Obtenção de inoculante e de coquetel enzimático lignocelulolítico a partir de comunidades microbianas termofílicas
Autor(a) principal: | |
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Data de Publicação: | 2012 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | LOCUS Repositório Institucional da UFV |
Texto Completo: | http://locus.ufv.br/handle/123456789/5349 |
Resumo: | Three thermophilic lignocellulolytic microbial communities were selected by enriched method with continual subcultivation at 55 °C. One community was selected from cow manure compost, another from decaying sugar cane bagasse, and the last one was obtained by mixing aliquots of the culture suspension from the first two. Those communities were able to degrade a cellulosic strip in three cultivation days. Evaluating the maximal day of CMCase and xylanase activity, it was observed that the mixed consortium showed the best results, with CMCase activity of 0.09 U mg-1 at the second day of cultivation, and xylanase activity of 2.86 U mg-1 at the fourth day. These enzymes were partially characterized with relation to temperature and pH of optimal activity. It was observed that CMCase showed the highest activity at 60 °C and pH 5.4, and kept 80% of its activity in a pH range of 4.5-6.5. On the other hand, the best activity for xylanase was verified at 65 °C, and in that same pH range, the enzyme kept 97% of its residual activity. Cell-free extract was concentrated by ultrafiltration. The enzymatic cocktail obtained showed CMCase activity 25-fold higher and xylanase activity 55-fold higher than the crude enzymatic extract. The cocktail was conserved by adding 50% glycerol. After storage for 60 days at 4 °C, xylanase kept 80% of the initial activity and CMCase didn t show loss of activity when kept at 25 °C for the same period. The mixed community cellular mass constitutes an inoculant able to maintain the cellulolytic phenotype after rapid freezing and storage at - 80 °C for 60 days. Fed-batch essay suggested that this community has potential to be manipulated in order to continuously hold the cellulolytic enzymes expression over time. The results evidenced the acquisition of an enzymatic cocktail from an inoculant which cellulolytic activity supported pH changes and optimal activity around 60 °C. |
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Souza, Robson de Assishttp://lattes.cnpq.br/5580102665962863Mantovani, Hilário Cuquettohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4727026Z7Kasuya, Maria Catarina Megumihttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4721444T5Passos, Flávia Maria Lopeshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781817D3Guimarães, Valéria Montezehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4798758T32015-03-26T13:51:56Z2012-08-232015-03-26T13:51:56Z2012-02-17SOUZA, Robson de Assis. Acquisition of an inoculant and a lignocellulolytic enzymatic cocktail from thermophilic microbial communities. 2012. 70 f. Dissertação (Mestrado em Associações micorrízicas; Bactérias láticas e probióticos; Biologia molecular de fungos de interesse) - Universidade Federal de Viçosa, Viçosa, 2012.http://locus.ufv.br/handle/123456789/5349Three thermophilic lignocellulolytic microbial communities were selected by enriched method with continual subcultivation at 55 °C. One community was selected from cow manure compost, another from decaying sugar cane bagasse, and the last one was obtained by mixing aliquots of the culture suspension from the first two. Those communities were able to degrade a cellulosic strip in three cultivation days. Evaluating the maximal day of CMCase and xylanase activity, it was observed that the mixed consortium showed the best results, with CMCase activity of 0.09 U mg-1 at the second day of cultivation, and xylanase activity of 2.86 U mg-1 at the fourth day. These enzymes were partially characterized with relation to temperature and pH of optimal activity. It was observed that CMCase showed the highest activity at 60 °C and pH 5.4, and kept 80% of its activity in a pH range of 4.5-6.5. On the other hand, the best activity for xylanase was verified at 65 °C, and in that same pH range, the enzyme kept 97% of its residual activity. Cell-free extract was concentrated by ultrafiltration. The enzymatic cocktail obtained showed CMCase activity 25-fold higher and xylanase activity 55-fold higher than the crude enzymatic extract. The cocktail was conserved by adding 50% glycerol. After storage for 60 days at 4 °C, xylanase kept 80% of the initial activity and CMCase didn t show loss of activity when kept at 25 °C for the same period. The mixed community cellular mass constitutes an inoculant able to maintain the cellulolytic phenotype after rapid freezing and storage at - 80 °C for 60 days. Fed-batch essay suggested that this community has potential to be manipulated in order to continuously hold the cellulolytic enzymes expression over time. The results evidenced the acquisition of an enzymatic cocktail from an inoculant which cellulolytic activity supported pH changes and optimal activity around 60 °C.Três comunidades microbianas lignocelulolíticas e termofílicas foram selecionadas por subcultivo em meio de enriquecimento a 55 °C. Uma comunidade foi selecionada a partir de esterco de gado em compostagem, outra de bagaço de cana em decomposição e a terceira foi obtida a partir da mistura de alíquotas do meio de cultivo das duas anteriores. Tais comunidades apresentavam a característica de decompor uma fita de celulose em três dias de cultivo. Ao se avaliar o dia de máxima atividade CMCase e xilanase em cada comunidade, verificou-se que os melhores resultados foram encontrados no consórcio misto, com atividade de 0,09 U mg-1 para CMCase no segundo dia de cultivo e 2,86 U mg-1 para xilanase no quarto dia. Essas enzimas foram parcialmente caracterizadas em relação à temperatura e pH ótimos de atuação. Verificou-se que CMCase apresentou maior atividade a 60 °C e pH 5,4, e manteve 80% de sua atividade numa faixa de pH de 4,5 a 6,5. Já para a xilanase, a temperatura ótima foi de 65 °C e nessa mesma faixa de pH manteve uma atividade residual de 97%. O extrato livre de células foi concentrado por ultrafiltração e obteve-se um coquetel enzimático com atividade de CMCase e xilanase maior que no extrato bruto cerca de 25 e 55 vezes, respectivamente. O coquetel foi conservado pela adição de 50% de glicerol. Após 60 dias de armazenamento a 4 °C, a xilanase manteve 80% de sua atividade inicial e a CMCase não apresentou perda de atividade quando mantida a 25 °C pelo mesmo período. A massa celular da comunidade mista constitui um inoculante capaz de manter o fenótipo celulolítico após congelamento rápido e armazenamento a - 80 °C por 60 dias. Ensaio em batelada alimentada mostrou que essa comunidade apresenta potencial para ser manipulada a fim de se manter continuamente a expressão de enzimas celulolíticas ao longo do tempo. Os resultados mostraram que foi possível obter um coquetel de enzimas a partir do inoculante, cuja atividade celulolítica tolerou variações de pH e temperatura ótima em torno de 60 °C.Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade Federal de ViçosaMestrado em Microbiologia AgrícolaUFVBRAssociações micorrízicas; Bactérias láticas e probióticos; Biologia molecular de fungos de interesseBiorrefinariaEnzimas lignocelulolíticasBagaço de cana-de-açúcarComunidade microbianaBiorefineryLignocellulolytic enzymesBagasse cane sugarMicrobial CommunityCNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::MICROBIOLOGIA AGRICOLAObtenção de inoculante e de coquetel enzimático lignocelulolítico a partir de comunidades microbianas termofílicasAcquisition of an inoculant and a lignocellulolytic enzymatic cocktail from thermophilic microbial communitiesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf807100https://locus.ufv.br//bitstream/123456789/5349/1/texto%20completo.pdf0492f1cb907f9d50a51882efd7f76fcdMD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain103222https://locus.ufv.br//bitstream/123456789/5349/2/texto%20completo.pdf.txt9c4f2450b4ad7e327fa22a1506a73de8MD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3661https://locus.ufv.br//bitstream/123456789/5349/3/texto%20completo.pdf.jpg48bceaffc656adf3c1602cd68b4da43eMD53123456789/53492016-04-10 23:18:18.502oai:locus.ufv.br:123456789/5349Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-11T02:18:18LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false |
dc.title.por.fl_str_mv |
Obtenção de inoculante e de coquetel enzimático lignocelulolítico a partir de comunidades microbianas termofílicas |
dc.title.alternative.eng.fl_str_mv |
Acquisition of an inoculant and a lignocellulolytic enzymatic cocktail from thermophilic microbial communities |
title |
Obtenção de inoculante e de coquetel enzimático lignocelulolítico a partir de comunidades microbianas termofílicas |
spellingShingle |
Obtenção de inoculante e de coquetel enzimático lignocelulolítico a partir de comunidades microbianas termofílicas Souza, Robson de Assis Biorrefinaria Enzimas lignocelulolíticas Bagaço de cana-de-açúcar Comunidade microbiana Biorefinery Lignocellulolytic enzymes Bagasse cane sugar Microbial Community CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::MICROBIOLOGIA AGRICOLA |
title_short |
Obtenção de inoculante e de coquetel enzimático lignocelulolítico a partir de comunidades microbianas termofílicas |
title_full |
Obtenção de inoculante e de coquetel enzimático lignocelulolítico a partir de comunidades microbianas termofílicas |
title_fullStr |
Obtenção de inoculante e de coquetel enzimático lignocelulolítico a partir de comunidades microbianas termofílicas |
title_full_unstemmed |
Obtenção de inoculante e de coquetel enzimático lignocelulolítico a partir de comunidades microbianas termofílicas |
title_sort |
Obtenção de inoculante e de coquetel enzimático lignocelulolítico a partir de comunidades microbianas termofílicas |
author |
Souza, Robson de Assis |
author_facet |
Souza, Robson de Assis |
author_role |
author |
dc.contributor.authorLattes.por.fl_str_mv |
http://lattes.cnpq.br/5580102665962863 |
dc.contributor.author.fl_str_mv |
Souza, Robson de Assis |
dc.contributor.advisor-co1.fl_str_mv |
Mantovani, Hilário Cuquetto |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4727026Z7 |
dc.contributor.advisor-co2.fl_str_mv |
Kasuya, Maria Catarina Megumi |
dc.contributor.advisor-co2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4721444T5 |
dc.contributor.advisor1.fl_str_mv |
Passos, Flávia Maria Lopes |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781817D3 |
dc.contributor.referee1.fl_str_mv |
Guimarães, Valéria Monteze |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4798758T3 |
contributor_str_mv |
Mantovani, Hilário Cuquetto Kasuya, Maria Catarina Megumi Passos, Flávia Maria Lopes Guimarães, Valéria Monteze |
dc.subject.por.fl_str_mv |
Biorrefinaria Enzimas lignocelulolíticas Bagaço de cana-de-açúcar Comunidade microbiana |
topic |
Biorrefinaria Enzimas lignocelulolíticas Bagaço de cana-de-açúcar Comunidade microbiana Biorefinery Lignocellulolytic enzymes Bagasse cane sugar Microbial Community CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::MICROBIOLOGIA AGRICOLA |
dc.subject.eng.fl_str_mv |
Biorefinery Lignocellulolytic enzymes Bagasse cane sugar Microbial Community |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::MICROBIOLOGIA AGRICOLA |
description |
Three thermophilic lignocellulolytic microbial communities were selected by enriched method with continual subcultivation at 55 °C. One community was selected from cow manure compost, another from decaying sugar cane bagasse, and the last one was obtained by mixing aliquots of the culture suspension from the first two. Those communities were able to degrade a cellulosic strip in three cultivation days. Evaluating the maximal day of CMCase and xylanase activity, it was observed that the mixed consortium showed the best results, with CMCase activity of 0.09 U mg-1 at the second day of cultivation, and xylanase activity of 2.86 U mg-1 at the fourth day. These enzymes were partially characterized with relation to temperature and pH of optimal activity. It was observed that CMCase showed the highest activity at 60 °C and pH 5.4, and kept 80% of its activity in a pH range of 4.5-6.5. On the other hand, the best activity for xylanase was verified at 65 °C, and in that same pH range, the enzyme kept 97% of its residual activity. Cell-free extract was concentrated by ultrafiltration. The enzymatic cocktail obtained showed CMCase activity 25-fold higher and xylanase activity 55-fold higher than the crude enzymatic extract. The cocktail was conserved by adding 50% glycerol. After storage for 60 days at 4 °C, xylanase kept 80% of the initial activity and CMCase didn t show loss of activity when kept at 25 °C for the same period. The mixed community cellular mass constitutes an inoculant able to maintain the cellulolytic phenotype after rapid freezing and storage at - 80 °C for 60 days. Fed-batch essay suggested that this community has potential to be manipulated in order to continuously hold the cellulolytic enzymes expression over time. The results evidenced the acquisition of an enzymatic cocktail from an inoculant which cellulolytic activity supported pH changes and optimal activity around 60 °C. |
publishDate |
2012 |
dc.date.available.fl_str_mv |
2012-08-23 2015-03-26T13:51:56Z |
dc.date.issued.fl_str_mv |
2012-02-17 |
dc.date.accessioned.fl_str_mv |
2015-03-26T13:51:56Z |
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 |
SOUZA, Robson de Assis. Acquisition of an inoculant and a lignocellulolytic enzymatic cocktail from thermophilic microbial communities. 2012. 70 f. Dissertação (Mestrado em Associações micorrízicas; Bactérias láticas e probióticos; Biologia molecular de fungos de interesse) - Universidade Federal de Viçosa, Viçosa, 2012. |
dc.identifier.uri.fl_str_mv |
http://locus.ufv.br/handle/123456789/5349 |
identifier_str_mv |
SOUZA, Robson de Assis. Acquisition of an inoculant and a lignocellulolytic enzymatic cocktail from thermophilic microbial communities. 2012. 70 f. Dissertação (Mestrado em Associações micorrízicas; Bactérias láticas e probióticos; Biologia molecular de fungos de interesse) - Universidade Federal de Viçosa, Viçosa, 2012. |
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http://locus.ufv.br/handle/123456789/5349 |
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por |
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Universidade Federal de Viçosa |
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Mestrado em Microbiologia Agrícola |
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UFV |
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BR |
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Associações micorrízicas; Bactérias láticas e probióticos; Biologia molecular de fungos de interesse |
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Universidade Federal de Viçosa |
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