Rivero, Cintia WandaGarcía, Natalia SoledadFernández Lucas, JesúsBetancor, LorenaRomanelli, Gustavo PabloTrelles, Jorge Abel2024-10-162024-10-162021Rivero CW, García NS, Fernández-Lucas J, Betancor L, Romanelli GP, Trelles JA. Green Production of Cladribine by Using Immobilized 2′-Deoxyribosyltransferase from Lactobacillus delbrueckii Stabilized through a Double Covalent/Entrapment Technology. Biomolecules 2021;11:657. https://doi.org/10.3390/biom1105065710.3390/biom11050657https://hdl.handle.net/20.500.14352/109033This research was mainly supported by Agencia Nacional de Promoción Científica y Tecnológica (PICT 2014–3438), Consejo Nacional de Investigaciones Científicas y Técnicas (PIP 2014- 11220130100805CO) and Universidad Nacional de Quilmes (PUNQ 1305/19, PPROF 2017-EXPTE.895/18)Nowadays, 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 2′-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 glutaraldehyde-activated 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.engAttribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/journal article2218-273Xhttps://doi.org/10.3390/biom11050657https://www.mdpi.com/2218-273X/11/5/657open access577.15577.260615.012Biomimetic silicaEnzyme immobilizationGlutaraldehydeEntrapmentCalcium alginateAntineoplastic drugBioquímica (Biología)Biología molecular (Biología)MedicamentosBiotecnología2403 Bioquímica2415 Biología Molecular2302.09 Enzimología3209.08 Preparación de Medicamentos