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A potential alternate substrate for orotidine-5′-monophosphate decarboxylase, 2- thio-orotidine-5′-monophosphate, was synthesized enzymatically and purified by a modification of a previous account (K. Shostak, and M. E. Jones 1992, Biochemistry31, 12155–12161). Characterization of the product was confirmed by mass spectrometry, 31 P NMR, and utilization by orotate phosphoribosyltransferase...