New protocol based on UHPLC-MS/MS for quantitation of metabolites in xylose-fermenting yeasts.

Detalhes bibliográficos
Autor(a) principal: CAMPOS, C. G.
Data de Publicação: 2017
Outros Autores: VERAS, H. C. T., RIBEIRO, J. A. de A., COSTA, P. P. K. G., ARAÚJO, K. P., RODRIGUES, C. M., ALMEIDA, J. R. M. de, ABDELNUR, P. V.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice)
Texto Completo: http://www.alice.cnptia.embrapa.br/alice/handle/doc/1075378
Resumo: Abstract: Xylose fermentation is a bottleneck in second-generation ethanol production. As such, a comprehensive understanding of xylose metabolism in naturally xylose-fermenting yeasts is essential for prospection and construction of recombinant yeast strains. The objective of the current study was to establish a reliable metabolomics protocol for quantification of key metabolites of xylose catabolism pathways in yeast, and to apply this protocol to Spathaspora arborariae. Ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) was used to quantify metabolites, and afterwards, sample preparation was optimized to examine yeast intracellular metabolites. S. arborariae was cultivated using xylose as a carbon source under aerobic and oxygen-limited conditions. Ion pair chromatography (IPC) and hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) were shown to efficiently quantify 14 and 5 metabolites, respectively, in a more rapid chromatographic protocol than previously described. Thirteen and eleven metabolites were quantified in S. arborariae under aerobic and oxygen-limited conditions, respectively. This targeted metabolomics protocol is shown here to quantify a total of 19 metabolites, including sugars, phosphates, coenzymes, monosaccharides, and alcohols, from xylose catabolism pathways (glycolysis, pentose phosphate pathway, and tricarboxylic acid cycle) in yeast. Furthermore, to our knowledge, this is the first time that intracellular metabolites have been quantified in S. arborariae after xylose consumption. The results indicated that fine control of oxygen levels during fermentation is necessary to optimize ethanol production by S. arborariae. The protocol presented here may be applied to other yeast species and could support yeast genetic engineering to improve second generation ethanol production. Graphical Abstract ?.
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spelling New protocol based on UHPLC-MS/MS for quantitation of metabolites in xylose-fermenting yeasts.Spathaspora arborariaeUHPLC-MS/MSXylose fermentationMass spectrometryMetabolomicsXyloseAbstract: Xylose fermentation is a bottleneck in second-generation ethanol production. As such, a comprehensive understanding of xylose metabolism in naturally xylose-fermenting yeasts is essential for prospection and construction of recombinant yeast strains. The objective of the current study was to establish a reliable metabolomics protocol for quantification of key metabolites of xylose catabolism pathways in yeast, and to apply this protocol to Spathaspora arborariae. Ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) was used to quantify metabolites, and afterwards, sample preparation was optimized to examine yeast intracellular metabolites. S. arborariae was cultivated using xylose as a carbon source under aerobic and oxygen-limited conditions. Ion pair chromatography (IPC) and hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) were shown to efficiently quantify 14 and 5 metabolites, respectively, in a more rapid chromatographic protocol than previously described. Thirteen and eleven metabolites were quantified in S. arborariae under aerobic and oxygen-limited conditions, respectively. This targeted metabolomics protocol is shown here to quantify a total of 19 metabolites, including sugars, phosphates, coenzymes, monosaccharides, and alcohols, from xylose catabolism pathways (glycolysis, pentose phosphate pathway, and tricarboxylic acid cycle) in yeast. Furthermore, to our knowledge, this is the first time that intracellular metabolites have been quantified in S. arborariae after xylose consumption. The results indicated that fine control of oxygen levels during fermentation is necessary to optimize ethanol production by S. arborariae. The protocol presented here may be applied to other yeast species and could support yeast genetic engineering to improve second generation ethanol production. Graphical Abstract ?.CHRISTIANE GONÇALVES CAMPOS, Universidade Federal de Goiás; HENRIQUE CÉSAR TEIXEIRA VERAS, Universidade de Brasília; JOSE ANTONIO DE AQUINO RIBEIRO, CNPAE; PATRICIA PINTO KALIL G COSTA, CNPAE; KATIÚSCIA PEREIRA ARAÚJO; CLENILSON MARTINS RODRIGUES, CNPAE; JOAO RICARDO MOREIRA DE ALMEIDA, CNPAE; PATRICIA VERARDI ABDELNUR, CNPAE.CAMPOS, C. G.VERAS, H. C. T.RIBEIRO, J. A. de A.COSTA, P. P. K. G.ARAÚJO, K. P.RODRIGUES, C. M.ALMEIDA, J. R. M. deABDELNUR, P. V.2021-04-06T18:33:52Z2021-04-06T18:33:52Z2017-09-112017info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlep. 2646-2657Journal of the American Society for Mass Spectrometry, v. 28, n. 12, p. 2646-2657, 2017.http://www.alice.cnptia.embrapa.br/alice/handle/doc/107537810.1007/s13361-017-1786-9enginfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice)instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa)instacron:EMBRAPA2021-04-06T18:34:03Zoai:www.alice.cnptia.embrapa.br:doc/1075378Repositório InstitucionalPUBhttps://www.alice.cnptia.embrapa.br/oai/requestopendoar:21542021-04-06T18:34:03falseRepositório InstitucionalPUBhttps://www.alice.cnptia.embrapa.br/oai/requestcg-riaa@embrapa.bropendoar:21542021-04-06T18:34:03Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa)false
dc.title.none.fl_str_mv New protocol based on UHPLC-MS/MS for quantitation of metabolites in xylose-fermenting yeasts.
title New protocol based on UHPLC-MS/MS for quantitation of metabolites in xylose-fermenting yeasts.
spellingShingle New protocol based on UHPLC-MS/MS for quantitation of metabolites in xylose-fermenting yeasts.
CAMPOS, C. G.
Spathaspora arborariae
UHPLC-MS/MS
Xylose fermentation
Mass spectrometry
Metabolomics
Xylose
title_short New protocol based on UHPLC-MS/MS for quantitation of metabolites in xylose-fermenting yeasts.
title_full New protocol based on UHPLC-MS/MS for quantitation of metabolites in xylose-fermenting yeasts.
title_fullStr New protocol based on UHPLC-MS/MS for quantitation of metabolites in xylose-fermenting yeasts.
title_full_unstemmed New protocol based on UHPLC-MS/MS for quantitation of metabolites in xylose-fermenting yeasts.
title_sort New protocol based on UHPLC-MS/MS for quantitation of metabolites in xylose-fermenting yeasts.
author CAMPOS, C. G.
author_facet CAMPOS, C. G.
VERAS, H. C. T.
RIBEIRO, J. A. de A.
COSTA, P. P. K. G.
ARAÚJO, K. P.
RODRIGUES, C. M.
ALMEIDA, J. R. M. de
ABDELNUR, P. V.
author_role author
author2 VERAS, H. C. T.
RIBEIRO, J. A. de A.
COSTA, P. P. K. G.
ARAÚJO, K. P.
RODRIGUES, C. M.
ALMEIDA, J. R. M. de
ABDELNUR, P. V.
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv CHRISTIANE GONÇALVES CAMPOS, Universidade Federal de Goiás; HENRIQUE CÉSAR TEIXEIRA VERAS, Universidade de Brasília; JOSE ANTONIO DE AQUINO RIBEIRO, CNPAE; PATRICIA PINTO KALIL G COSTA, CNPAE; KATIÚSCIA PEREIRA ARAÚJO; CLENILSON MARTINS RODRIGUES, CNPAE; JOAO RICARDO MOREIRA DE ALMEIDA, CNPAE; PATRICIA VERARDI ABDELNUR, CNPAE.
dc.contributor.author.fl_str_mv CAMPOS, C. G.
VERAS, H. C. T.
RIBEIRO, J. A. de A.
COSTA, P. P. K. G.
ARAÚJO, K. P.
RODRIGUES, C. M.
ALMEIDA, J. R. M. de
ABDELNUR, P. V.
dc.subject.por.fl_str_mv Spathaspora arborariae
UHPLC-MS/MS
Xylose fermentation
Mass spectrometry
Metabolomics
Xylose
topic Spathaspora arborariae
UHPLC-MS/MS
Xylose fermentation
Mass spectrometry
Metabolomics
Xylose
description Abstract: Xylose fermentation is a bottleneck in second-generation ethanol production. As such, a comprehensive understanding of xylose metabolism in naturally xylose-fermenting yeasts is essential for prospection and construction of recombinant yeast strains. The objective of the current study was to establish a reliable metabolomics protocol for quantification of key metabolites of xylose catabolism pathways in yeast, and to apply this protocol to Spathaspora arborariae. Ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) was used to quantify metabolites, and afterwards, sample preparation was optimized to examine yeast intracellular metabolites. S. arborariae was cultivated using xylose as a carbon source under aerobic and oxygen-limited conditions. Ion pair chromatography (IPC) and hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) were shown to efficiently quantify 14 and 5 metabolites, respectively, in a more rapid chromatographic protocol than previously described. Thirteen and eleven metabolites were quantified in S. arborariae under aerobic and oxygen-limited conditions, respectively. This targeted metabolomics protocol is shown here to quantify a total of 19 metabolites, including sugars, phosphates, coenzymes, monosaccharides, and alcohols, from xylose catabolism pathways (glycolysis, pentose phosphate pathway, and tricarboxylic acid cycle) in yeast. Furthermore, to our knowledge, this is the first time that intracellular metabolites have been quantified in S. arborariae after xylose consumption. The results indicated that fine control of oxygen levels during fermentation is necessary to optimize ethanol production by S. arborariae. The protocol presented here may be applied to other yeast species and could support yeast genetic engineering to improve second generation ethanol production. Graphical Abstract ?.
publishDate 2017
dc.date.none.fl_str_mv 2017-09-11
2017
2021-04-06T18:33:52Z
2021-04-06T18:33:52Z
dc.type.driver.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv Journal of the American Society for Mass Spectrometry, v. 28, n. 12, p. 2646-2657, 2017.
http://www.alice.cnptia.embrapa.br/alice/handle/doc/1075378
10.1007/s13361-017-1786-9
identifier_str_mv Journal of the American Society for Mass Spectrometry, v. 28, n. 12, p. 2646-2657, 2017.
10.1007/s13361-017-1786-9
url http://www.alice.cnptia.embrapa.br/alice/handle/doc/1075378
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv p. 2646-2657
dc.source.none.fl_str_mv reponame:Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice)
instname:Empresa Brasileira de Pesquisa Agropecuária (Embrapa)
instacron:EMBRAPA
instname_str Empresa Brasileira de Pesquisa Agropecuária (Embrapa)
instacron_str EMBRAPA
institution EMBRAPA
reponame_str Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice)
collection Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice)
repository.name.fl_str_mv Repositório Institucional da EMBRAPA (Repository Open Access to Scientific Information from EMBRAPA - Alice) - Empresa Brasileira de Pesquisa Agropecuária (Embrapa)
repository.mail.fl_str_mv cg-riaa@embrapa.br
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