Trichoderma koningiopsis 2OI2A1M: produção de enzimas do complexo hemicelulolítico e aplicação na sacarificação do bagaço de cevada

Detalhes bibliográficos
Autor(a) principal: Kerber, Caroline Marlene da Cruz
Data de Publicação: 2020
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações do UNIOESTE
Texto Completo: http://tede.unioeste.br/handle/tede/5691
Resumo: Filamentous fungi are excellent sources of industrial enzymes because of their high production and secretion capacity to the extracellular environment. Xylanases, cellulases, pectinases, amylases and invertases are examples of hydrolases, which have wide application in different industrial sectors. There are several uses of these enzymes, notably the food and beverage, animal feed, textile and pulp and paper industries. In addition, there is growing interest in the use of agricultural waste as a raw material in the production of bioethanol. Trichoderma fungi are well known and exploited for producing multiple enzymes for polysaccharide degradation, such as cellulose and hemicellulose. The Trichoderma koningiopsis species is reported as cosmopolitan but is still little explored in the biotechnological field. Thus, this study aimed to investigate the ability of T. koningiopsis 2OI2A1M fungus, previously isolated from the Paraná Atlantic Forest Biome, to produce enzymes that act on the depolymerization of lignocellulosic biomass found in agroindustrial byproducts, aiming at an application of these enzymes in saccharification of barley bagasse. The production of xylanase by T. koningiopsis 2OI2A1M fungus using different agroindustrial residues showed that barley bagasse provided higher enzymatic activity (83.91 U mL-1). Additionally, it has been found that kraft paper is capable of inducing xylanase production (51.93 U mL-1). Among the extracellular β-xylosidases and β- glucosidases, sorghum biomass and carboxymethylcellulose were the best inducer with 22.63 and 36.40 U mL-1, respectively. The crude extract xylanase of T. koningiopsis 2OI2A1M exhibited maximum activity pH and temperature at 5.0 and 60 °C, respectively. The xylanase activity remained totally preserved for 6 days of incubation at 4 °C, in the pH range 3.5-7.0, however, its activity was affected when incubated for more than 60 min at temperatures above 40 °C. The production of fermentable sugars from the enzymatic saccharification of barley bagasse, the in natura condition, yielded a higher concentration of glucose (5.9%), total reducing sugars (13.3%), and xylose (2.6%) after 48 h. The comparative analysis of the morphological structure of the barley bagasse by scanning electron microscopy demonstrated that the simultaneous action of T. koningiopsis 2OI2A1M enzymes does not require previous structural modifications in the barley bagasse for effectiveness in the saccharification process. Thus, the fungus T. koningiopsis 2OI2A1M has potential to produce enzymes that act on the depolymerization of lignocellulosic biomass and barley bagasse is an attractive candidate for a sustainable alternative for the production of fermentable sugars, a primary focus in the production of secondgeneration fuels such as bioethanol.
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spelling Kadowaki, Marina Kimikohttp://lattes.cnpq.br/1819723253019762Kadowaki, Marina Kimikohttp://lattes.cnpq.br/1819723253019762Maller, Alexandrehttp://lattes.cnpq.br/8153318875076127Heinen, Paulo Ricardohttp://lattes.cnpq.br/1492513157306177http://lattes.cnpq.br/3860746841962484Kerber, Caroline Marlene da Cruz2021-11-29T13:40:34Z2020-11-30KERBER, Caroline Marlene da Cruz. Trichoderma koningiopsis 2OI2A1M: produção de enzimas do complexo hemicelulolítico e aplicação na sacarificação do bagaço de cevada. 2020. 59 f. Dissertação (Mestrado em Ciências Farmacêuticas) - Universidade Estadual do Oeste do Paraná, Cascavel - PR.http://tede.unioeste.br/handle/tede/5691Filamentous fungi are excellent sources of industrial enzymes because of their high production and secretion capacity to the extracellular environment. Xylanases, cellulases, pectinases, amylases and invertases are examples of hydrolases, which have wide application in different industrial sectors. There are several uses of these enzymes, notably the food and beverage, animal feed, textile and pulp and paper industries. In addition, there is growing interest in the use of agricultural waste as a raw material in the production of bioethanol. Trichoderma fungi are well known and exploited for producing multiple enzymes for polysaccharide degradation, such as cellulose and hemicellulose. The Trichoderma koningiopsis species is reported as cosmopolitan but is still little explored in the biotechnological field. Thus, this study aimed to investigate the ability of T. koningiopsis 2OI2A1M fungus, previously isolated from the Paraná Atlantic Forest Biome, to produce enzymes that act on the depolymerization of lignocellulosic biomass found in agroindustrial byproducts, aiming at an application of these enzymes in saccharification of barley bagasse. The production of xylanase by T. koningiopsis 2OI2A1M fungus using different agroindustrial residues showed that barley bagasse provided higher enzymatic activity (83.91 U mL-1). Additionally, it has been found that kraft paper is capable of inducing xylanase production (51.93 U mL-1). Among the extracellular β-xylosidases and β- glucosidases, sorghum biomass and carboxymethylcellulose were the best inducer with 22.63 and 36.40 U mL-1, respectively. The crude extract xylanase of T. koningiopsis 2OI2A1M exhibited maximum activity pH and temperature at 5.0 and 60 °C, respectively. The xylanase activity remained totally preserved for 6 days of incubation at 4 °C, in the pH range 3.5-7.0, however, its activity was affected when incubated for more than 60 min at temperatures above 40 °C. The production of fermentable sugars from the enzymatic saccharification of barley bagasse, the in natura condition, yielded a higher concentration of glucose (5.9%), total reducing sugars (13.3%), and xylose (2.6%) after 48 h. The comparative analysis of the morphological structure of the barley bagasse by scanning electron microscopy demonstrated that the simultaneous action of T. koningiopsis 2OI2A1M enzymes does not require previous structural modifications in the barley bagasse for effectiveness in the saccharification process. Thus, the fungus T. koningiopsis 2OI2A1M has potential to produce enzymes that act on the depolymerization of lignocellulosic biomass and barley bagasse is an attractive candidate for a sustainable alternative for the production of fermentable sugars, a primary focus in the production of secondgeneration fuels such as bioethanol.Fungos filamentosos são excelentes fontes de enzimas industriais, em razão da elevada capacidade de produção e secreção dessas para o meio extracelular. Xilanases, celulases, pectinases, amilases e invertases são exemplos de hidrolases, as quais possuem ampla aplicação nos diferentes setores industriais. Diversas são as utilizações dessas enzimas, merecendo destaque as indústrias alimentícia e de bebidas, de ração animal, têxtil e de polpa e papel. Além disso, é crescente o interesse pela utilização de resíduos agrícolas como matéria-prima na produção de bioetanol. Os fungos do gênero Trichoderma são bem conhecidos e explorados por produzirem múltiplas enzimas para a degradação de polissacarídeos, como a celulose e a hemicelulose. A espécie Trichoderma koningiopsis é relatada como cosmopolita, porém, ainda é pouco explorada no campo biotecnológico. Dessa forma, este estudo teve como objetivo principal, investigar a habilidade do fungo T. koningiopsis 2OI2A1M, previamente isolado do Bioma da Mata Atlântica do Paraná, em produzir enzimas que atuem na despolimerização da biomassa lignocelulósica encontrada em subprodutos agroindustriais, visando uma aplicação dessas enzimas na sacarificação do bagaço de cevada. Ao avaliar-se a produção xilanásica pelo fungo T. koningiopsis 2OI2A1M, utilizando diferentes resíduos agroindustriais, verificou-se que o bagaço de cevada exibiu maior atividade enzimática (83,91 U mL-1). Adicionalmente, verificou-se que o papel kraft é capaz de induzir a produção de xilanase (51,93 U mL-1). Dentre as β-xilosidases e β-glicosidases extracelulares, o sorgo biomassa e a carboximetilcelulose foram os melhores indutores com 22,63 e 36,40 U mL-1, respectivamente. A xilanase do extrato bruto de T. koningiopsis 2OI2A1M exibiu pH e temperatura de máxima atividade em 5,0 e 60 °C, respectivamente. A atividade xilanásica manteve-se totalmente preservada por 6 dias de incubação a 4 °C, na faixa de pH 3,5-7,0, porém, a sua atividade foi afetada quando incubada por mais de 60 min em temperaturas superiores a 40 °C. Para a produção de açúcares fermentáveis a partir da sacarificação enzimática do bagaço de cevada, a condição in natura foi a melhor resposta para obtenção de glicose (5,9%), açúcares redutores totais (13,3%) e xilose (2,6%) após 48 h de hidrólise enzimática. A análise comparativa da estrutura morfológica do bagaço de cevada por microscopia eletrônica de varredura demonstrou que a ação conjunta das enzimas de T. koningiopsis 2OI2A1M não necessita de modificações estruturais no bagaço de cevada para eficácia no processo de sacarificação, portanto, sendo dispensável os pré-tratamentos químico ou físico do substrato. Diante do exposto, o fungo T. koningiopsis 2OI2A1M apresenta potencial em produzir enzimas que atuam na despolimerização da biomassa lignocelulósica e o bagaço de cevada torna-se uma alternativa sustentável para produção de açúcares fermentáveis, com foco primordial na produção de combustíveis de segunda geração, como o bioetanol.Submitted by Neusa Fagundes (neusa.fagundes@unioeste.br) on 2021-11-29T13:40:34Z No. of bitstreams: 2 Caroline_Kerber2020.pdf: 1480311 bytes, checksum: 558d9aefa0ada34a74d9c12bfbb3ef39 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Made available in DSpace on 2021-11-29T13:40:34Z (GMT). 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dc.title.por.fl_str_mv Trichoderma koningiopsis 2OI2A1M: produção de enzimas do complexo hemicelulolítico e aplicação na sacarificação do bagaço de cevada
dc.title.alternative.eng.fl_str_mv Trichoderma koningiopsis 2OI2A1M: enzyme production of the complex hemicellulolytic and application in the saccharification of barley bagasse
title Trichoderma koningiopsis 2OI2A1M: produção de enzimas do complexo hemicelulolítico e aplicação na sacarificação do bagaço de cevada
spellingShingle Trichoderma koningiopsis 2OI2A1M: produção de enzimas do complexo hemicelulolítico e aplicação na sacarificação do bagaço de cevada
Kerber, Caroline Marlene da Cruz
Açúcares fermentáveis
β-glicosidase
Conversão de biomassa
Endo-xilanase
Pré-tratamento
Screening enzimático
Fermentable sugars
β-glucosidase
Biomass conversion
Endoxylanase
Pretreatment
Enzymatic screening
CIENCIAS DA SAUDE::FARMACIA
title_short Trichoderma koningiopsis 2OI2A1M: produção de enzimas do complexo hemicelulolítico e aplicação na sacarificação do bagaço de cevada
title_full Trichoderma koningiopsis 2OI2A1M: produção de enzimas do complexo hemicelulolítico e aplicação na sacarificação do bagaço de cevada
title_fullStr Trichoderma koningiopsis 2OI2A1M: produção de enzimas do complexo hemicelulolítico e aplicação na sacarificação do bagaço de cevada
title_full_unstemmed Trichoderma koningiopsis 2OI2A1M: produção de enzimas do complexo hemicelulolítico e aplicação na sacarificação do bagaço de cevada
title_sort Trichoderma koningiopsis 2OI2A1M: produção de enzimas do complexo hemicelulolítico e aplicação na sacarificação do bagaço de cevada
author Kerber, Caroline Marlene da Cruz
author_facet Kerber, Caroline Marlene da Cruz
author_role author
dc.contributor.advisor1.fl_str_mv Kadowaki, Marina Kimiko
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/1819723253019762
dc.contributor.referee1.fl_str_mv Kadowaki, Marina Kimiko
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/1819723253019762
dc.contributor.referee2.fl_str_mv Maller, Alexandre
dc.contributor.referee2Lattes.fl_str_mv http://lattes.cnpq.br/8153318875076127
dc.contributor.referee3.fl_str_mv Heinen, Paulo Ricardo
dc.contributor.referee3Lattes.fl_str_mv http://lattes.cnpq.br/1492513157306177
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/3860746841962484
dc.contributor.author.fl_str_mv Kerber, Caroline Marlene da Cruz
contributor_str_mv Kadowaki, Marina Kimiko
Kadowaki, Marina Kimiko
Maller, Alexandre
Heinen, Paulo Ricardo
dc.subject.por.fl_str_mv Açúcares fermentáveis
β-glicosidase
Conversão de biomassa
Endo-xilanase
Pré-tratamento
Screening enzimático
topic Açúcares fermentáveis
β-glicosidase
Conversão de biomassa
Endo-xilanase
Pré-tratamento
Screening enzimático
Fermentable sugars
β-glucosidase
Biomass conversion
Endoxylanase
Pretreatment
Enzymatic screening
CIENCIAS DA SAUDE::FARMACIA
dc.subject.eng.fl_str_mv Fermentable sugars
β-glucosidase
Biomass conversion
Endoxylanase
Pretreatment
Enzymatic screening
dc.subject.cnpq.fl_str_mv CIENCIAS DA SAUDE::FARMACIA
description Filamentous fungi are excellent sources of industrial enzymes because of their high production and secretion capacity to the extracellular environment. Xylanases, cellulases, pectinases, amylases and invertases are examples of hydrolases, which have wide application in different industrial sectors. There are several uses of these enzymes, notably the food and beverage, animal feed, textile and pulp and paper industries. In addition, there is growing interest in the use of agricultural waste as a raw material in the production of bioethanol. Trichoderma fungi are well known and exploited for producing multiple enzymes for polysaccharide degradation, such as cellulose and hemicellulose. The Trichoderma koningiopsis species is reported as cosmopolitan but is still little explored in the biotechnological field. Thus, this study aimed to investigate the ability of T. koningiopsis 2OI2A1M fungus, previously isolated from the Paraná Atlantic Forest Biome, to produce enzymes that act on the depolymerization of lignocellulosic biomass found in agroindustrial byproducts, aiming at an application of these enzymes in saccharification of barley bagasse. The production of xylanase by T. koningiopsis 2OI2A1M fungus using different agroindustrial residues showed that barley bagasse provided higher enzymatic activity (83.91 U mL-1). Additionally, it has been found that kraft paper is capable of inducing xylanase production (51.93 U mL-1). Among the extracellular β-xylosidases and β- glucosidases, sorghum biomass and carboxymethylcellulose were the best inducer with 22.63 and 36.40 U mL-1, respectively. The crude extract xylanase of T. koningiopsis 2OI2A1M exhibited maximum activity pH and temperature at 5.0 and 60 °C, respectively. The xylanase activity remained totally preserved for 6 days of incubation at 4 °C, in the pH range 3.5-7.0, however, its activity was affected when incubated for more than 60 min at temperatures above 40 °C. The production of fermentable sugars from the enzymatic saccharification of barley bagasse, the in natura condition, yielded a higher concentration of glucose (5.9%), total reducing sugars (13.3%), and xylose (2.6%) after 48 h. The comparative analysis of the morphological structure of the barley bagasse by scanning electron microscopy demonstrated that the simultaneous action of T. koningiopsis 2OI2A1M enzymes does not require previous structural modifications in the barley bagasse for effectiveness in the saccharification process. Thus, the fungus T. koningiopsis 2OI2A1M has potential to produce enzymes that act on the depolymerization of lignocellulosic biomass and barley bagasse is an attractive candidate for a sustainable alternative for the production of fermentable sugars, a primary focus in the production of secondgeneration fuels such as bioethanol.
publishDate 2020
dc.date.issued.fl_str_mv 2020-11-30
dc.date.accessioned.fl_str_mv 2021-11-29T13:40:34Z
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.citation.fl_str_mv KERBER, Caroline Marlene da Cruz. Trichoderma koningiopsis 2OI2A1M: produção de enzimas do complexo hemicelulolítico e aplicação na sacarificação do bagaço de cevada. 2020. 59 f. Dissertação (Mestrado em Ciências Farmacêuticas) - Universidade Estadual do Oeste do Paraná, Cascavel - PR.
dc.identifier.uri.fl_str_mv http://tede.unioeste.br/handle/tede/5691
identifier_str_mv KERBER, Caroline Marlene da Cruz. Trichoderma koningiopsis 2OI2A1M: produção de enzimas do complexo hemicelulolítico e aplicação na sacarificação do bagaço de cevada. 2020. 59 f. Dissertação (Mestrado em Ciências Farmacêuticas) - Universidade Estadual do Oeste do Paraná, Cascavel - PR.
url http://tede.unioeste.br/handle/tede/5691
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