Inducible expression of human angiostatin by AOXI promoter in P. pastoris using high-density cell culture

文献类型： 外文期刊

第一作者： Zhang, Ai-Lian

作者： Zhang, Ai-Lian;Qu, Zhi;Yi, Guo-Hui;Su, Dong-Xiao;Tu, Fa-Zhi;Pan, Ying-Wen;Zhang, Tian-Yuan;Luo, Jin-Xian;Zhang, Tian-Yuan;Luo, Jin-Xian;Fu, Ce-Yi

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关键词： Inducible expression;Angiostatin;P. pastoris;High-density cell culture

期刊名称：MOLECULAR BIOLOGY REPORTS （ 影响因子：2.316； 五年影响因子：2.357 ）

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收录情况： SCI

摘要： A high-density cell culture method was successfully established in P. pastoris with the alcohol oxidase I (AOXI) promoter in order to produce large quantities of recombinant human angiostatin (AS) which has been reported to have antiangiogenic activity. A preliminary study on fermentation conditions in shaking flasks indicated that adequacy of biomass is beneficial to obtain more products. The fermentation was carried out in a 10 l bioreactor with 5 l modified growth medium recommended by Invitrogen at 30pC. The cells were first grown in glycerol-PTM4 trace salts for 24 h. When the cell density reached A = 125, methanol-PTM4 trace salts was added to induce the expression of AS. During the fermentation, dissolved oxygen level was maintained at 20-30%, pH was controlled at 5 by the addition of 7 M NHOH and the biomass was maintained at about A = 200. After 60 h of induction, the secreted AS was 153 mg/l. The recombinant AS inhibited the angiogenesis on CAM and suppressed the growth of B16 melanoma in C57BL/6J mice (P < 0.01).

分类号： Q7

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