响应面法优化重组大肠杆菌产ADI酶发酵培养基

Fermentation Medium Optimization for Arginine Deiminase Production from Recombinant Escherichia coli by Response Surface Methodology

在单因素优化的基础上,采用响应面分析方法对重组大肠杆菌生产精氨酸脱亚胺酶(ADI)的发酵培养基进行了优化。借助于SAS软件,结合Plackett-Burman和Box-Behnken实验设计对5种培养基组分进行优化。结果表明,最佳培养基组分为胰蛋白胨11.16 g/L,酵母提取物20 g/L,甘油6.3 g/L,磷酸氢二钾16.38 g/L,磷酸二氢钾2.31 g/L。采用优化后的培养基进行摇瓶发酵,ADI酶活达到5.7 U/m L发酵液,比初始培养基(3.72 U/m L发酵液)提高了1.53倍,比LB培养基(2.83 U/m L发酵液)提高了2.01倍。采用优化后的培养基在3 L发酵罐中进行发酵,ADI酶活达到15.17 U/m L发酵液,较LB培养基(4.32 U/m L发酵液)提高了3.51倍。

Response surface methodology was used to optimize the fermentation medium of recombinant E. coli for arginine deiminase production on the basis of single factor optimization. Five medium factors were optimized by SAS software combined with Plackett-Burman and Box-Behnken design. The optimized fermentation medium contains 11.16 g/L tryptone, 20 g/L yeast extract, 6.3 g/L glycerol, 16.38 g/L K2HPO4,and 2.31 g/L KH2PO4. Using the optimized medium,the activity of ADI reached 5.7 U/mL broth,which was 1.53-fold higher than that of in the initial medium （3.72 U/mL）and 2.01-fold of that in LB medium （2.83 U/mL）. In a 3-L bioreactor,ADI activity of 15.17 U/mL broth was attained using optimized medium, which was remarkably enhanced compared with that of LB medium （4.32 U/mL）.