基于关键因素调控发酵生产过氧化氢酶

Production of Catalase Based on Controlling of Key Factors

采用正交试验策略,分析了不同关键因素对重组E.coli发酵生产过氧化氢酶(CAT)的影响,确定CAT合成的最优摇瓶发酵条件为:甘油5 g/L、酵母粉35 g/L、初始pH 7.5、诱导温度为30℃。在3 L发酵罐发酵生产CAT过程中,当转速为300 r/min时,酶活力最高为201 17.2U/mL,最适发酵产酶初始甘油质量浓度为10 g/L。以0.67 g/(L·h)的速率进行流加甘油时,最高酶活达28 243.0 U/mL。在诱导开始时一次性补入16.7 g/L甘油,当发酵至47 h时CAT酶活达到最大值为50 369.5 U/mL,为优化前酶产量的2.5倍。

The effect of different key factors on production of catalase （CAT） in recombinant Escherichia coli was analyzed by orthogonal experiment.The optimum fermentation condition in shake flask was determined：glycerol 5 g/L,yeast extract 35 g/L,initial pH 7.5,and induction temperature 30 ℃.During production of CAT in 3 L fermentor,the highest activity reached 20 117.2 U/mL at the optimum stirring speed（300 r/min）.The optimum initial glycerol concentration was 10 g/L.The highest activity reached 28 243.0 U/mL when the speed of glycerol supplement was 0.67 g/（L·h）.When 16.7 g/L glycerol was supplemented at start induction,the CAT activity could reach 50 369.5 U/mL after incubation for 47 h,which was 2.5 fold of that before optimization.