重组大肠杆菌Trx-hPTH蛋白在双水相体系内的初步纯化

Partial Purification of Human Parathyroid Hormone 1-84 as a Thioredoxin Fusion Form Expressed by Recombinant Escherichia coli in Aqueous TwoPhase System

摘要: 用双水相体系萃取技术对重组大肠杆菌表达的人甲状旁腺激素融合蛋白Trx-hPTH进行了初步纯化，考察了盐的种类、PEG的平均分子量及浓度、MgSO4浓度、pH等因素对Trx-hPTH蛋白在PEG-MgSO4体系内分配行为的影响。结果表明：当双水相体系中w(MgSO4)=0.16，w(PEG 1000)=0.20，pH为8.0时，一步萃取后目标蛋白的分配系数为3.4，回收率为88.6%。

Abstract: The partitioning of Trx-hPTH(1-84) (human parathyroid hormone coupled with thioredoxin as a fusion partner) expressed by recombinant E. coli in the aqueous two-phase system(ATPS) comprised of PEG and salts was investigated. The effective factors on partition including the molecular weight and concentration of PEG, the types and concentration of salts and pH were evaluated. The maximum partitioning of the protein (K=3.4，Y=88.6%) was achieved in the ATPS when w(PEG 1000)=0.20 and w(MgSO4)=0.16 with the protein in the top PEG rich phase.