Several nucleoside antibiotics are structurally characterized by a 5″-amino-5″-deoxyribose (ADR) appended via a glycosidic bond to a high-carbon sugar nucleoside (5′S,6′S)-5′-C-glycyluridine (GlyU). GlyU is further modified with an N-alkylamine linker, the biosynthetic origin of which has yet to be established. By using a combination of feeding experiments with isotopically labeled precursors and characterization of recombinant proteins from multiple pathways, the biosynthetic mechanism for N-alkylamine installation for ADR–GlyU-containing nucleoside antibiotics has been uncovered. The data reveal S-adenosyl-l-methionine (AdoMet) as the direct precursor of the N-alkylamine, but, unlike conventional AdoMet- or decarboxylated AdoMet-dependent alkyltransferases, the reaction is catalyzed by a pyridoxal-5′-phosphate-dependent aminobutyryltransferase (ABTase) using a stepwise γ-replacement mechanism that couples γ-elimination of AdoMet with aza-γ-addition onto the disaccharide alkyl acceptor. In addition to using a conceptually different strategy for AdoMet-dependent alkylation, the newly discovered ABTases require a phosphorylated disaccharide alkyl acceptor, revealing a cryptic intermediate in the biosynthetic pathway.
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β-Hydroxylation of α-amino-β-hydroxylbutanoyl-glycyluridine catalyzed by a nonheme hydroxylase ensures the maturation of caprazamycin
Communications Chemistry Open Access 28 July 2022
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This work was supported in part by the National Institutes of Health (NIH, grant nos. AI087849 and GM115261), the National Center for Advancing Translational Sciences (grant nos. UL1TR000117 and UL1TR001998), the Deutsche Forschungsgemeinschaft (grant no. DU1095/5-1), the state of Lower Saxony (Lichtenberg doctoral fellowship (CaSuS program) for A.L.), the Konrad-Adenauer-Stiftung (doctoral fellowship for D.W.) and the Fonds der Chemischen Industrie (doctoral fellowship for G.N.). NMR (600 MHz) data were collected at the NMR facility of the Center for Environmental Systems Biochemistry supported in part by the NIH (grant no. DK097215). We thank the Department of Pharmaceutical Sciences and the College of Pharmacy for financial support for MS and NMR instrumentation.
The authors declare no competing interests.
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Cui, Z., Overbay, J., Wang, X. et al. Pyridoxal-5′-phosphate-dependent alkyl transfer in nucleoside antibiotic biosynthesis. Nat Chem Biol 16, 904–911 (2020). https://doi.org/10.1038/s41589-020-0548-3
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