| Abstract [eng] |
2-Oxoglutarate and Fe(II) dependent oxygenases catalyze a wide array of reactions that play a role in the biosynthesis of new compounds, catabolism, and regulation of cellular processes. This thesis delves into a metagenomic-derived 2OG/Fe(II) oxygenase with a novel activity of 2′ O methylated nucleoside demethylation at the 2′ O position. During this study, pH and thermal optimum, thermal stability, substrate specificity, and kinetic parameters of FJS demethylase were measured. Also, interaction between FJS demethylase and nucleic acids is investigated. Small nucleic acid fragments, bound to FJS, are extracted, moreover, EMSA results showed FJS-tRNA complexes. In addition to this, oxygenases with sequences homologous to that of FJS demethylase were studied. It is shown, that all three selected homologous 2OG/Fe(II) oxygenases are active in vivo and two of them are shown to catalyze the demethylation of 2′ O methylated nucleosides in vitro. |
