-treatment were applied, one acid, one alkaline and one hydrothermal. The conditions applied to each type of pretreatment were established from an experimental factorial design. In the case of chemical pretreatments, a 23 planning was used with three repetitions at the central point, varying the time (10, 40 and 70 min), temperature (40, 70 and 100 ⁰C) and reagent concentration (1, 2 and 3%). The hydrothermal pretreatment was carried out following an experimental factorial design of 22 with three replications in the central point, varying the time and temperature, using the same interval as that applied to the chemical pretreatments. Through fermentation tests (60% moisture, ambient temperature and spore concentration of 106 spores/mL), it was possible to obtain a maximum production of CMCase and FPase of 21.11 U/g and 1.29 U/g, respectively, in 216 h of fermentation, for cultivation carried out with wheat bran and corncob in the proportion of 50%. The -glucosidase activity reached its peak of 8.72 U/g in 216 h for cultivation carried out with pure wheat bran. In the results of enzymatic hydrolysis of corncob pretreated with H2SO4, a concentration of reducing sugars of 23.07 g/L of glucose was reached for hydrolysis with the enzymatic extract of Penicillium sp. FSDE15 and 47.62 g/L for hydrolysis using the commercial enzyme Celluclast®. For the corncob pretreated with NaOH, it was possible to obtain concentrations of 45.55 and 74.12 g/L of glucose for hydrolysis using the enzymatic extract and commercial cellulase, respectively. As for the hydrothermally pretreated corncob, 25.77 g/L of glucose were obtained for hydrolysis with the Penicillium extract and 21.34 g/L for the Celluclast® enzyme. All the best reducing sugar values were reached within 48 h of hydrolysis. These results, together with the study of scale-up for the production of bioethanol, demonstrate that the hydrothermally pretreated corncob, combined with the enzymatic extract produced by Penicillium sp. FSDE15, has great potential for the production of second generation ethanol.
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spelling Produção de celulases por Penicillium sp. FSDE15 e hidrólise enzimática do sabugo de milhoBiomassa lignocelulósicaFungo filamentosoEnzimasPré-tratamentoHidróliseLignocellulosic biomassFilamentous fungusEnzymesPretreatmentHydrolysisCNPQ::ENGENHARIAS::ENGENHARIA QUIMICAGiven the world scenario, where more and more waste is generated by the various sectors of industry and agriculture, special attention has been given to the reuse of these materials, such as corncob and straw and wheat bran. One of the alternatives is the production of products with high added value, such as enzymes, using these residues as substrates. Among the various enzymes, cellulases stand out (CMCases, FPases and -glucosidases), capable of converting part of the lignocellulosic biomass into fermentable sugars, which can be converted into cellulosic or second-generation ethanol. Therefore, this study aimed to produce cellulase enzymes through the filamentous fungus Penicillium sp. FSDE15, to carry out the enzymatic hydrolysis of the pretreated corncob aiming at the release of fermentable sugars for the production of second-generation ethanol. Initially, the pretreatment of the corncob was carried out. Three types of pre-treatment were applied, one acid, one alkaline and one hydrothermal. The conditions applied to each type of pretreatment were established from an experimental factorial design. In the case of chemical pretreatments, a 23 planning was used with three repetitions at the central point, varying the time (10, 40 and 70 min), temperature (40, 70 and 100 ⁰C) and reagent concentration (1, 2 and 3%). The hydrothermal pretreatment was carried out following an experimental factorial design of 22 with three replications in the central point, varying the time and temperature, using the same interval as that applied to the chemical pretreatments. Through fermentation tests (60% moisture, ambient temperature and spore concentration of 106 spores/mL), it was possible to obtain a maximum production of CMCase and FPase of 21.11 U/g and 1.29 U/g, respectively, in 216 h of fermentation, for cultivation carried out with wheat bran and corncob in the proportion of 50%. The -glucosidase activity reached its peak of 8.72 U/g in 216 h for cultivation carried out with pure wheat bran. In the results of enzymatic hydrolysis of corncob pretreated with H2SO4, a concentration of reducing sugars of 23.07 g/L of glucose was reached for hydrolysis with the enzymatic extract of Penicillium sp. FSDE15 and 47.62 g/L for hydrolysis using the commercial enzyme Celluclast®. For the corncob pretreated with NaOH, it was possible to obtain concentrations of 45.55 and 74.12 g/L of glucose for hydrolysis using the enzymatic extract and commercial cellulase, respectively. As for the hydrothermally pretreated corncob, 25.77 g/L of glucose were obtained for hydrolysis with the Penicillium extract and 21.34 g/L for the Celluclast® enzyme. All the best reducing sugar values were reached within 48 h of hydrolysis. These results, together with the study of scale-up for the production of bioethanol, demonstrate that the hydrothermally pretreated corncob, combined with the enzymatic extract produced by Penicillium sp. FSDE15, has great potential for the production of second generation ethanol.NenhumaDiante do cenário mundial, onde cada vez mais resíduos são gerados pelos diversos setores da indústria e da agricultura, uma atenção especial vem sendo dada ao reaproveitamento desses materiais, como sabugo e palha de milho e farelo de trigo. Uma das alternativas é a produção de produtos de alto valor agregado, como enzimas, a partir da utilização desses resíduos como substratos. Entre as diversas enzimas, destacam-se as celulases (CMCases, FPases e -glicosidases), capazes de converter parte da biomassa lignocelulósica em açúcares fermentescíveis, esses podem ser convertidos em etanol celulósico ou de segunda geração. Diante disso, este estudo teve como objetivo a produção de enzimas celulases, por meio do fungo filamentoso Penicillium sp. FSDE15, para realização da hidrólise enzimática do sabugo de milho pré-tratado visando a liberação de açúcares fermentescíveis para produção de etanol de segunda geração. Inicialmente foi realizado o pré-tratamento do sabugo de milho. Três tipos de pré-tratamento foram aplicados, sendo um ácido, um alcalino e um hidrotérmico. As condições aplicadas a cada tipo de pré-tratamento foram estabelecidas a partir de um planejamento fatorial experimental. No caso dos pré-tratamentos químicos, empregou-se um planejamento 23 com três repetições no ponto central, variando-se o tempo (10, 40 e 70 min), temperatura (40, 70 e 100 ⁰C) e concentração de reagente (1, 2 e 3%). O pré-tratamento hidrotérmico foi realizado seguindo um planejamento fatorial experimental de 22 com três repetições no ponto central, variando-se o tempo e a temperatura, sendo empregado o mesmo intervalo que o aplicado aos pré-tratamentos químicos. Por meio dos ensaios fermentativos (60% de umidade, temperatura ambiente e concentração de esporos de 106 esporos/mL), foi possível obter uma máxima produção de CMCase e FPase de 21,11 U/g e 1,29 U/g, respectivamente, em 216 h de fermentação, para cultivo realizado com farelo de trigo e sabugo de milho na proporção de 50%. A atividade -glicosidase atingiu seu pico de 8,72 U/g em 216 h para cultivo realizado com farelo de trigo puro. Nos resultados de hidrólise enzimática do sabugo pré-tratado com H2SO4, atingiu-se uma concentração de açúcares redutores (AR) de 23,07 g/L de glicose para a hidrólise com o extrato enzimático de Penicillium sp. FSDE15 e 47,62 g/L para a hidrólise utilizando a enzima comercial Celluclast®. Para o sabugo pré-tratado com NaOH, foi possível obter concentrações de 45,55 e 74,12 g/L de glicose, para as hidrólises utilizando o extrato enzimático e a celulase comercial, respectivamente. Já para o sabugo pré-tratado hidrotermicamente, obteve-se 25,77 g/L de glicose para a hidrólise com o extrato de Penicillium e 21,34 g/L para a enzima Celluclast®. Todos os melhores valores de açúcares redutores foram atingidos em 48 h de hidrólise. Esses resultados, juntamente com o estudo do aumento de escala para a produção de bioetanol, demonstram que o sabugo pré-tratado hidrotermicamente, aliado com o extrato enzimático produzido por Penicillium sp. FSDE15, possui grande potencial para a produção de etanol de segunda geração.Universidade Federal da ParaíbaBrasilEngenharia QuímicaPrograma de Pós-Graduação em Engenharia QuímicaUFPBSantos, Sharline Florentino de Melohttp://lattes.cnpq.br/4846443214585734Gonçalves, Laís Campos Teixeira de Carvalhohttp://lattes.cnpq.br/3411642497113875Santos, Felipe Augusto2021-10-13T13:43:45Z2021-06-152021-10-13T13:43:45Z2021-05-27info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesishttps://repositorio.ufpb.br/jspui/handle/123456789/21200porhttp://creativecommons.org/licenses/by-nd/3.0/br/info:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da UFPBinstname:Universidade Federal da Paraíba (UFPB)instacron:UFPB2022-08-09T17:34:03Zoai:repositorio.ufpb.br:123456789/21200Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufpb.br/PUBhttp://tede.biblioteca.ufpb.br:8080/oai/requestdiretoria@ufpb.br|| diretoria@ufpb.bropendoar:2022-08-09T17:34:03Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB)false
dc.title.none.fl_str_mv Produção de celulases por Penicillium sp. FSDE15 e hidrólise enzimática do sabugo de milho
title Produção de celulases por Penicillium sp. FSDE15 e hidrólise enzimática do sabugo de milho
spellingShingle Produção de celulases por Penicillium sp. FSDE15 e hidrólise enzimática do sabugo de milho
Santos, Felipe Augusto
Biomassa lignocelulósica
Fungo filamentoso
Enzimas
Pré-tratamento
Hidrólise
Lignocellulosic biomass
Filamentous fungus
Enzymes
Pretreatment
Hydrolysis
CNPQ::ENGENHARIAS::ENGENHARIA QUIMICA
title_short Produção de celulases por Penicillium sp. FSDE15 e hidrólise enzimática do sabugo de milho
title_full Produção de celulases por Penicillium sp. FSDE15 e hidrólise enzimática do sabugo de milho
title_fullStr Produção de celulases por Penicillium sp. FSDE15 e hidrólise enzimática do sabugo de milho
title_full_unstemmed Produção de celulases por Penicillium sp. FSDE15 e hidrólise enzimática do sabugo de milho
title_sort Produção de celulases por Penicillium sp. FSDE15 e hidrólise enzimática do sabugo de milho
author Santos, Felipe Augusto
author_facet Santos, Felipe Augusto
author_role author
dc.contributor.none.fl_str_mv Santos, Sharline Florentino de Melo
http://lattes.cnpq.br/4846443214585734
Gonçalves, Laís Campos Teixeira de Carvalho
http://lattes.cnpq.br/3411642497113875
dc.contributor.author.fl_str_mv Santos, Felipe Augusto
dc.subject.por.fl_str_mv Biomassa lignocelulósica
Fungo filamentoso
Enzimas
Pré-tratamento
Hidrólise
Lignocellulosic biomass
Filamentous fungus
Enzymes
Pretreatment
Hydrolysis
CNPQ::ENGENHARIAS::ENGENHARIA QUIMICA
topic Biomassa lignocelulósica
Fungo filamentoso
Enzimas
Pré-tratamento
Hidrólise
Lignocellulosic biomass
Filamentous fungus
Enzymes
Pretreatment
Hydrolysis
CNPQ::ENGENHARIAS::ENGENHARIA QUIMICA
description Given the world scenario, where more and more waste is generated by the various sectors of industry and agriculture, special attention has been given to the reuse of these materials, such as corncob and straw and wheat bran. One of the alternatives is the production of products with high added value, such as enzymes, using these residues as substrates. Among the various enzymes, cellulases stand out (CMCases, FPases and -glucosidases), capable of converting part of the lignocellulosic biomass into fermentable sugars, which can be converted into cellulosic or second-generation ethanol. Therefore, this study aimed to produce cellulase enzymes through the filamentous fungus Penicillium sp. FSDE15, to carry out the enzymatic hydrolysis of the pretreated corncob aiming at the release of fermentable sugars for the production of second-generation ethanol. Initially, the pretreatment of the corncob was carried out. Three types of pre-treatment were applied, one acid, one alkaline and one hydrothermal. The conditions applied to each type of pretreatment were established from an experimental factorial design. In the case of chemical pretreatments, a 23 planning was used with three repetitions at the central point, varying the time (10, 40 and 70 min), temperature (40, 70 and 100 ⁰C) and reagent concentration (1, 2 and 3%). The hydrothermal pretreatment was carried out following an experimental factorial design of 22 with three replications in the central point, varying the time and temperature, using the same interval as that applied to the chemical pretreatments. Through fermentation tests (60% moisture, ambient temperature and spore concentration of 106 spores/mL), it was possible to obtain a maximum production of CMCase and FPase of 21.11 U/g and 1.29 U/g, respectively, in 216 h of fermentation, for cultivation carried out with wheat bran and corncob in the proportion of 50%. The -glucosidase activity reached its peak of 8.72 U/g in 216 h for cultivation carried out with pure wheat bran. In the results of enzymatic hydrolysis of corncob pretreated with H2SO4, a concentration of reducing sugars of 23.07 g/L of glucose was reached for hydrolysis with the enzymatic extract of Penicillium sp. FSDE15 and 47.62 g/L for hydrolysis using the commercial enzyme Celluclast®. For the corncob pretreated with NaOH, it was possible to obtain concentrations of 45.55 and 74.12 g/L of glucose for hydrolysis using the enzymatic extract and commercial cellulase, respectively. As for the hydrothermally pretreated corncob, 25.77 g/L of glucose were obtained for hydrolysis with the Penicillium extract and 21.34 g/L for the Celluclast® enzyme. All the best reducing sugar values were reached within 48 h of hydrolysis. These results, together with the study of scale-up for the production of bioethanol, demonstrate that the hydrothermally pretreated corncob, combined with the enzymatic extract produced by Penicillium sp. FSDE15, has great potential for the production of second generation ethanol.
publishDate 2021
dc.date.none.fl_str_mv 2021-10-13T13:43:45Z
2021-06-15
2021-10-13T13:43:45Z
2021-05-27
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://repositorio.ufpb.br/jspui/handle/123456789/21200
url https://repositorio.ufpb.br/jspui/handle/123456789/21200
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv http://creativecommons.org/licenses/by-nd/3.0/br/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nd/3.0/br/
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Universidade Federal da Paraíba
Brasil
Engenharia Química
Programa de Pós-Graduação em Engenharia Química
UFPB
publisher.none.fl_str_mv Universidade Federal da Paraíba
Brasil
Engenharia Química
Programa de Pós-Graduação em Engenharia Química
UFPB
dc.source.none.fl_str_mv reponame:Biblioteca Digital de Teses e Dissertações da UFPB
instname:Universidade Federal da Paraíba (UFPB)
instacron:UFPB
instname_str Universidade Federal da Paraíba (UFPB)
instacron_str UFPB
institution UFPB
reponame_str Biblioteca Digital de Teses e Dissertações da UFPB
collection Biblioteca Digital de Teses e Dissertações da UFPB
repository.name.fl_str_mv Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB)
repository.mail.fl_str_mv diretoria@ufpb.br|| diretoria@ufpb.br
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