Biochemical and biophysical properties of a metagenome-derived GH5 endoglucanase displaying an unconventional domain architecture
Autor(a) principal: | |
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Data de Publicação: | 2017 |
Outros Autores: | , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1016/j.ijbiomac.2017.02.075 http://hdl.handle.net/11449/174298 |
Resumo: | Endoglucanases are key enzymes in the degradation of cellulose, the most abundant polymer on Earth. The aim of this work was to perform the biochemical and biophysical characterization of CelE2, a soil metagenome derived endoglucanase. CelE2 harbors a conserved domain from glycoside hydrolase family 5 (GH5) and a C-terminal domain with identity to Calx-beta domains. The recombinant CelE2 displayed preference for hydrolysis of oat beta-glucan, followed by lichenan and carboxymethyl cellulose. Optimum values of enzymatic activity were observed at 45 °C and pH 5.3, and CelE2 exhibited considerable thermal stability at 40 °C for up to 360 min. Regarding the cleavage pattern on polysaccharides, the release of oligosaccharides with a wide degree of polymerization indicated a characteristic of endoglucanase activity. Furthermore, the analysis of products generated from the cleavage of cellooligosaccharides suggested that CelE2 exhibited transglycosylation activity. Interestingly, the presence of CaCl2 positively affect CelE2, including in the presence of surfactants. SAXS experiments provided key information on the effect of CaCl2 on the stability of CelE2 and dummy atom and rigid-body models were generated. To the best of our knowledge this is the first biochemical and biophysical characterization of an endoglucanase from family GH5 displaying this unconventional modular organization. |
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Biochemical and biophysical properties of a metagenome-derived GH5 endoglucanase displaying an unconventional domain architectureCalx-beta domainGlycoside hydrolase family 5Lignocellulosic biomassEndoglucanases are key enzymes in the degradation of cellulose, the most abundant polymer on Earth. The aim of this work was to perform the biochemical and biophysical characterization of CelE2, a soil metagenome derived endoglucanase. CelE2 harbors a conserved domain from glycoside hydrolase family 5 (GH5) and a C-terminal domain with identity to Calx-beta domains. The recombinant CelE2 displayed preference for hydrolysis of oat beta-glucan, followed by lichenan and carboxymethyl cellulose. Optimum values of enzymatic activity were observed at 45 °C and pH 5.3, and CelE2 exhibited considerable thermal stability at 40 °C for up to 360 min. Regarding the cleavage pattern on polysaccharides, the release of oligosaccharides with a wide degree of polymerization indicated a characteristic of endoglucanase activity. Furthermore, the analysis of products generated from the cleavage of cellooligosaccharides suggested that CelE2 exhibited transglycosylation activity. Interestingly, the presence of CaCl2 positively affect CelE2, including in the presence of surfactants. SAXS experiments provided key information on the effect of CaCl2 on the stability of CelE2 and dummy atom and rigid-body models were generated. To the best of our knowledge this is the first biochemical and biophysical characterization of an endoglucanase from family GH5 displaying this unconventional modular organization.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE) Centro Nacional de Pesquisa em Energia e Materiais (CNPEM), Caixa Postal 6192Departamento de Bioquímica Instituto de Biologia (IB) Universidade Estadual de Campinas (UNICAMP) R. Monteiro Lobato, 255 – Cidade UniversitáriaPrograma de Pós Graduação em Biociências e Tecnologia de Produtos Bioativos (BTPB) Instituto de Biologia (IB) – CP 6109 Universidade Estadual de Campinas (UNICAMP)Departamento de Física e Biofísica Instituto de Biociências de Botucatu UNESP Univ Estadual Paulista, Distrito de Rubião Jr. s/nDepartamento de Física e Biofísica Instituto de Biociências de Botucatu UNESP Univ Estadual Paulista, Distrito de Rubião Jr. s/nCentro Nacional de Pesquisa em Energia e Materiais (CNPEM)Universidade Estadual de Campinas (UNICAMP)Universidade Estadual Paulista (Unesp)Pimentel, Agnes C.Ematsu, Gabriela C.G.Liberato, Marcelo V.Paixão, Douglas A.A.Franco Cairo, João Paulo L.Mandelli, FernandaTramontina, RobsonGandin, César A. [UNESP]de Oliveira Neto, Mario [UNESP]Squina, Fabio M.Alvarez, Thabata M.2018-12-11T17:10:19Z2018-12-11T17:10:19Z2017-06-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article384-393application/pdfhttp://dx.doi.org/10.1016/j.ijbiomac.2017.02.075International Journal of Biological Macromolecules, v. 99, p. 384-393.1879-00030141-8130http://hdl.handle.net/11449/17429810.1016/j.ijbiomac.2017.02.0752-s2.0-850146231522-s2.0-85014623152.pdfScopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengInternational Journal of Biological Macromolecules0,917info:eu-repo/semantics/openAccess2023-12-11T06:15:52Zoai:repositorio.unesp.br:11449/174298Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T20:02:14.790144Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Biochemical and biophysical properties of a metagenome-derived GH5 endoglucanase displaying an unconventional domain architecture |
title |
Biochemical and biophysical properties of a metagenome-derived GH5 endoglucanase displaying an unconventional domain architecture |
spellingShingle |
Biochemical and biophysical properties of a metagenome-derived GH5 endoglucanase displaying an unconventional domain architecture Pimentel, Agnes C. Calx-beta domain Glycoside hydrolase family 5 Lignocellulosic biomass |
title_short |
Biochemical and biophysical properties of a metagenome-derived GH5 endoglucanase displaying an unconventional domain architecture |
title_full |
Biochemical and biophysical properties of a metagenome-derived GH5 endoglucanase displaying an unconventional domain architecture |
title_fullStr |
Biochemical and biophysical properties of a metagenome-derived GH5 endoglucanase displaying an unconventional domain architecture |
title_full_unstemmed |
Biochemical and biophysical properties of a metagenome-derived GH5 endoglucanase displaying an unconventional domain architecture |
title_sort |
Biochemical and biophysical properties of a metagenome-derived GH5 endoglucanase displaying an unconventional domain architecture |
author |
Pimentel, Agnes C. |
author_facet |
Pimentel, Agnes C. Ematsu, Gabriela C.G. Liberato, Marcelo V. Paixão, Douglas A.A. Franco Cairo, João Paulo L. Mandelli, Fernanda Tramontina, Robson Gandin, César A. [UNESP] de Oliveira Neto, Mario [UNESP] Squina, Fabio M. Alvarez, Thabata M. |
author_role |
author |
author2 |
Ematsu, Gabriela C.G. Liberato, Marcelo V. Paixão, Douglas A.A. Franco Cairo, João Paulo L. Mandelli, Fernanda Tramontina, Robson Gandin, César A. [UNESP] de Oliveira Neto, Mario [UNESP] Squina, Fabio M. Alvarez, Thabata M. |
author2_role |
author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Centro Nacional de Pesquisa em Energia e Materiais (CNPEM) Universidade Estadual de Campinas (UNICAMP) Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Pimentel, Agnes C. Ematsu, Gabriela C.G. Liberato, Marcelo V. Paixão, Douglas A.A. Franco Cairo, João Paulo L. Mandelli, Fernanda Tramontina, Robson Gandin, César A. [UNESP] de Oliveira Neto, Mario [UNESP] Squina, Fabio M. Alvarez, Thabata M. |
dc.subject.por.fl_str_mv |
Calx-beta domain Glycoside hydrolase family 5 Lignocellulosic biomass |
topic |
Calx-beta domain Glycoside hydrolase family 5 Lignocellulosic biomass |
description |
Endoglucanases are key enzymes in the degradation of cellulose, the most abundant polymer on Earth. The aim of this work was to perform the biochemical and biophysical characterization of CelE2, a soil metagenome derived endoglucanase. CelE2 harbors a conserved domain from glycoside hydrolase family 5 (GH5) and a C-terminal domain with identity to Calx-beta domains. The recombinant CelE2 displayed preference for hydrolysis of oat beta-glucan, followed by lichenan and carboxymethyl cellulose. Optimum values of enzymatic activity were observed at 45 °C and pH 5.3, and CelE2 exhibited considerable thermal stability at 40 °C for up to 360 min. Regarding the cleavage pattern on polysaccharides, the release of oligosaccharides with a wide degree of polymerization indicated a characteristic of endoglucanase activity. Furthermore, the analysis of products generated from the cleavage of cellooligosaccharides suggested that CelE2 exhibited transglycosylation activity. Interestingly, the presence of CaCl2 positively affect CelE2, including in the presence of surfactants. SAXS experiments provided key information on the effect of CaCl2 on the stability of CelE2 and dummy atom and rigid-body models were generated. To the best of our knowledge this is the first biochemical and biophysical characterization of an endoglucanase from family GH5 displaying this unconventional modular organization. |
publishDate |
2017 |
dc.date.none.fl_str_mv |
2017-06-01 2018-12-11T17:10:19Z 2018-12-11T17:10:19Z |
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 |
http://dx.doi.org/10.1016/j.ijbiomac.2017.02.075 International Journal of Biological Macromolecules, v. 99, p. 384-393. 1879-0003 0141-8130 http://hdl.handle.net/11449/174298 10.1016/j.ijbiomac.2017.02.075 2-s2.0-85014623152 2-s2.0-85014623152.pdf |
url |
http://dx.doi.org/10.1016/j.ijbiomac.2017.02.075 http://hdl.handle.net/11449/174298 |
identifier_str_mv |
International Journal of Biological Macromolecules, v. 99, p. 384-393. 1879-0003 0141-8130 10.1016/j.ijbiomac.2017.02.075 2-s2.0-85014623152 2-s2.0-85014623152.pdf |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
International Journal of Biological Macromolecules 0,917 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
384-393 application/pdf |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1808129153560477696 |