运用高通量筛选技术优化红霉素A发酵的合成培养基

摘要

目的 设计并优化出一种适用于红霉素发酵的合成培养基.方法 利用单因素缺失实验设计来减少培养基组分,然后利用Plackett-Burman实验设计来优化组分浓度.上述实验都是采用高通量方法进行的.结果 最初选取了38种与生长和次级代谢相关的物质;然后通过2次单因素缺失实验将培养基组分减少到了19种;最后通过Plackett-Burman实验对19种组分的浓度进行了进一步优化,确定各物质的浓度.5L罐发酵结果表明本研究获得的优化后合成培养基生成的红霉素A产量是采用现有文献报道合成培养基得到红霉素A产量的13.6倍.结论 高通量筛选技术是一种快速有效的筛选方法,该方法适合于优化培养基的组分.采用本论文中得到合成培养基可以对红色糖多孢菌的胞内代谢特征和红霉素A合成的关键代谢因素进行深入分析,利用这些代谢特点和影响代谢的关键因素,本文可以更加理性地对红霉素A的发酵过程进行调控,进而提高工业红霉素A产量并降低生产成本.%Objective To design and optimize a synthetic medium for the production of erythromycin A.Methods Single factor deletion experiments were employed to reduce the components of the medium,then Plackett-Burman experiment was carried out to optimize the concentration of each component.All experiments were carried out by a high throughput method.Results The initial medium consists of 38 component related to cell growth and secondary metabolism.Two sequential single factor deletion experiments were conducted,and the number of component was reduced to 19.At last the Plackett-Burman experiment was carried out to optimize the final concentration of these components.The titer was 13.6 times higher using this optimized recipe than that of using original medium formula.Conclusion High through-put method is very efficient for synthetic medium optimization.In this study,we have a deeper understanding of the metabolism of the cell and the process of erythromycin production,thus it will help us promote the erythromycin A production and lower the costs.