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首页> 外文期刊>Journal of chromatography, B. Analytical technologies in the biomedical and life sciences >Purification of porcine pancreatic lipase by aqueous two-phase systems of polyethylene glycol and potassium phosphate
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Purification of porcine pancreatic lipase by aqueous two-phase systems of polyethylene glycol and potassium phosphate

机译：聚乙二醇和磷酸钾的水相两相体系纯化猪胰脂肪酶

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An aqueous two-phase system (ATPS) was applied for the purification of porcine pancreatic lipase (PPL) from crude PPL using polyethylene glycol (PEG) and potassium phosphate. Phase diagrams for polyethylene glycol (PEG) and potassium phosphate dibasic were determined at room temperature to find an operating region to first form the ATPS. The PPL was preferentially partitioned into the PEG-rich phase in systems with molecular weights of 1000 and 1500 and concentrated in the phosphate-rich phase in systems with PEG of 4000. Moreover, instead of tie line length (TLL), we used a stability ratio without NaCl in the system, and we first applied fluorescence spectroscopy for the protein conformational analysis of the ATPS. The molecular weight of the purified lipase was determined to be approximately 52. kDa by SDS-PAGE. The enzyme was efficiently purified in PEG 1500/potassium phosphate (17/13, %) at a pH of 7.0 at 4. °C. This system obtained an enzyme partition coefficient of 12.7, an extraction efficiency of 94.7% and a purification factor of approximately 4. These results demonstrate that the aqueous two-phase system is a highly efficient method for PPL purification.
机译：将水两相系统（ATPS）用于使用聚乙二醇（PEG）和磷酸钾从粗制PPL中纯化猪胰脂肪酶（PPL）。在室温下确定聚乙二醇（PEG）和磷酸氢二钾的相图，以找到首先形成ATPS的工作区域。在分子量为1000和1500的系统中，PPL优先分配到富含PEG的相中，而在PEG为4000的系统中则优先浓缩在富含磷酸盐的相中。在系统中没有NaCl的情况下，我们首先将荧光光谱用于ATPS的蛋白质构象分析。通过SDS-PAGE确定纯化的脂肪酶的分子量约为52.kDa。在4.°C的pH值为7.0的PEG 1500 /磷酸钾（17/13，％）中有效地纯化了该酶。该系统获得的酶分配系数为12.7，提取效率为94.7％，纯化系数约为4。这些结果表明，水相两相系统是一种高效的PPL纯化方法。

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