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首页> 外文期刊>Separation and Purification Technology >Purification of trypsinogen from bovine pancreas by combining aqueous two-phase partitioning and precipitation with charged flexible chain polymers
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Purification of trypsinogen from bovine pancreas by combining aqueous two-phase partitioning and precipitation with charged flexible chain polymers

机译：通过水相两相分配和沉淀与带电荷的柔性链聚合物的结合从牛胰腺中纯化胰蛋白酶原

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摘要

Studies on the partition and purification of trypsinogen (TRPz) - a precursor of trypsin - from pancreatic homogenate were performed in aqueous two-phase systems (ATPSs) formed by polyethylene glycol of molecular weight 1450 (PEG1450) and sodium citrate pH 8.20. Several variables such as tie line length (TLL), biomass composition and concentration and phase-volume ratio (V_r) were analyzed. Besides, a precipitation step with polyethyleneimine (PEI) - a positively charged flexible chain polymer - was assayed in order to improve the purification process. Nucleic acids were removed without a significant loss of TRPz content by applying a precipitation step on the homogenate with PEI 0.25% (w/w). ATPSs at long TLL showed TRPz recoveries (R_(TRPz)) and purification factors (PF) higher than those at a short TLL. An enhancement of both the TRPz recovery and PF was observed when the V_r was lowered to 0.5. The best selectivity conditions led to attain almost 5-fold purification and 84% yield.
机译：胰蛋白酶原的胰蛋白酶原（TRPz）的分离和纯化是在由分子量1450（PEG1450）和柠檬酸钠pH 8.20的聚乙二醇形成的水两相系统（ATPSs）中进行的。分析了一些变量，例如联系线长度（TLL），生物量组成，浓度和相体积比（V_r）。此外，测定了聚乙烯亚胺（PEI）（带正电荷的柔性链状聚合物）的沉淀步骤，以改善纯化工艺。通过在PEI为0.25％（w / w）的匀浆上进行沉淀步骤，可以去除核酸而不会显着降低TRPz含量。长TLL时的ATPS显示出TRPz回收率（R_（TRPz））和纯化因子（PF）高于短TLL时。当V_r降低到0.5时，观察到TRPz回收率和PF均提高。最佳的选择性条件导致获得几乎5倍的纯化和84％的收率。

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