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

文献类型： 外文期刊

作者： Zhang, Ai-Lian 1 ; Zhang, Tian-Yuan 2 ; Luo, Jin-Xian 2 ; Fu, Ce-Yi 4 ; Qu, Zhi 1 ; Yi, Guo-Hui 1 ; Su, Dong-Xiao 1 ; Tu, 1 ;

作者机构： 1.Chinese Acad Trop Agr Sci, Inst Trop Biosci & Biotechnol, Haikou 571101, Hainan, Peoples R China

2.Zhongshan Univ, Key Lab Gene Engn, Minist Educ, Guangzhou 510275, Guangdong, Peoples R China

3.Zhongshan Univ, Dept Biochem, Guangzhou 510275, Guangdong, Peoples R China

4.Hainan Prov Inst Drug Control, Haikou 570216, Hainan, Peoples R China

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).

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[2]Constitutive expression of barley alpha-amylase in Pichia pastoris by high-density cell culture. Liu, Z. W.,Yin, H. X.,Yi, X. P.,Zhang, A. L.,Zhang, Z. H.,Shen, J. C.,Chen, L. P.,Luo, J. X.,Zhang, T. Y.,Luo, J. X.,Zhang, T. Y.,Fu, C. Y..

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