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An Acidic Thermostable Recombinant Aspergillus nidulans Endoglucanase Is Active towards Distinct Agriculture Residues

Aspergillus nidulans is poorly exploited as a source of enzymes for lignocellulosic residues degradation for biotechnological purposes. This work describes the A. nidulans Endoglucanase A heterologous expression in Pichia pastoris, the purification and biochemical characterization of the recombinant...

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Autores principales: Tavares, Eveline Queiroz de Pinho, Rubini, Marciano Regis, Mello-de-Sousa, Thiago Machado, Duarte, Gilvan Caetano, de Faria, Fabrícia Paula, Ferreira Filho, Edivaldo Ximenes, Kyaw, Cynthia Maria, Silva-Pereira, Ildinete, Poças-Fonseca, Marcio Jose
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3723094/
https://www.ncbi.nlm.nih.gov/pubmed/23936633
http://dx.doi.org/10.1155/2013/287343
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author Tavares, Eveline Queiroz de Pinho
Rubini, Marciano Regis
Mello-de-Sousa, Thiago Machado
Duarte, Gilvan Caetano
de Faria, Fabrícia Paula
Ferreira Filho, Edivaldo Ximenes
Kyaw, Cynthia Maria
Silva-Pereira, Ildinete
Poças-Fonseca, Marcio Jose
author_facet Tavares, Eveline Queiroz de Pinho
Rubini, Marciano Regis
Mello-de-Sousa, Thiago Machado
Duarte, Gilvan Caetano
de Faria, Fabrícia Paula
Ferreira Filho, Edivaldo Ximenes
Kyaw, Cynthia Maria
Silva-Pereira, Ildinete
Poças-Fonseca, Marcio Jose
author_sort Tavares, Eveline Queiroz de Pinho
collection PubMed
description Aspergillus nidulans is poorly exploited as a source of enzymes for lignocellulosic residues degradation for biotechnological purposes. This work describes the A. nidulans Endoglucanase A heterologous expression in Pichia pastoris, the purification and biochemical characterization of the recombinant enzyme. Active recombinant endoglucanase A (rEG A) was efficiently secreted as a 35 kDa protein which was purified through a two-step chromatography procedure. The highest enzyme activity was detected at 50°C/pH 4. rEG A retained 100% of activity when incubated at 45 and 55°C for 72 h. Purified rEG A kinetic parameters towards CMC were determined as K (m) = 27.5 ± 4.33 mg/mL, V (max) = 1.185 ± 0.11 mmol/min, and 55.8 IU (international units)/mg specific activity. Recombinant P. pastoris supernatant presented hydrolytic activity towards lignocellulosic residues such as banana stalk, sugarcane bagasse, soybean residues, and corn straw. These data indicate that rEG A is suitable for plant biomass conversion into products of commercial importance, such as second-generation fuel ethanol.
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spelling pubmed-37230942013-08-09 An Acidic Thermostable Recombinant Aspergillus nidulans Endoglucanase Is Active towards Distinct Agriculture Residues Tavares, Eveline Queiroz de Pinho Rubini, Marciano Regis Mello-de-Sousa, Thiago Machado Duarte, Gilvan Caetano de Faria, Fabrícia Paula Ferreira Filho, Edivaldo Ximenes Kyaw, Cynthia Maria Silva-Pereira, Ildinete Poças-Fonseca, Marcio Jose Enzyme Res Research Article Aspergillus nidulans is poorly exploited as a source of enzymes for lignocellulosic residues degradation for biotechnological purposes. This work describes the A. nidulans Endoglucanase A heterologous expression in Pichia pastoris, the purification and biochemical characterization of the recombinant enzyme. Active recombinant endoglucanase A (rEG A) was efficiently secreted as a 35 kDa protein which was purified through a two-step chromatography procedure. The highest enzyme activity was detected at 50°C/pH 4. rEG A retained 100% of activity when incubated at 45 and 55°C for 72 h. Purified rEG A kinetic parameters towards CMC were determined as K (m) = 27.5 ± 4.33 mg/mL, V (max) = 1.185 ± 0.11 mmol/min, and 55.8 IU (international units)/mg specific activity. Recombinant P. pastoris supernatant presented hydrolytic activity towards lignocellulosic residues such as banana stalk, sugarcane bagasse, soybean residues, and corn straw. These data indicate that rEG A is suitable for plant biomass conversion into products of commercial importance, such as second-generation fuel ethanol. Hindawi Publishing Corporation 2013 2013-07-10 /pmc/articles/PMC3723094/ /pubmed/23936633 http://dx.doi.org/10.1155/2013/287343 Text en Copyright © 2013 Eveline Queiroz de Pinho Tavares et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Tavares, Eveline Queiroz de Pinho
Rubini, Marciano Regis
Mello-de-Sousa, Thiago Machado
Duarte, Gilvan Caetano
de Faria, Fabrícia Paula
Ferreira Filho, Edivaldo Ximenes
Kyaw, Cynthia Maria
Silva-Pereira, Ildinete
Poças-Fonseca, Marcio Jose
An Acidic Thermostable Recombinant Aspergillus nidulans Endoglucanase Is Active towards Distinct Agriculture Residues
title An Acidic Thermostable Recombinant Aspergillus nidulans Endoglucanase Is Active towards Distinct Agriculture Residues
title_full An Acidic Thermostable Recombinant Aspergillus nidulans Endoglucanase Is Active towards Distinct Agriculture Residues
title_fullStr An Acidic Thermostable Recombinant Aspergillus nidulans Endoglucanase Is Active towards Distinct Agriculture Residues
title_full_unstemmed An Acidic Thermostable Recombinant Aspergillus nidulans Endoglucanase Is Active towards Distinct Agriculture Residues
title_short An Acidic Thermostable Recombinant Aspergillus nidulans Endoglucanase Is Active towards Distinct Agriculture Residues
title_sort acidic thermostable recombinant aspergillus nidulans endoglucanase is active towards distinct agriculture residues
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3723094/
https://www.ncbi.nlm.nih.gov/pubmed/23936633
http://dx.doi.org/10.1155/2013/287343
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