Intracellular expression and purification of the Canstatin-N protein in Pichia pastoris

文献类型： 外文期刊

第一作者： Yin, Huixiang

作者： Yin, Huixiang;Liu, Zhenwang;Zhang, Ailian;Shen, Jincheng;Chen, Liping;Fu, Xian;Zhang, Zehua;Zhou, Bing;Fu, Ceyi;Zhang, Tianyuan;Luo, Jinxian;Zhang, Tianyuan;Luo, Jinxian

作者机构：

关键词： Bioactivity;Canstatin-n gene;Intracellular expression;Pichia pastoris;Purification

期刊名称：GENE （ 影响因子：3.688； 五年影响因子：3.329 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： Canstatin-N DNA fragment amplified from human genome was inserted into the MCS of pGAP9K*, an intracellular expression vector of Pichia pastoris, to generate pGAP9K*-can-N which was then transformed into P. pastoris GS115 by electroporation. A transformant was chosen as an engineering strain from the plate containing G418 (700μg/ml). d-sorbitol was selected as the only carbon source. The fermentation was carried out in a 50L bioreactor at a 20L working volume. After 48h fermentation with continuous feeding of 25% (w/v) d-sorbitol and 0.8% PTM4, the cell grew to A 600=178 and intracellularly expressed Canstatin-N reached 780mg/L. Snail enzyme was combined with water to crack P. pastoris and to release intracellular proteins. The purified recombinant Canstatin-N inhibited CAM angiogenesis and induced significant apoptosis of the human umbilical vein endothelial cell (EVC340).

分类号： R394

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