毕赤酵母工程菌发酵生产重组Cu/Zn-SOD的工艺研究

摘要

In this study,detailed investigations were performed on effective production in 5 L benchtop fermentor of a sika deer Cu/Zn-SOD by Pichia pastoris.The process parameters of recombinant SOD production were optimized to set up an appropriate fermentation procedure as follows:BSM medium,inoculum rate of 5％ (volume fraction),temperature of 35 ℃,agitation rate of 900 r · min-1 during the induction stage,pH value of 6.0,DO level above 20％ of the saturated value,and the cultivation period of 142 h.Under the optimal conditions,the SOD activity in the broth could reach as high as 102.789 kU · mL-1,which was increased by 3.5 folds.The results above established a highly effective fermentation procedure for industrial production of the recombinant sika deer Cu/Zn-SOD by Pichia pastoris that presented significant application prospect in future.%系统考察毕赤酵母工程菌GS115表达重组铜锌超氧化物歧化酶(Cu/Zn-SOD)的发酵工艺参数,优化后5L发酵罐上生产工艺为:采用BSM培养基,接种量5％(体积分数),发酵温度35℃,诱导期搅拌转速900r·min-1,pH值为6.0、维持溶氧(DO)水平为饱和值的20％以上.经过142 h发酵实验,SOD活力最高可达102.789 kU·mL-1,比优化前提高3.5倍.采用毕赤酵母工程菌能实现高效、大量制备优质动物来源的Cu/Zn-SOD重组蛋白,具有良好的产业化潜力和广泛的应用前景.