| Autor: |
Rivero CW; Laboratory of Sustainable Biotechnology (LIBioS), National University of Quilmes, Bernal B1876BXD, Argentina.; National Scientific and Technical Research Council (CONICET), Ciudad Autónoma de Buenos Aires C1425FQB, Argentina., García NS; Laboratory of Sustainable Biotechnology (LIBioS), National University of Quilmes, Bernal B1876BXD, Argentina., Fernández-Lucas J; Applied Biotechnology Group, Biomedical Science School, Universidad Europea de Madrid, 28670 Villaviciosa de Odón, Spain.; Research Group in Natural and Exact Sciences, GICNEX, Universidad de la Costa, CUC, Barranquilla 080002, Colombia., Betancor L; Biotechnology Laboratory, Faculty of Engineering, Universidad ORT Uruguay, Montevideo 11100, Uruguay., Romanelli GP; Center for Research and Development in Applied Sciences, National University of La Plata, La Plata B1900AJK, Argentina., Trelles JA; Laboratory of Sustainable Biotechnology (LIBioS), National University of Quilmes, Bernal B1876BXD, Argentina.; National Scientific and Technical Research Council (CONICET), Ciudad Autónoma de Buenos Aires C1425FQB, Argentina. |
| Abstrakt: |
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 ( Ld NDT) 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, SiG PEI 25000 - Ld NDT-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, SiG PEI 25000 - Ld NDT-Alg was successfully employed in the green production of cladribine at mg scale. |
