| 重组人胰岛素及C肽的分离纯化 |
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| 引用本文： | 冷雪,陈劲春,董鹏.重组人胰岛素及C肽的分离纯化[J].北京化工大学学报(自然科学版),2006,33(5):30-32. |
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| 作者姓名： | 冷雪 陈劲春 董鹏 |
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| 作者单位： | 北京化工大学，生命科学与技术学院，北京，100029 |
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| 摘 要： | 对基因工程构建的含重组人胰岛素和C肽基因的毕赤酵母工程菌进行诱导表达，工程菌用甲醇诱导72 h后，离心去除菌体，上清液经中空纤维膜超滤后SP-Sepharose Fast Flow阳离子交换柱层析，0~400 mmol/L NaCl梯度洗脱，得到纯度大于92％的重组人胰岛素原。经Lys-C酶和羧肽酶B酶切后，Sephadex G-25柱分离，得到了重组人胰岛素和C肽。
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| 关 键 词： | 重组人胰岛素 C肽 毕赤酵母 分离纯化 重组人胰岛素 C肽 毕赤酵母 分离纯化 |
| 收稿时间： | 2005-11-07 |
| 修稿时间： | 2005-11-07 |
Separation and purification of recombinant human insulin and C peptide expressed in the yeast Pichia pastoris |
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| LENG Xue,CHEN Jin-chun,DONG Peng.Separation and purification of recombinant human insulin and C peptide expressed in the yeast Pichia pastoris[J].Journal of Beijing University of Chemical Technology,2006,33(5):30-32. |
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| Authors: | LENG Xue CHEN Jin-chun DONG Peng |
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| Institution: | College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029 |
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| Abstract: | In order to obtain recombinant human insulin and C peptide expressed in the genetically engineered yeast Pichia pastoris, human proinsulin was prepared from the supernatant by SP-Sepharose Fast Flow ion-exchange chromatography after ultrafiltration through a hollow fiber membrane, and fractions eluted with 0~400 mmol/L NaCl. The purity of the recombinant human proinsulin was more than 92%, as shown by HPLC analysis. The proinsulin was digested by endoproteinase Lys-C and carboxypeptidase B and the digested sample was subsequently fractionated on sephadex G-25 gel chromatography to afford the human insulin and C peptide. |
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| Keywords: | recombinant human insulin C peptide Pichia pastoris separation purification |
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