BIOPROSPECÇÃO DE FUNGOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE LIPASES: POTENCIAL LIPOLÍTICO DE Trichoderma sp.
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações do UNICENTRO |
Texto Completo: | http://tede.unicentro.br:8080/jspui/handle/jspui/728 |
Resumo: | The soil from Araucaria forest had been the focus of few studies about its microbial diversity. The soil is a habitat that exhibits a large microbial diversity. Thus, it can be explored as a source of microorganisms that produce enzymes of industrial interest. Among of variousenzymes existent, the lipases (triacylglycerol-acid-hydrolases E.C.3.1.1.3) are extensively used, due to the wide range of reactions that they catalyze, as the total or partial hydrolysis of triacylglycerols (TAG), reactions of esterification, transesterification and inter-esterification of lipids. Therefore, the present study aimed to isolate different fungal strains from fragments of Araucaria Forest and evaluate the potential of lipase production by the fungi isolated. From the selection of the best producer strain, the Design Plackett-Burman was used, in order to select the variables that affect this process. After, the Central Composite Rotatable Design (CCRD) applicable to Response Surface Methodology (RSM) was employed, in order to establish the best conditions for the lipase production, using the castor bean waste as substrate. Finally, the enzyme produced under optimum conditions were biochemically characterized. The fungal isolation was conducted using the dilution series technique, allowing the achievement of 65 fungal strains. The lipase production capacity was verified in 17 of studied strains, in which a Trichoderma strain was the best producer. Tween 80, CaCl2, NH4Cl and yeast extract were selected as the variables that affect the lipase production process by Trichoderma sp. For this reason, these same variables were considered in DCCR. Under optimized conditions, the levels of lipases reached 64 U mL-1. Through the biochemical characterization, it was possible stablished that Trichoderma sp. lipases showed optimal activity in neutral pH, at 30 °C. In relation to the effect of various ions and substances on the lipase activity, 0.2 mM Na+2 activated these enzymes. In addition, an activator effect was verified in the presence of Triton X-100 surfactant. Among distinct organics solvents evaluated, only 1% toluene presented an activator effect on the produced enzymes. The results obtained in this study reveal the potential use of the castor bean waste as substrate for the lipase production. The employment of this waste on microbial enzymes production is very interesting, since economic and environmental benefits could be obtained from its uses. In addition, the screening studies presented here offer news perspectives, in order to promote the appreciation as well the rational exploration of Araucaria Forest fragments, priority areas for the Brazilian biodiversity conservation. |
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Knob, Adrianahttp://lattes.cnpq.br/1289646338155108Lima, Vanderlei Aparecido dehttp://lattes.cnpq.br/9090461949264421074.218.279-78http://lattes.cnpq.br/5803880589082611GUIMARÃES, MICHAEL WILIAN2017-06-26T12:50:15Z2016-02-26GUIMARÃES, MICHAEL WILIAN. BIOPROSPECÇÃO DE FUNGOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE LIPASES: POTENCIAL LIPOLÍTICO DE Trichoderma sp.. 2016. 82 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/728The soil from Araucaria forest had been the focus of few studies about its microbial diversity. The soil is a habitat that exhibits a large microbial diversity. Thus, it can be explored as a source of microorganisms that produce enzymes of industrial interest. Among of variousenzymes existent, the lipases (triacylglycerol-acid-hydrolases E.C.3.1.1.3) are extensively used, due to the wide range of reactions that they catalyze, as the total or partial hydrolysis of triacylglycerols (TAG), reactions of esterification, transesterification and inter-esterification of lipids. Therefore, the present study aimed to isolate different fungal strains from fragments of Araucaria Forest and evaluate the potential of lipase production by the fungi isolated. From the selection of the best producer strain, the Design Plackett-Burman was used, in order to select the variables that affect this process. After, the Central Composite Rotatable Design (CCRD) applicable to Response Surface Methodology (RSM) was employed, in order to establish the best conditions for the lipase production, using the castor bean waste as substrate. Finally, the enzyme produced under optimum conditions were biochemically characterized. The fungal isolation was conducted using the dilution series technique, allowing the achievement of 65 fungal strains. The lipase production capacity was verified in 17 of studied strains, in which a Trichoderma strain was the best producer. Tween 80, CaCl2, NH4Cl and yeast extract were selected as the variables that affect the lipase production process by Trichoderma sp. For this reason, these same variables were considered in DCCR. Under optimized conditions, the levels of lipases reached 64 U mL-1. Through the biochemical characterization, it was possible stablished that Trichoderma sp. lipases showed optimal activity in neutral pH, at 30 °C. In relation to the effect of various ions and substances on the lipase activity, 0.2 mM Na+2 activated these enzymes. In addition, an activator effect was verified in the presence of Triton X-100 surfactant. Among distinct organics solvents evaluated, only 1% toluene presented an activator effect on the produced enzymes. The results obtained in this study reveal the potential use of the castor bean waste as substrate for the lipase production. The employment of this waste on microbial enzymes production is very interesting, since economic and environmental benefits could be obtained from its uses. In addition, the screening studies presented here offer news perspectives, in order to promote the appreciation as well the rational exploration of Araucaria Forest fragments, priority areas for the Brazilian biodiversity conservation.O solo da Floresta Ombrófila Mista tem sido o foco de poucos estudos sobre sua diversidade microbiana. O solo é um habitat que apresenta uma ampla diversidade microbiana. Desta forma, pode ser explorado como uma fonte de microrganismos produtores de enzimas de interesse industrial. Dentre as diversas enzimas existentes, as lipases (triacilglicerol-acil-hidrolases E.C.3.1.1.3) são muito utilizadas, em virtude da grande gama de reações que podem catalisar, como a hidrólise total ou parcial de triacilgliceróis (TAG), além de reações de esterificação, transesterificação e interesterificação de lipídios. Diante disto, o presente estudo objetivou isolar diferentes linhagens fúngicas a partir de solo de remanescentes de Floresta Ombrófila Mista e avaliar o potencial de produção de lipases pelos fungos isolados. A partir da seleção da melhor linhagem produtora, o delineamento Plackett-Burman foi empregado, a fim de selecionar as variáveis que mais afetam este processo. Posteriormente, o Delineamento Central Composto Rotacional (DCCR) aplicável à Metodologia de Superfície de Resposta (RSM) foi empregado, no intuito de se estabelecer as condições ótimas para a produção dessas enzimas, utilizando a torta de mamona como substrato. Por fim, as lipases produzidas sob condições otimizadas foram bioquimicamente caracterizadas. O isolamento dos fungos foi realizado empregando-se a técnica de diluição sucessiva, permitindo a obtenção de 65 linhagens fúngicas. A capacidade de produção de lipases foi evidenciada em 17 das linhagens avaliadas, destacando-se como melhor produtor uma linhagem de Trichoderma sp. As variáveis concentração de Tween 80, CaCl2, NH4Cl e extrato de levedura foram selecionadas como as que mais afetam o processo de produção de lipases por Trichoderma sp. Por esta razão, estas mesmas variáveis foram consideradas no DCCR. Após a otimização, os níveis de produção atingidos corresponderam a 60,70 U/mL. Por meio da caracterização bioquímica realizada, constatou-se que as lipases de Trichoderma sp. apresentam atividade ótima em pH neutro e a 30 °C. Em relação ao efeito de diversos íons e substâncias sobre a atividade lipásica, verificou-se que o Na+2 na concentração de 0,2 mM promoveu a ativação destas enzimas. Ainda, um efeito ativador foi observado em presença do surfactante Triton X-100. Dentre os distintos solventes orgânicos avaliados apenas o tolueno a 1% ativou as enzimas produzidas. Os resultados obtidos neste trabalho revelam o potencial de utilização da torta de mamona como substrato para a produção de lipases. O emprego deste resíduo para a produção de enzimas microbianas se mostra bastante interessante, visto que tantos benefícios econômicos quanto ambientais poderão ser obtidos a partir de sua utilização. Em adição, os estudos de bioprospecção aqui apresentados oferecem novas perspectivas, a fim de promover a valorização bem como a exploração racional dos remanescentes de Floresta Ombrófila Mista, áreas prioritárias para a conservação da biodiversidade brasileira.Submitted by Fabiano Jucá (fjuca@unicentro.br) on 2017-06-26T12:50:15Z No. of bitstreams: 1 MICHAEL WILIAN GUIMARÃES.pdf: 2010475 bytes, checksum: 2a5b9b5f801aa01bea2c0addfb0513b8 (MD5)Made available in DSpace on 2017-06-26T12:50:15Z (GMT). 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dc.title.por.fl_str_mv |
BIOPROSPECÇÃO DE FUNGOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE LIPASES: POTENCIAL LIPOLÍTICO DE Trichoderma sp. |
title |
BIOPROSPECÇÃO DE FUNGOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE LIPASES: POTENCIAL LIPOLÍTICO DE Trichoderma sp. |
spellingShingle |
BIOPROSPECÇÃO DE FUNGOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE LIPASES: POTENCIAL LIPOLÍTICO DE Trichoderma sp. GUIMARÃES, MICHAEL WILIAN Bioprospecção Enzimas lipolíticas Delineamento Central Composto Rotacional Casca da torta de mamona screening lipolytic enzymes central composite rotatable design castor bean waste CIENCIAS BIOLOGICAS CIENCIAS BIOLOGICAS::BIOLOGIA GERAL CIENCIAS BIOLOGICAS::GENETICA CIENCIAS BIOLOGICAS::BOTANICA CIENCIAS BIOLOGICAS::ZOOLOGIA CIENCIAS BIOLOGICAS::ECOLOGIA CIENCIAS BIOLOGICAS::BIOQUIMICA CIENCIAS BIOLOGICAS::MICROBIOLOGIA |
title_short |
BIOPROSPECÇÃO DE FUNGOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE LIPASES: POTENCIAL LIPOLÍTICO DE Trichoderma sp. |
title_full |
BIOPROSPECÇÃO DE FUNGOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE LIPASES: POTENCIAL LIPOLÍTICO DE Trichoderma sp. |
title_fullStr |
BIOPROSPECÇÃO DE FUNGOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE LIPASES: POTENCIAL LIPOLÍTICO DE Trichoderma sp. |
title_full_unstemmed |
BIOPROSPECÇÃO DE FUNGOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE LIPASES: POTENCIAL LIPOLÍTICO DE Trichoderma sp. |
title_sort |
BIOPROSPECÇÃO DE FUNGOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE LIPASES: POTENCIAL LIPOLÍTICO DE Trichoderma sp. |
author |
GUIMARÃES, MICHAEL WILIAN |
author_facet |
GUIMARÃES, MICHAEL WILIAN |
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 |
Lima, Vanderlei Aparecido de |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://lattes.cnpq.br/9090461949264421 |
dc.contributor.authorID.fl_str_mv |
074.218.279-78 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/5803880589082611 |
dc.contributor.author.fl_str_mv |
GUIMARÃES, MICHAEL WILIAN |
contributor_str_mv |
Knob, Adriana Lima, Vanderlei Aparecido de |
dc.subject.por.fl_str_mv |
Bioprospecção Enzimas lipolíticas Delineamento Central Composto Rotacional Casca da torta de mamona |
topic |
Bioprospecção Enzimas lipolíticas Delineamento Central Composto Rotacional Casca da torta de mamona screening lipolytic enzymes central composite rotatable design castor bean waste CIENCIAS BIOLOGICAS CIENCIAS BIOLOGICAS::BIOLOGIA GERAL CIENCIAS BIOLOGICAS::GENETICA CIENCIAS BIOLOGICAS::BOTANICA CIENCIAS BIOLOGICAS::ZOOLOGIA CIENCIAS BIOLOGICAS::ECOLOGIA CIENCIAS BIOLOGICAS::BIOQUIMICA CIENCIAS BIOLOGICAS::MICROBIOLOGIA |
dc.subject.eng.fl_str_mv |
screening lipolytic enzymes central composite rotatable design castor bean waste |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS CIENCIAS BIOLOGICAS::BIOLOGIA GERAL CIENCIAS BIOLOGICAS::GENETICA CIENCIAS BIOLOGICAS::BOTANICA CIENCIAS BIOLOGICAS::ZOOLOGIA CIENCIAS BIOLOGICAS::ECOLOGIA CIENCIAS BIOLOGICAS::BIOQUIMICA CIENCIAS BIOLOGICAS::MICROBIOLOGIA |
description |
The soil from Araucaria forest had been the focus of few studies about its microbial diversity. The soil is a habitat that exhibits a large microbial diversity. Thus, it can be explored as a source of microorganisms that produce enzymes of industrial interest. Among of variousenzymes existent, the lipases (triacylglycerol-acid-hydrolases E.C.3.1.1.3) are extensively used, due to the wide range of reactions that they catalyze, as the total or partial hydrolysis of triacylglycerols (TAG), reactions of esterification, transesterification and inter-esterification of lipids. Therefore, the present study aimed to isolate different fungal strains from fragments of Araucaria Forest and evaluate the potential of lipase production by the fungi isolated. From the selection of the best producer strain, the Design Plackett-Burman was used, in order to select the variables that affect this process. After, the Central Composite Rotatable Design (CCRD) applicable to Response Surface Methodology (RSM) was employed, in order to establish the best conditions for the lipase production, using the castor bean waste as substrate. Finally, the enzyme produced under optimum conditions were biochemically characterized. The fungal isolation was conducted using the dilution series technique, allowing the achievement of 65 fungal strains. The lipase production capacity was verified in 17 of studied strains, in which a Trichoderma strain was the best producer. Tween 80, CaCl2, NH4Cl and yeast extract were selected as the variables that affect the lipase production process by Trichoderma sp. For this reason, these same variables were considered in DCCR. Under optimized conditions, the levels of lipases reached 64 U mL-1. Through the biochemical characterization, it was possible stablished that Trichoderma sp. lipases showed optimal activity in neutral pH, at 30 °C. In relation to the effect of various ions and substances on the lipase activity, 0.2 mM Na+2 activated these enzymes. In addition, an activator effect was verified in the presence of Triton X-100 surfactant. Among distinct organics solvents evaluated, only 1% toluene presented an activator effect on the produced enzymes. The results obtained in this study reveal the potential use of the castor bean waste as substrate for the lipase production. The employment of this waste on microbial enzymes production is very interesting, since economic and environmental benefits could be obtained from its uses. In addition, the screening studies presented here offer news perspectives, in order to promote the appreciation as well the rational exploration of Araucaria Forest fragments, priority areas for the Brazilian biodiversity conservation. |
publishDate |
2016 |
dc.date.issued.fl_str_mv |
2016-02-26 |
dc.date.accessioned.fl_str_mv |
2017-06-26T12:50:15Z |
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 |
GUIMARÃES, MICHAEL WILIAN. BIOPROSPECÇÃO DE FUNGOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE LIPASES: POTENCIAL LIPOLÍTICO DE Trichoderma sp.. 2016. 82 f. Dissertação (Programa de Pós-Graduação em Biologia Evolutiva - Mestrado) - Universidade Estadual do Centro-Oeste, Guarapuava - PR. |
dc.identifier.uri.fl_str_mv |
http://tede.unicentro.br:8080/jspui/handle/jspui/728 |
identifier_str_mv |
GUIMARÃES, MICHAEL WILIAN. BIOPROSPECÇÃO DE FUNGOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE LIPASES: POTENCIAL LIPOLÍTICO DE Trichoderma sp.. 2016. 82 f. Dissertação (Programa de Pós-Graduação em Biologia Evolutiva - Mestrado) - Universidade Estadual do Centro-Oeste, Guarapuava - PR. |
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Universidade Estadual do Centro-Oeste |
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Universidade Estadual do Centro-Oeste |
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