One-Pot, One-Step Production of Dietary Nucleotides by Magnetic Biocatalysts

The enzymatic synthesis of nucleotides offers several advantages over traditional multistep chemical methods, such as stereoselectivity, regioselectivity, enantioselectivity, simple downstream processing, and the use of mild reaction conditions. However, in order to scale up these bioprocesses, seve...

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Autores: Arco Arrieta, Jon del, Martínez Pascual, Sara, Clemente Suárez, Vicente Javier, Corral Pazos de Provens, Octavio Jorge, Jordaan, Justin, Hormigo Cisneros, Daniel, Perona Requena, Almudena, Fernández Lucas, Jesús
Tipo de recurso: artículo
Fecha de publicación:2018
País:España
Institución:Universidad Europea (UEM)
Repositorio:ABACUS. Repositorio de Producción Científica
Idioma:inglés
OAI Identifier:oai:abacus.universidadeuropea.com:11268/7614
Acceso en línea:http://hdl.handle.net/11268/7614
Access Level:acceso abierto
Palabra clave:Biotecnología alimentaria
Nucleótidos
Biotecnología
Industria alimentaria
Enzima
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spelling One-Pot, One-Step Production of Dietary Nucleotides by Magnetic BiocatalystsArco Arrieta, Jon delMartínez Pascual, SaraClemente Suárez, Vicente JavierCorral Pazos de Provens, Octavio JorgeJordaan, JustinHormigo Cisneros, DanielPerona Requena, AlmudenaFernández Lucas, JesúsBiotecnología alimentariaNucleótidosBiotecnologíaIndustria alimentariaEnzimaThe enzymatic synthesis of nucleotides offers several advantages over traditional multistep chemical methods, such as stereoselectivity, regioselectivity, enantioselectivity, simple downstream processing, and the use of mild reaction conditions. However, in order to scale up these bioprocesses, several drawbacks, such as the low enzyme stability and recycling, must be considered. Enzyme immobilization may overcome these cost-related problems by enhancing protein stability and facilitating the separation of products. In this regard, tetrameric hypoxanthine–guanine–xanthine phosphoribosyltransferase (HGXPRT) from Thermus thermophilus HB8 was covalently immobilized onto glutaraldehyde-activated MagReSyn®Amine magnetic iron oxide porous microparticles (MTtHGXPRT). In this context, two different strategies were followed: (a) an enzyme immobilization through its N-terminus residues at pH 8.5 (derivatives MTtHGXPRT1-3); and (b) a multipoint covalent immobilization through the surface lysine residues at pH 10 (derivatives MTtHGXPRT4-5). The immobilized derivatives of MTtHGXPRT3 (activity 1581 international units per gram of support, IU/g; retained activity 29%) and MTtHGXPRT5 (activity 1108 IU/g; retained activity 23%) displayed the best wet biocatalyst activity, and retained activity values in the enzymatic synthesis of inosine-50 -monophosphate (IMP). In addition, the dependence of the activities and stabilities of both derivatives on pH and temperature was tested, as well as their reusability potential. Taking these results into account, MTtHGXPRT3 was chosen as the best biocatalyst (negligible loss of activity at 60 ◦C during 24 h; reusable up to seven cycles). Finally, as proof of concept, the enzymatic production of dietary nucleotides from high concentrations of low soluble bases was achieved.20182018-11-2420182018-01-0120182018-01-01journal articlehttp://purl.org/coar/resource_type/c_6501info:eu-repo/semantics/articlehttp://hdl.handle.net/11268/7614reponame:ABACUS. Repositorio de Producción Científicainstname:Universidad Europea (UEM)Inglésengopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:abacus.universidadeuropea.com:11268/76142026-06-11T12:41:27Z
dc.title.none.fl_str_mv One-Pot, One-Step Production of Dietary Nucleotides by Magnetic Biocatalysts
title One-Pot, One-Step Production of Dietary Nucleotides by Magnetic Biocatalysts
spellingShingle One-Pot, One-Step Production of Dietary Nucleotides by Magnetic Biocatalysts
Arco Arrieta, Jon del
Biotecnología alimentaria
Nucleótidos
Biotecnología
Industria alimentaria
Enzima
title_short One-Pot, One-Step Production of Dietary Nucleotides by Magnetic Biocatalysts
title_full One-Pot, One-Step Production of Dietary Nucleotides by Magnetic Biocatalysts
title_fullStr One-Pot, One-Step Production of Dietary Nucleotides by Magnetic Biocatalysts
title_full_unstemmed One-Pot, One-Step Production of Dietary Nucleotides by Magnetic Biocatalysts
title_sort One-Pot, One-Step Production of Dietary Nucleotides by Magnetic Biocatalysts
dc.creator.none.fl_str_mv Arco Arrieta, Jon del
Martínez Pascual, Sara
Clemente Suárez, Vicente Javier
Corral Pazos de Provens, Octavio Jorge
Jordaan, Justin
Hormigo Cisneros, Daniel
Perona Requena, Almudena
Fernández Lucas, Jesús
author Arco Arrieta, Jon del
author_facet Arco Arrieta, Jon del
Martínez Pascual, Sara
Clemente Suárez, Vicente Javier
Corral Pazos de Provens, Octavio Jorge
Jordaan, Justin
Hormigo Cisneros, Daniel
Perona Requena, Almudena
Fernández Lucas, Jesús
author_role author
author2 Martínez Pascual, Sara
Clemente Suárez, Vicente Javier
Corral Pazos de Provens, Octavio Jorge
Jordaan, Justin
Hormigo Cisneros, Daniel
Perona Requena, Almudena
Fernández Lucas, Jesús
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv
dc.subject.none.fl_str_mv Biotecnología alimentaria
Nucleótidos
Biotecnología
Industria alimentaria
Enzima
topic Biotecnología alimentaria
Nucleótidos
Biotecnología
Industria alimentaria
Enzima
description The enzymatic synthesis of nucleotides offers several advantages over traditional multistep chemical methods, such as stereoselectivity, regioselectivity, enantioselectivity, simple downstream processing, and the use of mild reaction conditions. However, in order to scale up these bioprocesses, several drawbacks, such as the low enzyme stability and recycling, must be considered. Enzyme immobilization may overcome these cost-related problems by enhancing protein stability and facilitating the separation of products. In this regard, tetrameric hypoxanthine–guanine–xanthine phosphoribosyltransferase (HGXPRT) from Thermus thermophilus HB8 was covalently immobilized onto glutaraldehyde-activated MagReSyn®Amine magnetic iron oxide porous microparticles (MTtHGXPRT). In this context, two different strategies were followed: (a) an enzyme immobilization through its N-terminus residues at pH 8.5 (derivatives MTtHGXPRT1-3); and (b) a multipoint covalent immobilization through the surface lysine residues at pH 10 (derivatives MTtHGXPRT4-5). The immobilized derivatives of MTtHGXPRT3 (activity 1581 international units per gram of support, IU/g; retained activity 29%) and MTtHGXPRT5 (activity 1108 IU/g; retained activity 23%) displayed the best wet biocatalyst activity, and retained activity values in the enzymatic synthesis of inosine-50 -monophosphate (IMP). In addition, the dependence of the activities and stabilities of both derivatives on pH and temperature was tested, as well as their reusability potential. Taking these results into account, MTtHGXPRT3 was chosen as the best biocatalyst (negligible loss of activity at 60 ◦C during 24 h; reusable up to seven cycles). Finally, as proof of concept, the enzymatic production of dietary nucleotides from high concentrations of low soluble bases was achieved.
publishDate 2018
dc.date.none.fl_str_mv 2018
2018-11-24
2018
2018-01-01
2018
2018-01-01
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/11268/7614
url http://hdl.handle.net/11268/7614
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
eu_rights_str_mv openAccess
dc.source.none.fl_str_mv reponame:ABACUS. Repositorio de Producción Científica
instname:Universidad Europea (UEM)
instname_str Universidad Europea (UEM)
reponame_str ABACUS. Repositorio de Producción Científica
collection ABACUS. Repositorio de Producción Científica
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.301603

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