To improve the production and sugar-acid conversion rate of Escherichia coli fermented orotic acid, E. coli Ora-6 was used as the test strain, this study investigated the effect of several key B vitamins of VB1, VB3, VB5, VB6, VB7 and VB12 on the biomass, orotic acid production ,and sugar-acid conversion rate based on the original medium. The best combination addition was VB1 6.8 mg/L, VB3 4.8 mg/L, VB5 4 mg/L, VB6 3.4 mg/L, VB7 1.4 mg/L, VB12 4 mg/L. Under these conditions, the verification test in 5 L fermenter showed that the optimized medium biomass (OD600), orotic acid production, and sugar-acid conversion rate were 134.32, 121.41 g/L and 25.73%, an increase of 13.16%, 15.47%, and 12.96% respectively over the original medium. In addition, this study determined that the B vitamins were continuously fed from the 10th hour to the fermentation optimization strategy. The final biomass and production of orotic acid fermented in the 5 L fermenter reached 148.23 and 138.2 g/L, respectively. This study provides some references for the industrial fermentation of orotic acid.
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