title: Enzymatic synthesis of 2'-O-methylribonucleosides with a nucleoside hydrolase family enzyme from Lactobacillus buchneri LBK78.
abstract: 2'-O-Methylribonucleosides (2'-OMe-NRs) are promising raw materials for the production of nucleic acid drugs. We previously reported that LbNH, a nucleoside hydrolase from Lactobacillus buchneri LBK78 (NITE P-01581), was the first enzyme found to act on 2'-OMe-NRs. In the present study, we determined that LbNH also has the transribosylation activity between 2'-OMe-NRs and nucleobases, in addition to the hydrolyzing activity towards 2'-OMe-NRs. When 2'-O-methyluridine (2'-OMe-UR) and adenine were reacted with LbNH, 2'-O-methyladenosine (2'-OMe-AR) was produced. LbNH preferred purine nucleobases as its acceptor substrates for the transribosylation with 2'-OMe-UR as a donor substrate. Kinetic analysis of LbNH revealed that adenine behaved as a mixed inhibitor of the hydrolysis of 2'-OMe-UR. Under the optimal reaction conditions, the maximum molar yield of enzymatic 2'-OMe-AR produced reached 0.97% towards 2'-OMe-UR, corresponding to 0.16 g/L.
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan.
authors.forename: Yuuki
authors.initials: Y
authors.lastname: Mitsukawa
authors.affiliation: Industrial Microbiology, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Hibi
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan.
authors.forename: Narihiro
authors.initials: N
authors.lastname: Matsutani
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Horinouchi
authors.affiliation: Industrial Microbiology, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan.
authors.forename: Satomi
authors.initials: S
authors.lastname: Takahashi
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan. Electronic address: ogawa@kais.kyoto-u.ac.jp.
authors.forename: Jun
authors.initials: J
authors.lastname: Ogawa
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D055162:Biocatalysis; D006868:Hydrolysis; D007700:Kinetics; D007778:Lactobacillus / Q000201:enzymology*; D009699:N-Glycosyl Hydrolases / Q000378:metabolism*; D014529:Uridine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism
chemical_list: C024341:2'-O-methyladenosine; C052202:2'-O-methyluridine; D009699:N-Glycosyl Hydrolases; D000241:Adenosine; D014529:Uridine
keywords: 2′-O-Methylribonucleosides; Lactobacillus buchneri; Nucleic acid drugs; Nucleoside hydrolase; Transribosylation