重组人铁蛋白及牡蛎铁蛋白最佳表达条件探究  

The Optimal Expression Conditions of Recombinant Human Ferritin and Recombinant Oyster Ferritin

作　　者：程青丽 裴晨皓 冯晓文 卢知浩 李国明[1] 刘文颖[1] CHENG Qingli;PEI Chenhao;FENG Xiaowen;LU Zhihao;LI Guoming;LIU Wenying(China National Research Institute of Food&Fermentation Industry,Beijing Engineering Research Center of Protein and Functional Peptides,Beijing 100015,China;Beijing City University,Beijing 100083,China)

机构地区：[1]中国食品发酵工业研究院,北京市蛋白功能肽工程技术研究中心,北京100015 [2]北京城市学院,北京100083

出　　处：《食品科技》2021年第9期1-7,共7页Food Science and Technology

摘　　要：目的:探究铁蛋白最佳表达条件,用于开发具有靶向药物传递功能或者高效包被疫苗、营养物质等的纳米载体。方法:该实验对人体铁蛋白和牡蛎铁蛋白基因进行合成,分别通过单质粒表达和共转表达,结合前期经验选择特定培养条件,筛选出具有活性的目标蛋白。结果:人体重链铁蛋白及轻链铁蛋白在单转表达和共转表达系统下的最佳表达条件都分别是15℃诱导16 h和37℃诱导4 h,共转表达效果都不如单转表达。而牡蛎铁蛋白2种系统的最佳表达条件均为37℃诱导4 h,相比单转,共转的表达水平稍有下降但可溶性大幅上升,表达效果需要进一步纯化来评估。结论:对于人体重链铁蛋白及轻链铁蛋白,选择单转表达系统15℃诱导16 h生产所需蛋白较好;对于牡蛎铁蛋白,选择共转表达系统37℃诱导4 h生产所需蛋白较好,但具体大规模生产因培养条件、测定方法不同还需进一步考证这2种表达系统。Objective:To explore the optimal expression conditions of ferritin,and to develop nano carriers with targeting drug delivery function or high efficiency coating vaccines and nutrients.Methods:In this experiment,human ferritin and oyster ferritin were gene synthesised,respectively through the single expression and coexpression,combined with previous experience to select specific culture conditions of E.coli,and the target protein with activity were screened.Results:The optimal expression conditions of human heavy chain ferritin and light chain ferritin were induced at 15℃for 16 h and 37℃for 4 h,respectively.The optimal expression conditions of oyster ferritin in the two systems were 37℃induction for 4 h.Compared with the single expression,the expression level of coexpression was slightly decreased,but the solubility was increased significantly.The expression effect needs further purification to evaluate.Conclusion:For human heavy chain ferritin and light chain ferritin,it is better to select single expression system to induce 16 hours at 15℃;for oyster ferritin,it is better to choose coexpression system to induce 4 hours at 37℃.However.Due to different culture conditions and determination methods,the two expression systems need further study.

关 键 词：铁蛋白 纳米材料 大肠杆菌 单表达 共转表达 

分 类 号：TS201.2[轻工技术与工程—食品科学]