响应面法优化重组hCu,Zn-SOD改构体发酵培养基  

Optimization of Fermentation Cultivation Medium for Recombinant hCu,Zn-SOD Mutant Using Response Surface Methodology

作　　者：张月娟[1] 张琨[1] 万一[1] 

机构地区：[1]陕西省微生物研究所,陕西西安710043

出　　处：《生物技术》2011年第3期69-73,共5页Biotechnology

基　　金：中国科学院西部之光人才培养项目(2009DF03);陕西省科学院青年基金项目(2010K-18;2009K-28)资助

摘　　要：目的:优化已构建的重组hCu,Zn-SOD改构体基因工程菌的发酵培养基,提高重组hCu,Zn-SOD改构体活性蛋白产量。方法:单因素实验筛选发酵培养基的碳源和氮源,Plackett Burman设计筛选影响hCu,Zn-SOD活性的重要影响因子,最陡爬坡实验逼近重要影响因子的hCu,Zn-SOD活性的最大响应区域,Box-Behnken及响应分析法进行回归分析。结果:重组hCu,Zn-SOD改构体发酵培养基重要影响因子的最优取值为:酵母提取物7.464 6g/L,NaNO30.712 9g/L,Na2HPO4.12H2O30.487 6g/L,KH2PO44.183 0g/L,优化后的hCu,Zn-SOD活性是1 470 700U/L,较初始培养基提高了1.04倍。结论:响应面法优化重组hCu,Zn-SOD的发酵培养基提高了hCu,Zn-SOD的活性和产量,为重组hCu,Zn-SOD的工业化生产提供依据。Objective： Optimizing the fermentation cultivation medium for the E. coli strain of hCu, Zn - SOD mutant to increase hCu, Zn - SOD production. Method： The single - factor experiments were used to screen carbon source and nitrogen source. Then PIackett Burman design was undertaken to evaluate the effects of seven cultivation medium factors. The steepest ascent experiment was adopted to determine the optimal region of the critical factors. Finally, The response surface methodology was used to optimize the aboved critical factors. Resuit： The optimum levels of the the critical factors were yeast extract 7. 464 6g/L,NaNO3 0. 712 9g/L Na2HPO4·12H2030.487 6g/L, KH2PO4 4. 183 Og/L. The yield of hCu,Zn -SOD activity under the optimal cultivation medium could reach 1 470 700U/L, which was improved 1.04 times compared to that under original cultivation medium. Conclusion： The yeild and activity of rhCu, Zn - SOD was increased using response surface methodology optimizing fermentation cultivation medium , The result provided basis, for industrial production of rhCu, Zn - SOD.

关 键 词：超氧化物歧化酶 Plackett Burman设计 响应面法 发酵 

分 类 号：Q815[生物学—生物工程]