Fig. 5: Enzymatic cascade synthesis of ΨTP and dΨTP.
From: Reverse C-glycosidase reaction provides C-nucleotide building blocks of xenobiotic nucleic acids
Panels a and b show ΨTP and dΨTP, respectively. Pentoses (100 mM) were phosphorylated applying RbsK (15 µM for d-Rib; 30 µM for d-dRib) and pyruvate kinase (4 µM) using 100 mM PEP and 2 mM ATP as phosphate donors (see Supplementary Fig. 33). ΨMP synthesis was initiated by addition of 15 mM Ura, 1 mM MnCl2, YeiN (1 µM for d-Rib, 3 µM for d-dRib) and 10 mM HEPES buffer, resulting in a concentration of 25 mM pentose 5-phosphate (time point zero a, b. The arrow indicates the start of the phosphorylation reaction by addition of 15 mM PEP, 15 µM CMPK, and 1 mM ATP, resulting in a volume increase of 33% (n = 1).
