2′-Deoxyribosyltransferase from Leishmania mexicana, an efficient biocatalyst for one-pot, one-step synthesis of nucleosides from poorly soluble purine bases

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dc.creatorCrespo, N.
dc.creatorSánchez Murcia, Pedro Alejandro
dc.creatorGago, Federico
dc.creatorCejudo Sanches, J.
dc.creatorGalmes, M.A.
dc.creatorFernandez Lucas, Jesus
dc.creatorMancheño, José Miguel
dc.date2018-11-16T21:06:55Z
dc.date2018-11-16T21:06:55Z
dc.date2017
dc.date.accessioned2023-11-22T13:56:29Z
dc.date.available2023-11-22T13:56:29Z
dc.descriptionProcesses catalyzed by enzymes offer numerous advantages over chemical methods although in many occasions the stability of the biocatalysts becomes a serious concern. Traditionally, synthesis of nucleosides using poorly water-soluble purine bases, such as guanine, xanthine, or hypoxanthine, requires alkaline pH and/or high temperatures in order to solubilize the substrate. In this work, we demonstrate that the 2'-deoxyribosyltransferase from Leishmania mexicana (LmPDT) exhibits an unusually high activity and stability under alkaline conditions (pH 8-10) across a broad range of temperatures (30-70 °C) and ionic strengths (0-500 mM NaCl). Conversely, analysis of the crystal structure of LmPDT together with comparisons with hexameric, bacterial homologues revealed the importance of the relationships between the oligomeric state and the active site architecture within this family of enzymes. Moreover, molecular dynamics and docking approaches provided structural insights into the substrate-binding mode. Biochemical characterization of LmPDT identifies the enzyme as a type I NDT (PDT), exhibiting excellent activity, with specific activity values 100- and 4000-fold higher than the ones reported for other PDTs. Interestingly, LmPDT remained stable during 36 h at different pH values at 40 °C. In order to explore the potential of LmPDT as an industrial biocatalyst, enzymatic production of several natural and non-natural therapeutic nucleosides, such as vidarabine (ara A), didanosine (ddI), ddG, or 2'-fluoro-2'-deoxyguanosine, was carried out using poorly water-soluble purines. Noteworthy, this is the first time that the enzymatic synthesis of 2'-fluoro-2'-deoxyguanosine, ara G, and ara H by a 2'-deoxyribosyltransferase is reported.
dc.formatapplication/pdf
dc.identifierhttp://hdl.handle.net/11323/1163
dc.identifierDOI: 10.1007/s00253-017-8450-y
dc.identifierCorporación Universidad de la Costa
dc.identifierREDICUC - Repositorio CUC
dc.identifierhttps://repositorio.cuc.edu.co/
dc.identifier.urihttp://repository-salesiana.heoq.net/handle/123456789/316994
dc.languageeng
dc.publisherApplied Microbiology and Biotechnology
dc.rightsAtribución – No comercial – Compartir igual
dc.rightsinfo:eu-repo/semantics/openAccess
dc.rightshttp://purl.org/coar/access_right/c_abf2
dc.subject2′-deoxyribosyltransferase
dc.subjectEnzymatic Synthesis
dc.subjectIndustrial Biocatalyst
dc.subjectMolecular Docking
dc.subjectProtein Crystallography
dc.subjectPurine Nucleoside Analogues
dc.title2′-Deoxyribosyltransferase from Leishmania mexicana, an efficient biocatalyst for one-pot, one-step synthesis of nucleosides from poorly soluble purine bases
dc.typeArtículo de revista
dc.typehttp://purl.org/coar/resource_type/c_6501
dc.typeText
dc.typeinfo:eu-repo/semantics/article
dc.typeinfo:eu-repo/semantics/publishedVersion
dc.typehttp://purl.org/redcol/resource_type/ART
dc.typeinfo:eu-repo/semantics/acceptedVersion
dc.typehttp://purl.org/coar/version/c_ab4af688f83e57aa
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