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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...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Hindawi Publishing Corporation
2013
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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. |
format | Online Article Text |
id | pubmed-3723094 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Hindawi Publishing Corporation |
record_format | MEDLINE/PubMed |
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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