Biochemical characterization of an isolated 50 kDa beta-glucosidase from the thermophilic fungus Myceliophthora thermophila M.7.7

Detalhes bibliográficos
Autor(a) principal: Bonfá, Emily Colferai [UNESP]
Data de Publicação: 2018
Outros Autores: de Souza Moretti, Marcia Maria [UNESP], Gomes, Eleni [UNESP], Bonilla-Rodriguez, Gustavo Orlando [UNESP]
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1016/j.bcab.2018.01.008
http://hdl.handle.net/11449/179564
Resumo: This study characterized a 50 kDa β-glucosidase (BGL50) produced by the thermophilic fungus Myceliophthora thermophila M.7.7 in solid state cultivation using a mixture of (1:1) sugarcane bagasse and wheat bran. The crude extract zymogram showed two isoforms of β-glucosidase with approximately 50 and 200 kDa, which were separated by gel filtration chromatography. The characterization of BGL50 showed optimum activity at 60 °C and pH 5.0 when 4-nitrophenyl β-D-glucopyranoside (pNPG) was used as the substrate, whereas when using cellobiose, the highest activity was observed at 50 °C and pH 4.5. Several ions and reagents produced different effects on the enzyme activity depending on the substrate and there was complete inhibition with Cu2+ and Fe3+ for both substrates. In addition, nine phenolic compounds showed no inhibitory effects on the enzyme, a significant feature since β-glucosidase is used for the saccharification of lignocellulosic biomass that generates several phenolic compounds. Kinetic studies revealed competitive inhibition by glucose when pNPG was used, with a Ki value of 1.5 mM and a significantly lower Km (0.52 mM) than for cellobiose (8.50 mM). The thermodynamic parameters showed that BGL50 is very stable at 60 °C displaying a half-life of 855.6 min but it is easily denatured above this temperature. The results emphasize the importance of investigating potential β-glucosidases based on cellobiose instead of using only pNPG since, in the industrial process, the enzyme will act on this natural substrate. In addition, understanding the thermostability of the enzyme is an important contribution to enzyme technology.
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spelling Biochemical characterization of an isolated 50 kDa beta-glucosidase from the thermophilic fungus Myceliophthora thermophila M.7.7Beta-glucosidaseMyceliophthora thermophilaThermophilic fungusThis study characterized a 50 kDa β-glucosidase (BGL50) produced by the thermophilic fungus Myceliophthora thermophila M.7.7 in solid state cultivation using a mixture of (1:1) sugarcane bagasse and wheat bran. The crude extract zymogram showed two isoforms of β-glucosidase with approximately 50 and 200 kDa, which were separated by gel filtration chromatography. The characterization of BGL50 showed optimum activity at 60 °C and pH 5.0 when 4-nitrophenyl β-D-glucopyranoside (pNPG) was used as the substrate, whereas when using cellobiose, the highest activity was observed at 50 °C and pH 4.5. Several ions and reagents produced different effects on the enzyme activity depending on the substrate and there was complete inhibition with Cu2+ and Fe3+ for both substrates. In addition, nine phenolic compounds showed no inhibitory effects on the enzyme, a significant feature since β-glucosidase is used for the saccharification of lignocellulosic biomass that generates several phenolic compounds. Kinetic studies revealed competitive inhibition by glucose when pNPG was used, with a Ki value of 1.5 mM and a significantly lower Km (0.52 mM) than for cellobiose (8.50 mM). The thermodynamic parameters showed that BGL50 is very stable at 60 °C displaying a half-life of 855.6 min but it is easily denatured above this temperature. The results emphasize the importance of investigating potential β-glucosidases based on cellobiose instead of using only pNPG since, in the industrial process, the enzyme will act on this natural substrate. In addition, understanding the thermostability of the enzyme is an important contribution to enzyme technology.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)São Paulo State University (Unesp) Institute of Biosciences Humanities and Exact Sciences (Ibilce) São José do Rio Preto CampusSão Paulo State University (Unesp) Institute of Biosciences Humanities and Exact Sciences (Ibilce) São José do Rio Preto CampusFAPESP: 10/12624-0FAPESP: 11/23991-7Universidade Estadual Paulista (Unesp)Bonfá, Emily Colferai [UNESP]de Souza Moretti, Marcia Maria [UNESP]Gomes, Eleni [UNESP]Bonilla-Rodriguez, Gustavo Orlando [UNESP]2018-12-11T17:35:41Z2018-12-11T17:35:41Z2018-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article311-318application/pdfhttp://dx.doi.org/10.1016/j.bcab.2018.01.008Biocatalysis and Agricultural Biotechnology, v. 13, p. 311-318.1878-8181http://hdl.handle.net/11449/17956410.1016/j.bcab.2018.01.0082-s2.0-850416034772-s2.0-85041603477.pdfScopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBiocatalysis and Agricultural Biotechnology0,479info:eu-repo/semantics/openAccess2023-10-10T06:09:07Zoai:repositorio.unesp.br:11449/179564Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T14:33:44.784410Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Biochemical characterization of an isolated 50 kDa beta-glucosidase from the thermophilic fungus Myceliophthora thermophila M.7.7
title Biochemical characterization of an isolated 50 kDa beta-glucosidase from the thermophilic fungus Myceliophthora thermophila M.7.7
spellingShingle Biochemical characterization of an isolated 50 kDa beta-glucosidase from the thermophilic fungus Myceliophthora thermophila M.7.7
Bonfá, Emily Colferai [UNESP]
Beta-glucosidase
Myceliophthora thermophila
Thermophilic fungus
title_short Biochemical characterization of an isolated 50 kDa beta-glucosidase from the thermophilic fungus Myceliophthora thermophila M.7.7
title_full Biochemical characterization of an isolated 50 kDa beta-glucosidase from the thermophilic fungus Myceliophthora thermophila M.7.7
title_fullStr Biochemical characterization of an isolated 50 kDa beta-glucosidase from the thermophilic fungus Myceliophthora thermophila M.7.7
title_full_unstemmed Biochemical characterization of an isolated 50 kDa beta-glucosidase from the thermophilic fungus Myceliophthora thermophila M.7.7
title_sort Biochemical characterization of an isolated 50 kDa beta-glucosidase from the thermophilic fungus Myceliophthora thermophila M.7.7
author Bonfá, Emily Colferai [UNESP]
author_facet Bonfá, Emily Colferai [UNESP]
de Souza Moretti, Marcia Maria [UNESP]
Gomes, Eleni [UNESP]
Bonilla-Rodriguez, Gustavo Orlando [UNESP]
author_role author
author2 de Souza Moretti, Marcia Maria [UNESP]
Gomes, Eleni [UNESP]
Bonilla-Rodriguez, Gustavo Orlando [UNESP]
author2_role author
author
author
dc.contributor.none.fl_str_mv Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Bonfá, Emily Colferai [UNESP]
de Souza Moretti, Marcia Maria [UNESP]
Gomes, Eleni [UNESP]
Bonilla-Rodriguez, Gustavo Orlando [UNESP]
dc.subject.por.fl_str_mv Beta-glucosidase
Myceliophthora thermophila
Thermophilic fungus
topic Beta-glucosidase
Myceliophthora thermophila
Thermophilic fungus
description This study characterized a 50 kDa β-glucosidase (BGL50) produced by the thermophilic fungus Myceliophthora thermophila M.7.7 in solid state cultivation using a mixture of (1:1) sugarcane bagasse and wheat bran. The crude extract zymogram showed two isoforms of β-glucosidase with approximately 50 and 200 kDa, which were separated by gel filtration chromatography. The characterization of BGL50 showed optimum activity at 60 °C and pH 5.0 when 4-nitrophenyl β-D-glucopyranoside (pNPG) was used as the substrate, whereas when using cellobiose, the highest activity was observed at 50 °C and pH 4.5. Several ions and reagents produced different effects on the enzyme activity depending on the substrate and there was complete inhibition with Cu2+ and Fe3+ for both substrates. In addition, nine phenolic compounds showed no inhibitory effects on the enzyme, a significant feature since β-glucosidase is used for the saccharification of lignocellulosic biomass that generates several phenolic compounds. Kinetic studies revealed competitive inhibition by glucose when pNPG was used, with a Ki value of 1.5 mM and a significantly lower Km (0.52 mM) than for cellobiose (8.50 mM). The thermodynamic parameters showed that BGL50 is very stable at 60 °C displaying a half-life of 855.6 min but it is easily denatured above this temperature. The results emphasize the importance of investigating potential β-glucosidases based on cellobiose instead of using only pNPG since, in the industrial process, the enzyme will act on this natural substrate. In addition, understanding the thermostability of the enzyme is an important contribution to enzyme technology.
publishDate 2018
dc.date.none.fl_str_mv 2018-12-11T17:35:41Z
2018-12-11T17:35:41Z
2018-01-01
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.bcab.2018.01.008
Biocatalysis and Agricultural Biotechnology, v. 13, p. 311-318.
1878-8181
http://hdl.handle.net/11449/179564
10.1016/j.bcab.2018.01.008
2-s2.0-85041603477
2-s2.0-85041603477.pdf
url http://dx.doi.org/10.1016/j.bcab.2018.01.008
http://hdl.handle.net/11449/179564
identifier_str_mv Biocatalysis and Agricultural Biotechnology, v. 13, p. 311-318.
1878-8181
10.1016/j.bcab.2018.01.008
2-s2.0-85041603477
2-s2.0-85041603477.pdf
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Biocatalysis and Agricultural Biotechnology
0,479
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 311-318
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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