Experimental studies on phase equilibria of PEG-water-dextran/ammonium sulfate systems and partitioning of albumin into two water-continuous phases

The clouding points and equilibrium concentrations of PEG/dextran/water and PEG /ammonium sulfate/water systems were experimentally determined for different molecular weights of PEG by titration method and direct determination of concentrations. In phase diagrams of PEG-water with ammonium sulfate or dextran, the addition of salt or dextran induced the phase separation of PEG-rich phases near the PEG-lean phases having the different partitioning of PEG. The concentrations of PEG in PEG-rich phases increase as the amounts of PEG or salt and dextran increase, while the concentrations of PEG in aqueous media decrease in any cases. The higher the molecular weight of PEG has, the wider the two-phase regions are. In dextran DT10 systems, the partition coefficients of egg albumin into PEG-lean phases increase with concentrations of dextran and the molecular weight of PEG. In ammonium sulfate systems, the partitioning coefficients showed a maximum, having lower partitioning at the very high and low concentration of salts. It is also observed that as the amounts of albumin increase, the partitioning of albumin into PEG-lean phase increases.     

Improving glutathione extraction from crude yeast extracts by optimizing aqueous two-phase system composition and operation conditions

PEG-Dextran and PEG-salt aqueous two-phase systems (ATPS) have been applied to separate glutathione (GSH) from crude yeast extracts. Single-factor experiments were carried out to determine the important factors influencing the partition coefficient and extraction yield. The effect of PEG molecular weight, phase-forming components, PEG and Dextran concentration, pH value, and temperature on the GSH partitioning behavior in ATPS was investigated. Three factors, Dextran concentration, pH value, and temperature, were confirmed to have significant influence on the partition coefficient and extraction yield. These factors were further analyzed with the aid of central composite rotatable design and response surface methodology. The optimal conditions for GSH extraction in the PEGDextran system were determined, including PEG molecular weight 6,000, 10% PEG concentration, 14% Dextran concentration, pH 5.2, and temperature 32 °C. A high extraction yield (83.55%) of GSH from crude yeast extracts was achieved under these optimized conditions. This work is very helpful for developing one efficient and cost-effective process for the separation and purification of GSH from yeast broths.     

金属螯合双水相亲和分配技术分离纳豆激酶的研究

利用金属螯合亲和双水相分配技术对纳豆激酶的分离纯化进行了研究。考察了双水相系统、聚合物的分子量和浓度、亲和配基加入量、pH值、相比以及生物质加入量等因素对亲和分配的影响。结果表明,双聚合物系统比聚合物／无机盐系统更有利于纳豆激酶亲和分配;pH值和亲和配基加入量是影响分配的关键因素。优化的分配条件为：2．6％聚乙二醇,20．2％羟丙基淀粉,5％亲和配基PEG-IDA—Cu(Ⅱ),相比12,pH8．2,发酵液加入量15％。分配系统放大到100g,仍保持一致的酶活收率(90％)和纯化因子(2.0)。设计了两次分配分离流程,纯化因子达到3．52,总收率为81％。     

Response surface methodology for the evaluation of guanidine hydrochloride partitioning in polymer-salt aqueous two-phase system

The current study employed response surface methodology (RSM) with a face-centered central composite design (CCD) to indicate the essential variables on the partition coefficient of guanidine hydrochloride (GuHCl) in the poly (ethylene glycol) (PEG)-phosphate aqueous two-phase system (ATPS). To evaluate the partition coefficients of GuHCl in the mentioned ATPS, the pH (7.0, 8.5 and 10.0), GuHCl concentration (1.0, 3.5 and 6.0% w/w), PEG molecular weight (2,000, 4,000 and 6,000 gmol^?1) and PEG/potassium phosphate concentrations ratio were selected as independent variables. A quadratic model is suggested to find the impact of these variables. The suggested model has a strong harmony with the experimental data. The results of the model display that the GuHCl concentration and weight percent of the salt in feed have a large and small influence on the GuHCl partitioning.     

Partition of tannery wastewater proteins in aqueous two-phase poly (ethylene glycol)-magnesium sulfate systems: Effects of molecular weights and pH

The partitioning behavior of soluble proteins from tannery wastewater using aqueous two-phase system (ATPS) was investigated. An ATPS polyethylene glycol (PEG)/MgSO₄ was examined with regard to the effects of PEG molecular weight (MW) and concentration, MgSO₄ concentration, pH and NaCl concentration on protein partition and extraction. The partition coefficients measured for soluble proteins were proportional to the difference in PEG concentration between the phases. The MW and concentration of PEG were found to have significant effects on protein partition and extraction with low MW PEG4000 showing the best conditions for the partitioning of protein in PEG+MgSO₄+water system. Sulfate salt was chosen as the phase-forming salt because of its ability to promote hydrophobic difference between the phases. This system was operated at room temperature . Increase in pH of the system increases the partition coefficient of proteins from tannery wastewater. The addition of sodium chloride showed significant influence on the partition coefficient. ATPS comprising PEG4000-magnesium sulfate provided a means for the recovery of proteins from tannery wastewater. The maximum percentage yield of protein extracted is 82.68%.     

丙烯酰胺在聚乙二醇水溶液双水相聚合过程中的单体分配

用改进溴化法对丙烯酰胺（AM）在聚丙烯酰胺（PAM）-聚乙二醇（PEG）-H2O双水相体系中的分配进行了研究,分配系数随PEG浓度、分子量的增加而减少,随PAM浓度、分子量的增大而增大,而随温度的升高先减小后增大。并在此基础上,对AM在PEG水溶液双水相聚合过程中单体在两相的分配进行了研究,考察了PEG浓度、单体浓度、温度对聚合过程中单体分配的影响。     

Affinity Partitioning Of Enzymes Using Unbound Triazine Dyes In Peg/Phosphate System

Abstract

The partitioning of alcohol dehydrogenase glucose-6-phosphate dehydrogenase and hexokinase from pressed bakers' yeast was studied in polyethylene glycol (PEG)/potassium phosphate aqueous two-phase system using unbound reactive triazine dyes as affinity ligands. The various parameters investigated were ligand type and concentration, pH of the system, phase composition of the system, and molecular weight of PEG. It was found that the best ligands gave an increase in partition coefficient of 2 to 3 times over that in the absence of the ligand.     

Affinity Partitioning of Metal Ions in Aqueous Polyethylene Glycol/Salt Two-Phase Systems with PEG-Modified Chelators

Abstract

The systematic partitioning of metal ions in the presence of polyethylene glycol (PEG) chelate derivatives in PEG/salt two-phase systems is described. Results show that the partitioning is enhanced significantly, and this effect is apparently based on the characteristics of binding of the chelate and the nature of the metal ion in addition to the effect of the attached PEG molecule. Previous work with PEG-iminodiacetic acid has been extended with the chelating derivatives PEG-TED [tris(carboxymethyl)ethylene diamine], PEG-TREN-Cm [carboxy-methylated-tris(2-aminoethyl)amine], and PEG-ASP (aspartic acid). Their effectiveness to selectively partition the metal ions is demonstrated and discussed. The partition experiments were performed using two-phase systems of PEG (8000 MW), sodium sulfate, the PEG-chelating derivatives, and different concentrations of copper, nickel, and cobalt as model metal ions. Partition coefficients (ratio of top to bottom metal concentrations) greater than 1 were obtained for the metal ions with all the chelating derivatives tested. Selective enhancement of partitioning was observed in all cases. Based on the experimental results and theoretical principles, the method shows potential as an alternative mode to remove efficiently and selectively metal ions from solutions using aqueous two-phase systems and PEG-chelate derivatives.     

Partitioning of α-lactalbumin and β-lactoglobulin from cheese whey in aqueous two-phase systems containing poly (ethylene glycol) and sodium polyacrylate

Partitioning behavior of the whey proteins α-lactalbumin (α-la) and β-lactoglobulin (β-lg) in aqueous two-phase systems prepared with poly ethylene glycol (PEG) and sodium polyacrilate (NaPA) was investigated as a function of pH and polymer concentrations. It was observed that α-la concentrated in the PEG phase while β-lg concentrated in the NaPA phase. Response surface methodology was applied to optimize protein partitioning and to achieve the best conditions for their fractionation. Thermodynamic analysis based on isothermal titration microcalorimetry indicated that the partitioning of α-la was accompanied by endothermic heat and was entropically driven, while β-lg partitioning was accompanied by exothermic heat and was enthalpically driven at low polymer concentrations and entropically driven at high polymer concentrations. Purification and yield parameters were determined using fresh whey and the results allowed for conclusion of the great applicability of this new system for α-la and β-lg fractionation.     

聚合物－聚合物双水相系统相平衡计算

利用Ｃａｂｅｘｚａｓ模型，对１４个不同平均分子量的聚乙二醇－葡聚糖－水双水相系统相平衡作了计算。与文献值比较，结果尚可。引入相互作用参数后，得到了修正的Ｃａｂｅｚａｓ模型，用此模型关联上述各双水相系统相平衡，精度有显著提高。     

聚合物－聚合物双水相系统相平衡计算

利用Ｃａｂｅｘｚａｓ模型，对１４个不同平均分子量的聚乙二醇－葡聚糖－水双水相系统相平衡作了计算。与文献值比较，结果尚可。引入相互作用参数后，得到了修正的Ｃａｂｅｚａｓ模型，用此模型关联上述各双水相系统相平衡，精度有显著提高。     

Recovery of value‐added globular proteins from tannery wastewaters using PEG – salt aqueous two‐phase systems

Leather processing involves discharge of high‐value soluble globular proteins in the wastewater. The recovery of value‐added products from the wastewaters is gaining more importance in the context of recovery of wealth from waste. The recovery of these globular proteins from tannery wastewater was selected as a practical model system to study the implementation of polyethylene glycol (PEG)‐sulfate aqueous two‐phase systems (ATPS). The partition coefficient of bovine serum albumin is comparable to that of soluble proteins from tannery wastewaters. The influence of concentration of polymer, salt, pH and temperature on the partitioning of soluble proteins from tannery wastewaters has been studied. The PEG6000 + sodium sulfate + water system provide better partitioning of these soluble proteins as compared to PEG6000 + ammonium sulfate system. The maximum protein recovery yield for PEG6000 + sodium sulfate + water system at 20 °C is 92.75%. The influence of temperature indicates the recovery of proteins from tannery wastewater to be better at lower temperature. The findings of these studies raise the potential application of ATPS processes for protein recovery from complex biological systems. Copyright © 2006 Society of Chemical Industry     

Acid-base equilibria of sulfonephthalein dyes in aqueous polymer-surfactant media

The acid-base equilibria of three anionic sulfonephthalein dyes, viz., bromothymol blue (BTB), thymol blue (TB), and cresol red (CR), were studied spectroscopically in aqueous media containing the water-soluble noninonic polymers polyvinyl alcohol (PVA) and polyethylene glycol (PEG) in the presence of an anionic surfactant, sodium dodecyl sulfate (SDS). A partition equilibrium method was used to determine the equilibrium constant of partition of the dyes between the micellar pseudo phase and aqueous phase in the presence of PVA and PEG. The critical aggregation concentrations (CAC) of the surfactants in buffered aqueous systems containing the neutral polymers were also determined. The CAC of the polymer-surfactant systems were found to be lower than the critical micelle concentration of such systems in the absence of polymer, in otherwise identical conditions. The pH-dependent association constants, K _ass, of the sulfonephthalein dyes with the SDS-PVA system increased with the increase in molecular weight of the polymer. The interactions of the dyes with the buffered aqueous SDS-PVA and SDS-PEG systems were found to be endothermic and entropy oriented. In the polymer domain, the head group region of the micelles was more exposed at lower concentrations of the polymer, but under excess polymer concentration they were increasingly shielded, which impaired their electrostatic interaction with the dyes.     

蒲公英总黄酮在聚乙二醇-硫酸铵双水相体系中的分配与提取

蒲公英中内含的黄酮类物质具有较高药用价值,用双水相萃取法提取植物中有效成分是新型提取高附加值生物质的有效方法。本文考察了PEG/(NH4)2SO4双水相体系萃取分离蒲公英总黄酮时聚乙二醇相对分子质量、PEG质量分数、(NH4)2SO4质量分数、温度、pH值5个因素对分配行为的影响,并通过正交实验优化了工艺条件。结果表明最佳双水相提取工艺条件为：(NH4)2SO4质量分数18%,PEG1000质量分数23%,pH值5.34,提取温度25℃,NaCl盐的存在与否对萃取影响很小,蒲公英中总黄酮的提取率可达5.47%。因此,使用双水相法提取蒲公英总黄酮是该类提取技术中一种更加绿色环保和高效的方法。     

Medium composition effects on solute partitioning in solid–liquid two‐phase bioreactors

Biphasic systems such as two‐phase partitioning bioreactors (TPPBs) have been used to alleviate biological inhibition by sequestering inhibitory compounds within an immiscible phase. The use of solid polymer beads as this auxiliary phase provides a fully biocompatible alternative to potentially toxic organic solvents. While guidelines exist for the rational selection of the polymer phase, the effect of the aqueous phase composition on molecular sequestration has not been explored in the literature. This work aims to identify aspects of medium composition that influence the partitioning of target molecules into the sequestering phase. Using benzaldehyde as the target molecule and Hytrel G3548L (DuPont) as the polymer phase, pH, temperature, salt and glucose concentrations, as well as ethanol concentrations, were examined for their effects on the partition coefficient. pH and temperature were observed to have no significant effect on benzaldehyde partitioning. Salt and glucose additions increased the partition coefficient by 173% and 30%, respectively, compared with pure reverse osmosis (RO) water, while increasing ethanol concentration was found to decrease the partition coefficient from 44 ( ± 1.6) to 1 ( ± 0.3). Strategic changes to the aqueous phase can be made to improve affinity of the sequestering phase for target molecules. This provides a simple and cost‐effective method to potentially improve TPPB system performance. Copyright © 2010 Society of Chemical Industry     

蛋白质-磷酸钾-聚乙二醇双水相体系热力学研究

建立了由蛋白质、磷酸氢二钾、磷酸二氢钾、聚乙二醇所组成的双水相体系的平衡浓度分析测定方法,实验测定了6种蛋白质分子在25℃下的液液平衡分配数据,共23组.将Baskir等的吸附晶格模型和Pitzer的长程静电项相结合,扩展用于含电解质-高聚物双水相体系的热力学计算,仅用7组实验数据进行关联,得到2个吸附能参数,预测了其它16组蛋白质分配系数,结果满意。并提出了蛋白质分配系数与双水相成相物质在两相间浓度差的经验关系式。     

PEG/（NH4）2SO4双水相萃取分离茶氨酸的研究

采用聚乙二醇(PEG)/(NH4)2SO4双水相体系萃取分离生产茶多酚所得废液中的茶氨酸,考察了PEG分子量与含量、硫酸铵含量、pH、温度、加盐(KCl、KBr、KI)、茶氨酸含量对双水相及萃取分离茶氨酸的影响。结果表明,PEG/(NH4)2SO4双水相萃取分离茶氨酸的适宜条件是:PEG平均分子量为4 000,质量分数为10%,硫酸铵质量分数为15%,pH约为6,30℃。在此条件下,茶氨酸的分配系数K1=0.16,蛋白质的分配系数K2=0.28,糖类的分配系数K3=9.8,茶氨酸在下相的萃取率为89.5%,可以将茶多酚废液中的茶氨酸与糖类及其他有颜色的杂质分开。     

椰油丙基酰胺甜菜碱/聚乙二醇水溶液的分相研究

Phase separation behavior of cocamidopropyl betaine/water/polyethylene glycol (PEG) system was studied. The effects of concentration and molecular weight of PEG on the phase separation behavior were investigated. Clouding occurred when the con-centration of PEG was large enough in the betaine aqueous solution, and the concentration of PEG at cloud point decreased with the increase of PEG molecular weight for a constant betaine concentration. The bottom phase was the PEG-rich phase, and the upper phase was the betaine-rich phase. The volumetric ratio of PEG-rich phase to betaine-rich phase, at the same difference between the PEG concentration and the one at the cloud point, Ccp (0.1 g•ml－1), decreased as the PEG molecular weight increased and approached 1 for higher PEG molecular weight (about 20000), which was similar to the typical aqueous two-phase system. This volumetric ratio depended on the initial PEG concentration, but independent of PEG molecular weight. The concentration ratio of betaine to PEG in both phases depended on the Ccp, independent of PEG molecular weight.     

黄芩甙在EOPO/盐双水相系统中的分配系数测定及关联

The partition coefficients of baicalin were measured in ethylene oxide and propylene oxide(EOPO)/salt aqueous two-phase systems at 298.15K,It was found that most of baicalin partitioned into EOPO-rich phase.The partition coefficients of baicalin varied from 10 to 120.The effect of various factors,including tie-line lngth,salt composition,molecular weight of EOPO,and solution pH,on the partition behavior was investigated on EOPO/salt systems.Furthermore the partition coefficients of baicalin were correlated using the modified Diamond-Hsu model.Good agreement with experimental data is obtained.The average relative deviations are less than 5.0%.     

PEG-(NH_4)_2SO_4双水相萃取法提取壳聚糖酶的研究

采用PEG-(NH4)2SO4双水相体系直接从Bacillussp.LS发酵液上清液中分离壳聚糖酶。研究了体系中PEG分子量、PEG质量分数、(NH4)2SO4质量分数、NaCl质量分数和pH值对壳聚糖酶分配系数及萃取率的影响。结果表明,室温下双水相萃取最佳条件为:PEG600 20%、(NH4)2SO420%、NaCl 0.1%、pH值6.0,在此条件下壳聚糖酶分配系数达5.91,萃取率达88.7%。