重组AiiA蛋白可溶性表达及发酵条件优化  

Soluble Expression and Fermentation Condition Optimization of Recombined AiiA Protein

作　　者：杨彩云[1] 杜慈[1] 黄平[2] 温真[1] 毛惠民[1] 杨梅[1] 

机构地区：[1]福建师范大学生命科学学院,福建福州350108 [2]福建师范大学工业微生物教育部工程研究中心,福建福州350108

出　　处：《江西农业大学学报》2011年第6期1219-1227,共9页Acta Agriculturae Universitatis Jiangxiensis

基　　金：福建省科技厅(2009N0032);福建省教育厅(JA08041)

摘　　要：通过摇瓶培养确定重组工程菌E.coli BL21(DE3)-pET29a-aiiA表达AiiA蛋白可溶性表达的最优化条件,考察以牛肉膏蛋白胨为基础培养基添加不同种类的碳源、不同浓度的磷酸钾缓冲液(pH7.0)、微量元素、乙酸以及调整牛肉膏蛋白胨的比值对AiiA蛋白可溶性表达的影响,并用15 L发酵罐进行补料分批发酵,确定高密度发酵工艺条件。结果表明:优化发酵培养基为蔗糖10 g/L,牛肉膏7.8 g/L,蛋白胨31.2 g/L,NaCl 5 g/L,K2 HPO4.3H2 O 14 g/L,KH2 PO4 5.3 g/L,4 mL/L微量元素。优化后AiiA蛋白可溶性达4.31 mg/mL。在15 L发酵罐补料分批发酵培养中,诱导24 h时,发酵菌OD600达到18.5,可溶性蛋白表达量5.9 mg/mL,占AiiA总蛋白50.7%,实现了扩大培养。The optimum conditions for soluble AiiA protein expression of E.coli BL21（DE3）-pET29a-aiiA were determined by shaking culture.The effects of adding different varieties of carbon sources and trace elements,different concentration of potassium phosphate buffer solution（pH7.0）,trace elements,acetic acid into beef anointed medium,adjustmenting the ratio of beef extract to tryptone on AiiA protein soluble expression were investigated and the conditions for high density fermentation were determined through the experiment of 15L fermentor fed-batch culture.The results showed that the optimal components in the medium（g/L）were sucrose 10,beef extract 7.8 g,tryptone 31.2,NaCl 5,K2HPO4·3H2O 14,KH2PO4 5.3,and trace elements 4 mL/L.Under the optimal culture condition,the soluble AiiA protein reached 4.31 mg/mL.After fermented bacteria were induced for twenty-four hours in 15 L fermentor fed-batch culture with high-density,the OD600 of the fermented bacteria reached 18.5,the soluble AiiA protein was 5.9 mg/mL and accounted for 50.7% of the total protein making successful extended culture.

关 键 词：AiiA蛋白 补料分批发酵 条件优化 

分 类 号：TQ920.6[轻工技术与工程—发酵工程]