文献类型：期刊文献

中文题名：重组巴斯德毕赤酵母高密度培养中铵离子浓度的影响

英文题名：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：[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

年份：2004

卷号：30

期号：6

起止页码：723

中文期刊名：华东理工大学学报（自然科学版）

外文期刊名：Journal of East China University of Science and Technology

收录：CSTPCD;;EI(收录号：2005068830097);Scopus;北大核心:【北大核心2000】；CSCD:【CSCD2011_2012】；

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

语种：中文

中文关键词：巴斯德毕赤酵母;血管生长抑制素;发酵;铵离子浓度

外文关键词：Pichia pastoris; angiostatin; fermentation; ammonium concentration

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.