高密度培养大肠杆菌表达重组人胰高血糖素样肽-1

目的:高密度发酵培养表达重组人胰高血糖素样肽-1(hGLP-1).方法:将构建的重组质粒pGEX-hGLP-1转化大肠杆菌BL21(DE3),得到表达hGLP-1的工程菌株E.coli BL21(DE3)/pGEX-hGLP-1.在500 ml三角摇瓶中进行了诱导条件的摸索实验,之后用5 L自控发酵罐对工程菌进行高密度培养,以获取hGLP-1和谷胱甘肽S-转移酶融合蛋白(GST-hGLP-1).结果:采用分批培养和补料分批培养相结合的技术,控制碳源和氮源的补加,控制溶解氧的量,可使重组工程菌发酵光密度D600值达52,融合蛋白的表达量占菌体总蛋白量的28％,其含量达到2.1 g/L.用ESI质谱鉴定hGLP-1相对分子质量与理论值一致.结论:本研究为大规模生产重组hGLP-1奠定了基础.

Expression of recombinant human glucagon like peptide-1 by high density culture of E. coli

Objective: To express recombinant human glucagon like peptide 1 by high density fermentation culture of E. coli BL21(DE3). Methods: The recombinant plasmid pGEX hGLP 1 was constructed successfully and was transformed into E.coli BL21(DE3).The optimization of induction condition was carried out in 500 ml shake flasks,followed by 5 L fermentor with high cell density culture to produce recombinant hGLP 1 in E.coli BL21(DE3)/ pGEX hGLP 1. Results: Batch and fed batch culture by controlling carbon nitrogen concentration and keeping oxygen at 30% 40% were used to obtain the final cell density( D 600 =52)and fusion protein GST hGLP 1 (2.1 g/L). ESI MS showed that the relative molecular mass of hGLP 1 was the same as expected. Conclusion: This study provides a basic work for production of recombinant hGLP 1 in scale.