为提高大肠杆菌发酵乳清酸的产率和糖酸转化率,以大肠杆菌(Escherichia coli)Ora-6为供试菌株,在原有培养基的基础上,通过单因素试验及正交试验探究维生素B1、维生素B3、维生素B5、维生素B6、维生素B7、维生素B12几种关键B族维生素对菌体生长及产酸能力的影响,并确定最佳组合添加量为维生素B1 6.8 mg/L,维生素B3 4.8 mg/L,维生素B5 4 mg/L,维生素B6 3.4 mg/L,维生素B7 1.4 mg/L,维生素B12 4 mg/L。在此条件下,经5 L发酵罐验证对比,此时优化后的培养基菌体量(OD600)、乳清酸产量、糖酸转化率分别为134.32、121.41 g/L和25.73%,较原始培养基分别增长了13.16%、15.47%、12.96%。同时研究确定了B族维生素采取从10 h持续流加的发酵优化策略,使得5 L发酵罐发酵乳清酸最终菌体量和产量分别达到148.23、138.2 g/L,为工业化发酵生产乳清酸提供了参考依据。
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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