鲨鱼软骨活性段蛋白在大肠杆菌中的融合表达及培养条件的优化

采用三角摇瓶来摸索菌种BL21（DE3）/pET3C-aSCAIF表达目的蛋白的最优发酵条件,并在此基础上在Biotop CF-5L自动控制发酵罐中,利用补料分批培养技术,流加甘油,进行发酵培养.摇瓶培养最优条件：2%接种量、25 mL LB培养基、37℃培养、0.5%的甘油做碳源,接种后4 h加入终浓度为0.5 mmol/L的IPTG,诱导4 h后目的蛋白表达量最高.菌种在Biotop CF-5L自动控制发酵罐中的条件为：以10%的接种量接种到2.5 L发酵培养基中,设定溶氧30%,pH7.0,温度37℃,通气量3 L/min,流加甘油15 g,培养4 h后,加入终浓度为0.5 mmol/L的IPTG,诱导4 h后终止发酵,发酵罐中获得的菌体量为62.1 g.

The optimization of the recombiant shark cartilage activator's fermentation conditions

To express aSCAIF in BL21（DE3）/pET3C,the conditions for gene expression were optimized in flasks followed by culturing in Biotop CF-5L fermenter using fed-batch fermentation.It was found that the best expression could be achieved by growing of the cells which started from 2% inoculation,in 25 mL LB medium with 0.005 g/mL Glycerol as a carbon source at 37 ℃ for 4 hours and followed by additional 4 hours induction with 0.5 mmol/L IPTG.Large scale gene expression was performed in a Biotop CF-5L fermenter.Started with 10% inoculation,the cells were cultured in 2.5 L pH 7 fermentation medium supplemented with 30% DO and 15 g Glycerol feed at 37 ℃ for 4 hours,followed by 4 hours induction with 0.5 mmol/L IPTG.The final biomass was 62.1 g.