重组巴斯德毕赤酵母高密度培养中铵离子浓度的影响  

Effect of Ammonium Concentration on the Expression of Angiostatin in High Cell Density Culture of Recombinant Pichia pastoris

作　　者：谢静莉[1] 周庆玮[2] 杜鹏[2] 甘人宝[2] 叶勤[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237 [2]中国科学院上海生命科学研究院生物化学与细胞生物学研究所,上海200031

出　　处：《华东理工大学学报（自然科学版）》2004年第6期723-726,共4页Journal of East China University of Science and Technology

基　　金：教育部科学技术研究重点项目(99166);上海市重点学科基金

摘　　要：采用MutS表型的重组毕赤酵母生产血管生长抑制素,表达阶段流加甘油-甲醇混合碳源以提高菌体密度和血管生长抑制素的表达水平,菌体密度可达174g/L,约是表达阶段采用甲醇为单一碳源的发酵过程的3倍。菌体密度的提高导致表达阶段发酵液中铵离子浓度下降很快,当发酵液中的铵离子浓度低至40mmol/L时,影响了血管生长抑制素的表达。改变pH调节方式并在发酵后期添加25mmol/L(NH4)2SO4使发酵液中铵离子浓度维持在150mmol/L以上,血管生长抑制素的表达产量达到108mg/L。In fed-batch cultivation of recombinant Pichia pastoris (Mut^S phenotype), feeding with glycerol and methanol was conducted during the expression phase to enhance the cell growth and angiostatin expression. The cell density reached 174 g/L at the end of fermentation, which was about 3 fold of that obtained with methanol as the sole carbon source in the induction phase. High cell density resulted in a quick drop of ammonium concentration in the fermentation broth, and when the ammonium concentration was below 40 mmol/L, the production of angiostatin also decreased. The change of pH control strategy and addition of 25 mmol/L (NH_4)_2SO_4 were applied to release the effect caused by low ammonium concentration. The ammonium concentration was maintained above 150 mmol/L in the induction phase, and 108 mg/L angiostatin was achieved at the end of fermentation.

关 键 词：巴斯德毕赤酵母 血管生长抑制素 发酵 铵离子浓度 

分 类 号：TQ920.6[轻工技术与工程—发酵工程]