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:
- http://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 |
network_acronym_str |
RCUC2 |
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 |
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 |
http://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 |
http://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 |
https://repositorio.cuc.edu.co/bitstream/11323/1512/1/One-Pot%2c%20One-Step%20Production.pdf https://repositorio.cuc.edu.co/bitstream/11323/1512/2/license.txt https://repositorio.cuc.edu.co/bitstream/11323/1512/4/One-Pot%2c%20One-Step%20Production.pdf.jpg https://repositorio.cuc.edu.co/bitstream/11323/1512/5/One-Pot%2c%20One-Step%20Production.pdf.txt |
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MD5 MD5 MD5 MD5 |
repository.name.fl_str_mv |
Repositorio Universidad de La Costa |
repository.mail.fl_str_mv |
bdigital@metabiblioteca.com |
_version_ |
1808400096115556352 |
spelling |
Del Arco, Jon923cae3a1d238f02847617e53bdc6387Martínez Pascual, Sara9384f06e9eadebce91a5001f9b77a49eClemente Suárez, Vicente Javier4f367feceeed688412960a382c957dc4Corral Pazos de Provens, Octavio Jorge31f2317d6c71777eda712d065456b3b0Jordaan, Justin406cda6b514ffdd18ba17701827a23aeHormigo, Daniele41112d7ee54b8a5c0ffbe9440e5ed9fPerona, Almudena3b1f98d32d9e27b16f0155227b45512dFernandez Lucas, Jesus8a93c53d89f531d9974dd13de076402b2018-11-20T20:45:55Z2018-11-20T20:45:55Z2018-04-2720734344http://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.engCatalystsAtribución – 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/acceptedVersionORIGINALOne-Pot, One-Step Production.pdfOne-Pot, One-Step Production.pdfapplication/pdf1716504https://repositorio.cuc.edu.co/bitstream/11323/1512/1/One-Pot%2c%20One-Step%20Production.pdf063543ab5dddd524f3ef15ca8ea4a86dMD51open accessLICENSElicense.txtlicense.txttext/plain; charset=utf-81748https://repositorio.cuc.edu.co/bitstream/11323/1512/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52open accessTHUMBNAILOne-Pot, One-Step Production.pdf.jpgOne-Pot, One-Step Production.pdf.jpgimage/jpeg68771https://repositorio.cuc.edu.co/bitstream/11323/1512/4/One-Pot%2c%20One-Step%20Production.pdf.jpg2dbf1115c9afceb86591e904b72b3eb9MD54open accessTEXTOne-Pot, One-Step Production.pdf.txtOne-Pot, One-Step Production.pdf.txttext/plain54991https://repositorio.cuc.edu.co/bitstream/11323/1512/5/One-Pot%2c%20One-Step%20Production.pdf.txte2df8c42acebc0403579acdae8bd86a1MD55open access11323/1512oai:repositorio.cuc.edu.co:11323/15122023-12-14 14:18:53.819open accessRepositorio Universidad de La Costabdigital@metabiblioteca.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 |