基因重组酵母工程菌优化表达人源胶原蛋白  

Optimization of Fermentation Conditions of Genetically Engineered Pichia pastoris Expressing Recombinant Human-source Collagen

作　　者：刘斌[1] 郭红丽[1] 钮小松[1] 张小霞[1] 杨树林[1] 

机构地区：[1]南京理工大学环境与生物工程学院,江苏南京210094

出　　处：《南京理工大学学报》2011年第4期558-562,共5页Journal of Nanjing University of Science and Technology

基　　金：国家'863'计划资助项目(2008AA12A218);教育部博士点基金(20093219110013)

摘　　要：通过响应曲面法(RSM)优化巴氏毕赤酵母基因工程菌GS115/pPIC9KG6表达高亲水性重组人源胶原蛋白的摇瓶培养条件。采用Box-Behnken设计(BBD)考察初始诱导pH值、温度和甲醇添加量三个主要影响因子对重组人源胶原蛋白表达量的影响,利用DX7Trial软件设计实验方案,根据该方案设置不同培养条件进行实验并测定相关参数。响应面分析确定的最优因素水平组合为:初始诱导pH值为5.0,诱导温度为28℃,甲醇添加量为2.2%/24 h。通过响应面法优化发酵的内在因素水平,找出最佳条件,为利用其指导在发酵罐中进行高密度发酵生产人源胶原蛋白奠定基础。Response surface methodology（RSM）is used to optimize the fermentation conditions of genetically engineered Pichia pastoris（GS115/pPIC9KG6）expressing highly hydrophilic recombinant human-source collagen.Box-Behnken design（BBD）is introduced to estimate the impact of initial inducement pH,temperature and addition amount of methanol.An experiment is done by setting different culture conditions according to the scheme designed through the DX7Trial software,and the correlational parameters are determined.The RSM analysis result shows that the optimal scale of factors is：initial inducement pH 5.0,temperature 28 ℃,addition amount of methanol 2.2%/24 h.The experiment offers precise conditions for high density fermentation performed in fermenter.

关 键 词：巴氏毕赤酵母 重组人源胶原蛋白 响应曲面法 BOX-BEHNKEN设计 

分 类 号：Q815[生物学—生物工程]