摘要
为了探讨发酵条件对大肠杆菌表达人白细胞介素-11融合蛋白的影响，利用正交实验设计，对工程菌的生长条件和人白细胞介素-11融合蛋白表达进行优化。在摇瓶中研究了培养基中的葡萄糖、蛋白胨、酵母抽提物的浓度、pH及摇床转速、装液量、接种量等。确定了工程菌生长及表达的培养基和培养条件：葡萄糖10g/L，蛋白胨20g/L，酵母抽提物10g/L，pH7．5，接种量10％，装液量10％。摇床转速220r／min及诱导时间为4～5h。然后在BiofloⅢ-5L发酵罐中以优化的发酵条件进行了3批实验，结果表明：工程菌量达到55g／L（DCW），重组人白细胞介素-11融合蛋白表达量为33％左右，为进行中试研究奠定了理论基础。
The optimum fermentation conditions of the genetc engineeing E. coli （pET32/IL-11/BL（DE）21） expressing recombinant human interleukin-11 in the flask were studied by orthogonal experiment design, including the components of culture medium （glucose, tryptone, yeast extract and pH）, shaking speed, medium volume and inoculum concentration. The optimum conditions for cell growth and expression of rh-11 were as follows glucose 10g/L, tryptone 20g/L, yeast extract 10g/L, inoculum concentration 10%, shaking speed 220r/min, medium volume 10% and induction time 4-5h. Based on these data, fed-batch culture was carried out on BiofloⅢ-5L automatic fermentor. The dissolved oxygen was maintained about 20% ; the recombinant were harvested at 4h after induction of IPTG with three continuous batches. The average cell biomass was about 55g/L （DCW） and the average recombinant human Interleukin-11 fusion proton to total bacterium protein was about 33%. The results were helpful for the pilot-scale study of recombinant human interleukin-11 fermentation.
出处
《工业微生物》
CAS
CSCD
北大核心
2006年第1期38-42,共5页
Industrial Microbiology
