Preparation of Recombinant Human Insulin

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Preparation of Recombinant Human Insulin

更新时间：2023-11-28

- Preparation of Recombinant Human Insulin
- Advanced Engineering Sciences Vol. 39, Issue 4, Pages: 79-83(2007)
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- Published：2007，
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李晓红,朱惠玲,余蓉.重组人胰岛素制备工艺[J].工程科学与技术,2007,39(4):79-83.

Preparation of Recombinant Human Insulin. [J]. Advanced Engineering Sciences 39(4):79-83(2007)
李晓红,朱惠玲,余蓉.重组人胰岛素制备工艺[J].工程科学与技术,2007,39(4):79-83. DOI：

Preparation of Recombinant Human Insulin. [J]. Advanced Engineering Sciences 39(4):79-83(2007) DOI：

摘要

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

Abstract

Abstract: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.

关键词

(His)6-Arg-Arg-人胰岛素原重组人胰岛素大肠杆菌

Keywords

(His)6-Arg-Arg-human proinsulinrecombinant human insulinE.coli

references

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