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Título: | A Fungal Versatile GH10 Endoxylanase and Its Glycosynthase Variant: Synthesis of Xylooligosaccharides and Glycosides of Bioactive Phenolic Compounds |
Autor: | Pozo-Rodríguez, Ana CSIC ORCID; Méndez-Líter, Juan A. CSIC ; Eugenio, Laura I. de ; Nieto-Domínguez, Manuel; Calviño, Eva CSIC ORCID ; Cañada, F. Javier ; Santana, Andrés G. CSIC ORCID ; Díez, Jaime; Asensio, Juan L.; Barriuso, Jorge CSIC ORCID ; Prieto Orzanco, Alicia CSIC ORCID ; Martínez, María Jesús CSIC ORCID | Palabras clave: | Antioxidants Fungal enzyme Glycosylation Hydrolysis Oligosaccharides Protein engineering |
Fecha de publicación: | feb-2022 | Editor: | Multidisciplinary Digital Publishing Institute | Citación: | International Journal of Molecular Science 23 : 1383 (2022) | Resumen: | 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 het- erologously 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 arabi- noxylan. 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 application. | Versión del editor: | https://doi.org/10.3390/ijms23031383 | URI: | http://hdl.handle.net/10261/272197 | DOI: | 10.3390/ijms23031383 |
Aparece en las colecciones: | (IQOG) Artículos |
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ijms-23-01383.pdf | Artículo principal | 1,62 MB | Adobe PDF | Visualizar/Abrir |
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