Biochemical and biophysical properties of a metagenome-derived GH5 endoglucanase displaying an unconventional domain architecture

Detalhes bibliográficos
Autor(a) principal: Pimentel, Agnes C.
Data de Publicação: 2017
Outros Autores: 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.
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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spelling 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
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