摘要
对表达人骨形成蛋白 2成熟肽的基因工程大肠杆菌E .coliDH5α pDH B2 m在 5 0 0mL摇瓶中进行了培养条件的摸索实验 ,并在此基础上扩大至NBSBiofloIV 2 0L发酵罐 ,利用溶氧反馈 -分批补料培养技术 :在培养过程中保持适当的溶解氧 (4 0 % ) ,以溶氧值在线反馈控制搅拌速度及流加补料培养基 ,使细菌保持适当的比生长率 ,成功地进行了工程菌的高密度培养 ,最终菌体密度达OD60 0 =5 7,每升干菌量 2 2 .8g ,目的蛋白的表达量占细菌总蛋白的30 % ,人骨形成蛋白 2成熟肽的理论产率达到 3.5 9g L。
Optimization of cultivation condition of recombinant E.coli DH5α/pDH-B 2m and the condition suitable for expression of recombinant mature peptide of human bone morphogenetic protein-2 was carried out in 500mL shaking flasks and then transferred to NBS Bioflo IV, a 20L DO feed-back fed-batch culture system, to obtain rhBMP-2. The results indicate that keeping dissolved oxygen at 40% and controlling nutrient feeding rate with DO feed back strategy can obtain theoretically 3.59g recombinant mature peptide of hBMP-2 per liter of broth, the final cell density OD 600 reaches 57(22.8g dry cell weight/L), and the expression of rhBMP-2 amounts to 30% of the total protein in E. coli.
出处
《生物工程学报》
CAS
CSCD
北大核心
2002年第6期718-723,共6页
Chinese Journal of Biotechnology
基金
全军医药卫生科研基金重点课题 (No .912 0 0 4
9130 10
96z0 45 )
国家重点科技攻关项目 (No .85 72 2 0 1 0 8)~~
