摘要
为了研究重组人胸腺素α1(Tα1)工程菌的发酵工艺 ,在NBS MICROS15L自动控制发酵罐中 ,利用分批培养和补料分批培养技术培养含融合表达载体 pGEX Tα1的大肠杆菌DE3 (lys)。采用分批培养 ,得到的最终菌体密度OD60 0 为 8,GST Tα1融合蛋白的含量为 0 .1g L(发酵液 ) ;通过限制性流加补料 ,促进体系溶解氧能使发酵菌体密度及融合蛋白的含量显著提高 ,OD60 0 可达 3 2 ,融合蛋白的含量为 0 .7g L(发酵液 ) ,且融合蛋白主要以可溶形式表达 ,为后续蛋白纯化工作提供了便利。本研究确定了周期短、产率高且稳定的发酵工艺 。
To study the fermention of recombinant Thymos in 1(Tα1), E.coli DE3/pGEX -Tα1 was cultivated using batch and fed-batch culture in NBS- MICROS15L fermentor. In batch culture, the final cell density(OD 600 ) and t he amo unt of GST-Tα1 fusion protein was 8 and 0.1g/L respectively. In fed-batch cul ture, the final cell density(OD 600 ) and the amount of fusion protein increased to 32 and 0.7g/L respectively by means of controlling medium supplement and promoting dissolved oxygen.The GST-Tα1 fusion protein was expressed in a soluble way,thus facilita t ed further protein purification. We have established an easy and stable fermenta tion process with high level expression of target protein.
出处
《药物生物技术》
CAS
CSCD
2003年第2期68-71,共4页
Pharmaceutical Biotechnology
