Biochemical characterization of endoglucanases produced by Myceliophthora thermophila M.7.7 in solid-state culture

Detalhes bibliográficos
Autor(a) principal: Massarente, Vanessa Salto
Data de Publicação: 2020
Outros Autores: de Araujo Zanoni, Jéssica, Gomes, Eleni, 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.2020.101684
http://hdl.handle.net/11449/200794
Resumo: Cellulases have been investigated for their potential application in lignocellulosic residues, used to produce second-generation ethanol (2 GE), as these enzymes can hydrolyze cellulose and release glucose for alcoholic fermentation by yeasts. The aim of this project was to characterize the endoglucanases produced by the thermophilic fungus Myceliophthora thermophila M.7.7 in solid-state cultivation. Tests were carried out with the crude enzyme extract (CEE) to assess the effects of pH, temperature, and various chemicals including cations and phenolic compounds on endoglucanolytic activity. The molecular weight estimation was performed by polyacrylamide gel electrophoresis (SDS-PAGE), and it was possible to observe seven endoglucanases detected by zymography, with estimated Mw in the range between 26 and 82 kDa. The optimum pH for the CEE was 5.5 and the optimum incubation temperature was 70 °C. Concerning endoglucanase stability, the highest activities were achieved in the pH range of 6.0–6.5, and between temperatures of 30 and 40 °C up to 120 min. Among the tested reagents, Triton, dithiotreitol (DTT), and Isopropanol increased the enzymatic activity on the crude extract by about 10%. Among cations, only Sr2+ significantly increased the endoglucanases’ activity. Of the phenolic compounds, although all induced a decrease of the endoglucanolytic activity, none could nullify it after a pre-incubation for 24 h. The apparent kinetic parameters were determined using carboxymethylcellulose (CMC), being Vmax = 51.7 ± 2.7 μmol min−1 and Km = 0.99 ± 0.15 g% CMC (9.9 mg mL−1).
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spelling Biochemical characterization of endoglucanases produced by Myceliophthora thermophila M.7.7 in solid-state cultureBioenergyEndoglucanasesThermophilic fungiCellulases have been investigated for their potential application in lignocellulosic residues, used to produce second-generation ethanol (2 GE), as these enzymes can hydrolyze cellulose and release glucose for alcoholic fermentation by yeasts. The aim of this project was to characterize the endoglucanases produced by the thermophilic fungus Myceliophthora thermophila M.7.7 in solid-state cultivation. Tests were carried out with the crude enzyme extract (CEE) to assess the effects of pH, temperature, and various chemicals including cations and phenolic compounds on endoglucanolytic activity. The molecular weight estimation was performed by polyacrylamide gel electrophoresis (SDS-PAGE), and it was possible to observe seven endoglucanases detected by zymography, with estimated Mw in the range between 26 and 82 kDa. The optimum pH for the CEE was 5.5 and the optimum incubation temperature was 70 °C. Concerning endoglucanase stability, the highest activities were achieved in the pH range of 6.0–6.5, and between temperatures of 30 and 40 °C up to 120 min. Among the tested reagents, Triton, dithiotreitol (DTT), and Isopropanol increased the enzymatic activity on the crude extract by about 10%. Among cations, only Sr2+ significantly increased the endoglucanases’ activity. Of the phenolic compounds, although all induced a decrease of the endoglucanolytic activity, none could nullify it after a pre-incubation for 24 h. The apparent kinetic parameters were determined using carboxymethylcellulose (CMC), being Vmax = 51.7 ± 2.7 μmol min−1 and Km = 0.99 ± 0.15 g% CMC (9.9 mg mL−1).Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Graduate Program in MicrobiologyDepartment of BiologyDepartment of Chemistry and Environmental Sciences Biosciences Languages and Exact Sciences Institute São Paulo State University (IBILCE-UNESP)Department of Chemistry and Environmental Sciences Biosciences Languages and Exact Sciences Institute São Paulo State University (IBILCE-UNESP)Graduate Program in MicrobiologyUniversidade Estadual Paulista (Unesp)Massarente, Vanessa Saltode Araujo Zanoni, JéssicaGomes, EleniBonilla-Rodriguez, Gustavo Orlando [UNESP]2020-12-12T02:16:17Z2020-12-12T02:16:17Z2020-08-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://dx.doi.org/10.1016/j.bcab.2020.101684Biocatalysis and Agricultural Biotechnology, v. 27.1878-8181http://hdl.handle.net/11449/20079410.1016/j.bcab.2020.1016842-s2.0-85088381793Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBiocatalysis and Agricultural Biotechnologyinfo:eu-repo/semantics/openAccess2021-10-23T15:09:26Zoai:repositorio.unesp.br:11449/200794Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T19:00:33.445157Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Biochemical characterization of endoglucanases produced by Myceliophthora thermophila M.7.7 in solid-state culture
title Biochemical characterization of endoglucanases produced by Myceliophthora thermophila M.7.7 in solid-state culture
spellingShingle Biochemical characterization of endoglucanases produced by Myceliophthora thermophila M.7.7 in solid-state culture
Massarente, Vanessa Salto
Bioenergy
Endoglucanases
Thermophilic fungi
title_short Biochemical characterization of endoglucanases produced by Myceliophthora thermophila M.7.7 in solid-state culture
title_full Biochemical characterization of endoglucanases produced by Myceliophthora thermophila M.7.7 in solid-state culture
title_fullStr Biochemical characterization of endoglucanases produced by Myceliophthora thermophila M.7.7 in solid-state culture
title_full_unstemmed Biochemical characterization of endoglucanases produced by Myceliophthora thermophila M.7.7 in solid-state culture
title_sort Biochemical characterization of endoglucanases produced by Myceliophthora thermophila M.7.7 in solid-state culture
author Massarente, Vanessa Salto
author_facet Massarente, Vanessa Salto
de Araujo Zanoni, Jéssica
Gomes, Eleni
Bonilla-Rodriguez, Gustavo Orlando [UNESP]
author_role author
author2 de Araujo Zanoni, Jéssica
Gomes, Eleni
Bonilla-Rodriguez, Gustavo Orlando [UNESP]
author2_role author
author
author
dc.contributor.none.fl_str_mv Graduate Program in Microbiology
Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Massarente, Vanessa Salto
de Araujo Zanoni, Jéssica
Gomes, Eleni
Bonilla-Rodriguez, Gustavo Orlando [UNESP]
dc.subject.por.fl_str_mv Bioenergy
Endoglucanases
Thermophilic fungi
topic Bioenergy
Endoglucanases
Thermophilic fungi
description Cellulases have been investigated for their potential application in lignocellulosic residues, used to produce second-generation ethanol (2 GE), as these enzymes can hydrolyze cellulose and release glucose for alcoholic fermentation by yeasts. The aim of this project was to characterize the endoglucanases produced by the thermophilic fungus Myceliophthora thermophila M.7.7 in solid-state cultivation. Tests were carried out with the crude enzyme extract (CEE) to assess the effects of pH, temperature, and various chemicals including cations and phenolic compounds on endoglucanolytic activity. The molecular weight estimation was performed by polyacrylamide gel electrophoresis (SDS-PAGE), and it was possible to observe seven endoglucanases detected by zymography, with estimated Mw in the range between 26 and 82 kDa. The optimum pH for the CEE was 5.5 and the optimum incubation temperature was 70 °C. Concerning endoglucanase stability, the highest activities were achieved in the pH range of 6.0–6.5, and between temperatures of 30 and 40 °C up to 120 min. Among the tested reagents, Triton, dithiotreitol (DTT), and Isopropanol increased the enzymatic activity on the crude extract by about 10%. Among cations, only Sr2+ significantly increased the endoglucanases’ activity. Of the phenolic compounds, although all induced a decrease of the endoglucanolytic activity, none could nullify it after a pre-incubation for 24 h. The apparent kinetic parameters were determined using carboxymethylcellulose (CMC), being Vmax = 51.7 ± 2.7 μmol min−1 and Km = 0.99 ± 0.15 g% CMC (9.9 mg mL−1).
publishDate 2020
dc.date.none.fl_str_mv 2020-12-12T02:16:17Z
2020-12-12T02:16:17Z
2020-08-01
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.2020.101684
Biocatalysis and Agricultural Biotechnology, v. 27.
1878-8181
http://hdl.handle.net/11449/200794
10.1016/j.bcab.2020.101684
2-s2.0-85088381793
url http://dx.doi.org/10.1016/j.bcab.2020.101684
http://hdl.handle.net/11449/200794
identifier_str_mv Biocatalysis and Agricultural Biotechnology, v. 27.
1878-8181
10.1016/j.bcab.2020.101684
2-s2.0-85088381793
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Biocatalysis and Agricultural Biotechnology
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
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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