正交设计优化大肠杆菌表达LL-37-haFGF的摇瓶发酵条件  

Optimization of flask liquid fermentation for LL-37-haFGF expression in recom-binant Escherichia coli

作　　者：沈娟[1,2] 丁静 金小宝 朱家勇 

机构地区：[1]广东省生物活性药物研究重点实验室 [2]广东药学院生命科学与生物制药学院,广东广州510006

出　　处：《广东药学院学报》2012年第6期665-668,共4页Academic Journal of Guangdong College of Pharmacy

基　　金：广州市科技支撑计划项目(2010J-E411);广东省教育厅育苗计划(LYM11082)

摘　　要：目的在摇瓶发酵条件下,研究重组大肠杆菌诱导表达融合蛋白LL-37-haFGF的表达规律。方法以目的蛋白的相对表达量为评价指标,通过单因素分析和正交设计,从pH值、诱导时机、诱导时间、诱导剂浓度4个方面,对其发酵条件进行优化。结果影响LL-37-haFGF产生水平的主次因素顺序:诱导时机>pH值>诱导时间>诱导剂浓度。优化最佳工艺参数为:诱导时机为A600达0.6,pH值为7.5,IPTG诱导浓度为0.8 mmol/L,诱导时间为4 h,LL-37-haFGF表达量达53.7%。结论该表达条件的优化为LL-37-haFGF的进一步功能研究和应用奠定基础。Objective To optimize the expression of LL-37-haFGF fusion protein in Escherichia coli. Method The expression level of recombinant LL-37-haFGF was identified as the evaluation index. The influence of different culture conditions based on shake flask fermentation such as pH,inoculation time, IPTG concentration and induction time were investigated by using single factor experiment and orthogonal experiment. Result The importance of each factor was determined as inoculation time 〉 pH 〉 induction time 〉 IPTG concentration. The optimal conditions were pH of 7.5,A600 of 0.6, IPTC concentration of 0.8 mmol/L and induction time of 4 h. Under these conditions,LL-37-haFGF reached to the highest expression level,with the relative expression level of 53.7%. Conclusion This study was meaningful for the production and application of LL-37-haFGF for further research.

关 键 词：LL-37-haFGF 大肠杆菌 正交设计 摇瓶发酵 

分 类 号：TQ927[轻工技术与工程—发酵工程]