EcXyl43 β-xylosidase: molecular modeling, activity on natural and artificial substrates, and synergism with endoxylanases for lignocellulose deconstruction
Biomass hydrolysis constitutes a bottleneck for the biotransformation of lignocellulosic residues into bioethanol and high-value products. The efficient deconstruction of polysaccharides to fermentable sugars requires multiple enzymes acting concertedly. GH43 β-xylosidases are among the most interes...
| Autores principales: | , , , , , , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
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Springer
2018
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| Materias: | |
| Acceso en línea: | https://link.springer.com/article/10.1007%2Fs00253-018-9138-7 http://hdl.handle.net/20.500.12123/3683 https://doi.org/10.1007/s00253-018-9138-7 |
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| author | Ontañon, Ornella Mailén Ghio, Silvina Marrero Diaz De Vill, Rubén Piccinni, Florencia Elizabeth Talia, Paola Mónica Cerutti, Maria Laura Campos, Eleonora |
| author_browse | Campos, Eleonora Cerutti, Maria Laura Ghio, Silvina Marrero Diaz De Vill, Rubén Ontañon, Ornella Mailén Piccinni, Florencia Elizabeth Talia, Paola Mónica |
| author_facet | Ontañon, Ornella Mailén Ghio, Silvina Marrero Diaz De Vill, Rubén Piccinni, Florencia Elizabeth Talia, Paola Mónica Cerutti, Maria Laura Campos, Eleonora |
| author_sort | Ontañon, Ornella Mailén |
| collection | INTA Digital |
| description | Biomass hydrolysis constitutes a bottleneck for the biotransformation of lignocellulosic residues into bioethanol and high-value products. The efficient deconstruction of polysaccharides to fermentable sugars requires multiple enzymes acting concertedly. GH43 β-xylosidases are among the most interesting enzymes involved in hemicellulose deconstruction into xylose. In this work, the structural and functional properties of β-xylosidase EcXyl43 from Enterobacter sp. were thoroughly characterized. Molecular modeling suggested a 3D structure formed by a conserved N-terminal catalytic domain linked to an ancillary C-terminal domain. Both domains resulted essential for enzymatic activity, and the role of critical residues, from the catalytic and the ancillary modules, was confirmed by mutagenesis. EcXyl43 presented β-xylosidase activity towards natural and artificial substrates while arabinofuranosidase activity was only detected on nitrophenyl α-L-arabinofuranoside (pNPA). It hydrolyzed xylobiose and purified xylooligosaccharides (XOS), up to degree of polymerization 6, with higher activity towards longer XOS. Low levels of activity on commercial xylan were also observed, mainly on the soluble fraction. The addition of EcXyl43 to GH10 and GH11 endoxylanases increased the release of xylose from xylan and pre-treated wheat straw. Additionally, EcXyl43 exhibited high efficiency and thermal stability under its optimal conditions (40 °C, pH 6.5), with a half-life of 58 h. Therefore, this enzyme could be a suitable additive for hemicellulases in long-term hydrolysis reactions. Because of its moderate inhibition by monomeric sugars but its high inhibition by ethanol, EcXyl43 could be particularly more useful in separate hydrolysis and fermentation (SHF) than in simultaneous saccharification and co-fermentation (SSCF) or consolidated bioprocessing (CBP). |
| format | Artículo |
| id | INTA3683 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2018 |
| publishDateRange | 2018 |
| publishDateSort | 2018 |
| publisher | Springer |
| publisherStr | Springer |
| record_format | dspace |
| spelling | INTA36832018-10-23T18:44:49Z EcXyl43 β-xylosidase: molecular modeling, activity on natural and artificial substrates, and synergism with endoxylanases for lignocellulose deconstruction Ontañon, Ornella Mailén Ghio, Silvina Marrero Diaz De Vill, Rubén Piccinni, Florencia Elizabeth Talia, Paola Mónica Cerutti, Maria Laura Campos, Eleonora Hemicellulose Synergism Lignocellulose Hemicelulosa Sinergismo Lignocelulosa Bioetanol Bioethanol Biomass hydrolysis constitutes a bottleneck for the biotransformation of lignocellulosic residues into bioethanol and high-value products. The efficient deconstruction of polysaccharides to fermentable sugars requires multiple enzymes acting concertedly. GH43 β-xylosidases are among the most interesting enzymes involved in hemicellulose deconstruction into xylose. In this work, the structural and functional properties of β-xylosidase EcXyl43 from Enterobacter sp. were thoroughly characterized. Molecular modeling suggested a 3D structure formed by a conserved N-terminal catalytic domain linked to an ancillary C-terminal domain. Both domains resulted essential for enzymatic activity, and the role of critical residues, from the catalytic and the ancillary modules, was confirmed by mutagenesis. EcXyl43 presented β-xylosidase activity towards natural and artificial substrates while arabinofuranosidase activity was only detected on nitrophenyl α-L-arabinofuranoside (pNPA). It hydrolyzed xylobiose and purified xylooligosaccharides (XOS), up to degree of polymerization 6, with higher activity towards longer XOS. Low levels of activity on commercial xylan were also observed, mainly on the soluble fraction. The addition of EcXyl43 to GH10 and GH11 endoxylanases increased the release of xylose from xylan and pre-treated wheat straw. Additionally, EcXyl43 exhibited high efficiency and thermal stability under its optimal conditions (40 °C, pH 6.5), with a half-life of 58 h. Therefore, this enzyme could be a suitable additive for hemicellulases in long-term hydrolysis reactions. Because of its moderate inhibition by monomeric sugars but its high inhibition by ethanol, EcXyl43 could be particularly more useful in separate hydrolysis and fermentation (SHF) than in simultaneous saccharification and co-fermentation (SSCF) or consolidated bioprocessing (CBP). Instituto de Biotecnología Fil: Ontañon, Ornella Mailén. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ghio, Silvina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Suelos; Argentina Fil: Marrero Diaz de Villegas, Rubén. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Piccinni, Florencia Elizabeth. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Talia, Paola Mónica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Cerutti, Maria Laura. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Campos, Eleonora. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina 2018-10-23T18:23:22Z 2018-10-23T18:23:22Z 2018-08 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion https://link.springer.com/article/10.1007%2Fs00253-018-9138-7 http://hdl.handle.net/20.500.12123/3683 1432-0614 https://doi.org/10.1007/s00253-018-9138-7 eng info:eu-repo/semantics/restrictedAccess application/pdf Springer Applied Microbiology and Biotechnology 102 (16): 6959-6971 (Agosto 2018) |
| spellingShingle | Hemicellulose Synergism Lignocellulose Hemicelulosa Sinergismo Lignocelulosa Bioetanol Bioethanol Ontañon, Ornella Mailén Ghio, Silvina Marrero Diaz De Vill, Rubén Piccinni, Florencia Elizabeth Talia, Paola Mónica Cerutti, Maria Laura Campos, Eleonora EcXyl43 β-xylosidase: molecular modeling, activity on natural and artificial substrates, and synergism with endoxylanases for lignocellulose deconstruction |
| title | EcXyl43 β-xylosidase: molecular modeling, activity on natural and artificial substrates, and synergism with endoxylanases for lignocellulose deconstruction |
| title_full | EcXyl43 β-xylosidase: molecular modeling, activity on natural and artificial substrates, and synergism with endoxylanases for lignocellulose deconstruction |
| title_fullStr | EcXyl43 β-xylosidase: molecular modeling, activity on natural and artificial substrates, and synergism with endoxylanases for lignocellulose deconstruction |
| title_full_unstemmed | EcXyl43 β-xylosidase: molecular modeling, activity on natural and artificial substrates, and synergism with endoxylanases for lignocellulose deconstruction |
| title_short | EcXyl43 β-xylosidase: molecular modeling, activity on natural and artificial substrates, and synergism with endoxylanases for lignocellulose deconstruction |
| title_sort | ecxyl43 β xylosidase molecular modeling activity on natural and artificial substrates and synergism with endoxylanases for lignocellulose deconstruction |
| topic | Hemicellulose Synergism Lignocellulose Hemicelulosa Sinergismo Lignocelulosa Bioetanol Bioethanol |
| url | https://link.springer.com/article/10.1007%2Fs00253-018-9138-7 http://hdl.handle.net/20.500.12123/3683 https://doi.org/10.1007/s00253-018-9138-7 |
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