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- W2085194005 abstract "A general method for isotopic labeling of the purine base moiety of nucleotides and RNA has been developed through biochemical pathway engineering in vitro. A synthetic scheme was designed and implemented utilizing recombinant enzymes from the pentose phosphate and de novo purine synthesis pathways, with regeneration of folate, aspartate, glutamine, ATP, and NADPH cofactors, in a single-pot reaction. Syntheses proceeded quickly and efficiently in comparison to chemical methods with isolated yields up to 66% for 13C-, 15N-enriched ATP and GTP. The scheme is robust and flexible, requiring only serine, NH4+, glucose, and CO2 as stoichiometric precursors in labeled form. Using this approach, U-13C- GTP, U-13C,15N- GTP, 13C2,8- ATP, and U-15N- GTP were synthesized on a millimole scale, and the utility of the isotope labeling is illustrated in NMR spectra of HIV-2 transactivation region RNA containing 13C2,8-adenosine and 15N1,3,7,9,2-guanosine. Pathway engineering in vitro permits complex synthetic cascades to be effected, expanding the applicability of enzymatic synthesis." @default.
- W2085194005 created "2016-06-24" @default.
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- W2085194005 date "2008-08-01" @default.
- W2085194005 modified "2023-09-30" @default.
- W2085194005 title "Pathway Engineered Enzymatic <i>de Novo</i> Purine Nucleotide Synthesis" @default.
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- W2085194005 doi "https://doi.org/10.1021/cb800066p" @default.
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