Biochemical Characterization of Heat-Tolerant Recombinant lArabinose Isomerase from Enterococcus faecium DBFIQ E36 Strain with Feasible Applications in d-Tagatose Production
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | por |
Título da fonte: | Repositório Institucional da Universidade Federal do Ceará (UFC) |
Texto Completo: | http://www.repositorio.ufc.br/handle/riufc/62907 |
Resumo: | d-Tagatose is a ketohexose, which presents unique properties as a low-calorie functional sweetener possessing a sweet flavor profile similar to d-sucrose and having no aftertaste. Considered a generally recognized as safe (GRAS) substance by FAO/ WHO, d-tagatose can be used as an intermediate for the synthesis of other optically active compounds as well as an additive in detergent, cosmetic, and pharmaceutical formulations. This study reports important features for l-arabinose isomerase (EC 5.3.1.4) (L-AI) use in industry. We describe arabinose (araA) gene virulence analysis, gene isolation, sequencing, cloning, and heterologous overexpression of L-AI from the food-grade GRAS bacterium Enterococcus faecium DBFIQ E36 in Escherichia coli and assess biochemical properties of this recombinant enzyme. Recombinant L-AI (rL-AI) was one-step purified to homogeneity by Ni2+-agarose resin affinity chromatography and biochemical characterization revealed low identity with both thermophilic and mesophilic L-AIs but high degree of conservation in residues involved in substrate recognition. Optimal conditions for rL-AI activity were 50 °C, pH 5.5, and 0.3 mM Mn2+, exhibiting a low cofactor concentration requirement and an acidic optimum pH. Half-life at 45 °C and 50 °C were 1427 h and 11 h, respectively, and 21.5 h and 39.5 h at pH 4.5 and 5.6, respectively, showing the high stability of the enzyme in the presence of a metallic cofactor. Bioconversion yield for d-tagatose biosynthesis was 45% at 50 °C after 48 h. These properties highlight the technological potential of E. faecium rL-AI as biocatalyst for d-tagatose production |
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Biochemical Characterization of Heat-Tolerant Recombinant lArabinose Isomerase from Enterococcus faecium DBFIQ E36 Strain with Feasible Applications in d-Tagatose Productionl-Arabinose isomerased-TagatoseEnterococcus faeciumVirulence gene analysisd-Galactosed-Tagatose is a ketohexose, which presents unique properties as a low-calorie functional sweetener possessing a sweet flavor profile similar to d-sucrose and having no aftertaste. Considered a generally recognized as safe (GRAS) substance by FAO/ WHO, d-tagatose can be used as an intermediate for the synthesis of other optically active compounds as well as an additive in detergent, cosmetic, and pharmaceutical formulations. This study reports important features for l-arabinose isomerase (EC 5.3.1.4) (L-AI) use in industry. We describe arabinose (araA) gene virulence analysis, gene isolation, sequencing, cloning, and heterologous overexpression of L-AI from the food-grade GRAS bacterium Enterococcus faecium DBFIQ E36 in Escherichia coli and assess biochemical properties of this recombinant enzyme. Recombinant L-AI (rL-AI) was one-step purified to homogeneity by Ni2+-agarose resin affinity chromatography and biochemical characterization revealed low identity with both thermophilic and mesophilic L-AIs but high degree of conservation in residues involved in substrate recognition. Optimal conditions for rL-AI activity were 50 °C, pH 5.5, and 0.3 mM Mn2+, exhibiting a low cofactor concentration requirement and an acidic optimum pH. Half-life at 45 °C and 50 °C were 1427 h and 11 h, respectively, and 21.5 h and 39.5 h at pH 4.5 and 5.6, respectively, showing the high stability of the enzyme in the presence of a metallic cofactor. Bioconversion yield for d-tagatose biosynthesis was 45% at 50 °C after 48 h. These properties highlight the technological potential of E. faecium rL-AI as biocatalyst for d-tagatose productionMolecular Biotechnology2021-12-10T15:00:03Z2021-12-10T15:00:03Z2019info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfMANZO, Ricardo Martín et al. Biochemical Characterization of Heat-Tolerant Recombinant lArabinose Isomerase from Enterococcus faecium DBFIQ E36 Strain with Feasible Applications in d-Tagatose Production. Molecular Biotechnology, [s. l.], v. 61, n. 6, p. 385-399, 2019.http://www.repositorio.ufc.br/handle/riufc/62907Manzo, Ricardo MartínAntunes, André Saraiva Leão MarceloMendes, Jocélia de SousaHissa, Denise CavalcanteGonҫalves, Luciana Rocha BarrosMammarella, Enrique Joséinfo:eu-repo/semantics/openAccessporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFC2023-10-10T19:32:06Zoai:repositorio.ufc.br:riufc/62907Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2023-10-10T19:32:06Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false |
dc.title.none.fl_str_mv |
Biochemical Characterization of Heat-Tolerant Recombinant lArabinose Isomerase from Enterococcus faecium DBFIQ E36 Strain with Feasible Applications in d-Tagatose Production |
title |
Biochemical Characterization of Heat-Tolerant Recombinant lArabinose Isomerase from Enterococcus faecium DBFIQ E36 Strain with Feasible Applications in d-Tagatose Production |
spellingShingle |
Biochemical Characterization of Heat-Tolerant Recombinant lArabinose Isomerase from Enterococcus faecium DBFIQ E36 Strain with Feasible Applications in d-Tagatose Production Manzo, Ricardo Martín l-Arabinose isomerase d-Tagatose Enterococcus faecium Virulence gene analysis d-Galactose |
title_short |
Biochemical Characterization of Heat-Tolerant Recombinant lArabinose Isomerase from Enterococcus faecium DBFIQ E36 Strain with Feasible Applications in d-Tagatose Production |
title_full |
Biochemical Characterization of Heat-Tolerant Recombinant lArabinose Isomerase from Enterococcus faecium DBFIQ E36 Strain with Feasible Applications in d-Tagatose Production |
title_fullStr |
Biochemical Characterization of Heat-Tolerant Recombinant lArabinose Isomerase from Enterococcus faecium DBFIQ E36 Strain with Feasible Applications in d-Tagatose Production |
title_full_unstemmed |
Biochemical Characterization of Heat-Tolerant Recombinant lArabinose Isomerase from Enterococcus faecium DBFIQ E36 Strain with Feasible Applications in d-Tagatose Production |
title_sort |
Biochemical Characterization of Heat-Tolerant Recombinant lArabinose Isomerase from Enterococcus faecium DBFIQ E36 Strain with Feasible Applications in d-Tagatose Production |
author |
Manzo, Ricardo Martín |
author_facet |
Manzo, Ricardo Martín Antunes, André Saraiva Leão Marcelo Mendes, Jocélia de Sousa Hissa, Denise Cavalcante Gonҫalves, Luciana Rocha Barros Mammarella, Enrique José |
author_role |
author |
author2 |
Antunes, André Saraiva Leão Marcelo Mendes, Jocélia de Sousa Hissa, Denise Cavalcante Gonҫalves, Luciana Rocha Barros Mammarella, Enrique José |
author2_role |
author author author author author |
dc.contributor.author.fl_str_mv |
Manzo, Ricardo Martín Antunes, André Saraiva Leão Marcelo Mendes, Jocélia de Sousa Hissa, Denise Cavalcante Gonҫalves, Luciana Rocha Barros Mammarella, Enrique José |
dc.subject.por.fl_str_mv |
l-Arabinose isomerase d-Tagatose Enterococcus faecium Virulence gene analysis d-Galactose |
topic |
l-Arabinose isomerase d-Tagatose Enterococcus faecium Virulence gene analysis d-Galactose |
description |
d-Tagatose is a ketohexose, which presents unique properties as a low-calorie functional sweetener possessing a sweet flavor profile similar to d-sucrose and having no aftertaste. Considered a generally recognized as safe (GRAS) substance by FAO/ WHO, d-tagatose can be used as an intermediate for the synthesis of other optically active compounds as well as an additive in detergent, cosmetic, and pharmaceutical formulations. This study reports important features for l-arabinose isomerase (EC 5.3.1.4) (L-AI) use in industry. We describe arabinose (araA) gene virulence analysis, gene isolation, sequencing, cloning, and heterologous overexpression of L-AI from the food-grade GRAS bacterium Enterococcus faecium DBFIQ E36 in Escherichia coli and assess biochemical properties of this recombinant enzyme. Recombinant L-AI (rL-AI) was one-step purified to homogeneity by Ni2+-agarose resin affinity chromatography and biochemical characterization revealed low identity with both thermophilic and mesophilic L-AIs but high degree of conservation in residues involved in substrate recognition. Optimal conditions for rL-AI activity were 50 °C, pH 5.5, and 0.3 mM Mn2+, exhibiting a low cofactor concentration requirement and an acidic optimum pH. Half-life at 45 °C and 50 °C were 1427 h and 11 h, respectively, and 21.5 h and 39.5 h at pH 4.5 and 5.6, respectively, showing the high stability of the enzyme in the presence of a metallic cofactor. Bioconversion yield for d-tagatose biosynthesis was 45% at 50 °C after 48 h. These properties highlight the technological potential of E. faecium rL-AI as biocatalyst for d-tagatose production |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019 2021-12-10T15:00:03Z 2021-12-10T15:00:03Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
MANZO, Ricardo Martín et al. Biochemical Characterization of Heat-Tolerant Recombinant lArabinose Isomerase from Enterococcus faecium DBFIQ E36 Strain with Feasible Applications in d-Tagatose Production. Molecular Biotechnology, [s. l.], v. 61, n. 6, p. 385-399, 2019. http://www.repositorio.ufc.br/handle/riufc/62907 |
identifier_str_mv |
MANZO, Ricardo Martín et al. Biochemical Characterization of Heat-Tolerant Recombinant lArabinose Isomerase from Enterococcus faecium DBFIQ E36 Strain with Feasible Applications in d-Tagatose Production. Molecular Biotechnology, [s. l.], v. 61, n. 6, p. 385-399, 2019. |
url |
http://www.repositorio.ufc.br/handle/riufc/62907 |
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 |
application/pdf |
dc.publisher.none.fl_str_mv |
Molecular Biotechnology |
publisher.none.fl_str_mv |
Molecular Biotechnology |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da Universidade Federal do Ceará (UFC) instname:Universidade Federal do Ceará (UFC) instacron:UFC |
instname_str |
Universidade Federal do Ceará (UFC) |
instacron_str |
UFC |
institution |
UFC |
reponame_str |
Repositório Institucional da Universidade Federal do Ceará (UFC) |
collection |
Repositório Institucional da Universidade Federal do Ceará (UFC) |
repository.name.fl_str_mv |
Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC) |
repository.mail.fl_str_mv |
bu@ufc.br || repositorio@ufc.br |
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