BIOPROSPECÇÃO DE FUNGOS FILAMENTOSOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE POLIGALACTURONASES: POTENCIAL PECTINOLÍTICO DE Penicillium sp.
Autor(a) principal: | |
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Data de Publicação: | 2017 |
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/732 |
Resumo: | Pectinases have been extensively studied due to their wide variety of applications in the many industrial sectors. Among these enzymes, polygalacturonases (PG) are used mainly in the fruit juices clarification process. The selection of enzyme producing microorganisms becomes an important step in biotechnological processes. Thus, this study aimed to isolate several fungal strains and evaluate its potential production of PGs, using pineapple pell and passion fruit peel as substrates. Following the selection of the best producer strain, the Plackett-Burmann design was used in order to select the variables that most influence the production of these enzymes. Afther that, the optimal parameters for PGs production were stablishedd, using the Central Composite Rotary Design and the Response Surface Methodology. Finally, the PGs produced were biochemically characterized. As a result, 46 strains were isolated, showing, through the semi-quantitative test, the PG production hability for all strains evaluated. In the quantitative test, it was verified that the FX32 fungus was the best producer of PGs, corresponding to 6.12 U mL-1. Through the use of Plackett-Burmann design, it was found that the substrate concentration, pH and culture time were the parameters that significant influence PGs production. For these reason, these factors were considered as the independent variables in the Central Composite Rotary Design. Thus, the best conditions established for PGs production PG by Penicillium sp. were 2.9% of pineapple peel as substrate, culture pH of 7.8 and cultivation time of 8.6 days, corresponding to a production of 106.20 U mL-1. With regard to biochemical characteristics, it was verified that the enzymes produced by this fungal strain were more active at 50 °C and at pH 5.5. PGs were also very stable at pH 5.0 and 5.5, exibithing more than 80% of its initial activity under these conditions. Em relation to thermal stability, these enzymes were more stable at 45 °C. The effect of several ions and substances on the PG activity was studied. The PGs were activated by Mn2+, Ca2+ and mercaptoethanol and strong inhibited by Hg+. Finally, the hydrolysis of the pineapple peel was performed using Penicillium sp. and Aspergillus flavus crude extracts. A glucose yield equivalent to 67.42 mg g-1 substrate was achieved, under optimal hydrolysis conditions, corresponding to 15 hours of reaction, reactional pH 5.0 and reactional temperature of 50 °C. The pineapple peel was a good inducer for PGs production by Penicillium sp., as well as a promising substrate for glucose achievement through its enzymatic hydrolysis. This results provide important subsidies in order to infer industrial applications for the enzymes under study. |
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Knob, Adrianahttp://lattes.cnpq.br/1289646338155108075.938.129-10http://lattes.cnpq.br/4742847602269407WOUK, THAÍS2017-06-26T16:36:23Z2017-02-23WOUK, THAÍS. BIOPROSPECÇÃO DE FUNGOS FILAMENTOSOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE POLIGALACTURONASES: POTENCIAL PECTINOLÍTICO DE Penicillium sp.. 2017. 94 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/732Pectinases have been extensively studied due to their wide variety of applications in the many industrial sectors. Among these enzymes, polygalacturonases (PG) are used mainly in the fruit juices clarification process. The selection of enzyme producing microorganisms becomes an important step in biotechnological processes. Thus, this study aimed to isolate several fungal strains and evaluate its potential production of PGs, using pineapple pell and passion fruit peel as substrates. Following the selection of the best producer strain, the Plackett-Burmann design was used in order to select the variables that most influence the production of these enzymes. Afther that, the optimal parameters for PGs production were stablishedd, using the Central Composite Rotary Design and the Response Surface Methodology. Finally, the PGs produced were biochemically characterized. As a result, 46 strains were isolated, showing, through the semi-quantitative test, the PG production hability for all strains evaluated. In the quantitative test, it was verified that the FX32 fungus was the best producer of PGs, corresponding to 6.12 U mL-1. Through the use of Plackett-Burmann design, it was found that the substrate concentration, pH and culture time were the parameters that significant influence PGs production. For these reason, these factors were considered as the independent variables in the Central Composite Rotary Design. Thus, the best conditions established for PGs production PG by Penicillium sp. were 2.9% of pineapple peel as substrate, culture pH of 7.8 and cultivation time of 8.6 days, corresponding to a production of 106.20 U mL-1. With regard to biochemical characteristics, it was verified that the enzymes produced by this fungal strain were more active at 50 °C and at pH 5.5. PGs were also very stable at pH 5.0 and 5.5, exibithing more than 80% of its initial activity under these conditions. Em relation to thermal stability, these enzymes were more stable at 45 °C. The effect of several ions and substances on the PG activity was studied. The PGs were activated by Mn2+, Ca2+ and mercaptoethanol and strong inhibited by Hg+. Finally, the hydrolysis of the pineapple peel was performed using Penicillium sp. and Aspergillus flavus crude extracts. A glucose yield equivalent to 67.42 mg g-1 substrate was achieved, under optimal hydrolysis conditions, corresponding to 15 hours of reaction, reactional pH 5.0 and reactional temperature of 50 °C. The pineapple peel was a good inducer for PGs production by Penicillium sp., as well as a promising substrate for glucose achievement through its enzymatic hydrolysis. This results provide important subsidies in order to infer industrial applications for the enzymes under study.As pectinases têm sido alvo de muitos estudos devido a sua ampla variedade de aplicações nos mais diversos setores industriais. Dentre essas enzimas encontram-se as poligalacturonases, as quais são empregadas principalmente no processo na clarificação de sucos de frutas. A seleção de microrganismos produtores destas enzimas torna-se uma etapa importante em processos biotecnológicos. Deste modo, o objetivo desse estudo foi isolar diversas linhagens fúngicas e avaliar o seu potencial de produção de poligalacturonases, utilizando a casca de abacaxi e casca de maracujá como substrato. A partir da seleção da melhor linhagem produtora, o delineamento Plackett-Burmann foi empregado, a fim de selecionar as variáveis que mais influenciam a produção dessas enzimas. Posteriormente, os parâmetros ótimos para a produção de poligalacturonases foram verificados, empregando-se o Delineamento Central Composto Rotacional e a Metodologia de Superfície de Resposta. Por fim, as poligalacturonases produzidas foram caracterizadas bioquimicamente. Como resultado, 46 linhagens foram isoladas, evidenciando-se, por meio do teste semi-quantitativo, a capacidade de produção de poligalacturonases por todas as linhagens avaliadas. Já no teste quantitativo, verificou-se que o fungo FX32 foi o melhor produtor dessas enzimas utilizando casca de abacaxi como substrato, correspondendo a 6,12 U mL-1. A partir do delineamento Plackett-Burmann, constatou-se que a concentração do substrato, o pH e o tempo de cultivo foram os parâmetros que exerceram influência significativa sobre a produção de poligalacturonases. Por esta razão, esses fatores foram considerados como as variáveis independentes no Delineamento Central Composto Rotacional. Por meio de sua realização, estabeleceu-se as melhores condições para a produção de poligalacturonase por Penicillium sp. sendo elas 2,9% de casca de abacaxi como substrato, pH de cultivo de 7,8 e tempo de cultivo de 8,6 dias, correspondendo a uma produção de 106,20 U mL-1. Em relação as suas características bioquímicas, constatou-se que as enzimas produzidas por essa linhagem fúngica foram mais ativas a 50 °C e em pH 5,5. As poligalacturonases também foram muito estáveis em pH 5,0 e 5,5, onde mais de 80% de sua atividade inicial foi verificada. Em relação a estabilidade térmica, observou-se que essas enzimas foram mais estáveis a 45 ºC. O efeito de diversos íons e substâncias sobre a atividade poligalacturonásica também foi avaliado, evidenciando-se uma ativação enzimática na presença de íons Mn+2, Ca+2 e mercaptoetanol e uma forte inibição por Hg+. Por fim, a da hidrólise da casca de abacaxi foi realizada, utilizando-se os extratos brutos de Penicillium sp. e Aspergillus flavus. Uma liberação de glicose equivalente a 67,42 mg g-1 de substrato foi alcançada, após o estabelecimento das condições ótimas de hidrólise, correspondentes a 15 horas de reação, pH 5,0 e temperatura de 50 °C. A casca de abacaxi mostrou-se uma boa fonte indutora da produção de poligalacturonases por Penicillium sp., como também um substrato promissor para a obtenção de monômeros de glicose por meio de sua hidrólise enzimática. Os resultados obtidos nesse estudo fornecem informações importantes, a fim de inferir possíveis aplicações industriais para as enzimas em estudo.Submitted by Fabiano Jucá (fjuca@unicentro.br) on 2017-06-26T16:36:23Z No. of bitstreams: 1 THAÍS WOUK.pdf: 2521340 bytes, checksum: e63e59adabeb8b8a1a4b91667f75a979 (MD5)Made available in DSpace on 2017-06-26T16:36:23Z (GMT). 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dc.title.por.fl_str_mv |
BIOPROSPECÇÃO DE FUNGOS FILAMENTOSOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE POLIGALACTURONASES: POTENCIAL PECTINOLÍTICO DE Penicillium sp. |
title |
BIOPROSPECÇÃO DE FUNGOS FILAMENTOSOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE POLIGALACTURONASES: POTENCIAL PECTINOLÍTICO DE Penicillium sp. |
spellingShingle |
BIOPROSPECÇÃO DE FUNGOS FILAMENTOSOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE POLIGALACTURONASES: POTENCIAL PECTINOLÍTICO DE Penicillium sp. WOUK, THAÍS Pectinases sacarificação Delineamento Central Composto Rotacional substrato alternativo pectinases saccharification Central Composite Rotatable Design alternative substrate 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 FILAMENTOSOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE POLIGALACTURONASES: POTENCIAL PECTINOLÍTICO DE Penicillium sp. |
title_full |
BIOPROSPECÇÃO DE FUNGOS FILAMENTOSOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE POLIGALACTURONASES: POTENCIAL PECTINOLÍTICO DE Penicillium sp. |
title_fullStr |
BIOPROSPECÇÃO DE FUNGOS FILAMENTOSOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE POLIGALACTURONASES: POTENCIAL PECTINOLÍTICO DE Penicillium sp. |
title_full_unstemmed |
BIOPROSPECÇÃO DE FUNGOS FILAMENTOSOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE POLIGALACTURONASES: POTENCIAL PECTINOLÍTICO DE Penicillium sp. |
title_sort |
BIOPROSPECÇÃO DE FUNGOS FILAMENTOSOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE POLIGALACTURONASES: POTENCIAL PECTINOLÍTICO DE Penicillium sp. |
author |
WOUK, THAÍS |
author_facet |
WOUK, THAÍS |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Knob, Adriana |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/1289646338155108 |
dc.contributor.authorID.fl_str_mv |
075.938.129-10 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/4742847602269407 |
dc.contributor.author.fl_str_mv |
WOUK, THAÍS |
contributor_str_mv |
Knob, Adriana |
dc.subject.por.fl_str_mv |
Pectinases sacarificação Delineamento Central Composto Rotacional substrato alternativo |
topic |
Pectinases sacarificação Delineamento Central Composto Rotacional substrato alternativo pectinases saccharification Central Composite Rotatable Design alternative substrate 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 |
pectinases saccharification Central Composite Rotatable Design alternative substrate |
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 |
Pectinases have been extensively studied due to their wide variety of applications in the many industrial sectors. Among these enzymes, polygalacturonases (PG) are used mainly in the fruit juices clarification process. The selection of enzyme producing microorganisms becomes an important step in biotechnological processes. Thus, this study aimed to isolate several fungal strains and evaluate its potential production of PGs, using pineapple pell and passion fruit peel as substrates. Following the selection of the best producer strain, the Plackett-Burmann design was used in order to select the variables that most influence the production of these enzymes. Afther that, the optimal parameters for PGs production were stablishedd, using the Central Composite Rotary Design and the Response Surface Methodology. Finally, the PGs produced were biochemically characterized. As a result, 46 strains were isolated, showing, through the semi-quantitative test, the PG production hability for all strains evaluated. In the quantitative test, it was verified that the FX32 fungus was the best producer of PGs, corresponding to 6.12 U mL-1. Through the use of Plackett-Burmann design, it was found that the substrate concentration, pH and culture time were the parameters that significant influence PGs production. For these reason, these factors were considered as the independent variables in the Central Composite Rotary Design. Thus, the best conditions established for PGs production PG by Penicillium sp. were 2.9% of pineapple peel as substrate, culture pH of 7.8 and cultivation time of 8.6 days, corresponding to a production of 106.20 U mL-1. With regard to biochemical characteristics, it was verified that the enzymes produced by this fungal strain were more active at 50 °C and at pH 5.5. PGs were also very stable at pH 5.0 and 5.5, exibithing more than 80% of its initial activity under these conditions. Em relation to thermal stability, these enzymes were more stable at 45 °C. The effect of several ions and substances on the PG activity was studied. The PGs were activated by Mn2+, Ca2+ and mercaptoethanol and strong inhibited by Hg+. Finally, the hydrolysis of the pineapple peel was performed using Penicillium sp. and Aspergillus flavus crude extracts. A glucose yield equivalent to 67.42 mg g-1 substrate was achieved, under optimal hydrolysis conditions, corresponding to 15 hours of reaction, reactional pH 5.0 and reactional temperature of 50 °C. The pineapple peel was a good inducer for PGs production by Penicillium sp., as well as a promising substrate for glucose achievement through its enzymatic hydrolysis. This results provide important subsidies in order to infer industrial applications for the enzymes under study. |
publishDate |
2017 |
dc.date.accessioned.fl_str_mv |
2017-06-26T16:36:23Z |
dc.date.issued.fl_str_mv |
2017-02-23 |
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 |
WOUK, THAÍS. BIOPROSPECÇÃO DE FUNGOS FILAMENTOSOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE POLIGALACTURONASES: POTENCIAL PECTINOLÍTICO DE Penicillium sp.. 2017. 94 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/732 |
identifier_str_mv |
WOUK, THAÍS. BIOPROSPECÇÃO DE FUNGOS FILAMENTOSOS ISOLADOS DE REMANESCENTES DE FLORESTA OMBRÓFILA MISTA PRODUTORES DE POLIGALACTURONASES: POTENCIAL PECTINOLÍTICO DE Penicillium sp.. 2017. 94 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/732 |
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Universidade Estadual do Centro-Oeste |
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Universidade Estadual do Centro-Oeste |
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