Nucleoside 2'-Deoxyribosyltransferase from Psychrophilic Bacterium Bacillus psychrosaccharolyticus — Preparation of an Immobilized Biocatalyst for the Enzymatic Synthesis of Therapeutic Nucleosides

Impacto

Downloads

Downloads per month over past year

Fresco Taboada, Alba and Serra, Immacolata and Fernández Lucas, Jesús and Acebal Sarabia, Carmen and Arroyo, Miguel and Terreni, Marco and Mata Riesco, Isabel de la (2014) Nucleoside 2'-Deoxyribosyltransferase from Psychrophilic Bacterium Bacillus psychrosaccharolyticus — Preparation of an Immobilized Biocatalyst for the Enzymatic Synthesis of Therapeutic Nucleosides. Molecules, 19 (8). pp. 11231-11249. ISSN 1420-3049

[thumbnail of molecules-19-11231.pdf] PDF
Creative Commons Attribution.

958kB

Official URL: https://doi.org/10.3390/molecules190811231




Abstract

Nucleoside 2'-deoxyribosyltransferase (NDT) from the psychrophilic bacterium Bacillus psychrosaccharolyticus CECT 4074 has been cloned and produced for the first time. A preliminary characterization of the recombinant protein indicates that the enzyme is an NDT type II since it catalyzes the transfer of 2'-deoxyribose between purines and pyrimidines. The enzyme (BpNDT) displays a high activity and stability in a broad range of pH and temperature. In addition, different approaches for the immobilization of BpNDT onto several supports have been studied in order to prepare a suitable biocatalyst for the one-step industrial enzymatic synthesis of different therapeutic nucleosides. Best results were obtained by adsorbing the enzyme on PEI-functionalized agarose and subsequent cross-linking with aldehyde-dextran (20 kDa and 70% oxidation degree). The immobilized enzyme could be recycled for at least 30 consecutive cycles in the synthesis of 2'-deoxyadenosine from 2'-deoxyuridine and adenine at 37 °C and pH 8.0, with a 25% loss of activity. High conversion yield of trifluridine (64.4%) was achieved in 2 h when 20 mM of 2'-deoxyuridine and 10 mM 5-trifluorothymine were employed in the transglycosylation reaction catalyzed by immobilized BpNDT at 37 °C and pH 7.5.


Item Type:Article
Uncontrolled Keywords:nucleoside 2'-deoxyribosyltransferase; Bacillus psychrosaccharolyticus; enzyme immobilization; nucleoside synthesis; trifluridine
Subjects:Medical sciences > Biology > Molecular biology
ID Code:64396
Deposited On:16 Mar 2021 17:48
Last Modified:17 Mar 2021 08:41

Origin of downloads

Repository Staff Only: item control page