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Exploring the Molecular Basis for Substrate Affinity and Structural Stability in Bacterial GH39 β-Xylosidases
The glycoside hydrolase family 39 (GH39) is a functionally expanding family with limited understanding about the molecular basis for substrate specificity and extremophilicity. In this work, we demonstrate the key role of the positive-subsite region in modulating substrate affinity and how the lack...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7242879/ https://www.ncbi.nlm.nih.gov/pubmed/32500063 http://dx.doi.org/10.3389/fbioe.2020.00419 |
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author | de Morais, Mariana Abrahão Bueno Polo, Carla Cristina Domingues, Mariane Noronha Persinoti, Gabriela Felix Pirolla, Renan Augusto Siqueira de Souza, Flávio Henrique Moreira Correa, Jessica Batista de Lima dos Santos, Camila Ramos Murakami, Mário Tyago |
author_facet | de Morais, Mariana Abrahão Bueno Polo, Carla Cristina Domingues, Mariane Noronha Persinoti, Gabriela Felix Pirolla, Renan Augusto Siqueira de Souza, Flávio Henrique Moreira Correa, Jessica Batista de Lima dos Santos, Camila Ramos Murakami, Mário Tyago |
author_sort | de Morais, Mariana Abrahão Bueno |
collection | PubMed |
description | The glycoside hydrolase family 39 (GH39) is a functionally expanding family with limited understanding about the molecular basis for substrate specificity and extremophilicity. In this work, we demonstrate the key role of the positive-subsite region in modulating substrate affinity and how the lack of a C-terminal extension impacts on oligomerization and structural stability of some GH39 members. The crystallographic and SAXS structures of a new GH39 member from the phytopathogen Xanthomonas citri support the importance of an extended C-terminal to promote oligomerization as a molecular strategy to enhance thermal stability. Comparative structural analysis along with site-directed mutagenesis showed that two residues located at the positive-subsite region, Lys166 and Asp167, are critical to substrate affinity and catalytic performance, by inducing local changes in the active site for substrate binding. These findings expand the molecular understanding of the mechanisms involved in substrate recognition and structural stability of the GH39 family, which might be instrumental for biological insights, rational enzyme engineering and utilization in biorefineries. |
format | Online Article Text |
id | pubmed-7242879 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-72428792020-06-03 Exploring the Molecular Basis for Substrate Affinity and Structural Stability in Bacterial GH39 β-Xylosidases de Morais, Mariana Abrahão Bueno Polo, Carla Cristina Domingues, Mariane Noronha Persinoti, Gabriela Felix Pirolla, Renan Augusto Siqueira de Souza, Flávio Henrique Moreira Correa, Jessica Batista de Lima dos Santos, Camila Ramos Murakami, Mário Tyago Front Bioeng Biotechnol Bioengineering and Biotechnology The glycoside hydrolase family 39 (GH39) is a functionally expanding family with limited understanding about the molecular basis for substrate specificity and extremophilicity. In this work, we demonstrate the key role of the positive-subsite region in modulating substrate affinity and how the lack of a C-terminal extension impacts on oligomerization and structural stability of some GH39 members. The crystallographic and SAXS structures of a new GH39 member from the phytopathogen Xanthomonas citri support the importance of an extended C-terminal to promote oligomerization as a molecular strategy to enhance thermal stability. Comparative structural analysis along with site-directed mutagenesis showed that two residues located at the positive-subsite region, Lys166 and Asp167, are critical to substrate affinity and catalytic performance, by inducing local changes in the active site for substrate binding. These findings expand the molecular understanding of the mechanisms involved in substrate recognition and structural stability of the GH39 family, which might be instrumental for biological insights, rational enzyme engineering and utilization in biorefineries. Frontiers Media S.A. 2020-05-15 /pmc/articles/PMC7242879/ /pubmed/32500063 http://dx.doi.org/10.3389/fbioe.2020.00419 Text en Copyright © 2020 Morais, Polo, Domingues, Persinoti, Pirolla, de Souza, Correa, dos Santos and Murakami. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Bioengineering and Biotechnology de Morais, Mariana Abrahão Bueno Polo, Carla Cristina Domingues, Mariane Noronha Persinoti, Gabriela Felix Pirolla, Renan Augusto Siqueira de Souza, Flávio Henrique Moreira Correa, Jessica Batista de Lima dos Santos, Camila Ramos Murakami, Mário Tyago Exploring the Molecular Basis for Substrate Affinity and Structural Stability in Bacterial GH39 β-Xylosidases |
title | Exploring the Molecular Basis for Substrate Affinity and Structural Stability in Bacterial GH39 β-Xylosidases |
title_full | Exploring the Molecular Basis for Substrate Affinity and Structural Stability in Bacterial GH39 β-Xylosidases |
title_fullStr | Exploring the Molecular Basis for Substrate Affinity and Structural Stability in Bacterial GH39 β-Xylosidases |
title_full_unstemmed | Exploring the Molecular Basis for Substrate Affinity and Structural Stability in Bacterial GH39 β-Xylosidases |
title_short | Exploring the Molecular Basis for Substrate Affinity and Structural Stability in Bacterial GH39 β-Xylosidases |
title_sort | exploring the molecular basis for substrate affinity and structural stability in bacterial gh39 β-xylosidases |
topic | Bioengineering and Biotechnology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7242879/ https://www.ncbi.nlm.nih.gov/pubmed/32500063 http://dx.doi.org/10.3389/fbioe.2020.00419 |
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