Development of a high efficient biocatalyst by oriented covalent immobilization of a novel recombinant 2′-N-deoxyribosyltransferase from Lactobacillus animalis
| Autor: | Mariana B. Méndez, Jorge A. Trelles, Jose M. Guisan, Cintia W. Rivero, Fernando López-Gallego |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
Models
Molecular 0301 basic medicine STABILIZATION Immobilized enzyme Protein Conformation GLYOXYL AGAROSE FLOXURIDINE MOLECULAR CLONING Bioengineering INGENIERÍAS Y TECNOLOGÍAS Molecular cloning 01 natural sciences Applied Microbiology and Biotechnology Biotecnología Industrial 03 medical and health sciences chemistry.chemical_compound Bacterial Proteins Transferases Enzyme Stability Cloning Molecular Bioprocess chemistry.chemical_classification BIOCATALYSIS 010405 organic chemistry Chemistry General Medicine Enzymes Immobilized Bioprocesamiento Tecnológico Biocatálisis Fermentación Combinatorial chemistry Environmentally friendly Recombinant Proteins 0104 chemical sciences Lactobacillus 030104 developmental biology Enzyme Biocatalysis Covalent bond Derivative (chemistry) Biotechnology |
| Popis: | The 2′-N-deoxyribosyltransferases [NDT; EC 2.4.2.6] are a group of enzymes widely used as biocatalysts for nucleoside biosynthesis. In this work, the molecular cloning, expression and purification of a novel NDT from Lactobacillus animalis (LaNDT) have been reported. On the other hand, biocatalyst stability has been significantly enhanced by multipoint covalent immobilization using a hetero-functional support activated with nickel-chelates and glyoxyl groups. The immobilized enzyme could be reused for more than 300 h and stored during almost 3 months without activity loss. Besides, the obtained derivative (Ni2+-Gx-LaNDT) was able to biosynthesize 88 mg floxuridine/g biocatalyst after 1 h of reaction. In this work, a green bioprocess by employing an environmentally friendly methodology was developed, which allowed the obtaining of a compound with proven anti-tumor activity. Therefore, the obtained enzymatic biocatalyst meets the requirements of high activity, stability, and short reaction times needed for low-cost production in a future preparative application. Fil: Mendez, Mariana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Investigación en Biotecnología Sustentable; Argentina Fil: Rivero, Cintia Wanda. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Investigación en Biotecnología Sustentable; Argentina Fil: López Gallego, Fernando. Basque Foundation for Science; España. Heterogeneous Biocatalysis Group; España Fil: Guisán, José M.. Consejo Superior de Investigaciones Científicas; España Fil: Trelles, Jorge Abel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Investigación en Biotecnología Sustentable; Argentina |
| Databáze: | OpenAIRE |
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