响应面法优化工程菌E.coli HBVA发酵培养基  

Optimization of Fermentation Medium for Engineering Bacterium E.coli HBVA by Response Surface Methodology

作　　者：王宇[1] 吴昊[1] 朱晓明[1] 张亮[1] 徐元基[1] 张巍[1] 杜芝燕[1] 肖毅[1] 王琳[1,2] 李盼[1] 侯沙沙[1] 于继云[1] 阎瑾琦[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京100850 [2]解放军316医院,北京100093

出　　处：《生物技术》2013年第4期64-70,共7页Biotechnology

基　　金：国家自然科学基金项目("慢性乙肝新型可复制型DNA疫苗的免疫增效策略研究";31100655)资助

摘　　要：目的:对携带乙型肝炎DNA疫苗质粒pSVK-HBVA的工程菌发酵培养基进行优化。方法:采用单因素法比较不同碳源、氮源以及无机盐对发酵产量的影响;利用响应面法对各因素最佳水平及交互作用进行进一步研究。结果:单次单因子法优化结果显示,LB为最适基础培养基,最佳碳源和氮源分别是甘油和国产的眎蛋白胨;PB实验结果表明,影响质粒产量的3个主要影响因子为眎蛋白胨、甘油、NH4Cl,经过最陡爬坡和CCD优化后的发酵培养基,质粒的容积产率达到13.61 mg/L,是基础培养基LB发酵产量的2倍以上。结论:优化了工程菌E.coli HBVA的发酵培养基,提高了质粒pSVK-HBVA的产量。Objective： To optimize the fermentation medium for engineering bacterium that carries DNA vaccine plasmid pSVK - HBVA for HBV. Method： The one - factor - at - a - time experiment was used to screen the carbon sources and nitrogen sources and inorganic salt; The Response Surface Methodology （RSM） was used to investigate the optimal level of each variable and the interaction between two varia- bles. Result： The one - factor - at - a - time experiments demonstrated that Luria - Bertani （LB） was the optimal basic medium. The opti- mal carbon source and nitrogen source were glycerol and home - made proteose peptone, respectively. Based on the Plackett - Burman de- sign （PB）, proteose peptone, glycerol and NH4C1 were identified as the significant variables; these variables were further optimized by the steepest ascent （descent） method and central composite design （CCD）. Growth medium optimization in 500 mL shake flasks by re- sponse surface methodology （RSM） resulted in a maximum volumetric yield of 13.61 mg/L, which was approximately 2. 5 times higher than that obtained from the basic medium （LB）. Conclusion： In this work, the medium optimization for engineering bacterium E. coli HB- VA was employed, and successfully improved the production of pSVK - HBVA.

关 键 词：DNA疫苗 发酵 培养基优化 响应面 

分 类 号：Q93-335[生物学—微生物学]