作　　者： ; ; ; ;

机构地区： 嘉应学院生命科学学院

出　　处： 《工业微生物》 2004年第3期16-19,共4页

摘　　要： 在反应器中研究了转人TNF α基因鱼腥藻 712 0 (Anabaenasp .PCC712 0 ,pDC TNF)的培养。结果表明气升式反应器适合于转基因鱼腥藻的培养。气升式反应器中通气量和光照是主要的影响因素 ,观察到 1L罐中最适通气量为 6 0～ 75L/h ,最适光照强度为 12 0 0lx ,此时在 2 5℃混养 ,光照时间 /黑暗时间为 12h/ 12h ,15d生物量干重大于 3g/L ,TNF表达水平约占总可溶蛋白的2 2 % ,达到了摇瓶培养水平。实验发现添加维生素B130 0 μg/L、B12 2 0 0 μg/L和生物素 4 μg/L时 ,生产周期为 12d ,缩短 2 0 % ,表达水平相同。培养过程通入含有 5 %CO2 的空气 ,能促进生长 ,缩短生产周期 ,但收获生物量不受影响。从添加维生素和通入CO2 的培养结果证明反应器中培养时 ,光照是限制性因素 ,当反应器系统一定时 ,最终生物量有一个最大值 ,如需进一步提高产量 ,必须设法改变光照系统。 The culture of recombinant Anabaena sp. PCC7120 harboring pDC-TNF was studied in bioreactor. It was determined that the recombinant Anabaena sp. PCC7120 grown better in air-lift bioreactor than that in bubble column bioreactor. In air-lift bioreactor, aeration rate and light were primarily effective factors. When aeration rate was 60～75L/hr,light intensity was 1200lx,light/dark cycle was 12h/12h,temperature was 25℃, cell density and TNF expression in the bioreactor were over 3g/L and about 22% respectively, and it was similar to that in shake flask. Adding vitamin B 1 (300μg/L), B 12 (200μg/L) and biotin(4μg/L) to medium,production period in bioreactor was 12 days. It was 20% shorter than that cultured in medium without vitamin. The biomass was greater than that cultured in medium without vitamin for 15 days. TNF expression level remained the original level. When air contained 5% CO 2,the cell growth was promoted, and culture period was shorten. Light is a limited factor when anabaena was cultured in bioreactor. In order to improve the biomass, light specific surface area must be extended.

关 键 词： 生物反应器 转基因 鱼腥藻 基因工程 表达

领　　域： [生物学] [生物学]