人脑钠素基因工程菌的发酵条件及产物纯化研究

为进一步提高工程菌的下游纯化效率，本文利用高效表达质粒载体将多拷贝脑钠素基因转人宿主菌E．xoliM5219，摸索最佳发酵条件，热诱导获较高表达（30％）:通过超声裂解收获目的蛋白包涵体，采用制备性电泳成功获得纯度大于95％的5BNP融合蛋白；经BNP／SKATOLE裂解成单体后测得有舒张血管活性。该纯化路线为后续工作提供了线索。

Research on Fermetation and Product Purification of hBNPEngineering Bacteria

In order to facilitate downstream purification of the engineering bactena, by means of molecular cloning tech.niqUes, we cloned the tandemly repeated ceding seqUence of SBNP into plashhed PMS3lb -- c. Host E. colt M5219 wasthen ti'ansformed and induced by temperature to express protein of purPOse. Proper ferment condihons were detelmined,which led to a high expression over 30%. SBNP fusion protein which was pIDduced as inclusion they (IB) wasseparated by breaking bacteria enough sonication. IB was properly dissolved in the presence of SM Urea. By use of initiahve preparative elecrmphoresis, SBNP fusion protein was obtained with a purity Of over 95%. When broken byBNPS/Skatole into slope BNP, and diluted with renatUration soluhon so that folding process can be initiated, the pacified product was highly effective in relating whhit aortic stLrip.