Abstract: In order to study the β-nicotinamide mononucleotide-enriched fruit-vegetable Jiaosu and evaluate its bioactivity. Cucumber powder, broccoli powder, banana powder and apple powder were used as substrates, and the process parameters were optimized by multi-species co-fermentation to investigate the effects of strain combinations, fermentation time, temperature and precursor addition on β-NMN enrichment. At last, the enzymatic activities of superoxide dismutase, lipase, and amylase in the Jiaosu, as well as the hydroxyl radical scavenging characteristics and total antioxidant capacity, were determined. Results demonstrated that synergistic co-fermentation employing Rhizopus oryzae, Monascus purpureus, yeast, and Lactobacillus plantarum strains yielded maximal β-NMN production under the optimal conditions, as the mass ratio of cucumber powder, broccoli powder, banana powder, apple powder was 2:2:1:1 (w/w), and cultured at 25 ℃ for 60 h. Under this condition, the content of β-NMN in the fermentation broth was 808.8 μg/L, which was 20.86 times higher than that before fermentation. The contents of total polyphenols, reducing sugar and protein after fermentation were 0.3425, 1.5556 and 3.7986 mg/mL, respectively. In addition, the total superoxide dismutase activity, lipase activity and amylase activity in the jiaosu powder were 50.9917, 9.5104 and 13.7088 U/mL, respectively, which were 1.98 and 2.05 times higher than the hydroxyl radical inhibition ability and total antioxidant capacity (FRAP) of the fruit and vegetable stock solution before fermentation. The results of this study provide a new reference for the development of β-NMN-rich jiaosu and their applications in the fields of food, medicine and cosmetics.