Nicotinamide adenine dinucleotide (NAD), a cofactor for hundreds of metabolic reactions in all cell types, plays an essential role in metabolism, DNA repair, and aging. However, how NAD metabolism is impacted by the environment remains unclear. Here, we report an unexpected trans-kingdom cooperation between bacteria and mammalian cells wherein bacteria contribute to host NAD biosynthesis. Bacteria confer resistance to inhibitors of NAMPT, the rate-limiting enzyme in the amidated NAD salvage pathway, in cancer cells and xenograft tumors. Mechanistically, a microbial nicotinamidase (PncA) that converts nicotinamide to nicotinic acid, a precursor in the alternative deamidated NAD salvage pathway, is necessary and sufficient for this protective effect. Using stable isotope tracing and microbiota-depleted mice, we demonstrate that this bacteria-mediated deamidation contributes substantially to the NAD-boosting effect of oral nicotinamide and nicotinamide riboside supplementation in several tissues. Collectively, our findings reveal an important role of bacteria-enabled deamidated pathway in host NAD metabolism. Display omitted •Bacteria confer host cells with resistance to NAMPT inhibitors (NAMPTis)•Bacteria produce deamidated NAD precursors and prevent NAD depletion•Bacteria rescue NAMPTi-induced toxicity through nicotinamidase PncA•Oral NAM and NR boost in vivo NAD largely via microbiota-dependent deamidated pathway Shats et al. describe an unexpected trans-kingdom interaction between bacteria and mammals, wherein bacteria contribute to mammalian host NAD biosynthesis through a microbial nicotinamidase (PncA). This bacteria/gut microbiota-mediated facilitation of the deamidated NAD biosynthesis is important for the efficacy of commercial oral NAD-boosting supplements in mice.