Hypoxanthine-guanine phosphoribosyltransferase/adenylate kinase from Zobellia galactanivorans: A bifunctional catalyst for the synthesis of nucleoside-5′-mono-, di- and triphosphates
In our search for novel biocatalysts for the synthesis of nucleic acid derivatives, we found a good candidate in a putative dual-domain hypoxanthine-guanine phosphoribosyltransferase (HGPRT)/adenylate kinase (AMPK) from Zobellia galactanivorans (ZgHGPRT/AMPK). In this respect, we report for the firs...
| Autores: | , , , , , , , |
|---|---|
| Tipo de recurso: | artículo |
| Fecha de publicación: | 2020 |
| País: | España |
| Institución: | Universidad Autónoma de Madrid |
| Repositorio: | Biblos-e Archivo. Repositorio Institucional de la UAM |
| Idioma: | inglés |
| OAI Identifier: | oai:repositorio.uam.es:10486/708834 |
| Acceso en línea: | http://hdl.handle.net/10486/708834 https://dx.doi.org/10.3389/fbioe.2020.00677 |
| Access Level: | acceso abierto |
| Palabra clave: | Biocatalysts Biomolecules Nucleic Acids Positive Ions Recombinant Proteins Biología y Biomedicina / Biología |
| id |
ES_ab1fa7e2a11fb8fe0b58d357c34522cb |
|---|---|
| oai_identifier_str |
oai:repositorio.uam.es:10486/708834 |
| network_acronym_str |
ES |
| network_name_str |
España |
| repository_id_str |
|
| spelling |
Hypoxanthine-guanine phosphoribosyltransferase/adenylate kinase from Zobellia galactanivorans: A bifunctional catalyst for the synthesis of nucleoside-5′-mono-, di- and triphosphatesAcosta, JavierArco, Jon delPozo, María Luisa delHerrera-Tapias, BeliñaClemente-Suárez, Vicente JavierBerenguer Carlos, JoséHidalgo Huertas, AurelioFernández-Lucas, JesúsBiocatalystsBiomoleculesNucleic AcidsPositive IonsRecombinant ProteinsBiología y Biomedicina / BiologíaIn our search for novel biocatalysts for the synthesis of nucleic acid derivatives, we found a good candidate in a putative dual-domain hypoxanthine-guanine phosphoribosyltransferase (HGPRT)/adenylate kinase (AMPK) from Zobellia galactanivorans (ZgHGPRT/AMPK). In this respect, we report for the first time the recombinant expression, production, and characterization of a bifunctional HGPRT/AMPK. Biochemical characterization of the recombinant protein indicates that the enzyme is a homodimer, with high activity in the pH range 6-7 and in a temperature interval from 30 to 80°C. Thermal denaturation experiments revealed that ZgHGPRT/AMPK exhibits an apparent unfolding temperature (Tm) of 45°C and a retained activity of around 80% when incubated at 40°C for 240 min. This bifunctional enzyme shows a dependence on divalent cations, with a remarkable preference for Mg2+ and Co2+ as cofactors. More interestingly, substrate specificity studies revealed ZgHGPRT/AMPK as a bifunctional enzyme, which acts as phosphoribosyltransferase or adenylate kinase depending upon the nature of the substrate. Finally, to assess the potential of ZgHGPRT/AMPK as biocatalyst for the synthesis of nucleoside-5′-mono, di- and triphosphates, the kinetic analysis of both activities (phosphoribosyltransferase and adenylate kinase) and the effect of water-miscible solvents on enzyme activity were studiedBIO2016-77031-RFrontiers MediaDepartamento de Biología MolecularFacultad de Ciencias20202020-06-24research articlehttp://purl.org/coar/resource_type/c_2df8fbb1VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10486/708834https://dx.doi.org/10.3389/fbioe.2020.00677reponame:Biblos-e Archivo. Repositorio Institucional de la UAMinstname:Universidad Autónoma de MadridInglésengopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:repositorio.uam.es:10486/7088342026-06-23T12:46:27Z |
| dc.title.none.fl_str_mv |
Hypoxanthine-guanine phosphoribosyltransferase/adenylate kinase from Zobellia galactanivorans: A bifunctional catalyst for the synthesis of nucleoside-5′-mono-, di- and triphosphates |
| title |
Hypoxanthine-guanine phosphoribosyltransferase/adenylate kinase from Zobellia galactanivorans: A bifunctional catalyst for the synthesis of nucleoside-5′-mono-, di- and triphosphates |
| spellingShingle |
Hypoxanthine-guanine phosphoribosyltransferase/adenylate kinase from Zobellia galactanivorans: A bifunctional catalyst for the synthesis of nucleoside-5′-mono-, di- and triphosphates Acosta, Javier Biocatalysts Biomolecules Nucleic Acids Positive Ions Recombinant Proteins Biología y Biomedicina / Biología |
| title_short |
Hypoxanthine-guanine phosphoribosyltransferase/adenylate kinase from Zobellia galactanivorans: A bifunctional catalyst for the synthesis of nucleoside-5′-mono-, di- and triphosphates |
| title_full |
Hypoxanthine-guanine phosphoribosyltransferase/adenylate kinase from Zobellia galactanivorans: A bifunctional catalyst for the synthesis of nucleoside-5′-mono-, di- and triphosphates |
| title_fullStr |
Hypoxanthine-guanine phosphoribosyltransferase/adenylate kinase from Zobellia galactanivorans: A bifunctional catalyst for the synthesis of nucleoside-5′-mono-, di- and triphosphates |
| title_full_unstemmed |
Hypoxanthine-guanine phosphoribosyltransferase/adenylate kinase from Zobellia galactanivorans: A bifunctional catalyst for the synthesis of nucleoside-5′-mono-, di- and triphosphates |
| title_sort |
Hypoxanthine-guanine phosphoribosyltransferase/adenylate kinase from Zobellia galactanivorans: A bifunctional catalyst for the synthesis of nucleoside-5′-mono-, di- and triphosphates |
| dc.creator.none.fl_str_mv |
Acosta, Javier Arco, Jon del Pozo, María Luisa del Herrera-Tapias, Beliña Clemente-Suárez, Vicente Javier Berenguer Carlos, José Hidalgo Huertas, Aurelio Fernández-Lucas, Jesús |
| author |
Acosta, Javier |
| author_facet |
Acosta, Javier Arco, Jon del Pozo, María Luisa del Herrera-Tapias, Beliña Clemente-Suárez, Vicente Javier Berenguer Carlos, José Hidalgo Huertas, Aurelio Fernández-Lucas, Jesús |
| author_role |
author |
| author2 |
Arco, Jon del Pozo, María Luisa del Herrera-Tapias, Beliña Clemente-Suárez, Vicente Javier Berenguer Carlos, José Hidalgo Huertas, Aurelio Fernández-Lucas, Jesús |
| author2_role |
author author author author author author author |
| dc.contributor.none.fl_str_mv |
Departamento de Biología Molecular Facultad de Ciencias |
| dc.subject.none.fl_str_mv |
Biocatalysts Biomolecules Nucleic Acids Positive Ions Recombinant Proteins Biología y Biomedicina / Biología |
| topic |
Biocatalysts Biomolecules Nucleic Acids Positive Ions Recombinant Proteins Biología y Biomedicina / Biología |
| description |
In our search for novel biocatalysts for the synthesis of nucleic acid derivatives, we found a good candidate in a putative dual-domain hypoxanthine-guanine phosphoribosyltransferase (HGPRT)/adenylate kinase (AMPK) from Zobellia galactanivorans (ZgHGPRT/AMPK). In this respect, we report for the first time the recombinant expression, production, and characterization of a bifunctional HGPRT/AMPK. Biochemical characterization of the recombinant protein indicates that the enzyme is a homodimer, with high activity in the pH range 6-7 and in a temperature interval from 30 to 80°C. Thermal denaturation experiments revealed that ZgHGPRT/AMPK exhibits an apparent unfolding temperature (Tm) of 45°C and a retained activity of around 80% when incubated at 40°C for 240 min. This bifunctional enzyme shows a dependence on divalent cations, with a remarkable preference for Mg2+ and Co2+ as cofactors. More interestingly, substrate specificity studies revealed ZgHGPRT/AMPK as a bifunctional enzyme, which acts as phosphoribosyltransferase or adenylate kinase depending upon the nature of the substrate. Finally, to assess the potential of ZgHGPRT/AMPK as biocatalyst for the synthesis of nucleoside-5′-mono, di- and triphosphates, the kinetic analysis of both activities (phosphoribosyltransferase and adenylate kinase) and the effect of water-miscible solvents on enzyme activity were studied |
| publishDate |
2020 |
| dc.date.none.fl_str_mv |
2020 2020-06-24 |
| dc.type.none.fl_str_mv |
research article http://purl.org/coar/resource_type/c_2df8fbb1 VoR http://purl.org/coar/version/c_970fb48d4fbd8a85 |
| dc.type.openaire.fl_str_mv |
info:eu-repo/semantics/article |
| format |
article |
| dc.identifier.none.fl_str_mv |
http://hdl.handle.net/10486/708834 https://dx.doi.org/10.3389/fbioe.2020.00677 |
| url |
http://hdl.handle.net/10486/708834 https://dx.doi.org/10.3389/fbioe.2020.00677 |
| 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.format.none.fl_str_mv |
application/pdf |
| dc.publisher.none.fl_str_mv |
Frontiers Media |
| publisher.none.fl_str_mv |
Frontiers Media |
| dc.source.none.fl_str_mv |
reponame:Biblos-e Archivo. Repositorio Institucional de la UAM instname:Universidad Autónoma de Madrid |
| instname_str |
Universidad Autónoma de Madrid |
| reponame_str |
Biblos-e Archivo. Repositorio Institucional de la UAM |
| collection |
Biblos-e Archivo. Repositorio Institucional de la UAM |
| repository.name.fl_str_mv |
|
| repository.mail.fl_str_mv |
|
| _version_ |
1869416241565794304 |
| score |
15,301603 |