Nicotinamide bacterial synthesis

Synthesis and purification of 2 dNAD+. We used p-NMN+ adenyl transferase to synthesize [adenylate- P]2 dNAD+ from [a-3 ]2 dATP. Residual 2 dATP was removed by affinity chromatography on a boronate gel as described elsewhere (14). The elimination of contaminating P-NMN+ (Fig. lA), which also boimd to the boronate gel, was facilitated by its conversion to nicotinamide ribose by treatment with bacterial alkaline phosphatase (Fig. IB). Figure 1C shows that following preparative HPLC, a single peak corresponding to pure 2 dNAD was detected. It is important to note that all of the 32p radiolabel co-eluated with this peak. 

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