嗜热四膜虫的培养条件优化及其扩大培养研究  

Optimization of Culture Conditions for Large Scale Cultivation of Tetrahymena thermophila in Bioreactor

作　　者：钟秋萍[1,2] 李青[3] 袁冬霞[1] 周志刚[3] 缪炜[1] 

机构地区：[1]中国科学院水生生物研究所水生生物多样性与保护重点实验室,武汉430072 [2]中国科学院大学,北京100049 [3]中国农业科学院饲料研究所,北京100081

出　　处：《基因组学与应用生物学》2014年第2期322-330,共9页Genomics and Applied Biology

基　　金：中国科学院知识创新工程重要方向项目(KSCX2-EW-G-6-4);湖北省自然科学基金杰出青年项目(ZRZ0251)共同资助

摘　　要：嗜热四膜虫是一种良好的异源基因真核表达系统。本研究的目的是建立经济高效的四膜虫扩大培养技术,为四膜虫大量表达生产产物奠定基础。我们以嗜热四膜虫为实验材料,选择价格相对便宜的GSPP培养基,然后对此培养基进行初始pH值、装液量、起始接种密度和转速等条件的优化,进而进行发酵罐分批扩大培养和发酵罐补料扩大培养的研究。我们的研究结果表明:新选择的培养基GSPP的价格是常规培养基SPP价格的1/33,大大节省了成本;以GSPP为培养基,通过摇瓶实验获得的优化发酵条件:初始pH值为7.0、装液量为50%、起始接种密度为6 250 cells/mL和转速为100 r/min。发酵罐分批扩大培养时,获得的最大细胞密度为1.22×106 cells/mL,最大细胞干重2.15 g/L;发酵罐补料扩大培养时,获得的最大细胞密度为2.05×106 cells/mL、最大细胞干重为4.40 g/L。本研究以相对廉价的培养基实现了四膜虫的经济、高密度扩大培养,为四膜虫高效表达产物及工业化生产奠定了坚实的基础。Tetrahymena thermophila is a free-living unicelluar model organism which has long been taken as an excellent heterologous gene expression system in eukaryotic cell. In order to reduce the cost of the culture medium and find the optimal culture conditions for the large scale cultivation of T. thermophila cells in bioreactor, we developed a fermentation medium with lower cost and optimized the culture conditions. Our results revealed that the cost of the newly developed GSPP medium was reduced to 1/33 of the general SPP medium. Additionally, the optimal initial pH value for the large scale cultivation in GSPP medium is 7.0, the liquid volume is 50% of the container, the initial cell inoculum denisty is 6 250 cells/mL, and the rotation speed is 100 r/min. Moreover, by growing Tetrahymena cells in bioreactor, we found that the maximal cell concentration and dry weight produced in batch fermentation can achieve 1.22×10^6cells/mL and 2.15 g/L, respectively. However, in fed-batch fermentation, the maximal cell concentration and dry weight is increased to 2.05×10^6cells/mL and 4.4 g/L, respectively. Therefore, our studies will provide important resources for the low-cost and high-efficient large scale cultivation of high density T. thermophila cells in bioreactor, and also pave the way for using T. thermophila as an eukaryotic expression system in biopharmaceuticals in industry.

关 键 词：嗜热四膜虫 培养条件 优化 扩大培养 补料分批培养 

分 类 号：Q95-3[生物学—动物学]