大肠埃希菌中普卡那肽的重组表达  

Recombinant Expression of Plecanatide in Escherichia coli

作　　者：李晓婉 吴勇[3] 黄宗庆 赵文杰 冯军 LI Xiaowan;WU Yong;HUANG Zongqing;ZHAO Wenjie;FENG Jun(China State Institute of Pharmaceutical Industry,Shanghai 201203;School of Pharmacy,Fudan University,Shanghai 200433;Shanghai Duomirui Bio-Technology Co.,Ltd.,Shanghai 201203)

机构地区：[1]中国医药工业研究总院,上海201203 [2]复旦大学药学院,上海200433 [3]上海多米瑞生物技术有限公司,上海201203

出　　处：《中国医药工业杂志》2020年第10期1282-1287,共6页Chinese Journal of Pharmaceuticals

摘　　要：建立了一种重组表达获得普卡那肽的方法。通过对本实验室构建的重组大肠埃希菌gp55-SUMO-SP304菌株进行高密度发酵和高压匀浆破胞,获得了gp55-SUMO-SP304包涵体,再通过酸切和酶切去除gp55-SUMO-SP304包涵体蛋白的融合标签,采用超滤和反相浓缩获得普卡那肽及其异构体。结果显示,每升gp55-SUMO-SP304菌株发酵液可获得包涵体蛋白33.4 g,去除融合标签后可产生普卡那肽148.47 mg。A method for acquisition plecanatide by recombinant expression was established in this paper.Through high-density fermentation and high-pressure homogenization of the recombinant Escherichia coli gp55-SUMOSP304 strain constructed by our laboratory,the gp55-SUMO-SP304 inclusion bodies were obtained.The fusion tag of gp55-SUMO-SP304 inclusion body protein could be removed by acid digestion and restriction enzyme digestion,and plecanatide and its isomers were obtained by ultrafiltration and reverse-phase concentration.The results showed that 33.4 g of inclusion body protein per liter could be obtained in fermentation broth of gp55-SUMO-SP304 strains,and 148.47 mg of plecanatide could be produced after removing the fusion tag.

关 键 词：重组表达 普卡那肽 包涵体蛋白 融合标签 

分 类 号：Q936[生物学—微生物学] Q816