CpG基序重组菌发酵培养基的优化

Optimization of the Fermentation Medium of the Strain with the CpG Motifs Recombinant Plasmid

目的:为提高CpG基序重组菌的发酵水平,降低CpG重组质粒的生产成本,运用响应面法优化CpG重组菌发酵培养基。方法:利用Plackett-Burman试验筛选出影响发酵水平的3个最显著因素:糖蜜、玉米浆和工业蛋白胨。然后用响应面设计试验并优化得到最显著因素的最佳浓度,并进一步通过发酵试验验证优化后的培养基。结果:得到一组具有较高菌浓及质粒产量且价格低廉的发酵用培养基:糖蜜4.5g/L,玉米浆8.5g/L,工业蛋白胨8.5g/L,甘油10mL/L,KH2PO41.5g/L,K2HPO42.3g/L,MgSO4.7H2O 0.25g/L,在此培养条件下,OD600实测值为0.6771,理论值为0.6643,两者接近,与标准LB培养基相比,质粒产量提高了15%左右。结论:最终筛选到的培养基具有较高的性价比。

Objective:In order to improve the production of the CpG motifs recombinant stain and lower the cost,the response surface method was applied to optimize the fermentation medium.Method:Firstly,Plackett-Burman design was undertaken to determine the three significant factors affecting strain production as follows:molasses,corn syrup and peptone.the response surface design was used to optimize the three critical factors and the optimum levels,furthermore the fermentation was testified the optimum medium.Result:The optimum fermentation medium was 4.5g/L molasses,8.5g/L corn syrup,8.5g/L peptone,10mL/L glycerol,1.5g/L KH2PO4,2.3g/L K2HPO4,0.25g/L MgSO4·7H2O.Under such conditions,the predicted OD600 was 0.664 3,which was in excellent agreement with the experimental result 0.6771,about 15% higher than the plasmid production under the initial LB medium.Conclusion:The final optimum medium was identified to make lower costs and higher yield of strain and plasmid yield.