毕赤酵母植酸酶基因工程菌高密度培养及诱导表达  

High-level recombinant phytase production by high cell density fermentation of Pichia pastoris

作　　者：王志林[1] 陈庄[1] 朱选[1] 蒋宗勇[1] 

机构地区：[1]广东省农科院畜牧研究所,广东广州510640

出　　处：《广东农业科学》2009年第10期158-161,共4页Guangdong Agricultural Sciences

基　　金：广东省自然科学基金(990508);广东省重点科技项目(2KM02505G)

摘　　要：研究了毕赤酵母基因工程菌高密度发酵培养方式和植酸酶基因高效表达策略。采用逐步增加流加方式进行高密度培养,结果发酵液中细胞干重达到64 g/L。通过在线检测细胞消耗甲醇的MSUR,估算细胞在单位时间内对甲醇需求量,从而确定甲醇流加速率,采用此种方法可使重组毕赤酵母高水平表达植酸酶,最终发酵上清液中总蛋白量为8.58 g/L,植酸酶活力达到853 U/mL。A Pichia pastoris high cell density fermentation and a high-level expression strategy of phytase gene were studied. High cell density fermentation was carried out by a fed-batch fermentation process in stepwise increase in feed rate, and a dry cell weight of 64 g/L was achieved. Based on the on-line measurement of the maximum substrate uptake rate（MSUR）, the methanol feeding rate was determined, and high-level phytase was produced by recombinant Pichia pastoris. At the end of the induction phase the total supernant protein was 8.58 g/L, and the phytase activity reached 853 U/mL.

关 键 词：毕赤酵母基因工程菌 高密度培养 高效表达 

分 类 号：Q93-335[生物学—微生物学]