Expression of an extremophilic xylanase in Nicotiana benthamiana and its use for the production of prebiotic xylooligosaccharides
A gene construct encoding a xylanase, which is active in extreme conditions of temperature and alkaline pH (90 °C, pH 10.5), has been transitorily expressed with high efficiency in Nicotiana benthamiana using a viral vector. The enzyme, targeted to the apoplast, accumulates in large amounts in plant tissues in as little as 7 days after inoculation, without detrimental effects on plant growth. The properties of the protein produced by the plant, in terms of resistance to temperature, pH, and enzymatic activity, are equivalent to those observed when Escherichia coli is used as a host. Purification of the plant-produced recombinant xylanase is facilitated by exporting the protein to the apoplastic space. The production of this xylanase by N. benthamiana, which avoids the hindrances derived from the use of E. coli, namely, intracellular production requiring subsequent purification, represents an important step for potential applications in the food industry in which more sustainable and green products are continuously demanded. As an example, the use of the enzyme producing prebiotic xylooligosdaccharides from xylan is here reported.
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2022-09-21
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dig-iata-es-10261-2806572024-05-16T20:38:47Z Expression of an extremophilic xylanase in Nicotiana benthamiana and its use for the production of prebiotic xylooligosaccharides Talens Perales, David Nicolau Sanus, María Polaina Molina, Julio Daròs Arnau, José Antonio Ministerio de Ciencia e Innovación (España) European Commission 0000-0002-8693-4239 0000-0002-9963-5131 0000-0001-9912-0640 0000-0002-6535-2889 A gene construct encoding a xylanase, which is active in extreme conditions of temperature and alkaline pH (90 °C, pH 10.5), has been transitorily expressed with high efficiency in Nicotiana benthamiana using a viral vector. The enzyme, targeted to the apoplast, accumulates in large amounts in plant tissues in as little as 7 days after inoculation, without detrimental effects on plant growth. The properties of the protein produced by the plant, in terms of resistance to temperature, pH, and enzymatic activity, are equivalent to those observed when Escherichia coli is used as a host. Purification of the plant-produced recombinant xylanase is facilitated by exporting the protein to the apoplastic space. The production of this xylanase by N. benthamiana, which avoids the hindrances derived from the use of E. coli, namely, intracellular production requiring subsequent purification, represents an important step for potential applications in the food industry in which more sustainable and green products are continuously demanded. As an example, the use of the enzyme producing prebiotic xylooligosdaccharides from xylan is here reported. This work was supported by grant PID2020-114691RB-I00 from the Spanish Ministerio de Ciencia e Innovación, through the Agencia Estatal de Investigación (co-financed European Regional Development Fund), and by the Bio Based Industries Joint Undertaking, under the European Union’s Horizon 2020 research and innovation program (Project WOODZYMES, Grant Agreement H2020-BBI-JU-792070). M.N.-S. is the recipient of a predoctoral contract from the Spanish Ministerio de Ciencia e Innovación (PRE2018-084771). Peer reviewed 2022-10-06T14:03:59Z 2022-10-06T14:03:59Z 2022-09-21 artículo http://purl.org/coar/resource_type/c_6501 Scientific reports 12(1): 15743 (2022) http://hdl.handle.net/10261/280657 10.1038/s41598-022-19774-5 2045-2322 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100004837 36131073 2-s2.0-85138290438 https://api.elsevier.com/content/abstract/scopus_id/85138290438 en #PLACEHOLDER_PARENT_METADATA_VALUE# #PLACEHOLDER_PARENT_METADATA_VALUE# info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2020-114691RB-I00/ES/BIOTECNOLOGIA DE VIRUS DE PLANTAS: VECTORES VIRALES Y ESTRATEGIAS DE RESISTENCIA/ info:eu-repo/grantAgreement/EC/H2020/792070 Scientific reports Publisher's version https://doi.org/10.1038/s41598-022-19774-5 Sí open Springer Nature |
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A gene construct encoding a xylanase, which is active in extreme conditions of temperature and alkaline pH (90 °C, pH 10.5), has been transitorily expressed with high efficiency in Nicotiana benthamiana using a viral vector. The enzyme, targeted to the apoplast, accumulates in large amounts in plant tissues in as little as 7 days after inoculation, without detrimental effects on plant growth. The properties of the protein produced by the plant, in terms of resistance to temperature, pH, and enzymatic activity, are equivalent to those observed when Escherichia coli is used as a host. Purification of the plant-produced recombinant xylanase is facilitated by exporting the protein to the apoplastic space. The production of this xylanase by N. benthamiana, which avoids the hindrances derived from the use of E. coli, namely, intracellular production requiring subsequent purification, represents an important step for potential applications in the food industry in which more sustainable and green products are continuously demanded. As an example, the use of the enzyme producing prebiotic xylooligosdaccharides from xylan is here reported. |
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Ministerio de Ciencia e Innovación (España) |
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Ministerio de Ciencia e Innovación (España) Talens Perales, David Nicolau Sanus, María Polaina Molina, Julio Daròs Arnau, José Antonio |
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Talens Perales, David Nicolau Sanus, María Polaina Molina, Julio Daròs Arnau, José Antonio |
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Talens Perales, David Nicolau Sanus, María Polaina Molina, Julio Daròs Arnau, José Antonio Expression of an extremophilic xylanase in Nicotiana benthamiana and its use for the production of prebiotic xylooligosaccharides |
author_sort |
Talens Perales, David |
title |
Expression of an extremophilic xylanase in Nicotiana benthamiana and its use for the production of prebiotic xylooligosaccharides |
title_short |
Expression of an extremophilic xylanase in Nicotiana benthamiana and its use for the production of prebiotic xylooligosaccharides |
title_full |
Expression of an extremophilic xylanase in Nicotiana benthamiana and its use for the production of prebiotic xylooligosaccharides |
title_fullStr |
Expression of an extremophilic xylanase in Nicotiana benthamiana and its use for the production of prebiotic xylooligosaccharides |
title_full_unstemmed |
Expression of an extremophilic xylanase in Nicotiana benthamiana and its use for the production of prebiotic xylooligosaccharides |
title_sort |
expression of an extremophilic xylanase in nicotiana benthamiana and its use for the production of prebiotic xylooligosaccharides |
publisher |
Springer Nature |
publishDate |
2022-09-21 |
url |
http://hdl.handle.net/10261/280657 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100004837 https://api.elsevier.com/content/abstract/scopus_id/85138290438 |
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