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A Fungal Versatile GH10 Endoxylanase and Its Glycosynthase Variant: Synthesis of Xylooligosaccharides and Glycosides of Bioactive Phenolic Compounds

The study of endoxylanases as catalysts to valorize hemicellulosic residues and to obtain glycosides with improved properties is a topic of great industrial interest. In this work, a GH10 β-1,4-endoxylanase (XynSOS), from the ascomycetous fungus Talaromyces amestolkiae, has been heterologously produ...

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Autores principales: Pozo-Rodríguez, Ana, Méndez-Líter, Juan A., de Eugenio, Laura I., Nieto-Domínguez, Manuel, Calviño, Eva, Cañada, Francisco Javier, Santana, Andrés G., Díez, Jaime, Asensio, Juan L., Barriuso, Jorge, Prieto, Alicia, Martínez, María Jesús
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8836076/
https://www.ncbi.nlm.nih.gov/pubmed/35163307
http://dx.doi.org/10.3390/ijms23031383
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author Pozo-Rodríguez, Ana
Méndez-Líter, Juan A.
de Eugenio, Laura I.
Nieto-Domínguez, Manuel
Calviño, Eva
Cañada, Francisco Javier
Santana, Andrés G.
Díez, Jaime
Asensio, Juan L.
Barriuso, Jorge
Prieto, Alicia
Martínez, María Jesús
author_facet Pozo-Rodríguez, Ana
Méndez-Líter, Juan A.
de Eugenio, Laura I.
Nieto-Domínguez, Manuel
Calviño, Eva
Cañada, Francisco Javier
Santana, Andrés G.
Díez, Jaime
Asensio, Juan L.
Barriuso, Jorge
Prieto, Alicia
Martínez, María Jesús
author_sort Pozo-Rodríguez, Ana
collection PubMed
description The study of endoxylanases as catalysts to valorize hemicellulosic residues and to obtain glycosides with improved properties is a topic of great industrial interest. In this work, a GH10 β-1,4-endoxylanase (XynSOS), from the ascomycetous fungus Talaromyces amestolkiae, has been heterologously produced in Pichia pastoris, purified, and characterized. rXynSOS is a highly glycosylated monomeric enzyme of 53 kDa that contains a functional CBM1 domain and shows its optimal activity on azurine cross-linked (AZCL)–beechwood xylan at 70 °C and pH 5. Substrate specificity and kinetic studies confirmed its versatility and high affinity for beechwood xylan and wheat arabinoxylan. Moreover, rXynSOS was capable of transglycosylating phenolic compounds, although with low efficiencies. For expanding its synthetic capacity, a glycosynthase variant of rXynSOS was developed by directed mutagenesis, replacing its nucleophile catalytic residue E236 by a glycine (rXynSOS-E236G). This novel glycosynthase was able to synthesize β-1,4-xylooligosaccharides (XOS) of different lengths (four, six, eight, and ten xylose units), which are known to be emerging prebiotics. rXynSOS-E236G was also much more active than the native enzyme in the glycosylation of a broad range of phenolic compounds with antioxidant properties. The interesting capabilities of rXynSOS and its glycosynthase variant make them promising tools for biotechnological applications.
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spelling pubmed-88360762022-02-12 A Fungal Versatile GH10 Endoxylanase and Its Glycosynthase Variant: Synthesis of Xylooligosaccharides and Glycosides of Bioactive Phenolic Compounds Pozo-Rodríguez, Ana Méndez-Líter, Juan A. de Eugenio, Laura I. Nieto-Domínguez, Manuel Calviño, Eva Cañada, Francisco Javier Santana, Andrés G. Díez, Jaime Asensio, Juan L. Barriuso, Jorge Prieto, Alicia Martínez, María Jesús Int J Mol Sci Article The study of endoxylanases as catalysts to valorize hemicellulosic residues and to obtain glycosides with improved properties is a topic of great industrial interest. In this work, a GH10 β-1,4-endoxylanase (XynSOS), from the ascomycetous fungus Talaromyces amestolkiae, has been heterologously produced in Pichia pastoris, purified, and characterized. rXynSOS is a highly glycosylated monomeric enzyme of 53 kDa that contains a functional CBM1 domain and shows its optimal activity on azurine cross-linked (AZCL)–beechwood xylan at 70 °C and pH 5. Substrate specificity and kinetic studies confirmed its versatility and high affinity for beechwood xylan and wheat arabinoxylan. Moreover, rXynSOS was capable of transglycosylating phenolic compounds, although with low efficiencies. For expanding its synthetic capacity, a glycosynthase variant of rXynSOS was developed by directed mutagenesis, replacing its nucleophile catalytic residue E236 by a glycine (rXynSOS-E236G). This novel glycosynthase was able to synthesize β-1,4-xylooligosaccharides (XOS) of different lengths (four, six, eight, and ten xylose units), which are known to be emerging prebiotics. rXynSOS-E236G was also much more active than the native enzyme in the glycosylation of a broad range of phenolic compounds with antioxidant properties. The interesting capabilities of rXynSOS and its glycosynthase variant make them promising tools for biotechnological applications. MDPI 2022-01-26 /pmc/articles/PMC8836076/ /pubmed/35163307 http://dx.doi.org/10.3390/ijms23031383 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Pozo-Rodríguez, Ana
Méndez-Líter, Juan A.
de Eugenio, Laura I.
Nieto-Domínguez, Manuel
Calviño, Eva
Cañada, Francisco Javier
Santana, Andrés G.
Díez, Jaime
Asensio, Juan L.
Barriuso, Jorge
Prieto, Alicia
Martínez, María Jesús
A Fungal Versatile GH10 Endoxylanase and Its Glycosynthase Variant: Synthesis of Xylooligosaccharides and Glycosides of Bioactive Phenolic Compounds
title A Fungal Versatile GH10 Endoxylanase and Its Glycosynthase Variant: Synthesis of Xylooligosaccharides and Glycosides of Bioactive Phenolic Compounds
title_full A Fungal Versatile GH10 Endoxylanase and Its Glycosynthase Variant: Synthesis of Xylooligosaccharides and Glycosides of Bioactive Phenolic Compounds
title_fullStr A Fungal Versatile GH10 Endoxylanase and Its Glycosynthase Variant: Synthesis of Xylooligosaccharides and Glycosides of Bioactive Phenolic Compounds
title_full_unstemmed A Fungal Versatile GH10 Endoxylanase and Its Glycosynthase Variant: Synthesis of Xylooligosaccharides and Glycosides of Bioactive Phenolic Compounds
title_short A Fungal Versatile GH10 Endoxylanase and Its Glycosynthase Variant: Synthesis of Xylooligosaccharides and Glycosides of Bioactive Phenolic Compounds
title_sort fungal versatile gh10 endoxylanase and its glycosynthase variant: synthesis of xylooligosaccharides and glycosides of bioactive phenolic compounds
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8836076/
https://www.ncbi.nlm.nih.gov/pubmed/35163307
http://dx.doi.org/10.3390/ijms23031383
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