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:
Del Arco, Jon
Martínez Pascual, Sara
Clemente Suárez, Vicente Javier
Corral Pazos de Provens, Octavio Jorge
Jordaan, Justin
Hormigo, Daniel
Perona, Almudena
Fernandez Lucas, Jesus
Tipo de recurso:
Article of journal
Fecha de publicación:
2018
Institución:
Corporación Universidad de la Costa
Repositorio:
REDICUC - Repositorio CUC
Idioma:
eng
OAI Identifier:
oai:repositorio.cuc.edu.co:11323/1512
Acceso en línea:
https://hdl.handle.net/11323/1512
https://repositorio.cuc.edu.co/
Palabra clave:
Dietary nucleotides
Enzyme immobilization
Green process
Phosphoribosyltransferases
Rights
openAccess
License
Atribución – No comercial – Compartir igual
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network_name_str REDICUC - Repositorio CUC
repository_id_str
dc.title.eng.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
Dietary nucleotides
Enzyme immobilization
Green process
Phosphoribosyltransferases
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.fl_str_mv Del Arco, Jon
Martínez Pascual, Sara
Clemente Suárez, Vicente Javier
Corral Pazos de Provens, Octavio Jorge
Jordaan, Justin
Hormigo, Daniel
Perona, Almudena
Fernandez Lucas, Jesus
dc.contributor.author.spa.fl_str_mv Del Arco, Jon
Martínez Pascual, Sara
Clemente Suárez, Vicente Javier
Corral Pazos de Provens, Octavio Jorge
Jordaan, Justin
Hormigo, Daniel
Perona, Almudena
Fernandez Lucas, Jesus
dc.subject.eng.fl_str_mv Dietary nucleotides
Enzyme immobilization
Green process
Phosphoribosyltransferases
topic Dietary nucleotides
Enzyme immobilization
Green process
Phosphoribosyltransferases
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-5′-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.accessioned.none.fl_str_mv 2018-11-20T20:45:55Z
dc.date.available.none.fl_str_mv 2018-11-20T20:45:55Z
dc.date.issued.none.fl_str_mv 2018-04-27
dc.type.spa.fl_str_mv Artículo de revista
dc.type.coar.fl_str_mv http://purl.org/coar/resource_type/c_2df8fbb1
dc.type.coar.spa.fl_str_mv http://purl.org/coar/resource_type/c_6501
dc.type.content.spa.fl_str_mv Text
dc.type.driver.spa.fl_str_mv info:eu-repo/semantics/article
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dc.identifier.issn.spa.fl_str_mv 20734344
dc.identifier.uri.spa.fl_str_mv https://hdl.handle.net/11323/1512
dc.identifier.instname.spa.fl_str_mv Corporación Universidad de la Costa
dc.identifier.reponame.spa.fl_str_mv REDICUC - Repositorio CUC
dc.identifier.repourl.spa.fl_str_mv https://repositorio.cuc.edu.co/
identifier_str_mv 20734344
Corporación Universidad de la Costa
REDICUC - Repositorio CUC
url https://hdl.handle.net/11323/1512
https://repositorio.cuc.edu.co/
dc.language.iso.none.fl_str_mv eng
language eng
dc.rights.spa.fl_str_mv Atribución – No comercial – Compartir igual
dc.rights.accessrights.spa.fl_str_mv info:eu-repo/semantics/openAccess
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rights_invalid_str_mv Atribución – No comercial – Compartir igual
http://purl.org/coar/access_right/c_abf2
eu_rights_str_mv openAccess
dc.publisher.spa.fl_str_mv Catalysts
institution Corporación Universidad de la Costa
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spelling Del Arco, JonMartínez Pascual, SaraClemente Suárez, Vicente JavierCorral Pazos de Provens, Octavio JorgeJordaan, JustinHormigo, DanielPerona, AlmudenaFernandez Lucas, Jesus2018-11-20T20:45:55Z2018-11-20T20:45:55Z2018-04-2720734344https://hdl.handle.net/11323/1512Corporación Universidad de la CostaREDICUC - Repositorio CUChttps://repositorio.cuc.edu.co/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-5′-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). 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