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oai:docta.ucm.es:20.500.14352/343202024-03-04T13:19:24Zcom_20.500.14352_14col_20.500.14352_15

Fresco Taboada, Alba Serra, Immacolata Fernández Lucas, Jesús Acebal Sarabia, Carmen Arroyo, Miguel Terreni, Marco De La Mata Riesco, Mª Isabel 2023-06-19T13:43:16Z 2023-06-19T13:43:16Z 2014-07-31 1420-3049 10.3390/molecules190811231 https://hdl.handle.net/20.500.14352/34320 https://doi.org/10.3390/molecules190811231 https://www.mdpi.com/1420-3049/19/8/11231 Nucleoside 2'-deoxyribosyltransferase (NDT) from the psychrophilic bacterium Bacillus psychrosaccharolyticus CECT 4074 has been cloned and produced for the first time. A preliminary characterization of the recombinant protein indicates that the enzyme is an NDT type II since it catalyzes the transfer of 2'-deoxyribose between purines and pyrimidines. The enzyme (BpNDT) displays a high activity and stability in a broad range of pH and temperature. In addition, different approaches for the immobilization of BpNDT onto several supports have been studied in order to prepare a suitable biocatalyst for the one-step industrial enzymatic synthesis of different therapeutic nucleosides. Best results were obtained by adsorbing the enzyme on PEI-functionalized agarose and subsequent cross-linking with aldehyde-dextran (20 kDa and 70% oxidation degree). The immobilized enzyme could be recycled for at least 30 consecutive cycles in the synthesis of 2'-deoxyadenosine from 2'-deoxyuridine and adenine at 37 °C and pH 8.0, with a 25% loss of activity. High conversion yield of trifluridine (64.4%) was achieved in 2 h when 20 mM of 2'-deoxyuridine and 10 mM 5-trifluorothymine were employed in the transglycosylation reaction catalyzed by immobilized BpNDT at 37 °C and pH 7.5. eng https://creativecommons.org/licenses/by/3.0/es/ open access Atribución 3.0 España Nucleoside 2'-Deoxyribosyltransferase from Psychrophilic Bacterium Bacillus psychrosaccharolyticus — Preparation of an Immobilized Biocatalyst for the Enzymatic Synthesis of Therapeutic Nucleosides journal article