Nicotinamide adenine dinucleotide is an ubiquitous cofactor in cells. Non-natural cofactor or biomimetic cofactors have attracted great attention on account of their low cost, high stability and the application of efficient synthesis. Nicotinamide cytosine dinucleotide is reported as a non-natural cofactor that can be synthesized in vivo, and metabolic systems have been built using NCD as the cofactor. In this study, we investigated the biological effects of this non-natural cofactor by conducting proteomics analysis.