Isolamento de fungos filamentosos produtores de inulinase

Detalhes bibliográficos
Autor(a) principal: Castanon, Francisco Singulani
Data de Publicação: 2013
Tipo de documento: Dissertação
Idioma: por
Título da fonte: LOCUS Repositório Institucional da UFV
Texto Completo: http://locus.ufv.br/handle/123456789/5363
Resumo: The inulinases present a great variety of applications, such as the production of fructose syrup, bioethanol and fructooligosaccharides. The industrial versatility presented by these enzymes drives the discovery of new sources of microbial inulinase. In this study, 48 fungi were isolated from the rhizosphere of yacon tuber (Polymnia sanchifolia). The criteria to screening the fungi producers of inulinase were: growth in inulin as unique carbon source and enzyme activitiy by the DNS method. Six promising fungi were selected for the analysis of halo formation by inulin hydrolysis and molecular identification using the ITS rDNA region. For the following studies, the fungus Fusarium oxysporum F6 was selected for having the best production of inulinase and the conditions of inulinase production by this fungus were studied. The best enzyme production occurred at 72 h of growth in minimal medium with 2% inulin. Through the response surface methodology using solid-state fermentation, it was possible to estimate that the increase of the inulin concentration affected the inulinase production by this fungus, reaching a production of up to 2.2 U/ml of inulinase. Optimal activity was observed at 50 °C and pH 5.5. The thin layer chromatography showed that the enzyme has endo-inulinase activity. Comparison of the amino acid sequence of F. oxysporum f. sp. cubense 4 with other fungal inulinases reinforce this characteristic, given the presence of conserved regions with the sequences of other endo-inulinase. The phylogenetic analysis also showed a similarity between the amino acid sequences encoding fungal endo-inulinase. This study demonstrated that the yacon tuber is a good source for the isolation of fungal inulinase producers because a reasonable number of isolates were obtained in trials using this source. The fungus Fusarium oxysporum F6 showed the greatest potential for inulinase production, mainly in solid state fermentation. The fungus Staphylotrichum boninense was also isolated and inulinase production of this fungus will be studied in more detail, due to the reports of their differential production of secondary metabolites by this fungus.
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spelling Castanon, Francisco Singulanihttp://lattes.cnpq.br/6750687954545773Kasuya, Maria Catarina Megumihttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4721444T5Queiroz, Marisa Vieira dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4785812Z5Bazzolli, Denise Mara Soareshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4761710D6Ribon, Andréa de Oliveira Barroshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4727026E62015-03-26T13:51:58Z2014-10-072015-03-26T13:51:58Z2013-09-06CASTANON, Francisco Singulani. Isolation of filamentous fungi producing inulinase. 2013. 55 f. Dissertação (Mestrado em Associações micorrízicas; Bactérias láticas e probióticos; Biologia molecular de fungos de interesse) - Universidade Federal de Viçosa, Viçosa, 2013.http://locus.ufv.br/handle/123456789/5363The inulinases present a great variety of applications, such as the production of fructose syrup, bioethanol and fructooligosaccharides. The industrial versatility presented by these enzymes drives the discovery of new sources of microbial inulinase. In this study, 48 fungi were isolated from the rhizosphere of yacon tuber (Polymnia sanchifolia). The criteria to screening the fungi producers of inulinase were: growth in inulin as unique carbon source and enzyme activitiy by the DNS method. Six promising fungi were selected for the analysis of halo formation by inulin hydrolysis and molecular identification using the ITS rDNA region. For the following studies, the fungus Fusarium oxysporum F6 was selected for having the best production of inulinase and the conditions of inulinase production by this fungus were studied. The best enzyme production occurred at 72 h of growth in minimal medium with 2% inulin. Through the response surface methodology using solid-state fermentation, it was possible to estimate that the increase of the inulin concentration affected the inulinase production by this fungus, reaching a production of up to 2.2 U/ml of inulinase. Optimal activity was observed at 50 °C and pH 5.5. The thin layer chromatography showed that the enzyme has endo-inulinase activity. Comparison of the amino acid sequence of F. oxysporum f. sp. cubense 4 with other fungal inulinases reinforce this characteristic, given the presence of conserved regions with the sequences of other endo-inulinase. The phylogenetic analysis also showed a similarity between the amino acid sequences encoding fungal endo-inulinase. This study demonstrated that the yacon tuber is a good source for the isolation of fungal inulinase producers because a reasonable number of isolates were obtained in trials using this source. The fungus Fusarium oxysporum F6 showed the greatest potential for inulinase production, mainly in solid state fermentation. The fungus Staphylotrichum boninense was also isolated and inulinase production of this fungus will be studied in more detail, due to the reports of their differential production of secondary metabolites by this fungus.As inulinases apresentam uma variedade de aplicações, como a produção de xarope de frutose, bioetanol e frutooligossarídeos. A versatilidade industrial apresentada por estas enzimas impulsiona a descoberta de novas fontes de inulinases microbianas. Neste estudo, 48 fungos foram isolados a partir da rizosfera do tubérculo yacon (Polymnia sanchifolia). Os isolados passaram por uma triagem, em que os critérios avaliados foram o crescimento em meio mínimo contendo 1% de inulina como única fonte de carbono e a atividade enzimática pelo método de DNS. Os seis fungos mais promissores foram selecionados para a análise da formação de halo de hidrólise da inulina e identificação molecular utilizando a região ITS rDNA. Para os estudos seguintes, o fungo Fusarium oxysporum F6 foi selecionado por apresentar a melhor produção de inulinase e as condições de produção de inulinase por esse fungo foram estudadas. A melhor produção da enzima ocorreu em 72 h de crescimento em meio mínimo e em 2% de inulina. Através da metodologia de superfície de resposta, utilizando a fermentação em estado sólido, foi possível verificar que o aumento na concentração de inulina afetou positivamente a produção de inulinase por esse fungo, alcançando uma produção de até 2,2 U/mL de inulinase. A atividade ótima foi verificada na temperatura de 50 °C e pH 5,5. A cromatografia de camada delgada mostrou que a enzima apresenta atividade de endo-inulinase. Comparações de uma sequência de aminoácidos de F. oxysporum f. sp. cubense 4 com outras inulinases fúngicas reforçaram essa característica, devido à presença de regiões conservadas com as sequências de outras endo-inulinases. A análise filogenética também demonstrou a proximidade entre as sequências de aminoácidos que codificam endo-inulinases fúngicas. Este trabalho demonstrou que o tubérculo yacon é uma boa fonte para o isolamento de fungos produtores de inulinase, pois um razoável número de isolados foi obtido nas triagens utilizando essa fonte. O fungo Fusarium oxysporum F6 apresentou o maior potencial de produção de inulinase, principalmente em fermentação no estado sólido. O fungo Staphylotrichum boninense também foi isolado e a produção de inulinase desse fungo ainda será estudada mais detalhadamente, devido aos relatos de sua produção diferenciada de metabólitos secundários.Conselho Nacional de Desenvolvimento Científico e Tecnológicoapplication/pdfporUniversidade Federal de ViçosaMestrado em Microbiologia AgrícolaUFVBRAssociações micorrízicas; Bactérias láticas e probióticos; Biologia molecular de fungos de interesseYaconInulinaIlunilaseFusarium oxyporumYaconInulinIlunilaseFusarium oxyporumCNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::MICROBIOLOGIA AGRICOLAIsolamento de fungos filamentosos produtores de inulinaseIsolation of filamentous fungi producing inulinaseinfo: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/pdf1021310https://locus.ufv.br//bitstream/123456789/5363/1/texto%20completo.pdf0c045c919e03b4c610ced66f97c4c031MD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain97017https://locus.ufv.br//bitstream/123456789/5363/2/texto%20completo.pdf.txt0358c788b3a896e229e2d672cc94fcf3MD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3511https://locus.ufv.br//bitstream/123456789/5363/3/texto%20completo.pdf.jpgb55fd3bc2dd19609e73ac49fcf7da1f1MD53123456789/53632016-04-10 23:19:47.047oai:locus.ufv.br:123456789/5363Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-11T02:19:47LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.por.fl_str_mv Isolamento de fungos filamentosos produtores de inulinase
dc.title.alternative.eng.fl_str_mv Isolation of filamentous fungi producing inulinase
title Isolamento de fungos filamentosos produtores de inulinase
spellingShingle Isolamento de fungos filamentosos produtores de inulinase
Castanon, Francisco Singulani
Yacon
Inulina
Ilunilase
Fusarium oxyporum
Yacon
Inulin
Ilunilase
Fusarium oxyporum
CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::MICROBIOLOGIA AGRICOLA
title_short Isolamento de fungos filamentosos produtores de inulinase
title_full Isolamento de fungos filamentosos produtores de inulinase
title_fullStr Isolamento de fungos filamentosos produtores de inulinase
title_full_unstemmed Isolamento de fungos filamentosos produtores de inulinase
title_sort Isolamento de fungos filamentosos produtores de inulinase
author Castanon, Francisco Singulani
author_facet Castanon, Francisco Singulani
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/6750687954545773
dc.contributor.author.fl_str_mv Castanon, Francisco Singulani
dc.contributor.advisor-co1.fl_str_mv Kasuya, Maria Catarina Megumi
dc.contributor.advisor-co1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4721444T5
dc.contributor.advisor-co2.fl_str_mv Queiroz, Marisa Vieira de
dc.contributor.advisor-co2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4785812Z5
dc.contributor.advisor1.fl_str_mv Bazzolli, Denise Mara Soares
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4761710D6
dc.contributor.referee1.fl_str_mv Ribon, Andréa de Oliveira Barros
dc.contributor.referee1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4727026E6
contributor_str_mv Kasuya, Maria Catarina Megumi
Queiroz, Marisa Vieira de
Bazzolli, Denise Mara Soares
Ribon, Andréa de Oliveira Barros
dc.subject.por.fl_str_mv Yacon
Inulina
Ilunilase
Fusarium oxyporum
topic Yacon
Inulina
Ilunilase
Fusarium oxyporum
Yacon
Inulin
Ilunilase
Fusarium oxyporum
CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::MICROBIOLOGIA AGRICOLA
dc.subject.eng.fl_str_mv Yacon
Inulin
Ilunilase
Fusarium oxyporum
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::MICROBIOLOGIA AGRICOLA
description The inulinases present a great variety of applications, such as the production of fructose syrup, bioethanol and fructooligosaccharides. The industrial versatility presented by these enzymes drives the discovery of new sources of microbial inulinase. In this study, 48 fungi were isolated from the rhizosphere of yacon tuber (Polymnia sanchifolia). The criteria to screening the fungi producers of inulinase were: growth in inulin as unique carbon source and enzyme activitiy by the DNS method. Six promising fungi were selected for the analysis of halo formation by inulin hydrolysis and molecular identification using the ITS rDNA region. For the following studies, the fungus Fusarium oxysporum F6 was selected for having the best production of inulinase and the conditions of inulinase production by this fungus were studied. The best enzyme production occurred at 72 h of growth in minimal medium with 2% inulin. Through the response surface methodology using solid-state fermentation, it was possible to estimate that the increase of the inulin concentration affected the inulinase production by this fungus, reaching a production of up to 2.2 U/ml of inulinase. Optimal activity was observed at 50 °C and pH 5.5. The thin layer chromatography showed that the enzyme has endo-inulinase activity. Comparison of the amino acid sequence of F. oxysporum f. sp. cubense 4 with other fungal inulinases reinforce this characteristic, given the presence of conserved regions with the sequences of other endo-inulinase. The phylogenetic analysis also showed a similarity between the amino acid sequences encoding fungal endo-inulinase. This study demonstrated that the yacon tuber is a good source for the isolation of fungal inulinase producers because a reasonable number of isolates were obtained in trials using this source. The fungus Fusarium oxysporum F6 showed the greatest potential for inulinase production, mainly in solid state fermentation. The fungus Staphylotrichum boninense was also isolated and inulinase production of this fungus will be studied in more detail, due to the reports of their differential production of secondary metabolites by this fungus.
publishDate 2013
dc.date.issued.fl_str_mv 2013-09-06
dc.date.available.fl_str_mv 2014-10-07
2015-03-26T13:51:58Z
dc.date.accessioned.fl_str_mv 2015-03-26T13:51:58Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
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dc.identifier.citation.fl_str_mv CASTANON, Francisco Singulani. Isolation of filamentous fungi producing inulinase. 2013. 55 f. Dissertação (Mestrado em Associações micorrízicas; Bactérias láticas e probióticos; Biologia molecular de fungos de interesse) - Universidade Federal de Viçosa, Viçosa, 2013.
dc.identifier.uri.fl_str_mv http://locus.ufv.br/handle/123456789/5363
identifier_str_mv CASTANON, Francisco Singulani. Isolation of filamentous fungi producing inulinase. 2013. 55 f. Dissertação (Mestrado em Associações micorrízicas; Bactérias láticas e probióticos; Biologia molecular de fungos de interesse) - Universidade Federal de Viçosa, Viçosa, 2013.
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