Metabolômica de leveduras fermentadoras de xilose na identificação de alvos metabólicos para aumento da produção de etanol 2g
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| Hlavní autor: | |
|---|---|
| Datum vydání: | 2019 |
| Médium: | Doctoral thesis |
| Jazyk: | por |
| Zdroj: | Repositório Institucional da UFG |
| Download full: | http://repositorio.bc.ufg.br/tede/handle/tede/9653 |
Shrnutí: | One of the challenges in the production of second generation (2G) ethanol is in the development of efficient yeasts to convert xylose into ethanol in the process of fermenting sugars. Thus, the comprehensive analysis of xylose fermenting yeast metabolism is essential to facilitate the identification of the limiting factors in the conversion metabolism of this pentose and, thus, help in the construction of more efficient genetically modified strains. The objective of this work was to use an advanced analytical tool, the metabolomics, to quantify the main metabolites related to the xylose to ethanol conversion pathways in four species of yeast xyloses: Scheffersomyces stipitis, Spathaspora passalidarum, Spathaspora arborariae and Candida Tenuis. The yeasts were grown in xylose under two different growth conditions, aerobic and microaerobic. The preparation of the sample for metabolic analysis included the steps of quenching and extraction of the metabolites, for which the protocols of ice cold methanol and boiling ethanol, were used, respectively. The quantification of the metabolites was performed by ultra high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/ MS), using ion-pair chromatography and hydrophilic interaction liquid chromatography. The data obtained in the quantification were processed and treated through analysis of variance (ANOVA) using RStudio software. A total of eleven metabolites of the xylose metabolism pathway were quantified and validated through metabolic flow analysis (MFA) with R2>90. These metabolites were used to construct the metabolic flow of the three species of yeasts (Scheffersomyces stipitis, Spathaspora passalidarum and Spathaspora arborariae) analyzed. The comparison of molecular targets in yeast xylose showed that the S. stipitis and S. passalidarum species have the best yields of ethanol when grown with limited oxygen. The metabolomic analysis performed in this study may help in the genetic improvement of these microorganisms and in the production of 2G ethanol. |
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| author | Cavalcanti, Christiane Gonçalves Campos |
| author_browse | Cavalcanti, Christiane Gonçalves Campos |
| author_facet | Cavalcanti, Christiane Gonçalves Campos |
| author_role | author |
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| collection | Repositório Institucional da UFG |
| contributor_str_mv | Abdelnur, Patrícia Verardi Rodrigue, Clenilson Martins Abdelnur, Patrícia Verardi Vaz, Boniek Gontijo Côrrea, Mauro Vicentini Parachin, Nádia Skorupa Silva, Caio de Oliveira Gorgulho |
| dc.contributor.advisor-co1.fl_str_mv | Rodrigue, Clenilson Martins |
| dc.contributor.advisor-co1Lattes.fl_str_mv | http://lattes.cnpq.br/9689635455677053 |
| dc.contributor.advisor1.fl_str_mv | Abdelnur, Patrícia Verardi |
| dc.contributor.advisor1Lattes.fl_str_mv | http://lattes.cnpq.br/0386805742968055 |
| dc.contributor.author.fl_str_mv | Cavalcanti, Christiane Gonçalves Campos |
| dc.contributor.authorLattes.fl_str_mv | http://lattes.cnpq.br/4601394638111230 |
| dc.contributor.referee1.fl_str_mv | Abdelnur, Patrícia Verardi |
| dc.contributor.referee2.fl_str_mv | Vaz, Boniek Gontijo |
| dc.contributor.referee3.fl_str_mv | Côrrea, Mauro Vicentini |
| dc.contributor.referee4.fl_str_mv | Parachin, Nádia Skorupa |
| dc.contributor.referee5.fl_str_mv | Silva, Caio de Oliveira Gorgulho |
| dc.date.accessioned.fl_str_mv | 2019-05-28T12:23:18Z |
| dc.date.issued.fl_str_mv | 2019-02-18 |
| dc.format.none.fl_str_mv | application/pdf |
| dc.identifier.citation.fl_str_mv | CAMPOS, C. G. Metabolômica de leveduras fermentadoras de xilose na identificação de alvos metabólicos para aumento da produção de etanol 2g. 2019. 120 f. Tese (Doutorado em Química) - Universidade Federal de Goiás, Goiânia, 2019. |
| dc.identifier.uri.fl_str_mv | http://repositorio.bc.ufg.br/tede/handle/tede/9653 |
| dc.language.iso.fl_str_mv | por |
| dc.publisher.country.fl_str_mv | Brasil |
| dc.publisher.department.fl_str_mv | Instituto de Química - IQ (RG) |
| dc.publisher.initials.fl_str_mv | UFG |
| dc.publisher.none.fl_str_mv | Universidade Federal de Goiás |
| dc.publisher.program.fl_str_mv | Programa de Pós-graduação em Química (IQ) |
| dc.relation.cnpq.fl_str_mv | 1143264510063203452 |
| dc.relation.confidence.fl_str_mv | 600 600 600 600 |
| dc.relation.department.fl_str_mv | 7826066743741197278 |
| dc.relation.program.fl_str_mv | 663693921325415158 |
| dc.relation.sponsorship.fl_str_mv | 2075167498588264571 |
| dc.rights.driver.fl_str_mv | http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
| dc.source.none.fl_str_mv | reponame:Repositório Institucional da UFG instname:Universidade Federal de Goiás (UFG) instacron:UFG |
| dc.subject.cnpq.fl_str_mv | QUIMICA ANALITICA::INSTRUMENTACAO ANALITICA |
| dc.subject.eng.fl_str_mv | Metabolomics Mass spectrometry Liquid chromatography Biotechnology Yeasts |
| dc.subject.por.fl_str_mv | Metabolômica Espectrometria de massas Cromatografia líquida Biotecnologia e leveduras |
| dc.title.alternative.eng.fl_str_mv | Metabolomics of xylose fermenting yeasts in the identification of metabolic targets to enhance ethanol 2g production |
| dc.title.eng.fl_str_mv | Metabolômica de leveduras fermentadoras de xilose na identificação de alvos metabólicos para aumento da produção de etanol 2g |
| dc.type.driver.fl_str_mv | info:eu-repo/semantics/doctoralThesis |
| dc.type.status.fl_str_mv | info:eu-repo/semantics/publishedVersion |
| description | One of the challenges in the production of second generation (2G) ethanol is in the development of efficient yeasts to convert xylose into ethanol in the process of fermenting sugars. Thus, the comprehensive analysis of xylose fermenting yeast metabolism is essential to facilitate the identification of the limiting factors in the conversion metabolism of this pentose and, thus, help in the construction of more efficient genetically modified strains. The objective of this work was to use an advanced analytical tool, the metabolomics, to quantify the main metabolites related to the xylose to ethanol conversion pathways in four species of yeast xyloses: Scheffersomyces stipitis, Spathaspora passalidarum, Spathaspora arborariae and Candida Tenuis. The yeasts were grown in xylose under two different growth conditions, aerobic and microaerobic. The preparation of the sample for metabolic analysis included the steps of quenching and extraction of the metabolites, for which the protocols of ice cold methanol and boiling ethanol, were used, respectively. The quantification of the metabolites was performed by ultra high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/ MS), using ion-pair chromatography and hydrophilic interaction liquid chromatography. The data obtained in the quantification were processed and treated through analysis of variance (ANOVA) using RStudio software. A total of eleven metabolites of the xylose metabolism pathway were quantified and validated through metabolic flow analysis (MFA) with R2>90. These metabolites were used to construct the metabolic flow of the three species of yeasts (Scheffersomyces stipitis, Spathaspora passalidarum and Spathaspora arborariae) analyzed. The comparison of molecular targets in yeast xylose showed that the S. stipitis and S. passalidarum species have the best yields of ethanol when grown with limited oxygen. The metabolomic analysis performed in this study may help in the genetic improvement of these microorganisms and in the production of 2G ethanol. |
| eu_rights_str_mv | openAccess |
| format | doctoralThesis |
| id | UFG-2_edcf28682cae3bba8ab805b27ca4e109 |
| identifier_str_mv | CAMPOS, C. G. Metabolômica de leveduras fermentadoras de xilose na identificação de alvos metabólicos para aumento da produção de etanol 2g. 2019. 120 f. Tese (Doutorado em Química) - Universidade Federal de Goiás, Goiânia, 2019. |
| instacron_str | UFG |
| institution | UFG |
| instname_str | Universidade Federal de Goiás (UFG) |
| language | por |
| network_acronym_str | UFG-2 |
| network_name_str | Repositório Institucional da UFG |
| oai_identifier_str | oai:repositorio.bc.ufg.br:tede/9653 |
| publishDate | 2019 |
| publishDateSort | 2019 |
| publisher.none.fl_str_mv | Universidade Federal de Goiás |
| reponame_str | Repositório Institucional da UFG |
| repository.mail.fl_str_mv | grt.bc@ufg.br |
| repository.name.fl_str_mv | Repositório Institucional da UFG - Universidade Federal de Goiás (UFG) |
| repository_id_str | oai:repositorio.bc.ufg.br:tede/1234 |
| rights_invalid_str_mv | http://creativecommons.org/licenses/by-nc-nd/4.0/ |
| spelling | Abdelnur, Patrícia Verardihttp://lattes.cnpq.br/0386805742968055Rodrigue, Clenilson Martinshttp://lattes.cnpq.br/9689635455677053Abdelnur, Patrícia VerardiVaz, Boniek GontijoCôrrea, Mauro VicentiniParachin, Nádia SkorupaSilva, Caio de Oliveira Gorgulhohttp://lattes.cnpq.br/4601394638111230Cavalcanti, Christiane Gonçalves Campos2019-05-28T12:23:18Z2019-02-18CAMPOS, C. G. Metabolômica de leveduras fermentadoras de xilose na identificação de alvos metabólicos para aumento da produção de etanol 2g. 2019. 120 f. Tese (Doutorado em Química) - Universidade Federal de Goiás, Goiânia, 2019.http://repositorio.bc.ufg.br/tede/handle/tede/9653One of the challenges in the production of second generation (2G) ethanol is in the development of efficient yeasts to convert xylose into ethanol in the process of fermenting sugars. Thus, the comprehensive analysis of xylose fermenting yeast metabolism is essential to facilitate the identification of the limiting factors in the conversion metabolism of this pentose and, thus, help in the construction of more efficient genetically modified strains. The objective of this work was to use an advanced analytical tool, the metabolomics, to quantify the main metabolites related to the xylose to ethanol conversion pathways in four species of yeast xyloses: Scheffersomyces stipitis, Spathaspora passalidarum, Spathaspora arborariae and Candida Tenuis. The yeasts were grown in xylose under two different growth conditions, aerobic and microaerobic. The preparation of the sample for metabolic analysis included the steps of quenching and extraction of the metabolites, for which the protocols of ice cold methanol and boiling ethanol, were used, respectively. The quantification of the metabolites was performed by ultra high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/ MS), using ion-pair chromatography and hydrophilic interaction liquid chromatography. The data obtained in the quantification were processed and treated through analysis of variance (ANOVA) using RStudio software. A total of eleven metabolites of the xylose metabolism pathway were quantified and validated through metabolic flow analysis (MFA) with R2>90. These metabolites were used to construct the metabolic flow of the three species of yeasts (Scheffersomyces stipitis, Spathaspora passalidarum and Spathaspora arborariae) analyzed. The comparison of molecular targets in yeast xylose showed that the S. stipitis and S. passalidarum species have the best yields of ethanol when grown with limited oxygen. The metabolomic analysis performed in this study may help in the genetic improvement of these microorganisms and in the production of 2G ethanol.Um dos desafios na produção do etanol de segunda geração (2G) está no desenvolvimento de leveduras eficientes para converter xilose em etanol no processo de fermentação de açúcares. Desta forma, a análise abrangente do metabolismo de leveduras fermentadoras de xilose é essencial para facilitar a identificação dos fatores limitantes no metabolismo de conversão desta pentose e, assim, auxiliar na construção de linhagens modificadas geneticamente mais eficientes. O objetivo deste trabalho foi utilizar uma ferramenta analítica avançada, a metabolômica, para quantificar os principais metabólitos relacionados às vias de conversão da xilose a etanol em quatro espécies de leveduras fermentadoras de xiloses: Scheffersomyces stipitis, Spathaspora passalidarum, Spathaspora arborariae e Candida Tenuis. As leveduras foram cultivadas em xilose sob duas diferentes condições de crescimento, aeróbica e microaeróbica. O preparo de amostra para análise metabolômica incluiu as etapas de quenching e extração dos metabólitos, e para tal foram utilizados os protocolos de metanol gelado e etanol fervente, respectivamente. A quantificação dos metabólitos foi realizada por cromatografia líquida de ultra alta eficiência acoplada à espectrometria de massas tandem (UHPLC-MS/MS), utilizando cromatografia de par iônico e a cromatografia líquida de interação hidrofílica. Os dados obtidos na quantificação foram processados e tratados através da análise de variância (ANOVA) utilizando o software RStudio. Um total de onze metabólitos da via do metabolismo da xilose foram quantificados e validados através da análise de fluxo metabólico (MFA) com R2>90. Estes metabólitos foram utilizados para a construção do fluxo metabólico das três espécies de leveduras analisadas (Scheffersomyces stipitis, Spathaspora passalidarum e Spathaspora arborariae). A comparação de alvos moleculares nas leveduras fermentadoras de xilose mostrou que as espécies S. stipitis e S. passalidarum tem os melhores rendimentos de produção de etanol quando crescidas com oxigênio limitado. A análise metabolômica realizada neste estudo poderá auxiliar no melhoramento genético destes microrganismos e na produção de etanol 2G.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESapplication/pdfhttp://repositorio.bc.ufg.br/tede/retrieve/54812/Tese%20-%20Christiane%20Gon%c3%a7alves%20Campos%20Cavalcanti%20-%202019.pdf.jpgporUniversidade Federal de GoiásPrograma de Pós-graduação em Química (IQ)UFGBrasilInstituto de Química - IQ (RG)http://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessMetabolômicaEspectrometria de massasCromatografia líquidaBiotecnologia e levedurasMetabolomicsMass spectrometryLiquid chromatographyBiotechnologyYeastsQUIMICA ANALITICA::INSTRUMENTACAO ANALITICAMetabolômica de leveduras fermentadoras de xilose na identificação de alvos metabólicos para aumento da produção de etanol 2gMetabolomics of xylose fermenting yeasts in the identification of metabolic targets to enhance ethanol 2g productioninfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis663693921325415158600600600600782606674374119727811432645100632034522075167498588264571reponame:Repositório Institucional da UFGinstname:Universidade Federal de Goiás (UFG)instacron:UFGLICENSElicense.txtlicense.txttext/plain; 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| spellingShingle | Metabolômica de leveduras fermentadoras de xilose na identificação de alvos metabólicos para aumento da produção de etanol 2g Cavalcanti, Christiane Gonçalves Campos Metabolômica Espectrometria de massas Cromatografia líquida Biotecnologia e leveduras Metabolomics Mass spectrometry Liquid chromatography Biotechnology Yeasts QUIMICA ANALITICA::INSTRUMENTACAO ANALITICA |
| status_str | publishedVersion |
| title | Metabolômica de leveduras fermentadoras de xilose na identificação de alvos metabólicos para aumento da produção de etanol 2g |
| title_full | Metabolômica de leveduras fermentadoras de xilose na identificação de alvos metabólicos para aumento da produção de etanol 2g |
| title_fullStr | Metabolômica de leveduras fermentadoras de xilose na identificação de alvos metabólicos para aumento da produção de etanol 2g |
| title_full_unstemmed | Metabolômica de leveduras fermentadoras de xilose na identificação de alvos metabólicos para aumento da produção de etanol 2g |
| title_short | Metabolômica de leveduras fermentadoras de xilose na identificação de alvos metabólicos para aumento da produção de etanol 2g |
| title_sort | Metabolômica de leveduras fermentadoras de xilose na identificação de alvos metabólicos para aumento da produção de etanol 2g |
| topic | Metabolômica Espectrometria de massas Cromatografia líquida Biotecnologia e leveduras Metabolomics Mass spectrometry Liquid chromatography Biotechnology Yeasts QUIMICA ANALITICA::INSTRUMENTACAO ANALITICA |
| url | http://repositorio.bc.ufg.br/tede/handle/tede/9653 |