Resumen:
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 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.