重组人胰岛素制备工艺
Preparation of Recombinant Human Insulin
作者：李晓红(四川大学 华西药学院,四川 成都 610041)；朱惠玲(四川大学 华西药学院,四川 成都 610041)；余蓉(四川大学 华西药学院,四川 成都 610041)
Author：(West China School of Pharmacy, Sichuan Univ., Chengdu 610041,China)；(West China School of Pharmacy, Sichuan Univ., Chengdu 610041,China)；(West China School of Pharmacy, Sichuan Univ., Chengdu 610041,China)
收稿日期：2006-01-20 年卷（期）页码：2007,39(4):79-83
期刊名称：工程科学与技术
Journal Name：Advanced Engineering Sciences
关键字：(His)6-Arg-Arg-人胰岛素原；重组人胰岛素；大肠杆菌
Key words：(His)6-Arg-Arg-human proinsulin;recombinant human insulin;E.coli
基金项目：科技部创新基金资助项目（06C26215100476）
中文摘要
为了提高重组人胰岛素的表达量，用E.coli发酵表达(His)6-Arg-Arg-人胰岛素原[(His)6-Arg-Arg-human proinsulin，RRhPI]，并对影响RRhPI高效表达的各主要因素以及下游纯化工艺的各关键步骤进行优化。研究表明，RRhPI E.coli在最优培养基:5 g/L胰蛋白胨，2 g/L谷氨酸，7 g/L酵母浸膏，0.5 g/L酸铵，2.5 g/L葡萄糖，2.7 g/L甘油,100 mg/L氨苄青霉素,50 mg/L卡那霉素，pH 7.2的条件下，用发酵罐进行发酵,在
英文摘要
To obtain recombinant human insulin(rhI), (His)6-Arg-Arg-human proinsulin (RRhPI) was expressed by Escherichia coli(E.coli). In the optimum medium of 5 g/L tryptone，2 g/L glutamic acid，7 g/L yeast extract，0.5 g/L ammonium sulfate，2.5 g/L glucose，2.7 g/L glycerol,100 mg/L ampicillin,50 mg/L kanamycin，and pH 7.2, and with the addition of 0.5 mmol/L IPTG at intermediate exponential phase and induction for 4 h , 51.2 g wet E.coli and 16.2 g inclusion body were harvested from 1 L medium in fermenter. The results showed that certain concentration of β-mercaptoethanol in inclusion body solution and oxidized glutathione in refolding solution were helpful to RRhPI refolding. With the well optimized technology, 74 mg rhI was obtained from 1 L culture medium.
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