Culturas oxidadoras de metano provenientes de ambientes aquáticos brasileiros e seu potencial para a produção de polihidroxialcanoatos
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=6616534 https://repositorio.unifesp.br/handle/11600/52569 |
Resumo: | Microorganisms play a key role in the methane cycle. Methanogenic arches produce most of the planet's biogenic methane in anaerobic environments. Methanotrophic aerobic bacteria use methane as a source of carbon and energy, being responsible for the consumption of gas in terrestrial and aquatic environments, as well as having potential for the production of polyhydroxyalkanoates (PHA). This study aimed to study enriched metanotrophic cultures of Brazilian aquatic environments, aiming to promote and characterize the production of PHB from methane. For this, an approach was that combines dependent and independent of cultivation techniques dependent were adopted. Cultures of interest were identified by sequencing the new generation of the 16S RNAr gene on the Illumina MiSeq platform. The detection and quantification of methanotrophic indicator genes and PHA production (pmoA and phaC, respectively) were done by the PCR technique. Nile red staining and fluorescence microscopy were used as indicators of the production of polyhydroxybutyrates. The results indicated the presence of pmoA and phaC genes in all cultures analyzed. The initial Nile red assessment indicated the presence of PHAs associated with methane consumption in the cultures of interest. The sequencing results indicate the dominance of the methanotrophic genera Methylocystis (38.8% - 75%) in three of the five established cultures. One of the cultures, RB-5, was selected for a stirred vial production assay. The accumulation of polymers was quantified by means of gas chromatography. Growth curves were constructed for the determination of methanotrophic activity. The results for the production assay show a low accumulation of the poly-3-hydroxybutyrate and poly-3-hydroxyvalerate polymers by the RB-5 culture. It is expected that this work will contribute to the subsequent realization of tests aiming the optimization of polymer production by the RB-5 culture. |
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Culturas oxidadoras de metano provenientes de ambientes aquáticos brasileiros e seu potencial para a produção de polihidroxialcanoatosPHBPHAMethanotrophic bacteriaBiopolymersMethanePHBPHABactérias metanotróficasBiopolímerosMetanoMicroorganisms play a key role in the methane cycle. Methanogenic arches produce most of the planet's biogenic methane in anaerobic environments. Methanotrophic aerobic bacteria use methane as a source of carbon and energy, being responsible for the consumption of gas in terrestrial and aquatic environments, as well as having potential for the production of polyhydroxyalkanoates (PHA). This study aimed to study enriched metanotrophic cultures of Brazilian aquatic environments, aiming to promote and characterize the production of PHB from methane. For this, an approach was that combines dependent and independent of cultivation techniques dependent were adopted. Cultures of interest were identified by sequencing the new generation of the 16S RNAr gene on the Illumina MiSeq platform. The detection and quantification of methanotrophic indicator genes and PHA production (pmoA and phaC, respectively) were done by the PCR technique. Nile red staining and fluorescence microscopy were used as indicators of the production of polyhydroxybutyrates. The results indicated the presence of pmoA and phaC genes in all cultures analyzed. The initial Nile red assessment indicated the presence of PHAs associated with methane consumption in the cultures of interest. The sequencing results indicate the dominance of the methanotrophic genera Methylocystis (38.8% - 75%) in three of the five established cultures. One of the cultures, RB-5, was selected for a stirred vial production assay. The accumulation of polymers was quantified by means of gas chromatography. Growth curves were constructed for the determination of methanotrophic activity. The results for the production assay show a low accumulation of the poly-3-hydroxybutyrate and poly-3-hydroxyvalerate polymers by the RB-5 culture. It is expected that this work will contribute to the subsequent realization of tests aiming the optimization of polymer production by the RB-5 culture.Os microorganismos têm papel chave no ciclo do metano. Arqueias metanogênicas produzem a maior parte do metano biogênico do planeta, em ambientes anaeróbios. As bactérias metanotróficas aeróbias utilizam o metano como fonte de carbono e energia, sendo responsáveis pelo consumo do gás em ambientes terrestres e aquáticos, além de possuírem potencial para a produção de polihidroxialcanoatos (PHA). Este estudo teve como objetivo estudar culturas metanotróficas enriquecidas de ambientes aquáticos brasileiros, visando promover e caracterizar a produção de PHB a partir de metano. Para isso, foi adotada uma abordagem que alia técnicas dependentes e independentes de cultivo. As culturas de interesse foram identificadas através do sequenciamento de nova geração do gene RNAr 16S em plataforma Illumina MiSeq. A detecção e quantificação de genes indicadores de metanotrofia e produção de PHA (pmoA e phaC, respectivamente) foram feitas por meio da técnica PCR. A coloração com vermelho do Nilo e a microscopia de fluorescência foram utilizadas como indicadores da produção de polihidroxibutiratos. Os resultados indicaram a presença dos genes pmoA e phaC em todas as culturas analisadas. A avaliação inicial com vermelho do Nilo indicou a presença de PHA’s associada ao consumo de metano nos cultivos de interesse. Os resultados referentes ao sequenciamento indicam a dominânica do gênero metanotrófico Methylocystis (38,8% - 75,%) em três, dos cinco cultivos estabelecidos. Um dos cultivos, RB-5 foi selecionado para a realização de um ensaio de produção em frasco agitado. A acumulação de polímeros foi quantificada por meio de cromatografia gasosa. Foram construídas curvas de crescimento para a determinação da atividade metanotrófica. Os resultados referentes ao ensaio de produção evidenciam uma baixa acumulação dos polímeros poli-3-hidroxibutirato e poli-3-hidroxivalerato pela cultura RB-5. Espera-se que o trabalho contribua para a posterior realização de ensaios visando a otimização da produção de polímeros pela cultura RB-5.Dados abertos - Sucupira - Teses e dissertações (2018)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade Federal de São PauloNakayama, Cristina Rossi [UNIFESP]Universidade Federal de São Paulo (UNIFESP)Soares, Juliana Aparecida Bomjardim [UNIFESP]2020-03-25T11:44:03Z2020-03-25T11:44:03Z2018-10-05info:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/publishedVersion94 p.application/pdfhttps://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=66165342018-0510.pdfhttps://repositorio.unifesp.br/handle/11600/52569porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP2024-08-02T17:06:26Zoai:repositorio.unifesp.br/:11600/52569Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestbiblioteca.csp@unifesp.bropendoar:34652024-08-02T17:06:26Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.none.fl_str_mv |
Culturas oxidadoras de metano provenientes de ambientes aquáticos brasileiros e seu potencial para a produção de polihidroxialcanoatos |
title |
Culturas oxidadoras de metano provenientes de ambientes aquáticos brasileiros e seu potencial para a produção de polihidroxialcanoatos |
spellingShingle |
Culturas oxidadoras de metano provenientes de ambientes aquáticos brasileiros e seu potencial para a produção de polihidroxialcanoatos Soares, Juliana Aparecida Bomjardim [UNIFESP] PHB PHA Methanotrophic bacteria Biopolymers Methane PHB PHA Bactérias metanotróficas Biopolímeros Metano |
title_short |
Culturas oxidadoras de metano provenientes de ambientes aquáticos brasileiros e seu potencial para a produção de polihidroxialcanoatos |
title_full |
Culturas oxidadoras de metano provenientes de ambientes aquáticos brasileiros e seu potencial para a produção de polihidroxialcanoatos |
title_fullStr |
Culturas oxidadoras de metano provenientes de ambientes aquáticos brasileiros e seu potencial para a produção de polihidroxialcanoatos |
title_full_unstemmed |
Culturas oxidadoras de metano provenientes de ambientes aquáticos brasileiros e seu potencial para a produção de polihidroxialcanoatos |
title_sort |
Culturas oxidadoras de metano provenientes de ambientes aquáticos brasileiros e seu potencial para a produção de polihidroxialcanoatos |
author |
Soares, Juliana Aparecida Bomjardim [UNIFESP] |
author_facet |
Soares, Juliana Aparecida Bomjardim [UNIFESP] |
author_role |
author |
dc.contributor.none.fl_str_mv |
Nakayama, Cristina Rossi [UNIFESP] Universidade Federal de São Paulo (UNIFESP) |
dc.contributor.author.fl_str_mv |
Soares, Juliana Aparecida Bomjardim [UNIFESP] |
dc.subject.por.fl_str_mv |
PHB PHA Methanotrophic bacteria Biopolymers Methane PHB PHA Bactérias metanotróficas Biopolímeros Metano |
topic |
PHB PHA Methanotrophic bacteria Biopolymers Methane PHB PHA Bactérias metanotróficas Biopolímeros Metano |
description |
Microorganisms play a key role in the methane cycle. Methanogenic arches produce most of the planet's biogenic methane in anaerobic environments. Methanotrophic aerobic bacteria use methane as a source of carbon and energy, being responsible for the consumption of gas in terrestrial and aquatic environments, as well as having potential for the production of polyhydroxyalkanoates (PHA). This study aimed to study enriched metanotrophic cultures of Brazilian aquatic environments, aiming to promote and characterize the production of PHB from methane. For this, an approach was that combines dependent and independent of cultivation techniques dependent were adopted. Cultures of interest were identified by sequencing the new generation of the 16S RNAr gene on the Illumina MiSeq platform. The detection and quantification of methanotrophic indicator genes and PHA production (pmoA and phaC, respectively) were done by the PCR technique. Nile red staining and fluorescence microscopy were used as indicators of the production of polyhydroxybutyrates. The results indicated the presence of pmoA and phaC genes in all cultures analyzed. The initial Nile red assessment indicated the presence of PHAs associated with methane consumption in the cultures of interest. The sequencing results indicate the dominance of the methanotrophic genera Methylocystis (38.8% - 75%) in three of the five established cultures. One of the cultures, RB-5, was selected for a stirred vial production assay. The accumulation of polymers was quantified by means of gas chromatography. Growth curves were constructed for the determination of methanotrophic activity. The results for the production assay show a low accumulation of the poly-3-hydroxybutyrate and poly-3-hydroxyvalerate polymers by the RB-5 culture. It is expected that this work will contribute to the subsequent realization of tests aiming the optimization of polymer production by the RB-5 culture. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-10-05 2020-03-25T11:44:03Z 2020-03-25T11:44:03Z |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=6616534 2018-0510.pdf https://repositorio.unifesp.br/handle/11600/52569 |
url |
https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=6616534 https://repositorio.unifesp.br/handle/11600/52569 |
identifier_str_mv |
2018-0510.pdf |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
94 p. application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de São Paulo |
publisher.none.fl_str_mv |
Universidade Federal de São Paulo |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UNIFESP instname:Universidade Federal de São Paulo (UNIFESP) instacron:UNIFESP |
instname_str |
Universidade Federal de São Paulo (UNIFESP) |
instacron_str |
UNIFESP |
institution |
UNIFESP |
reponame_str |
Repositório Institucional da UNIFESP |
collection |
Repositório Institucional da UNIFESP |
repository.name.fl_str_mv |
Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP) |
repository.mail.fl_str_mv |
biblioteca.csp@unifesp.br |
_version_ |
1814268382810210304 |