摘要
通过对发酵基质和发酵关键参数的优化，确定了发酵培养基的磷酸盐浓度为０．１５Ｍ，甘油浓度为１．２ｍｌ／Ｌ，补料中甘油浓度为２０ｍｌ／Ｌ，发酵过程中溶解氧控制在３０％～６０％，ｐＨ控制在６．８５左右。在５Ｌ在ＮＢＳ－Ｂｉｏｆｌｏ３０００型自动控制发酵罐中采取加速补料的补料分批培养，重组大肠杆菌ＹＫ５３７／ｐＳＢ－ＴＫ经１０ｈ３０°Ｃ培养和５ｈ４２°Ｃ诱导培养，最终密度达到６０ＯＤ６００，ｒｈＴＮＦα－ＤＫ２的表达量占菌体总蛋白的５０％以上，每升发酵液纯化可得到近２ｇ的ｒｈＴＮＦα－ＤＫ２。
he fermentation medium and other essential parameters were optimized. It was determined that the concentration of phosphate and glycerol were 0.15 M and 1.2 ml/L in the fermentation medium, and 20 ml/L in the supplied medium respectively. In the process, dissovled oxygen concentration was kept at 30-60%, pH at 6.85, with accelerated fedbatch cultivation in NBS Bioflo3000 type autocontrolled fermentor.After 10 h cultivation at 30°C and 5 h induction cultivation at 42°C, the final cell density reached 60 OD600, rhTNFaDK2 was more than 50% of the total amount of protein in E.coli, near 2 g pure rhTNFαDK2 per liter broth was obtained.
出处
《工业微生物》
CAS
CSCD
1998年第2期20-25,共6页
Industrial Microbiology
关键词
补料分批培养
人肿瘤坏死因子
高密度培养
Fedbatch culture, Human tumor necroses factor, High density cell culture
