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Biblioteca(s): |
Embrapa Agroenergia. |
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Data corrente: |
23/03/2020 |
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Data da última atualização: |
27/04/2020 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
SILVA, C. de O. G.; TEIXEIRA, S. T.; RODRIGUES, K. B.; SOUZA, A. A.; MONCLARO, A. V.; MENDES, T. D.; RIBEIRO, J. A. de A.; SIQUEIRA, F. G. de; FAVARO, L. C. de L.; ABDELNUR, P. V. |
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Afiliação: |
Caio de Oliveira Gorgulho Silva; Tallyta Santos Teixeira, Universidade Federal de Tocantins; Kelly Barreto Rodrigues; Amanda Araujo Souza; Antonielle Vieira Monclaro; THAIS DEMARCHI MENDES, CNPAE; JOSE ANTONIO DE AQUINO RIBEIRO, CNPAE; FELIX GONCALVES DE SIQUEIRA, CNPAE; LEIA CECILIA DE LIMA FAVARO, CNPAE; PATRICIA VERARDI ABDELNUR, CNPAE. |
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Título: |
Combination of MALDI-TOF MS and UHPLC-ESI-MS for the characterization of lytic polysaccharide monooxygenase activity. |
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Ano de publicação: |
2020 |
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Fonte/Imprenta: |
Analytical Methods, n. 2, 2020. |
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Idioma: |
Inglês |
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Conteúdo: |
Lytic polysaccharide monooxygenases (LPMOs) are redox enzymes of high biotechnological interest due to their capacity to degrade recalcitrant polysaccharides, such as cellulose, by an oxidative mechanism. The characterization of LPMOs is challenging since they generate a variety of catalytic products which include native oligosaccharides (non-oxidized) and oligosaccharides oxidized at the reducing end (C1 position), the non-reducing end (C4 position), or both ends, with different degrees of polymerization. Moreover, oxidized products exist in equilibrium with their hydrated forms, which further complicates their identification. The lack of commercial analytical standards for all these possible forms of oxidized oligosaccharides and the low concentration of these products make LPMO functional characterization dependent on advanced mass spectrometry techniques capable of identifying the profile of oxidized products. Here, a new approach for the characterization of cellulose-active LPMOs based on the combination of MALDI-TOF MS and hydrophilic interaction UHPLC-ESI-MS was proposed and optimized. The LPMO TrAA9A from Trichoderma reesei was used as a model enzyme to develop and test the MS methods. MALDI-TOF MS and UHPLC-ESI-MS methods were both capable of identifying putative C1, C4 and C1/C4 oxidized cello-oligosaccharides as well as their native counterparts generated after cellulose treatment with LPMO, which allows their utilization to characterize type I (C1-oxidizer), type II (C4-oxidizer) and type III (C1- and C4-oxidizer) LPMOs. Moreover, both methods were complementary since MALDI-TOF MS was capable of detecting oligosaccharides with higher degrees of polymerization (DP3?DP10), while UHPLC-ESI-MS allowed the evaluation of smaller oligosaccharides (DP1?DP5). The combined use of both methods offers a comprehensive description of LPMO catalytic products. MenosLytic polysaccharide monooxygenases (LPMOs) are redox enzymes of high biotechnological interest due to their capacity to degrade recalcitrant polysaccharides, such as cellulose, by an oxidative mechanism. The characterization of LPMOs is challenging since they generate a variety of catalytic products which include native oligosaccharides (non-oxidized) and oligosaccharides oxidized at the reducing end (C1 position), the non-reducing end (C4 position), or both ends, with different degrees of polymerization. Moreover, oxidized products exist in equilibrium with their hydrated forms, which further complicates their identification. The lack of commercial analytical standards for all these possible forms of oxidized oligosaccharides and the low concentration of these products make LPMO functional characterization dependent on advanced mass spectrometry techniques capable of identifying the profile of oxidized products. Here, a new approach for the characterization of cellulose-active LPMOs based on the combination of MALDI-TOF MS and hydrophilic interaction UHPLC-ESI-MS was proposed and optimized. The LPMO TrAA9A from Trichoderma reesei was used as a model enzyme to develop and test the MS methods. MALDI-TOF MS and UHPLC-ESI-MS methods were both capable of identifying putative C1, C4 and C1/C4 oxidized cello-oligosaccharides as well as their native counterparts generated after cellulose treatment with LPMO, which allows their utilization to characterize type I (C1-oxidizer), type... Mostrar Tudo |
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Palavras-Chave: |
Caracterização de LPMOs. |
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Categoria do assunto: |
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Marc: |
LEADER 02626naa a2200241 a 4500
001 2121326
005 2020-04-27
008 2020 bl uuuu u00u1 u #d
100 1 $aSILVA, C. de O. G.
245 $aCombination of MALDI-TOF MS and UHPLC-ESI-MS for the characterization of lytic polysaccharide monooxygenase activity.$h[electronic resource]
260 $c2020
520 $aLytic polysaccharide monooxygenases (LPMOs) are redox enzymes of high biotechnological interest due to their capacity to degrade recalcitrant polysaccharides, such as cellulose, by an oxidative mechanism. The characterization of LPMOs is challenging since they generate a variety of catalytic products which include native oligosaccharides (non-oxidized) and oligosaccharides oxidized at the reducing end (C1 position), the non-reducing end (C4 position), or both ends, with different degrees of polymerization. Moreover, oxidized products exist in equilibrium with their hydrated forms, which further complicates their identification. The lack of commercial analytical standards for all these possible forms of oxidized oligosaccharides and the low concentration of these products make LPMO functional characterization dependent on advanced mass spectrometry techniques capable of identifying the profile of oxidized products. Here, a new approach for the characterization of cellulose-active LPMOs based on the combination of MALDI-TOF MS and hydrophilic interaction UHPLC-ESI-MS was proposed and optimized. The LPMO TrAA9A from Trichoderma reesei was used as a model enzyme to develop and test the MS methods. MALDI-TOF MS and UHPLC-ESI-MS methods were both capable of identifying putative C1, C4 and C1/C4 oxidized cello-oligosaccharides as well as their native counterparts generated after cellulose treatment with LPMO, which allows their utilization to characterize type I (C1-oxidizer), type II (C4-oxidizer) and type III (C1- and C4-oxidizer) LPMOs. Moreover, both methods were complementary since MALDI-TOF MS was capable of detecting oligosaccharides with higher degrees of polymerization (DP3?DP10), while UHPLC-ESI-MS allowed the evaluation of smaller oligosaccharides (DP1?DP5). The combined use of both methods offers a comprehensive description of LPMO catalytic products.
653 $aCaracterização de LPMOs
700 1 $aTEIXEIRA, S. T.
700 1 $aRODRIGUES, K. B.
700 1 $aSOUZA, A. A.
700 1 $aMONCLARO, A. V.
700 1 $aMENDES, T. D.
700 1 $aRIBEIRO, J. A. de A.
700 1 $aSIQUEIRA, F. G. de
700 1 $aFAVARO, L. C. de L.
700 1 $aABDELNUR, P. V.
773 $tAnalytical Methods$gn. 2, 2020.
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Embrapa Agroenergia (CNPAE) |
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| Registros recuperados : 123 | |
| 22. |  | COUTO, J. M. C.; GODOY, R. C. B. de; BENASSI, M. de T. Determinação do tempo de cozimento de pinhões em função da textura do produto final. In: EVENTO DE INICIAÇÃO CIENTÍFICA DA EMBRAPA FLORESTAS, 12., 2013, Colombo. Anais. Colombo: Embrapa Florestas, 2013. (Embrapa Florestas. Documentos, 253). Editores técnicos: Marcílio José Thomazini, Elenice Fritzsons, Patrícia Raquel Silva, Guilherme Schnell e Schuhli, Denise Jeton Cardoso, Luziane Franciscon. EVINCI. Resumos.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Florestas. |
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| 25. | | JOCOSK, L.; COUTO, J. M. C.; GODOY, R. C. B. de. Caracterização biométrica de pinhões coletados em diferentes épocas. In: EVENTO DE INICIAÇÃO CIENTÍFICA DA EMBRAPA FLORESTAS, 12., 2013, Colombo. Anais. Colombo: Embrapa Florestas, 2013. (Embrapa Florestas. Documentos, 253). Editores técnicos: Marcílio José Thomazini, Elenice Fritzsons, Patrícia Raquel Silva, Guilherme Schnell e Schuhli, Denise Jeton Cardoso, Luziane Franciscon. EVINCI. Resumos.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Florestas. |
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| 34. | | GODOY, R. C. B. de; PEREIRA, L. O.; SEOANE, C. E. S.; RETT, H. T. Juçara (Euterpe edulis M.): importância ecológica e alimentícia. Colombo: Embrapa Florestas, 2022. 27 p. (Embrapa Florestas. Documentos, 372). Selo ODS 2, 8, 9, 12, 15 e 17.| Biblioteca(s): Embrapa Florestas. |
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| 37. | | CARMO, A. A. O. do; GODOY, R. C. B. de; SOUZA, L. A. de. Morfologia dos órgãos vegetativos e reprodutivos de Araucaria angustifolia (Bert.) O. Ktze (Araucariaceae). In: SOUSA, V. A. de; FRITZSONS, E.; PINTO JUNIOR, J. E.; AGUIAR, A. V. de (ed.). Araucária: pesquisa e desenvolvimento no Brasil. Brasília, DF: Embrapa, 2021. cap. 2, p. 35-45.| Tipo: Capítulo em Livro Técnico-Científico |
| Biblioteca(s): Embrapa Florestas. |
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| 38. | | ZORTÉA-GUIDOLIN, M. E. B.; CARVALHO, C. W. P. de; GODOY, R. C. B. de. Potencial de uso do pinhão da araucária no desenvolvimento de produtos de conveniência por extrusão termoplástica. In: SEMINÁRIO SUL-BRASILEIRO SOBRE A SUSTENTABILIDADE DA ARAUCÁRIA, 3., 2018, Passo Fundo. Uso sustentável Produção Inovação Educação Legislação Conservação: anais. Passo Fundo: Livraria e editora Werlang, 2018. p. 24-27. p. 24-27 Organizadores Cristiano Roberto Buzatto, Nêmora Pauletti Prestes, Jaime Martinez, Alexandre Augusto Nienow.| Tipo: Artigo em Anais de Congresso |
| Biblioteca(s): Embrapa Agroindústria de Alimentos; Embrapa Florestas. |
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| 40. | | LAZZAROTTO, M.; RIBEIRO, L. S.; CORDOBA, L. do P.; GODOY, R. C. B. de; SCHNITZLER, E. Thermal tool in the evaluation of the "pinhão'' (seed of Araucaria angustifolia) starch aimed agroindustrial process. Brazilian Journal of Thermal Analysis, v. 7, p. 144-147, 2015. Disponível na internet. Edição dos Anais do 7º Simpósio de Análise Térmica, 2015, Bauru.| Tipo: Artigo em Anais de Congresso |
| Biblioteca(s): Embrapa Florestas. |
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| Registros recuperados : 123 | |
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