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不同甲醇流加策略对重组毕赤酵母高密度发酵生产水蛭素的影响

Effects of Different Methanol Feeding Strategy on Hirudin Production in High-density Fermentation by Recombinant Pichia pastoris

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【作者】 周祥山范卫民张元兴

【Author】 ZHOU Xiang-Shan FAN Wei-Min ZHANG Yuan-Xing * (State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China)

【机构】 华东理工大学生物反应器工程国家重点实验室华东理工大学生物反应器工程国家重点实验室 上海200237上海200237上海200237

【摘要】 考察了不同甲醇流加策略对毕赤酵母高密度发酵生产水蛭素的影响。溶氧控制法不能有效地防止甲醇的过量流加。气相色谱离线检测法虽然防止了甲醇流加过量 ,但甲醇浓度的波动较大。利用甲醇传感器在线检测控制甲醇的流加可维持较恒定的甲醇浓度。在流加甲醇的同时 ,以限制性速率流加甘油可以增加表达期间的能量供应 ,提高产物的表达量。经优化后 ,采用甲醇甘油混合流加时细胞干重达到 16 2g L ,水蛭素活性达到 2 4×10 4ATU mL ,即 1 7g L。

【Abstract】 Four different methanol feeding modes were evaluated in the hirudin producton in high-density fermentation by Pichia pastoris. It was difficult to avoid methanol excessive in the broth with the feeding strategy only based on DO level. On the other hand, the fluctuation in methanol concentration was observed with methanol feeding strategy by off-line gas chromatography. However, the stable methanol concentration was perfectly achieved by the on-line monitoring with methanol sensor. The supply of energy was improved by feeding glycerol at a limited rate as well as methanol in the induction phase. Therefore, the high cell dry weight (162g/L) and high hirudin activity (2.4×10 4ATU/mL or 1.7 g/L) was obtained in the fed-batch fermentation of recombinant Pichia pastoris by methanol-glycerol mixed feeding.

【关键词】 毕赤酵母水蛭素甲醇流加发酵
【Key words】 hirudinPichia pastorismethanolfed-batchfermentation
  • 【文献出处】 生物工程学报 ,Chinese Journal of Biotechnology , 编辑部邮箱 ,2002年03期
  • 【分类号】Q939.9
  • 【被引频次】70
  • 【下载频次】657
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