ISOLAMENTO E BIOPROSPECÇÃO DE FUNGOS TERMOFÍLICOS ISOLADOS DA FLORESTA OMBRÓFILA MISTA: PRODUÇÃO DE XILANASES POR Aspergillus fumigatus
Ano de defesa: | 2020 |
---|---|
Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | |
Tipo de documento: | Dissertação |
Tipo de acesso: | Acesso aberto |
Idioma: | por |
Instituição de defesa: |
Universidade Estadual do Centro-Oeste
|
Programa de Pós-Graduação: |
Programa de Pós-Graduação em Biologia Evolutiva (Mestrado)
|
Departamento: |
Unicentro::Departamento de Ciências Agrárias e Ambientais
|
País: |
Brasil
|
Palavras-chave em Português: | |
Palavras-chave em Inglês: | |
Área do conhecimento CNPq: | |
Link de acesso: | http://tede.unicentro.br:8080/jspui/handle/jspui/1613 |
Resumo: | The study of fungal biodiversity found in forest fragments is of great importance. Through knowledge of this biodiversity and the bioprospecting of biotechnological products, we seek to obtain subsidies to promote the conservation and valorisation of these remaining fragments. This work aimed to isolate thermophilous filamentous fungi from remnants of Araucaria Forest (AF), and to perform a bioprospecting for obtention of fungal strains xylanase producers. The objective was also to establish the optimal parameters for the production of these enzymes and to characterize them biochemically. In order to optimize production, the respective fungus was grown in Vogel liquid medium, in stationary condition at 45 ° C, for incubation periods and conditions specified by the Central Rotational Compound Design (DCCR) from planning 2 3 , where there was variation in cultivation time, pH of the medium and percentage of carbon source. In addition, the enzymes, in the non-purified form, were biochemically characterized in relation to the optimal temperature and pH of performance, thermal stability, stability against pH. The effect of adding ions and other substances to the reaction medium was also investigated. A total of 15 strains were isolated from AF soil and litter samples. Among these, the PJ10 strain distinghishes itself by the levels of xylanases produced when it grown in the presence of corn straw. The respective strain was molecularly characterized as belonging to the species Aspergillus fumigatus. Based on its growth potential under different temperatures, the respective strain was classified as thermotolerant. The highest levels of xilanase production were obtained when the fungus was grown for 72 hours, in the presence of 2% corn straw, at pH 9.0 and at 45 ° C, reaching 78,17 U/mL-1. Xylanases were more active at 65 ° C and at pH 6.0. They were not very stable at 60, 65 and 70 °C. Furthermore, the enzyme was stable in the pH range of 3.0 to 8.0, and it was activated by Mn2+ ,Cu2+, Mg2+ and β-mercaptoethanol and inhibited by Hg+ and SDS. From the results presented, it appears that the remaining fragments of AF exhibit great potential as sources of potentially new biological resources, such as the xylanase-producing thermophilic fungal strain described in the present investigation. |
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Knob, Adrianahttp://lattes.cnpq.br/1289646338155108Beitel, Susan Michelzhttp://lattes.cnpq.br/5929035791324142095.014.089-90http://lattes.cnpq.br/6921371166803184MARMENTINI, JÉSSICA2021-05-12T11:41:20Z2020-04-29MARMENTINI, JÉSSICA. ISOLAMENTO E BIOPROSPECÇÃO DE FUNGOS TERMOFÍLICOS ISOLADOS DA FLORESTA OMBRÓFILA MISTA: PRODUÇÃO DE XILANASES POR Aspergillus fumigatus. 2020. 89 f. Dissertação (Programa de Pós-Graduação em Biologia Evolutiva - Mestrado) - Universidade Estadual do Centro-Oeste, Guarapuava - PR.http://tede.unicentro.br:8080/jspui/handle/jspui/1613The study of fungal biodiversity found in forest fragments is of great importance. Through knowledge of this biodiversity and the bioprospecting of biotechnological products, we seek to obtain subsidies to promote the conservation and valorisation of these remaining fragments. This work aimed to isolate thermophilous filamentous fungi from remnants of Araucaria Forest (AF), and to perform a bioprospecting for obtention of fungal strains xylanase producers. The objective was also to establish the optimal parameters for the production of these enzymes and to characterize them biochemically. In order to optimize production, the respective fungus was grown in Vogel liquid medium, in stationary condition at 45 ° C, for incubation periods and conditions specified by the Central Rotational Compound Design (DCCR) from planning 2 3 , where there was variation in cultivation time, pH of the medium and percentage of carbon source. In addition, the enzymes, in the non-purified form, were biochemically characterized in relation to the optimal temperature and pH of performance, thermal stability, stability against pH. The effect of adding ions and other substances to the reaction medium was also investigated. A total of 15 strains were isolated from AF soil and litter samples. Among these, the PJ10 strain distinghishes itself by the levels of xylanases produced when it grown in the presence of corn straw. The respective strain was molecularly characterized as belonging to the species Aspergillus fumigatus. Based on its growth potential under different temperatures, the respective strain was classified as thermotolerant. The highest levels of xilanase production were obtained when the fungus was grown for 72 hours, in the presence of 2% corn straw, at pH 9.0 and at 45 ° C, reaching 78,17 U/mL-1. Xylanases were more active at 65 ° C and at pH 6.0. They were not very stable at 60, 65 and 70 °C. Furthermore, the enzyme was stable in the pH range of 3.0 to 8.0, and it was activated by Mn2+ ,Cu2+, Mg2+ and β-mercaptoethanol and inhibited by Hg+ and SDS. From the results presented, it appears that the remaining fragments of AF exhibit great potential as sources of potentially new biological resources, such as the xylanase-producing thermophilic fungal strain described in the present investigation.O estudo da biodiversidade fúngica encontrada em fragmentos florestais é de grande importância. Busca-se, por meio do conhecimento dessa biodiversidade e da bioprospecção de produtos biotecnológicos, obter subsídios que promovam a conservação e valorização desses remanescentes. Este trabalho teve como objetivo isolar fungos filamentosos termófilos de remanescentes de Floresta Ombrófila Mista (FOM), e realizar uma bioprospecção quanto ao seu potencial de produção de xilanases. Objetivouse, ainda, estabelecer os parâmetros ótimos para a produção dessas enzimas e caracterizá-las bioquimicamente. A fim de otimizar a produção, o fungo obtido da bioprospecção foi cultivado em meio líquido de Vogel, em condição estacionária a 45 °C, por períodos de incubação e condições especificadas pelo Delineamento Central Composto Rotacional (DCCR), a partir de planejamento 23 , variando-se o tempo de cultivo, pH do meio e porcentagem da fonte de carbono. Em adição, as enzimas do extrato bruto foram bioquimicamente caracterizadas em relação à temperatura e o pH ótimos de atuação, estabilidade térmica e estabilidade frente ao pH. O efeito da adição de íons e de outras substâncias no meio reacional também foi investigado. Um total de 15 linhagens foram isoladas a partir de amostras de solo e de serapilheira. Dentre estas, a linhagem PJ10 se destacou pelos níveis de xilanases produzidos quando crescida na presença de palha de milho. A respectiva linhagem foi molecularmente caracterizada como pertencente à espécie Aspergillus fumigatus. Com base em seu potencial de crescimento sob temperaturas distintas, a respectiva linhagem foi classificada como termotolerante. Os maiores níveis de produção foram obtidos quando o fungo foi cultivado por 72 horas, em presença de 2% de palha de milho, em pH 9,0 e a 45 °C, alcançando 78,17 U mL-1. As xilanases foram mais ativas a 65 °C e em pH 6,0, mostraram-se pouco estáveis em 60, 65 e 70 °C. Ainda, a enzima foi estável na faixa de pH de 3,0 a 8,0, sendo ativada por Mn+2, Cu+2, Mg+2 e βmercaptoetanol e inibida por Hg+ e SDS. Pelos resultados apresentados, constata-se que os remanescentes de FOM exibem grande potencial como fontes de recursos biológicos potencialmente novos, como a linhagem fúngica termofílica produtora de xilanases descrita na presente investigação.Submitted by Fabiano Jucá (fjuca@unicentro.br) on 2021-05-12T11:41:20Z No. of bitstreams: 1 Dissertação Jéssica Marmentini versão final.pdf: 1376848 bytes, checksum: d61b726077cffbd361e2fc3fb950e03f (MD5)Made available in DSpace on 2021-05-12T11:41:20Z (GMT). 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dc.title.por.fl_str_mv |
ISOLAMENTO E BIOPROSPECÇÃO DE FUNGOS TERMOFÍLICOS ISOLADOS DA FLORESTA OMBRÓFILA MISTA: PRODUÇÃO DE XILANASES POR Aspergillus fumigatus |
title |
ISOLAMENTO E BIOPROSPECÇÃO DE FUNGOS TERMOFÍLICOS ISOLADOS DA FLORESTA OMBRÓFILA MISTA: PRODUÇÃO DE XILANASES POR Aspergillus fumigatus |
spellingShingle |
ISOLAMENTO E BIOPROSPECÇÃO DE FUNGOS TERMOFÍLICOS ISOLADOS DA FLORESTA OMBRÓFILA MISTA: PRODUÇÃO DE XILANASES POR Aspergillus fumigatus MARMENTINI, JÉSSICA Fragmentos florestais Mata Atlântica fungos filamentosos termófilos enzimas xilanolíticas Forest fragments Atlantic Forest filamentous fungi thermophiles xylanolitic enzymes CIENCIAS BIOLOGICAS::BIOLOGIA GERAL CIENCIAS BIOLOGICAS::BIOQUIMICA CIENCIAS BIOLOGICAS::BOTANICA CIENCIAS BIOLOGICAS::ECOLOGIA CIENCIAS BIOLOGICAS::GENETICA CIENCIAS BIOLOGICAS::MICROBIOLOGIA CIENCIAS BIOLOGICAS::ZOOLOGIA CIENCIAS BIOLOGICAS |
title_short |
ISOLAMENTO E BIOPROSPECÇÃO DE FUNGOS TERMOFÍLICOS ISOLADOS DA FLORESTA OMBRÓFILA MISTA: PRODUÇÃO DE XILANASES POR Aspergillus fumigatus |
title_full |
ISOLAMENTO E BIOPROSPECÇÃO DE FUNGOS TERMOFÍLICOS ISOLADOS DA FLORESTA OMBRÓFILA MISTA: PRODUÇÃO DE XILANASES POR Aspergillus fumigatus |
title_fullStr |
ISOLAMENTO E BIOPROSPECÇÃO DE FUNGOS TERMOFÍLICOS ISOLADOS DA FLORESTA OMBRÓFILA MISTA: PRODUÇÃO DE XILANASES POR Aspergillus fumigatus |
title_full_unstemmed |
ISOLAMENTO E BIOPROSPECÇÃO DE FUNGOS TERMOFÍLICOS ISOLADOS DA FLORESTA OMBRÓFILA MISTA: PRODUÇÃO DE XILANASES POR Aspergillus fumigatus |
title_sort |
ISOLAMENTO E BIOPROSPECÇÃO DE FUNGOS TERMOFÍLICOS ISOLADOS DA FLORESTA OMBRÓFILA MISTA: PRODUÇÃO DE XILANASES POR Aspergillus fumigatus |
author |
MARMENTINI, JÉSSICA |
author_facet |
MARMENTINI, JÉSSICA |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Knob, Adriana |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/1289646338155108 |
dc.contributor.advisor-co1.fl_str_mv |
Beitel, Susan Michelz |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://lattes.cnpq.br/5929035791324142 |
dc.contributor.authorID.fl_str_mv |
095.014.089-90 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/6921371166803184 |
dc.contributor.author.fl_str_mv |
MARMENTINI, JÉSSICA |
contributor_str_mv |
Knob, Adriana Beitel, Susan Michelz |
dc.subject.por.fl_str_mv |
Fragmentos florestais Mata Atlântica fungos filamentosos termófilos enzimas xilanolíticas |
topic |
Fragmentos florestais Mata Atlântica fungos filamentosos termófilos enzimas xilanolíticas Forest fragments Atlantic Forest filamentous fungi thermophiles xylanolitic enzymes CIENCIAS BIOLOGICAS::BIOLOGIA GERAL CIENCIAS BIOLOGICAS::BIOQUIMICA CIENCIAS BIOLOGICAS::BOTANICA CIENCIAS BIOLOGICAS::ECOLOGIA CIENCIAS BIOLOGICAS::GENETICA CIENCIAS BIOLOGICAS::MICROBIOLOGIA CIENCIAS BIOLOGICAS::ZOOLOGIA CIENCIAS BIOLOGICAS |
dc.subject.eng.fl_str_mv |
Forest fragments Atlantic Forest filamentous fungi thermophiles xylanolitic enzymes |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS::BIOLOGIA GERAL CIENCIAS BIOLOGICAS::BIOQUIMICA CIENCIAS BIOLOGICAS::BOTANICA CIENCIAS BIOLOGICAS::ECOLOGIA CIENCIAS BIOLOGICAS::GENETICA CIENCIAS BIOLOGICAS::MICROBIOLOGIA CIENCIAS BIOLOGICAS::ZOOLOGIA CIENCIAS BIOLOGICAS |
description |
The study of fungal biodiversity found in forest fragments is of great importance. Through knowledge of this biodiversity and the bioprospecting of biotechnological products, we seek to obtain subsidies to promote the conservation and valorisation of these remaining fragments. This work aimed to isolate thermophilous filamentous fungi from remnants of Araucaria Forest (AF), and to perform a bioprospecting for obtention of fungal strains xylanase producers. The objective was also to establish the optimal parameters for the production of these enzymes and to characterize them biochemically. In order to optimize production, the respective fungus was grown in Vogel liquid medium, in stationary condition at 45 ° C, for incubation periods and conditions specified by the Central Rotational Compound Design (DCCR) from planning 2 3 , where there was variation in cultivation time, pH of the medium and percentage of carbon source. In addition, the enzymes, in the non-purified form, were biochemically characterized in relation to the optimal temperature and pH of performance, thermal stability, stability against pH. The effect of adding ions and other substances to the reaction medium was also investigated. A total of 15 strains were isolated from AF soil and litter samples. Among these, the PJ10 strain distinghishes itself by the levels of xylanases produced when it grown in the presence of corn straw. The respective strain was molecularly characterized as belonging to the species Aspergillus fumigatus. Based on its growth potential under different temperatures, the respective strain was classified as thermotolerant. The highest levels of xilanase production were obtained when the fungus was grown for 72 hours, in the presence of 2% corn straw, at pH 9.0 and at 45 ° C, reaching 78,17 U/mL-1. Xylanases were more active at 65 ° C and at pH 6.0. They were not very stable at 60, 65 and 70 °C. Furthermore, the enzyme was stable in the pH range of 3.0 to 8.0, and it was activated by Mn2+ ,Cu2+, Mg2+ and β-mercaptoethanol and inhibited by Hg+ and SDS. From the results presented, it appears that the remaining fragments of AF exhibit great potential as sources of potentially new biological resources, such as the xylanase-producing thermophilic fungal strain described in the present investigation. |
publishDate |
2020 |
dc.date.issued.fl_str_mv |
2020-04-29 |
dc.date.accessioned.fl_str_mv |
2021-05-12T11:41:20Z |
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 |
MARMENTINI, JÉSSICA. ISOLAMENTO E BIOPROSPECÇÃO DE FUNGOS TERMOFÍLICOS ISOLADOS DA FLORESTA OMBRÓFILA MISTA: PRODUÇÃO DE XILANASES POR Aspergillus fumigatus. 2020. 89 f. Dissertação (Programa de Pós-Graduação em Biologia Evolutiva - Mestrado) - Universidade Estadual do Centro-Oeste, Guarapuava - PR. |
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MARMENTINI, JÉSSICA. ISOLAMENTO E BIOPROSPECÇÃO DE FUNGOS TERMOFÍLICOS ISOLADOS DA FLORESTA OMBRÓFILA MISTA: PRODUÇÃO DE XILANASES POR Aspergillus fumigatus. 2020. 89 f. Dissertação (Programa de Pós-Graduação em Biologia Evolutiva - Mestrado) - Universidade Estadual do Centro-Oeste, Guarapuava - PR. |
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http://tede.unicentro.br:8080/jspui/handle/jspui/1613 |
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Universidade Estadual do Centro-Oeste |
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