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Cloning, expression and biochemical characterization of xanthine and adenine phosphoribosyltransferases from thermus thermophilus HB8

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dc.contributor.author Arco Arrieta, Jon del
dc.contributor.author Martínez, María
dc.contributor.author Donday, Manuel
dc.contributor.author Clemente Suárez, Vicente Javier
dc.contributor.author Fernández Lucas, Jesús
dc.date.accessioned 2017-06-27T09:19:09Z
dc.date.available 2017-06-27T09:19:09Z
dc.date.issued 2018
dc.identifier.citation Del Arco, J., Martínez, M., Donday, M., Clemente-Suárez, V. J., & Fernández-Lucas, J. (2018). Cloning, expression and biochemical characterization of xanthine and adenine phosphoribosyltransferases from thermus thermophilus HB8. Biocatalysis and Biotransformation, 36(3), 216-223. doi: 10.1080/10242422.2017.1313837 spa
dc.identifier.issn 10242422
dc.identifier.uri http://hdl.handle.net/11268/6505
dc.description.abstract Purine phosphoribosyltransferases, purine PRTs, are essential enzymes in the purine salvage pathway of living organisms. They are involved in the formation of C-N glycosidic bonds in purine nucleosides-50-monophosphate (NMPs) through the transfer of the 5-phosphoribosyl group from 5-phospho-a-D-ribosyl-1-pyrophosphate (PRPP) to purine nucleobases in the presence of Mg2þ. Herein, we report a simple and thermostable process for the one-pot, one-step synthesis of some purine NMPs using xanthine phosphoribosyltransferase, XPRT or adenine phosphoribosyltransferase, APRT2, from Thermus thermophilus HB8. In this sense, the cloning, expression and purification of TtXPRT and TtAPRT2 is described for the first time. Both genes, xprt and aprt2 were expressed as his-tagged enzymes in E. coli BL21(DE3) and purified by a heat-shock treatment, followed by Ni-affinity chromatography and a final, polishing gel-filtration chromatography. Biochemical characterization revealed TtXPRT as a tetramer and TtAPRT2 as a dimer. In addition, both enzymes displayed a strong temperature dependence (relative activity >75% in a temperature range from 70 to 90 C), but they also showed very different behaviour under the influence of pH. While TtXPRT is active in a pH range from 5 to 7, TtAPRT2 has a high dependence of alkaline conditions, showing highest activity values in a pH range from 8 to 10. Finally, substrate specificity studies were performed in order to explore their potential as industrial biocatalyst for NMPs synthesis. spa
dc.description.sponsorship European University of Madrid and Santander Foundation spa
dc.language.iso eng spa
dc.title Cloning, expression and biochemical characterization of xanthine and adenine phosphoribosyltransferases from thermus thermophilus HB8 spa
dc.type article spa
dc.description.impact 1.627 JCR (2018) Q3, 121/162 Biotechnology & Applied Microbiology; Q4, 252/299 Biochemistry & Molecular Biology spa
dc.description.impact 0.299 SJR (2018) Q3, 184/342 Biotechnology; Q4, 347/462 Biochemistry, 44/60 Catalysis spa
dc.description.impact No data IDR 2018 spa
dc.identifier.doi 10.1080/10242422.2017.1313837
dc.rights.accessRights closedAccess spa
dc.subject.uem Enzimas - Biotecnología spa
dc.subject.unesco Enzima spa
dc.subject.unesco Biotecnología spa
dc.description.filiation UEM spa
dc.peerreviewed Si spa


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