摘要
为确立肝素酶Ⅰ的高效生产工艺,利用麦芽糖结合蛋白(MBP)与肝素酶Ⅰ融合性能,通过构建相应的表达质粒pMHS,在大肠杆菌方面实现了肝素酶Ⅰ可溶性表达。通过对LB培养基摇瓶培养E.coliTB1(pMHS)的诱导时机、诱导剂用量以及添加葡萄糖、酵母提取物、乙醇、氯霉素和卡那霉素等一系列培养条件的优化,确定了该可溶性肝素酶融合蛋白MBP-hepA的最佳生产条件。
In order to achieve high production of heparinase Ⅰ , which is the most useful enzyme in production of LMWH （Low molecular weight heparin）,the plasmid pMHS with recombinant heparinase Ⅰ fused to the Cterminal of maltose binding protein （MBP） was constructed and the soluble expression in E. coli was realized. Based on the constructed vector, optimization of the cultivation conditions using LB medium was carried out, including the induction time, the concentration of the inducer, the additions of glucose, yeast extact, ethanol, chloromycetin （Cm） and kanamycin （Kam）
出处
《生物加工过程》
CAS
CSCD
2006年第3期28-32,共5页
Chinese Journal of Bioprocess Engineering
基金
自然科学基金重点项目(20336010)资助
关键词
肝素酶
大肠杆菌
麦芽糖结合蛋白
优化
可溶性表达
heparinase
E. coli
maltose binding protein （MBP）
optimization
soluble expression
