摘要
在5L发酵罐中进行了重组毕赤酵母高密度培养表达猪α干扰素（IFN-α）的研究.在诱导阶段，高且稳定的摄氧速度（OUR）是实现毕赤酵母流加培养高效表达IFN-α的最关键因素，利用多变量在线测量可有效地帮助寻找高且稳定的OUR的形成条件.结果表明，高且稳定的OUR[约200-280mmol/（L·h）]可通过适度控制混加期及诱导期的菌体比增殖速度和甲醇浓度（约10g/L）而得以实现.在此条件下，猪α干扰素最高活性可达6.6×10^6IU/mL，是较低OUR的罐发酵及摇瓶条件下的10-300倍.
Expression of porcine interferon-or （plFN-ct） under high density recombinant Pichia pastoris cultivation was conducted in a 5 L batch-scaled bioreactor. The results indicated that a high and stable oxygen uptake rate （OUR） during induction phase was the most important factor in achievement of effective pIFN-α production. On-line measurements of multi-variables would help to interpret the reasons for the formation of high and stable OUR. The high and stable OUR [200-280 mmol/（L·h）] could be realized by adjusting specific growth rate with a proper substrate feeding strategy during transient phase, followed by controlling methanol concentration at a moderate level （around 10 g/L） during the induction phase. Under high and stable OUR condition, the highest pIFN-α antiviral activity reached 6.6×10^6 IU/mL, which was about 10-300 folds of those obtained under lower OUR in the same bioreactor and the shaking flask experiments.
出处
《过程工程学报》
CAS
CSCD
北大核心
2009年第3期563-567,共5页
The Chinese Journal of Process Engineering
基金
国家重点基础研究发展规划(973)基金资助项目(编号:2007CB714303)
国家高技术研究发展计划(863)基金资助项目(编号:2006AA020101)
国家自然科学基金资助项目(编号:60604028)
上海科学技术委员会农业科技重点基金资助项目(编号:073919108)
关键词
毕赤酵母
表达
流加培养
摄氧速率
猪Α干扰素
Pichiapastoris
expression
fed-batch culture
oxygen uptake rate
porcine interferon-α
