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...
- 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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oai:repositorio.cuc.edu.co:11323/1512 |
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REDICUC - Repositorio CUC |
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|
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 |
dc.type.redcol.spa.fl_str_mv |
http://purl.org/redcol/resource_type/ART |
dc.type.version.spa.fl_str_mv |
info:eu-repo/semantics/acceptedVersion |
format |
http://purl.org/coar/resource_type/c_6501 |
status_str |
acceptedVersion |
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 |
dc.rights.coar.spa.fl_str_mv |
http://purl.org/coar/access_right/c_abf2 |
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 |
bitstream.url.fl_str_mv |
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1811760768849281024 |
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). Finally, as proof of concept, the enzymatic production of dietary nucleotides from high concentrations of low soluble bases was achieved.Del Arco, Jon-c5ed68af-857c-4b28-99a7-33a4254ed926-0Martínez Pascual, Sara-30107093-0d62-402f-be21-5cdce39381a4-0Clemente Suárez, Vicente Javier-0000-0002-2397-2801-600Corral Pazos de Provens, Octavio Jorge-8fc346ae-5cc4-4214-acf1-96eeba116366-0Jordaan, Justin-57160430-2621-4b93-b018-4d7d12540701-0Hormigo, Daniel-915adee9-292d-4836-8d92-b79ac158d173-0Perona, Almudena-d6388df9-bb67-4ff9-be56-ab05771f1650-0Fernandez Lucas, Jesus-3f36c351-7522-42ea-8605-cd7e804a6387-0engCatalystsAtribución – No comercial – Compartir igualinfo:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2Dietary nucleotidesEnzyme immobilizationGreen processPhosphoribosyltransferasesOne-pot, one-step production of dietary nucleotides by magnetic biocatalystsArtículo de revistahttp://purl.org/coar/resource_type/c_6501http://purl.org/coar/resource_type/c_2df8fbb1Textinfo:eu-repo/semantics/articlehttp://purl.org/redcol/resource_type/ARTinfo:eu-repo/semantics/acceptedVersionPublicationORIGINALOne-Pot, One-Step Production.pdfOne-Pot, One-Step Production.pdfapplication/pdf1716504https://repositorio.cuc.edu.co/bitstreams/fdc09488-2514-45d6-8e0c-95defe7331fb/download063543ab5dddd524f3ef15ca8ea4a86dMD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748https://repositorio.cuc.edu.co/bitstreams/c483cff9-815f-4fd1-b335-87891ea97435/download8a4605be74aa9ea9d79846c1fba20a33MD52THUMBNAILOne-Pot, One-Step Production.pdf.jpgOne-Pot, One-Step Production.pdf.jpgimage/jpeg68771https://repositorio.cuc.edu.co/bitstreams/1400c3a9-4727-431f-94cc-683819c5b703/download2dbf1115c9afceb86591e904b72b3eb9MD54TEXTOne-Pot, One-Step Production.pdf.txtOne-Pot, One-Step Production.pdf.txttext/plain54991https://repositorio.cuc.edu.co/bitstreams/6dc45355-30e3-4d19-8bdc-020cbe17a06c/downloade2df8c42acebc0403579acdae8bd86a1MD5511323/1512oai:repositorio.cuc.edu.co:11323/15122024-09-17 11:03:42.478open.accesshttps://repositorio.cuc.edu.coRepositorio de la Universidad de la Costa CUCrepdigital@cuc.edu.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 |