title: One-step enzymatic synthesis of nucleosides from low water-soluble purine bases in non-conventional media.
abstract: The effect of several water-miscible cosolvents on activity and stability of soluble and immobilized 2'-deoxyribosyltransferase from Lactobacillus reuteri on Sepabeads® has been studied in order to establish optimal conditions for enzymatic synthesis of nucleosides using purine bases with low solubility in aqueous buffer. As a rule of thumb, there was a general reduction of soluble enzyme activity when cosolvent content was gradually increased in reaction medium. In contrast, immobilized enzyme activity was enhanced 1.2-1.4-fold at 20% of methanol, ethanol, 2-propanol, diethylene glycol, and acetone; and at 10% and 30% acetonitrile. Likewise, highest increased activity (1.8-fold) was also obtained in presence of 20% acetonitrile. Immobilized enzyme was successfully used in the synthesis of 2'-deoxyxanthosine and 2'-deoxyguanosine using 2'-deoxyuridine as sugar donor and the corresponding poor water-soluble base in the presence of 30% of methanol, ethanol, 2-propanol, ethylene glycol, acetonitrile, and DMSO, giving high nucleoside yields at 4h.
authors.affiliation: Departamento de Bioquímica y Biología Molecular I, Facultad de Ciencias Biológicas, Universidad Complutense de Madrid, Madrid, Spain.
authors.forename: Jesús
authors.initials: J
authors.lastname: Fernández-Lucas
authors.forename: Alba
authors.initials: A
authors.lastname: Fresco-Taboada
authors.forename: Isabel
authors.initials: I
authors.lastname: de la Mata
authors.forename: Miguel
authors.initials: M
authors.lastname: Arroyo
mesh_terms: D055162:Biocatalysis / Q000187:drug effects; D004789:Enzyme Activation / Q000187:drug effects; D004795:Enzyme Stability / Q000187:drug effects; D004800:Enzymes, Immobilized / Q000378:metabolism; D007778:Lactobacillus / Q000201:enzymology*; D009705:Nucleosides / Q000378:metabolism*; D010430:Pentosyltransferases / Q000378:metabolism*; D011687:Purines / Q000378:metabolism*; D012997:Solvents / Q000494:pharmacology*; D013696:Temperature; D014867:Water / Q000737:chemistry*
chemical_list: D004800:Enzymes, Immobilized; D009705:Nucleosides; D011687:Purines; D012997:Solvents; D014867:Water; D010430:Pentosyltransferases; C021888:nucleoside deoxyribosyltransferase; C030985:purine