稀释复性法体外复性重组牛凝血酶原_2包涵体.pdfVIP

19 2 No.2 Vol.19 2005 4 Journal of Chemical Engineering of Chinese Universities Apr. 2005 1003-9015(2005)02-0228-05 -2 , , , (, 310027) -2 -2 α- -2 E.coli 2.5mol⋅L− 1 0.7 Triton X-100 8 mol⋅L− 1 0.3 mol⋅L− 1 DTT 2.7 mol⋅L− 1 0.6 mmol⋅L− 1 GSSGGSSG/GSH 0.90.1% PEG6000 0.5 mol⋅L− 1 L-ArgpH 7.4 Na PO 3 4 -2 Echis Carinatus Snake Venom α- 2948 U⋅mL− 1-2 -2 Q51 Q513.2 Q556.9 A Renaturation of Recombinant Bovine Prethrombin-2 in vitro from Inclusion Bodies in E.coli CHEN Jian-liang, GUAN Yi-xin, CUI Zhi-fang, YAO Shan-jing (Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, China) Abstract: Prethrombin-2 is the smallest single-chain precursor to thrombin and has the same molecular weight with thrombin. The recombinant bovine prethrombin-2 was expressed and overproduced in E.coli BL21(DE3) as intracellular insoluble inclusion bodies. After washed by 2.5mol⋅L− 1 urea and 0.7 Triton X-100, solubilized by 8 mol ⋅L− 1 urea and 0.3 mol ⋅L− 1 DTT, the key parameters in the refolding process, such as GSSG concentration, GSSG/GSH, urea concentration, L-Arg concentration and operation temperat