NAD⁺ is well known as a crucial cofactor in the redox balance of metabolism. Moreover, NAD⁺ is degraded in ADP-ribosyl transfer reactions, which are important components of multitudinous signalling reactions. These include reactions linked to DNA repair and aging. In the present study, using the concept of EFMs (elementary flux modes), we established all of the potential routes in a network describing NAD⁺ biosynthesis and degradation. All known biosynthetic pathways, which include de novo synthesis starting from tryptophan as well as the classical Preiss–Handler pathway and NAD⁺ synthesis from other vitamin precursors, were detected as EFMs. Moreover, several EFMs were found that degrade NAD⁺, represent futile cycles or have other functionalities. The systematic analysis and comparison of the networks specific for yeast and humans document significant differences between species with regard to the use of precursors, biosynthetic routes and NAD⁺-dependent signalling.