Biochemical characterization of an isolated 50 kDa beta-glucosidase from the thermophilic fungus Myceliophthora thermophila M.7.7
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Outros Autores: | , , |
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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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 |
|
_version_ |
1808128378475118592 |