Wanda Rivero, CintiaSoledad García, NataliaFernández Lucas, JesúsBetancor, LorenaRomanelli, Gustavo PabloTrelles, Jorge Abel2023-06-042023-06-042021Wanda Rivero, C., Soledad García, N., Fernández-Lucas, J., Betancor, L., Romanelli, G. P., & Trelles, J. A. (2021). Green Production of Cladribine by Using Immobilized 2′-Deoxyribosyltransferase from Lactobacillus delbrueckii Stabilized through a Double Covalent/Entrapment Technology. Biomolecules, 11(5), 657. https://doi.org/10.3390/biom110506572218-273Xhttp://hdl.handle.net/11268/12104Nowadays, enzyme-mediated processes offer an eco-friendly and efficient alternative to the traditional multistep and environmentally harmful chemical processes. Herein we report the enzymatic synthesis of cladribine by a novel 20 -deoxyribosyltransferase (NDT)-based combined biocatalyst. To this end, Lactobacillus delbrueckii NDT (LdNDT) was successfully immobilized through a two-step immobilization methodology, including a covalent immobilization onto glutaraldehydeactivated biomimetic silica nanoparticles followed by biocatalyst entrapment in calcium alginate. The resulting immobilized derivative, SiGPEI 25000-LdNDT-Alg, displayed 98% retained activity and was shown to be active and stable in a broad range of pH (5–9) and temperature (30–60 ◦C), but also displayed an extremely high reusability (up to 2100 reuses without negligible loss of activity) in the enzymatic production of cladribine. Finally, as a proof of concept, SiGPEI 25000-LdNDT-Alg was successfully employed in the green production of cladribine at mg scale.spaAtribución 4.0 Internacionalhttp://creativecommons.org/licenses/by/4.0/journal article10.3390/biom11050657open accessBiología molecularEnzimaBioquímica