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Molecules 2014, 19(8), 11231-11249; doi:10.3390/molecules190811231
Article

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

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Received: 11 June 2014; in revised form: 23 July 2014 / Accepted: 24 July 2014 / Published: 31 July 2014
(This article belongs to the Special Issue Enzyme Immobilization)
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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.
Keywords: nucleoside 2'-deoxyribosyltransferase; Bacillus psychrosaccharolyticus; enzyme immobilization; nucleoside synthesis; trifluridine nucleoside 2'-deoxyribosyltransferase; Bacillus psychrosaccharolyticus; enzyme immobilization; nucleoside synthesis; trifluridine
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Fresco-Taboada, A.; Serra, I.; Fernández-Lucas, J.; Acebal, C.; Arroyo, M.; Terreni, M.; de la Mata, I. Nucleoside 2'-Deoxyribosyltransferase from Psychrophilic Bacterium Bacillus psychrosaccharolyticus — Preparation of an Immobilized Biocatalyst for the Enzymatic Synthesis of Therapeutic Nucleosides. Molecules 2014, 19, 11231-11249.

AMA Style

Fresco-Taboada A, Serra I, Fernández-Lucas J, Acebal C, Arroyo M, Terreni M, de la Mata I. Nucleoside 2'-Deoxyribosyltransferase from Psychrophilic Bacterium Bacillus psychrosaccharolyticus — Preparation of an Immobilized Biocatalyst for the Enzymatic Synthesis of Therapeutic Nucleosides. Molecules. 2014; 19(8):11231-11249.

Chicago/Turabian Style

Fresco-Taboada, Alba; Serra, Immacolata; Fernández-Lucas, Jesús; Acebal, Carmen; Arroyo, Miguel; Terreni, Marco; de la Mata, Isabel. 2014. "Nucleoside 2'-Deoxyribosyltransferase from Psychrophilic Bacterium Bacillus psychrosaccharolyticus — Preparation of an Immobilized Biocatalyst for the Enzymatic Synthesis of Therapeutic Nucleosides." Molecules 19, no. 8: 11231-11249.



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