PEG4000/柠檬酸钠和PEG8000/磷酸钠双水相萃取牛血清白蛋白

Hawraa A. Mahdi, K. Hameed, Abdul-Jabbar A. Ali
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摘要

水两相系统(ATPS)或液-液萃取在生物技术中用于从原料中回收有价值的化合物。在双水相体系中,许多因素影响分配系数K(即上相与下相的蛋白质浓度之比)和回收率(Rec%)。本研究采用聚乙二醇4000/柠檬酸钠(SC)和聚乙二醇8000/磷酸钠(SPH)两种ATPS体系提取牛血清白蛋白(BSA)。在整个研究过程中,通过温度、聚乙二醇(PEG4000和PEG8000)浓度、柠檬酸钠或磷酸钠浓度、pH和氯化钠作为支撑剂的加入等五个参数的影响,研究了纯(BSA)在atp中的Rec%和K的行为。这些因素对牛血清白蛋白的回收率及其分配系数均有不同程度的显著影响。本研究中影响最大的变量是两种体系的PEG浓度。除PEG浓度外,NaCl的稳定作用也是关键因素。随着聚乙二醇浓度的提高,生物分子与聚乙二醇的相互作用变得更加疏水。在31℃、PEG4000浓度为1.5g/10 ml、柠檬酸钠浓度为2.7 g/10 ml、pH为10、NaCl浓度为0.5 M的条件下,PEG4000/SC体系的回收率为98.99%,分配系数为97.69。在31℃、PEG8000浓度1.5 g/10 ml、磷酸钠浓度2.4 g/10 ml、pH 10、NaCl浓度0.5 M条件下,PEG8000/SPH体系的回收率和分配系数分别为98.93%和92.12。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Extraction of Bovine Serum Albumin by Aqueous Two-Phase System Using PEG4000/Sodium Citrate and PEG8000/Sodium Phosphate
Aqueous Two Phase System (ATPS) or liquid-liquid extraction is used in biotechnology to recover valuable compounds from raw sources. In Aqueous Two-Phase Systems, many factors influence the Partition coefficient, K, (which is the ratio of protein concentration in the top phase to that in the bottom phase) and the Recovery percentage (Rec%). In this research, two systems of ATPS were used: first, polyethylene glycol (PEG) 4000/Sodium citrate (SC), and the second, PEG8000/ Sodium phosphate (SPH), for the extraction of Bovine Serum Albumin (BSA). The behavior of Rec% and K of pure (BSA) in ATPS has been investigated throughout the study by the effects of five parameters: temperature, concentration of polyethylene glycol (PEG4000 and PEG8000), the concentration of Sodium citrate or Sodium phosphate, pH, and the addition of sodium chloride as a supporting agent. The recovery percentage of BSA and its partition coefficient are significantly influenced by these factors to various degrees. The most influential variable in this study is PEG concentration for both systems. In addition to the PEG concentration, the stabilizing impact of NaCl is a crucial factor. The interaction between biomolecules and PEG gets more hydrophobic as the PEG concentration is raised. In the first system (PEG4000/SC), the maximum recovery percentage and partition coefficient were 98.99% and 97.69, respectively, at 31°C, PEG4000 concentration 1.5g/10 ml, Sodium citrate concentration 2.7 g/10 ml, pH 10, and 0.5 M NaCl concentration. While in the second system (PEG8000/SPH), the maximum recovery percentage and partition coefficient was 98.93% and 92.12, respectively, at 31oC, PEG8000 concentration 1.5 g/10 ml, Sodium phosphate concentration 2.4 g/10 ml, pH 10, and concentration of NaCl 0.5 M.
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