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›› 2009, Vol. 9 ›› Issue (3): 563-567.

• 生化工程专栏 • 上一篇    下一篇

多变量在线测量条件下的猪α干扰素高效表达

金虎 徐俊 高敏杰 段作营 李震 刘立明 史仲平   

  1. 江南大学生物工程学院工业生物技术教育部重点实验室 江南大学生物工程学院工业生物技术教育部重点实验室 江南大学工业生物技术教育部重点实验室 江南大学生物工程学院工业生物技术教育部重点实验室 上海市农业科学院畜牧兽医研究所 江南大学生物工程学院环境生物技术实验室
  • 收稿日期:2009-01-05 修回日期:2009-03-27 出版日期:2009-06-20 发布日期:2009-06-20
  • 通讯作者: 金虎

Effective Expression of Porcine Interferon-a with On-line Multi-variable Monitoring

JIN Hu GAO Min-jie XU Jun DUAN Zuo-ying LIU Li-ming LI Zhen SHI zhong-ping   

  1. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University The Key Laboratory of Industrial Biotechnology, Southern Yangtze University Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University Animal Husbandry and Veterinary Research Institute, Shanghai Academy of Agricultural Science, Shanghai Municipal Key Laboratory of Agri-Genetics and Breeding Lab of Biotechnology, School of Biotechnology, Southern Yangtze University
  • Received:2009-01-05 Revised:2009-03-27 Online:2009-06-20 Published:2009-06-20
  • Contact: JIN Hu

摘要: 在5 L发酵罐中进行了重组毕赤酵母高密度培养表达猪a干扰素(IFN-a)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-a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200~280 mmol/(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10 g/L)而得以实现. 在此条件下,猪a干扰素最高活性可达6.6×106 IU/mL,是较低OUR的罐发酵及摇瓶条件下的10~300倍.

关键词: 毕赤酵母, 表达, 流加培养, 摄氧速率, 猪a干扰素

Abstract: Expression of porcine interferon-a (pIFN-a) 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-a 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-a antiviral activity reached 6.6×106 IU/mL, which was about 10~300 folds of those obtained under lower OUR in the same bioreactor and the shaking flask experiments.

Key words: Pichia pastoris, expression, fed-batch culture, oxygen uptake rate, porcine interferon-a

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