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Molecules 2018, 23(11), 2904;

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

Department of Molecular Microbiology and Biotechnology, Institute of Biochemistry, Life Sciences Center, Vilnius University, LT-10257 Vilnius, Lithuania
Department of Chemistry and Bioengineering, Vilnius Gediminas Technical University, LT-10223 Vilnius, Lithuania
Author to whom correspondence should be addressed.
Received: 2 October 2018 / Revised: 30 October 2018 / Accepted: 4 November 2018 / Published: 7 November 2018
(This article belongs to the Section Chemical Biology)
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Ribose methylation is among the most ubiquitous modifications found in RNA. 2′-O-methyluridine is found in rRNA, snRNA, snoRNA and tRNA of Archaea, Bacteria, and Eukaryota. Moreover, 2′-O-methylribonucleosides are promising starting materials for the production of nucleic acid-based drugs. Despite the countless possibilities of practical use for the metabolic enzymes associated with methylated nucleosides, there are very few reports regarding the metabolic fate and enzymes involved in the metabolism of 2′-O-alkyl nucleosides. The presented work focuses on the cellular degradation of 2′-O-methyluridine. A novel enzyme was found using a screening strategy that employs Escherichia coli uracil auxotroph and the metagenomic libraries. A 2′-O-methyluridine hydrolase (RK9NH) has been identified together with an aldolase (RK9DPA)—forming a part of a probable gene cluster that is involved in the degradation of 2′-O-methylated nucleosides. The RK9NH is functional in E. coli uracil auxotroph and in vitro. The RK9NH nucleoside hydrolase could be engineered to enzymatically produce 2′-O-methylated nucleosides that are of great demand as raw materials for production of nucleic acid-based drugs. Moreover, RK9NH nucleoside hydrolase converts 5-fluorouridine, 5-fluoro-2′-deoxyuridine and 5-fluoro-2′-O-methyluridine into 5-fluorouracil, which suggests it could be employed in cancer therapy. View Full-Text
Keywords: metagenomics; nucleoside hydrolase; 2′-O-methyluridine; 2′-O-alkyl nucleosides; 5-fluorouracil metagenomics; nucleoside hydrolase; 2′-O-methyluridine; 2′-O-alkyl nucleosides; 5-fluorouracil

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Aučynaitė, A.; Rutkienė, R.; Tauraitė, D.; Meškys, R.; Urbonavičius, J. Identification of a 2′-O-Methyluridine Nucleoside Hydrolase Using the Metagenomic Libraries. Molecules 2018, 23, 2904.

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