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oai:docta.ucm.es:20.500.14352/1099602025-03-18T12:49:21Zcom_20.500.14352_14col_20.500.14352_15

Characterization of an atypical, thermostable, organic solvent- and acid-tolerant 2′-deoxyribosyltransferase from Chroococcidiopsis thermalis Del Arco, Jon Sánchez-Murcia, Pedro Alejandro Mancheño Gómez, José Miguel Gago, Federico Fernández Lucas, Jesús 577.15 579.8 579.22.08 Enzymatic synthesis Nucleoside analogues Nucleoside 2′-deoxyribosyltransferase Extremophiles Homology modeling Bioquímica (Biología) Microbiología (Biología) Biotecnología 2403 Bioquímica 2414 Microbiología 2302.09 Enzimología 2302.23 Ácidos Nucleicos In our search for thermophilic and acid-tolerant nucleoside 2′-deoxyribosyltransferases (NDTs), we found a good candidate in an enzyme encoded by Chroococcidiopsis thermalis PCC 7203 (CtNDT). Biophysical and biochemical characterization revealed CtNDT as a homotetramer endowed with good activity and stability at both high temperatures (50–100 °C) and a wide range of pH values (from 3 to 7). CtNDT recognizes purine bases and their corresponding 2′-deoxynucleosides but is also proficient using cytosine and 2′-deoxycytidine as substrates. These unusual features preclude the strict classification of CtNDT as either a type I or a type II NDT and further suggest that this simple subdivision may need to be updated in the future. Our findings also hint at a possible link between oligomeric state and NDT’s substrate specificity. Interestingly from a practical perspective, CtNDT displays high activity (80–100%) in the presence of several water-miscible co-solvents in a proportion of up to 20% and was successfully employed in the enzymatic production of several therapeutic nucleosides such as didanosine, vidarabine, and cytarabine. Ministerio de Economía y Competitividad (España) Ministerio de Ciencia e Innovación (España) Universidad Europea de Madrid Fundación Santander Depto. de Bioquímica y Biología Molecular Fac. de Ciencias Biológicas Fac. de Ciencias Químicas TRUE pub 2024-11-04T16:41:35Z 2024-11-04T16:41:35Z 2018 journal article VoR https://hdl.handle.net/20.500.14352/109960 0175-7598 10.1007/s00253-018-9134-y 1432-0614 eng info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/SAF2015-64629-C2-2-R/CARACTERIZACION Y BLOQUEO MEDIANTE PEPTIDOS Y MOLECULAS PEQUEÑAS DE PROTEINAS DIANA IMPLICADAS EN LA PROLIFERACION DE MICROORGANISMOS PATOGENOS Y CELUAS CANCEROSAS info:eu-repo/grantAgreement/Universidad Europea de Madrid//2017%2FUEM23/ES info:eu-repo/grantAgreement/Fundación Santander//SAN151610/ES Del Arco, J., Sánchez-Murcia, P. A., Mancheño, J. M., Gago, F., & Fernández-Lucas, J. (2018). Characterization of an atypical, thermostable, organic solvent- and acid-tolerant 2′-deoxyribosyltransferase from Chroococcidiopsis thermalis. Applied Microbiology and Biotechnology, 102(16), 6947-6957. https://doi.org/10.1007/S00253-018-9134-Y restricted access application/pdf Springer Nature