工程菌LacU V5par8egf产hEGF发酵方法的比较

The studies on hEGF productions by recombinant E.Coli harboring expression LacU V5par8egf in batch fermentation

目的 :明确培养条件对工程菌 L ac U V5 par8egf表达水平和质粒稳定性的影响。方法 :采用优化 MBL培养基 ,利用摇瓶发酵研究工程菌 L ac U V5 par8egf的最优培养条件 ,并与 2 L发酵罐上的发酵进行比较。结果 :摇瓶发酵的最优培养条件为 :温度 32℃ ,接种量 10 % ,种子液 OD5 5 0 1.0～ 3.0 ,装液量 10 % ,摇床转速 2 2 0 rpm;摇瓶发酵的最优诱导条件是在对数生长中后期诱导 ,IPTG最适浓度为 0 .2 m M。 2 L 发酵罐发酵与摇瓶发酵相比 ,发酵周期缩短了约三分之一 ,h EGF表达水平接近摇瓶中发酵水平 ,达到 2 7.7m g/L。结论 :利用摇瓶最优培养条件 ,较好实现了重组 h

OBJECTIVE:Clarifying the fermentation conditions of hEGF production by recombinant E.coli harboring expression LacUV5par8egf and plasmid stability was our objective.METHODS:The detalied optimized fermentation conditions in the shaken culture in MBL medium had been studied.The fermentation in shaken flask and 2L bench top fermentor had been compared too.RESULTS:In the shaken culture,the detailed optimized fermentation conditions were obtained as following:temperature 32℃,inoculum volume 10%(V/V),inoculum age OD 550 1.0～3.0,medium volume10%(V/V),rotating speed 220 rpm.It was also shown that induction conditions could affect hEGF accumulation obviously.The maximum hEGF production was attained when the induction was done at middle or late logarithmic phase.The addition of 0.2 mM IPTG led to the highest hEGF accumulation.The fermentation time in 2L bench top fermentors was two thirds of the time in shaken culture,But hEGF in concentration of 27.7 mg/L was almost equal to which in shaken culture.CONCLUSION:The enlarged fermentation from shaken flask to fermentor was successful.