Fresco Taboada, AlbaSerra, ImmacolataFernández Lucas, JesúsAcebal Sarabia, CarmenArroyo, MiguelTerreni, MarcoDe La Mata Riesco, Mª Isabel2023-06-192023-06-192014-07-311420-304910.3390/molecules190811231https://hdl.handle.net/20.500.14352/34320Nucleoside 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.engAtribución 3.0 Españajournal articlehttps://doi.org/10.3390/molecules190811231https://www.mdpi.com/1420-3049/19/8/11231open accessnucleoside 2'-deoxyribosyltransferaseBacillus psychrosaccharolyticusenzyme immobilizationnucleoside synthesistrifluridineBiología molecular (Biología)2415 Biología Molecular