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Biblioteca(s): |
Embrapa Agroenergia. |
Data corrente: |
23/03/2020 |
Data da última atualização: |
27/04/2020 |
Tipo da produção científica: |
Artigo em Periódico Indexado |
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. |
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. |
Título: |
Combination of MALDI-TOF MS and UHPLC-ESI-MS for the characterization of lytic polysaccharide monooxygenase activity. |
Ano de publicação: |
2020 |
Fonte/Imprenta: |
Analytical Methods, n. 2, 2020. |
Idioma: |
Inglês |
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 |
Palavras-Chave: |
Caracterização de LPMOs. |
Categoria do assunto: |
-- |
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 : 16 | |
2. | | TEIXEIRA, T. S.; BARROS, J. S. de; MACÊDO, J. K. A.; RODRIGUES, K. B.; FAVARO, L. C. de L.; SIQUEIRA, F. G. de; ABDELNUR, P. V. Avaliação do perfil de oligossacarídeos produzidos por celulose do tipo PASC em diferentes tampões por MALDI-TOF MS. In: ENCONTRO DE PESQUISA E INOVAÇÃO DA EMBRAPA AGROENERGIA, 4.,2017, Brasília, DF. Anais... Brasília, DF: Embrapa, 2017. p. 141-147.Tipo: Artigo em Anais de Congresso |
Biblioteca(s): Embrapa Agroenergia. |
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4. | | GUIMARÃES, J. P. S. M.; RODRIGUES, K. B.; SOUZA, B. G. de; BRANDÃO, L. T. D.; QUECINE, M. C.; FAVARO, L. C. de L. Characterization of Bacillus, Paenibacillus, Peribacillus, and Alkalihalobacillus strains for indole-3-acetic acid production. In: INTERNATIONAL SYMPOSIUM OF MICROBIOLOGY AND BIOTECHNOLOGY, 6., 2023, Viçosa, MG. Biotecnologia Microbiana Soluções para os grandes desafios globais: anais. Viçosa, MG: Universidade Federal de Viçosa, 2023. Resumo.Tipo: Resumo em Anais de Congresso |
Biblioteca(s): Embrapa Agroenergia. |
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5. | | SANTOS, F. P. dos; RODRIGUES, K. B.; SANTANA, M. W. P. R. de; SALUM, T. F. C.; FAVARO, L. C. de L.; BRASIL, B. dos S. A. F. Expressão heteróloga de monoxigenases de polissacarídeos bacterianas em Pichia pastoris. In: ENCONTRO DE PESQUISA E INOVAÇÃO DA EMBRAPA AGROENERGIA, 3., 2016, Brasília, DF. Anais ... Brasília, DF: Embrapa, 2016. p. 27-35.Tipo: Artigo em Anais de Congresso |
Biblioteca(s): Embrapa Agroenergia. |
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6. | | GREGORINE, L. F.; MONCLARO, A. V.; SILVA, C. de O. G.; RODRIGUES, K. B.; PEIXOTO, J. S. G.; ABDELNUR, P. V.; FAVARO, L. C. de L. Heterologous expression of a new lytic polysaccharide monooxygenase from Hahella ganghwensis and their functional characterization. In: SIMPÓSIO NACIONAL DE BIOPROCESSOS, 22.; SIMPÓSIO DE HIDRÓLISE ENZIMÁTICA DE BIOMASSA, 13., 2019, Uberlandia, MG. [Anais ...]. São Paulo: Associação Brasileira de Engenharia Química, 2019.Tipo: Artigo em Anais de Congresso |
Biblioteca(s): Embrapa Agroenergia. |
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7. | | NUNES, L. S.; FREIRE, Í. A.; RODRIGUES, K. B.; MIDORIKAWA, G. E. O.; QUECINE, M. C.; FAVARO, L. C. de L. In vitro assessment of inorganic phosphate-solubilizing potential of Bacillus, Paenibacillus, Peribacillus, and Alkalihalobacillus strains. In: INTERNATIONAL SYMPOSIUM OF MICROBIOLOGY AND BIOTECHNOLOGY, 6., 2023, Viçosa, MG. Biotecnologia Microbiana Soluções para os grandes desafios globais: anais. Viçosa, MG: Universidade Federal de Viçosa, 2023. Resumo.Tipo: Resumo em Anais de Congresso |
Biblioteca(s): Embrapa Agroenergia. |
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8. | | SILVA, C. de O. G.; TEIXEIRA, T. S.; RIBEIRO, J. A. de A.; RODRIGUES, K. B.; ANASTÁCIO, G. S.; SOUZA, A. A.; MENDES, T. D.; SALUM, T. F. C.; FAVARO, L. C. de L.; ABDELNUR, P. V. avaliação da atividade da LPMO Cel61A de trichoderma reesei sobre celulose por espectrometria de massas. In: ENCONTRO DE PESQUISA E INOVAÇÃO DA EMBRAPA AGROENERGIA, 5., 2018, Brasília, DF. Anais ... Brasília, DF: Embrapa Agroenergia, 2018. p. 25.Tipo: Resumo em Anais de Congresso |
Biblioteca(s): Embrapa Agroenergia. |
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9. | | SANTANA, M. W. P. R. de; RODRIGUES, K. B.; SANTOS, F. P. dos; MENDES, T. D.; DAMASO, M. C. T.; FAVARO, L. C. de L.; SIQUEIRA, F. G. de. Clonagem e expressão de genes de monoxigenases de polissacarídeos líticas (LPMOs) de fungos em Pichia pastoris. In: ENCONTRO DE PESQUISA E INOVAÇÃO DA EMBRAPA AGROENERGIA, 3., 2016, Brasília, DF. Anais ... Brasília, DF: Embrapa, 2016. p. 36-47.Tipo: Artigo em Anais de Congresso |
Biblioteca(s): Embrapa Agroenergia. |
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10. | | 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. Combination of MALDI-TOF MS and UHPLC-ESI-MS for the characterization of lytic polysaccharide monooxygenase activity. Analytical Methods, n. 2, 2020.Tipo: Artigo em Periódico Indexado | Circulação/Nível: A - 2 |
Biblioteca(s): Embrapa Agroenergia. |
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11. | | PEIXOTO, J. de S. G.; RODRIGUES, K. B.; MENINO, G. S.; MENDES, T. D.; GOMES, H. A. R.; LOPES, R. S.; GONCALVES, S. B.; SILVA, F. M.; FAVARO, L. C. de L. Produção de uma Beta-expansina de cana-de-açúcar por Komagataella phaffii. In: ENCONTRO DE PESQUISA E INOVAÇÃO DA EMBRAPA AGROENERGIA, 5., 2018, Brasília, DF. Anais ... Brasília, DF: Embrapa Agroenergia, 2018. p. 37.Tipo: Resumo em Anais de Congresso |
Biblioteca(s): Embrapa Agroenergia. |
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12. | | RODRIGUES, K. B.; MACÊDO, J. K. A.; TEIXEIRA, T.; BARROS, J. S.; ARAÚJO, A. C. B.; SANTOS, F. P.; QUIRINO, B. F.; BRASIL, B. dos S. A. F.; SALUM, T. F. C.; ABDELNUR, P. V.; FAVARO, L. C. de L. Recombinant expression of Thermobifida fusca E7 LPMO in Pichia pastoris and Escherichia coli and their functional characterization. Carbohydrate Research, v. 448, p. 175-181, 2017.Tipo: Artigo em Periódico Indexado | Circulação/Nível: A - 2 |
Biblioteca(s): Embrapa Agroenergia. |
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13. | | RODRIGUES, K. B.; MACÊDO, J. K. A.; BARROS, J. S.; ARAÚJO, A. C. B.; SANTOS, F. P.; QUIRINO, B. F.; BRASIL, B. dos S. A. F.; SALUM, T. F. C.; ABDELNUR, P. V.; FAVARO, L. C. de L. Recombinant expression of Thermobifida fusca E7 LPMO in Pichia pastoris and Escherichia coli and their funstional characterization. In: SYMPOSIUM ON THE CHEMISTRY, BIOLOGY AND APPLICATION OF LYTIC POLYSACCHARIDE MONOOXYGENASES, 2016, Copenhagen, Denmark. [Proceedings ...]. Hellerup: Novo Nordisk Fonden, 2016. Não paginado. Resumo nº 20.Tipo: Artigo em Anais de Congresso |
Biblioteca(s): Embrapa Agroenergia. |
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14. | | RODRIGUES, K. B.; SOUZA, A. A.; PEIXOTO, J. de S. G.; SILVA, C. de O. G.; ANDREANI, L.; MENDES, T. D.; VALADARES, L. F.; ABDELNUR, P. V.; SILVA, F. M.; GONCALVES, S. B.; RODRIGUES, D. de S.; SALUM, T. F. C.; DAMASO, M. C. T.; FAVARO, L. C. de L. Produção heteróloga e caracterização funcional das expansinas de cana-de-açúcar SacEXP4 e SacEXP7. In: ENCONTRO DE PESQUISA E INOVAÇÃO DA EMBRAPA AGROENERGIA, 6., 2020, Brasília, DF. Anais... Brasília, DF: Embrapa, 2020. p. 215-223 il.Tipo: Artigo em Anais de Congresso |
Biblioteca(s): Embrapa Agroenergia. |
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15. | | SILVA, C. de O. G.; RODRIGUES, K. B.; SOUZA, A. A.; MONCLARO, A. V.; ANASTÁCIO, G. S.; MENDES, T. D.; GONCALVES, S. B.; SALUM, T. F. C.; RODRIGUES, D. de S.; DAMASO, M. C. T.; ABDELNUR, P. V.; FAVARO, L. C. de L. Produção heteróloga e caracterização funcional de mono-oxigenases líticas de polissacarídeos de fungos e bactérias. In: ENCONTRO DE PESQUISA E INOVAÇÃO DA EMBRAPA AGROENERGIA, 6., 2020, Brasília, DF. Anais... Brasília, DF: Embrapa, 2020. p. 224-232 il.Tipo: Artigo em Anais de Congresso |
Biblioteca(s): Embrapa Agroenergia. |
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16. | | FAVARO, L. C. de L.; RODRIGUES, K. B.; MACEDO, J. K. A.; LOPES, R. S.; SANTOS, F. P.; SANTANA, M. W. P. R.; ARAUJO, A. C. B.; MENDES, T. D.; GONCALVES, S. B.; SALUM, T. F. C.; SIQUEIRA, F. G. de; DAMASO, M. C. T.; QUIRINO, B. F.; BRASIL, B. dos S. A. F.; RODRIGUES, D. de S.; ABDELNUR, P. V.; MELO, F. L. Recombinant expression and functional characterization of LPMOs and expansin-like proteins. In: Workshop on Second Generation Bioethanol, 2016, Campinas, SP. Abstract book ? Campinas, SP: CTBE, 2016. p. 30.Tipo: Resumo em Anais de Congresso |
Biblioteca(s): Embrapa Agroenergia. |
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Registros recuperados : 16 | |
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Nenhum registro encontrado para a expressão de busca informada. |
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