重组大肠杆菌产融合蛋白AG的发酵优化-《集美大学学报（自然科学版）》

文章信息/Info

Title:: Fermentation Condition Optimization of Recombinant Escherichia coli Producing Recombinant Protein AG

作者:: 蓝萌萌1; 黄玲珑1; 倪辉2; 李志朋1; 3; 姜泽东1; 3; 朱艳冰1; 3; (1.集美大学海洋食品与生物工程学院，福建厦门 361021；2.厦门海洋职业技术学院，福建厦门 361102；3.福建省食品微生物与酶工程重点实验室，福建厦门 361021)

Author(s):: LAN Mengmeng1; HUANG Linglong1; NI Hui2; LI Zhipeng1; 3; JIANG Zedong1; 3; ZHU Yanbing1; 3; (1.College of Ocean Food and Biological Engineering,Jimei University,Xiamen 361021,China;2.Xiamen Ocean Vocational College,Xiamen 361102,China;3.Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering,Xiamen 361021,China)

Keywords:: recombinant protein AG; fermentation optimization; recombinant E.coli; medium component; biomass

摘要:: 对产融合蛋白AG的重组大肠杆菌BL21(DE3)/pET-28a-rPAG进行发酵优化以提高其表达量。从筛选后的TB培养基出发，通过单因素实验对TB培养基的组分和摇瓶发酵的培养条件进行优化，确定了最适的培养基组分，即酵母浸粉为24 g/L、大豆蛋白胨为12 g/L、蔗糖为8 g/L、KH2PO4为2.31 g/L、K2HPO4为 12.54 g/L。在此基础上，利用单因素实验获得了最佳的培养条件，即培养基初始pH值为7.0、接种量为5%、装液量为40%、菌液A(600)=0.8时加入0.3 mmol/L IPTG、30 ℃诱导发酵8 h。优化培养基组分和发酵条件后，重组大肠杆菌的A(600)值是优化前的3.58倍，融合蛋白AG相对含量是优化前的2.92倍。在30 L发酵罐进行初步放大验证，诱导发酵8 h后，融合蛋白AG相对含量最高，A(600)值最高达8.688，湿质量达到18.56 g/L。

Abstract:: Fermentation of the recombinant Escherichia coli BL21(DE3)/pET-28a-rPAG was optimized to improve its production of recombinant protein AG.The components of the TB medium and the culture conditions of shake flask fermentation were optimized by single-factor experiments based on the screened TB medium.The optimal medium composition was determined as 24 g/L yeast extract,12 g/L soy peptone,8 g/L sucrose,2.31 g/L KH2PO4,12.54 g/L K2HPO4.On this basis,the optimal culture conditions were initial medium pH of 7.0,an inoculation volume of 5%,a liquid volume of 40% ,and the addition of 0.3 mmol/L IPTG when the A(600) value of the culture reached 0.8,followed by induction at 30 ℃ for 8 h.After optimizing,the A(600) of recombinant E.coli increased by 3.58 times,and the relative content of the fusion protein AG increased by 2.92 times.A preliminary scale-up verification was conducted in a 30 L fermenter.After 8 h of induction fermentation,the relative content of the recombinant protein AG was the highest,with the A(600) value reaching up to 8.688 and the wet weight reaching 18.56 g/L.