Enzymatic synthesis of 2′- O -methylribonucleosides with a nucleoside hydrolase family enzyme from Lactobacillus buchneri LBK78

Characterization of an atypical, thermostable, organic solvent- and acid-tolerant 2′-deoxyribosyltransferase from Chroococcidiopsis thermalis

“…The enzymatic synthesis of nucleoside analogues by transglycosylation reactions is an interesting and sustainable alternative to traditional multistep chemical methods (Iglesias et al 2015;Lapponi et al 2016;Mikhailopulo 2007). In this regard, nucleoside 2′deoxyribosyltransferases (NDTs) (Fresco-Taboada et al 2013), nucleoside phosphorylases (Lewkowicz and Iribarren 2006), and some nucleoside hydrolases (Mitsukawa et al 2017) have been used as valuable catalysts in the synthesis of many different modified nucleosides.…”

Identification of a 2′-O-Methyluridine Nucleoside Hydrolase Using the Metagenomic Libraries

“…Most unexpectedly, RK9NH is not similar to a 2′- O -methylribonucleoside hydrolase from Lactobacillus buchneri (see Figure 3 ). This enzyme was discovered recently [ 27 ] and is already being used for the enzymatic synthesis of 2′- O -methylribonucleosides [ 28 ]. The phylogenetic analysis placed the RK9NH and the 2′- O -methylribonucleoside hydrolase from L. buchneri enzymes into separate groups ( Figure 2 ).…”

“…Hence, nucleic acid drugs comprising modified DNA or RNA oligomers are the next-generation medicine with various applications such as antisense drugs, ribozymes, small interfering RNA drugs and aptamers [ 45 ]. Enzymatic synthesis of methylated nucleosides has only recently been achieved [ 28 , 29 ], which means that the RK9NH enzyme is the gateway to a new and promising field of biocatalysis. In addition, since 5-fluoro-2′- O -methyluridine is a good substrate of RK9NH hydrolase, a novel prodrug/enzyme combination might be considered as an addition to the existing systems generating 5-fluorouracil, a well-known anticancer compound [ 46 , 47 , 48 ].…”

“…Solvent system used for analysis of uracil, 5-fluorouracil, uridine, 2′-deoxyuridine, 5-fluorouridine, 5-fluoro-2′-deoxyuridine: methanol/chloroform (1/9, v / v ). Solvent system used for analysis of 2′- O -methyluridine and 5-fluoro-2′- O -methyluridine: ethyl acetate/methanol/0.1 N hydrochloric acid (15/2/2, v / v / v ) [ 28 ]. The spots were detected and visualized under the 254 nm UV light.…”

“…The ribose 2′- O -methylation increases the hydrophobicity of nucleotides and protects them against the action of nucleases [ 25 , 26 ], which is a desired trait, especially for, e.g., DNA aptamers [ 24 ]. Despite the countless possibilities of practical use for the metabolic enzymes associated with methylated nucleosides, there are very few reports regarding enzymes involved in the metabolism of 2′- O -alkyl nucleosides [ 27 , 28 , 29 ]. The presented work focuses on the cellular degradation of 2′- O -methyluridine that is found naturally in rRNA, snRNA, snoRNA and tRNA of Archaea , Eubacteria , and Eukaryota [ 1 ].…”