人内皮细胞抑制生长素在毕赤酵母中的高效表达  

High Expression of Human Endostatin in Pichia pastoris

作　　者：苏志坚[1] 吴晓萍[1] 郑青[1] 冯雅[2] 于平野[2] 曲红艳[2] 刘太胜[2] 李校堃[1] 

机构地区：[1]暨南大学药学院,广州510630 [2]暨南大学医药生物技术研究开发中心,广州510632

出　　处：《中国生物工程杂志》2004年第5期47-51,共5页China Biotechnology

基　　金：国家"973"计划资助项目 (G19990 5 42 0 4)

摘　　要：根据人胶原蛋白ⅩⅧNC结构域C末端编码内皮细胞抑制生长素 (Endostatin)的成熟多肽序列 ,人工合成了由毕赤酵母 (Pichiapastoris)中偏爱密码子组成的内皮细胞抑制生长素基因序列。该基因以N端融合的方式正确插入毕赤酵母诱导型表达载体pPICZαA中。通过电激将线性化的重组质粒转化到毕赤酵母SMD1 1 68细胞中 ,筛选获得表达具有生物活性的内皮细胞生长抑制素的高产菌株。在摇瓶发酵水平上 ,产量达到 80mg L。而利用生物反应器进行高密度发酵 ,Endostatin的产量可达 1 2 5mg L。The gene of Endostatin that consisted of 20 codons of preference in Pichia pastoris was artificial synthesized according to its mature peptide sequence in human collagenⅩⅧ.The artificial gene was cloned into methanol inducible expression vector pPICZαA.The linearized recombinant plasmid was transformed into chromosome of Pichia pastoris SMD1168 strain by electroporation.5 recombinant yeast strains were fermented in flasks and 1 recombinant yeast strain P.pastoris Endo V was performed for high cell density fermentation.The expression of Endostatin in supernate was 80mg/L and 125mg/L,respectively.After purification,the Endostatin could restrain the proliferation of endothelium cells in vitro. ;

关 键 词：人工合成 人内皮细胞生长抑制素 毕赤酵母 高密度发酵 癌症 

分 类 号：TQ920.1[轻工技术与工程—发酵工程] Q78[生物学—分子生物学]