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Biochemical Characterization, Thermal Stability, and Partial Sequence of a Novel Exo-Polygalacturonase from the Thermophilic Fungus Rhizomucor pusillus A13.36 Obtained by Submerged Cultivation
This work reports the production of an exo-polygalacturonase (exo-PG) by Rhizomucor pusillus A13.36 in submerged cultivation (SmC) in a shaker at 45°C for 96 h. A single pectinase was found and purified in order to analyze its thermal stability, by salt precipitation and hydrophobic interaction chro...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Hindawi Publishing Corporation
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5153499/ https://www.ncbi.nlm.nih.gov/pubmed/28025649 http://dx.doi.org/10.1155/2016/8653583 |
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author | Trindade, Lucas Vinícius Desagiacomo, Carla Polizeli, Maria de Lourdes Teixeira de Moraes Damasio, André Ricardo de Lima Lima, Aline Margarete Furuyama Gomes, Eleni Bonilla-Rodriguez, Gustavo Orlando |
author_facet | Trindade, Lucas Vinícius Desagiacomo, Carla Polizeli, Maria de Lourdes Teixeira de Moraes Damasio, André Ricardo de Lima Lima, Aline Margarete Furuyama Gomes, Eleni Bonilla-Rodriguez, Gustavo Orlando |
author_sort | Trindade, Lucas Vinícius |
collection | PubMed |
description | This work reports the production of an exo-polygalacturonase (exo-PG) by Rhizomucor pusillus A13.36 in submerged cultivation (SmC) in a shaker at 45°C for 96 h. A single pectinase was found and purified in order to analyze its thermal stability, by salt precipitation and hydrophobic interaction chromatography. The pectinase has an estimated Mw of approximately 43.5–47 kDa and optimum pH of 4.0 but is stable in pH ranging from 3.5 to 9.5 and has an optimum temperature of 61°C. It presents thermal stability between 30 and 60°C, has 70% activation in the presence of Ca(2+), and was tested using citrus pectin with a degree of methyl esterification (DE) of 26%. E (a(d)) for irreversible denaturation was 125.5 kJ/mol with positive variations of entropy and enthalpy for that and ΔG ((d)) values were around 50 kJ/mol. The hydrolysis of polygalacturonate was analyzed by capillary electrophoresis which displayed a pattern of sequential hydrolysis (exo). The partial identification of the primary sequence was done by MS MALDI-TOF and a comparison with data banks showed the highest identity of the sequenced fragments of exo-PG from R. pusillus with an exo-pectinase from Aspergillus fumigatus. Pectin hydrolysis showed a sigmoidal curve for the Michaelis-Menten plot. |
format | Online Article Text |
id | pubmed-5153499 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Hindawi Publishing Corporation |
record_format | MEDLINE/PubMed |
spelling | pubmed-51534992016-12-26 Biochemical Characterization, Thermal Stability, and Partial Sequence of a Novel Exo-Polygalacturonase from the Thermophilic Fungus Rhizomucor pusillus A13.36 Obtained by Submerged Cultivation Trindade, Lucas Vinícius Desagiacomo, Carla Polizeli, Maria de Lourdes Teixeira de Moraes Damasio, André Ricardo de Lima Lima, Aline Margarete Furuyama Gomes, Eleni Bonilla-Rodriguez, Gustavo Orlando Biomed Res Int Research Article This work reports the production of an exo-polygalacturonase (exo-PG) by Rhizomucor pusillus A13.36 in submerged cultivation (SmC) in a shaker at 45°C for 96 h. A single pectinase was found and purified in order to analyze its thermal stability, by salt precipitation and hydrophobic interaction chromatography. The pectinase has an estimated Mw of approximately 43.5–47 kDa and optimum pH of 4.0 but is stable in pH ranging from 3.5 to 9.5 and has an optimum temperature of 61°C. It presents thermal stability between 30 and 60°C, has 70% activation in the presence of Ca(2+), and was tested using citrus pectin with a degree of methyl esterification (DE) of 26%. E (a(d)) for irreversible denaturation was 125.5 kJ/mol with positive variations of entropy and enthalpy for that and ΔG ((d)) values were around 50 kJ/mol. The hydrolysis of polygalacturonate was analyzed by capillary electrophoresis which displayed a pattern of sequential hydrolysis (exo). The partial identification of the primary sequence was done by MS MALDI-TOF and a comparison with data banks showed the highest identity of the sequenced fragments of exo-PG from R. pusillus with an exo-pectinase from Aspergillus fumigatus. Pectin hydrolysis showed a sigmoidal curve for the Michaelis-Menten plot. Hindawi Publishing Corporation 2016 2016-11-29 /pmc/articles/PMC5153499/ /pubmed/28025649 http://dx.doi.org/10.1155/2016/8653583 Text en Copyright © 2016 Lucas Vinícius Trindade et al. https://creativecommons.org/licenses/by/4.0/ 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 Trindade, Lucas Vinícius Desagiacomo, Carla Polizeli, Maria de Lourdes Teixeira de Moraes Damasio, André Ricardo de Lima Lima, Aline Margarete Furuyama Gomes, Eleni Bonilla-Rodriguez, Gustavo Orlando Biochemical Characterization, Thermal Stability, and Partial Sequence of a Novel Exo-Polygalacturonase from the Thermophilic Fungus Rhizomucor pusillus A13.36 Obtained by Submerged Cultivation |
title | Biochemical Characterization, Thermal Stability, and Partial Sequence of a Novel Exo-Polygalacturonase from the Thermophilic Fungus Rhizomucor pusillus A13.36 Obtained by Submerged Cultivation |
title_full | Biochemical Characterization, Thermal Stability, and Partial Sequence of a Novel Exo-Polygalacturonase from the Thermophilic Fungus Rhizomucor pusillus A13.36 Obtained by Submerged Cultivation |
title_fullStr | Biochemical Characterization, Thermal Stability, and Partial Sequence of a Novel Exo-Polygalacturonase from the Thermophilic Fungus Rhizomucor pusillus A13.36 Obtained by Submerged Cultivation |
title_full_unstemmed | Biochemical Characterization, Thermal Stability, and Partial Sequence of a Novel Exo-Polygalacturonase from the Thermophilic Fungus Rhizomucor pusillus A13.36 Obtained by Submerged Cultivation |
title_short | Biochemical Characterization, Thermal Stability, and Partial Sequence of a Novel Exo-Polygalacturonase from the Thermophilic Fungus Rhizomucor pusillus A13.36 Obtained by Submerged Cultivation |
title_sort | biochemical characterization, thermal stability, and partial sequence of a novel exo-polygalacturonase from the thermophilic fungus rhizomucor pusillus a13.36 obtained by submerged cultivation |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5153499/ https://www.ncbi.nlm.nih.gov/pubmed/28025649 http://dx.doi.org/10.1155/2016/8653583 |
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