Procion Yellow HE-3G Research Articles

Investigation of the usability of zinc ferrite nanoparticles synthesized by microwave assisted combustion method as photocatalyst for removal of organic dyes from wastewaters

In this study, zinc ferrite nanoparticles, which has an important place among the spinel ferrite structured nanomaterials due to its unique properties, were synthesized by microwave-assisted combustion method and later were used as photocatalysts in the removal of dyestuffs by photocatalytic degradation method from wastewaters of textile industry. In the synthesis studies, it was determined that the microwave effect alone was not sufficient to complete the transformation. Heat treatment application is envisaged to solve this problem and in order to determine the optimum heat treatment temperature, the sample produced by microwave effect were subjected to heat treatment at 300℃, 400℃, 500℃, 600℃, 700℃, 800℃ and 900℃, respectively. It has been observed that the heat treatment has a significant effect on the crystal structure of the particles and 700℃ has been determined as the optimum heat treatment temperature. The data obtained showed that, under these conditions, the zinc ferrite nanoparticles were successfully synthesized and the powder produced completely consisted of nano-sized particles. Moreover, results showed that the synhesized zinc ferrite nanoparticles has a saturation magnetization value sufficient to separate them from the aqueous medium. Finally, zinc ferrite nanoparticles produced under optimum conditions were used as photocatalysts in the removal of textile dye known as Procion Yellow HE-3G from wastewater by photocatalytic oxidation. In photocatalysis experiments, it was observed that synthesized zinc ferrite nanoparticles provided very high removal efficiencies as photocatalysts and almost all of the dye content in the solution could be removed.

The effects of fuel type onto the structural, morphological, magnetic and photocatalytic properties of nanoparticles in the synthesis of cobalt ferrite nanoparticles with microwave assisted combustion method

In this experimental study, the synthesis of cobalt ferrite nanoparticles, one of the most important types of ferromagnetic spinel ferrite materials was realized by using a method known as microwave assisted combustion method. The effects of the substance used as fuel on the structural, morphological, magnetic and photocatalytic properties of synthesized nanoparticles were investigated. Firstly, samples were prepared using urea, glycine and citric acid as fuel and then the samples were comprehensively characterized by advanced analysis techniques such as XRD, FT-IR, TEM, SEM, VSM, BET. Moreover, the usability of the obtained nanoparticles in photocatalytic degradation process of Procion Yellow H-E3G textile dye as photocatalyst was investigated. The results obtained indicate that the fuel used is highly effective on various properties of the synthesized materials. From the experiments performed to evaluate the photocatalytic performances of the nanoparticles, it was observed that each of the three different fuel matters used for the synthesis of nanoparticles were highly effective on the photocatalytic degradation of the organic pollutant. Furthermore, almost all the dye content in solution was completely removed at the end of the experimental period which lasted for 90 min.

Modification of polystyrenic matrices for the purification of proteins: III. Effects of poly(vinyl alcohol) modification on the characteristics of protein adsorption on conventional and perfusion polystyrenic matrices

Poly(styrene–divinylbenzene) (PS–DVB) chromatography matrices, CG1000sd 20–50 μm (TosoHaas), PLRP4000s 15–25 μm, PLRP4000s 50–70 μm (Polymer Laboratories) have been modified by the adsorption and crosslinking of poly(vinyl alcohol) (PVA) to create a matrix suitable for the attachment of dye ligands. The adsorption capacities of lysozyme and HSA on these Procion Yellow HE-3G dyed PVA modified PS–DVB matrices were measured at various flow-rates and the capacities were compared with a Procion Yellow HE-3G dyed OH-activated POROS 20, 20-μm matrix (PerSeptive Biosystems). The adsorption of small proteins was not hindered by the smaller pores of the CG1000sd beads, but as protein size increased, and at high flow-rates, a high mass transfer rate became more dependent on large pore size and small particle diameter.

Expanded bed affinity chromatography of dehydrogenases from bakers' yeast using dye–ligand perfluoropolymer supports

Malate dehydrogenase (MDH) and glucose 6-phosphate dehydrogenase (G6PDH) have been partially purified from preparations of homogenized yeast cells using Procion Yellow H-E3G and Procion Red H-E7B, respectively, immobilized on solid perfluoropolymer supports in an expanded bed. A series of pilot experiments were carried out in small packed beds using clarified homogenate to determine the optimal elution conditions for both MDH and G6PDH. Selective elution of MDH using NADH was effective but the yields obtained were dependent on the concentration of NADH used. Selective elution was found to be most effective when a low concentration of NaCl (0.1 M) was present. MDH could be recovered in 84% yield with a purification factor of 94 when this strategy was adopted. In the case of G6PDH, specific elution using NADP(+) was successful in purifying G6PDH 178-fold in 96% yield. The dynamic capacity of both affinity supports was estimated by frontal analysis, in an expanded bed with unclarified homogenate, and corresponded to 17 U MDH/mL of settled Procion Yellow H-E3G perfluoropolymer support and 7.7 U H6PDH/mL of settled Procion Red H-E7B perfluoropolymer support. Expanded bed affinity chromatography of MDH resulted in an eluted fraction containing 89% of the applied activity with a purification factor of 113. Expanded bed affinity chromatography of G6PDH resulted in an eluted fraction containing 84% of the applied activity with a purification factor of 172. With both enzymes, the overall recovery of enzyme activity was greater than 94%, showing that the expanded bed approach to purification was nondenaturing.

Isolation and biochemical characterization of highly purified Escherichia coli molecular chaperone Cpn60 (GroEL) by affinity chromatography and urea-induced monomerization

Isolated Escherichia coli molecular chaperone Cpn60 (GroEL) has been further purified from tightly bound substrate polypeptides by two different procedures: (i) group-specific affinity chromatography by using the triazine dye Procion yellow HE-3G as affinity ligand, and (ii) urea-induced monomerization and subsequent chromatography. Procion yellow binds specifically to aromatic amino-acid side chains present in the majority of proteins, but has no affinity to GroEL because of its low content of aromatic residues. Some GroEL-bound polypeptides are buried within the aqueous cavity of the GroEL oligomer, whereas others are exposed on its surface and available for affinity-ligand interactions and the complex is thereby retarded on Procion yellow columns. Pure substrate-free GroEL was obtained after ion-exchange chromatography of GroEL monomers followed by reassembly of the purified monomers into functional GroEL oligomers. The final preparation contained no substrate polypeptides bound to GroEL as judged by electrophoretic analysis and lack of tryptophan fluorescence. GroEL preparations also displayed two equally strong bands on native electrophoresis suggesting the presence of two conformers. Monomers of GroEl showed heterogeneity with respect to isoelectric point and molecular mass when analysed by MALDI-MS and electrophoresis under native and denaturing conditions respectively. By use of MALDI-MS, highly accurate molecular masses of wild-type and a truncated form of GroEL were determined and verified, by comparison with their respective gene sequences.

Interactions in affinity partition studied using fluorescence spectroscopy

Fluorescence titration has been used to determine the binding constant and number of binding sites for the textile triazine dye Procion Yellow HE-3G to lactate dehydrogenase from rabbit muscle (E.C. 1.1.1.27). Triazine dye was either free in solution or attached to one of the polymer carriers, polyethylene glycol or dextran. Titrations were performed in solutions of buffer, dextran, and polyethylene glycol. Aqueous two-phase systems composed of polyethylene glycol and dextran were prepared and the binding constant and number of binding sites for ligand polyethylene glycol-Procion Yellow to lactate dehydrogenase were determined in both upper and lower phases of these systems. Affinity partition of lactate dehydrogenase in a PEG-dextran system was also performed using PEG-Procion Yellow as ligand, and partition coefficients of lactate dehydrogenase showed good agreement with theoretical partition coefficients calculated from the binding constant and number of binding sites obtained from fluorescence titration. The advantage of using fluorescence titration to determine affinity of a polymer ligand for a protein is that measurement of binding strength can be made in the actual environment encountered by protein-ligand complex during the purification process.

Interactions of human alkaline phosphatase isoenzymes with triazine dyes using affinity partitioning, affinity chromatography and difference spectroscopy

Aqueous two-phase systems consisting of dextran, polyethylene glycol and dye-liganded polyethylene glycol were employed to investigate the affinity partitioning behaviour of isoenzymes of human alkaline phosphatase. Whereas in the system without a dye ligand the partition coefficients of the isoenzymes from human intestine and placenta were identical, the isoenzyme from human liver showed a significantly lower partition coefficient under the same conditions. After addition of dye-liganded polyethylene glycol two groups of dyes possessing substantial affinities to the isoenzymes were found. One, represented by Procion Yellow HE-3G, interacts specifically with the active centre of the isoenzymes. Differences in the affinity of the isoenzymes towards the individual dye ligands are caused only by the carbohydrate content, especially by the terminal sialic acid residues. The other group of dye ligands, represented by Procion Navy MX-RB, binds obviously in a more complex fashion involving other binding sites, which are only present in alkaline phosphatase of human liver. Procion Navy MX-RB was found to function as a suitable affinity ligand for the separation of human liver alkaline phosphatase from the other isoenzymes. Differences in the primary structure of two allelic forms of human placental alkaline phosphatase [(SS) and (F)] are not recognized in aqueous two-phase systems with or without dye-liganded polyethylene glycol.

Subfractions of membranes from calf brain synaptosomes obtained and studied by liquid—liquid partitioning

Synaptosomes isolated from calf brain cortex were lysed and fragmented by Yeda press treatment. The obtained membranes have previously been fractionated in a counter-current distribution process using a liquid—liquid two-phase system consisting of water, dextran, Ficoll and poly(ethylene glycol) [ J. Chromatogr., 358 (1986) 147]. Using the fact that there are discrete membrane populations, a rapid preparative method for isolation of the two main fractions is presented in the present work, as well as a subfractionation of one of them using liquid—liquid extraction with dextran-bound Procion yellow HE-3G. The content of several membrane constituents, i.e. protein, acetylcholinesterase, succinate dehydrogenase and ATPase, as well as opiate binding, were determined for the three fractions. Counter-current distribution of the fractions elucidates their heterogeneity and the effectiveness of the purification.

Heterogeneity of synaptosomal membrane preparations from different regions of calf brain studied by partitioning and counter-current distribution

1. 1. Membranes obtained by lysis and Yeda-press treatment of synaptosomes (nerve endings) from cortex, caudateus nucleus, and hippocampal region of calf brain have been studied by partitioning within a liquid-liquid aqueous two-phase system consisting of water, dextran, Ficoll, and poly(ethylene glycol). 2. 2. The partitioning of membranes was sensitive to the presence of a dextran-bound dye, Procion yellow HE-3G, in the lower phase. 3. 3. The two-phase system was used for counter-current distribution to study the heterogeneity of the synaptic membranes from the three regions of the brain and to separate the membranes into fractions. 4. 4. The obtained counter-current distribution profiles strongly depended on the region of the brain from which the membranes were isolated. 5. 5. The membrane fractions obtained showed marked differences in their SDS electrophoresis pattern.

Combination of polymer-bound charged groups and affinity ligands for extraction of enzymes by partitioning in aqueous two-phase systems

The partition of glucose-6-phosphate dehydrogenase in a water—dextran—polyethylene glycol two-phase system was influenced by the mutual addition of polymers carrying charged groups and affinity ligands. These polymers included DEAE-dextran, dextran sulphate, ligand—dextran, ligand—polyethylene glycol, ligand—Ficoll and ligand—DEAE-dextran, where the ligands used were the dyes Cibacron Blue F3G-A and Procion Yellow HE-3G. Some of these two-phase systems were also used for the counter-current distribution of yeast proteins, demonstrating the separation of several enzymes.

Affinity liquid‐liquid extraction of lactate dehydrogenase on a large scale

Rapid liquid-liquid extraction of lactate dehydrogenase from muscle by using a low-cost aqueous bipolymer two-phase system was achieved by using a centrifugal separator. Extraction of the target enzyme into the upper phase was enhanced by including the dye Procion yellow HE-3G (bound to polyethylene glycol). The dye acted as an affinity ligand for the enzyme. The isolation of the enzyme was carried out either by using a cell extract or by homogenizing the muscle directly in the system. The latter approach reduced the preparation time with a factor of 0.25. The two methods gave, respectively, 310 and 360 kU lactate dehydrogenase/kg muscle (measured at 22 degrees C). By using a small centrifugal separator, Alfa Laval LAPX 202, 3-5 kg muscle could be processed/h in a 30-L, two-phase system.

Effect of polymer structure on affinity partitioning of lactate dehydrogenase in polymer—water two-phase systems

Lactate dehydrogenase from rabbit muscle (EC 1.1.1.27) was partitioned between the liquid phases of aqueous two-phase systems containing two kinds of polymers. The partition was influenced by the use of a dye ligand, Procion Yellow HE3G, bound to a polymer. The ligand-carrying polymers were poly(ethyleneglycol), poly(vinyl alcohol), ethylhydroxyethylcellulose, hydroxypropyl-starch, Ficoll and dextran. Their influence on the partition of lactate dehydrogenase was studied in systems composed of the following pairs of polymers: dextran—poly(ethylene glycol), dextran—poly(vinyl alcohol), dextran—Ficoll and hydroxypropyl-starch—poly(ethylene glycol). The ratio between the change in the logarithmic partition coefficient of the enzyme, Δ log K, caused by the ligand and the logarithm of the partition coefficient of the ligand—polymer, log K L—polymer, was used as a measure of the number of ligand polymers bound per enzyme molecule under saturation conditions. Possible explanations for the observed effects are discussed.

Affinity partitioning of enzymes using dextran-bound procion yellow HE-3G : Influence of dye-ligand density

The dye Procion Yellow HE-3G was bound to dextran of molecular weight 70 000 and the partitioning of this dye-polymer within an aqueous two-phase system containing dextran and poly(ethylene glycol) was studied as function of ligand density, polymer concentration, type of salt, concentration of salt and concentration of dye-dextran. Even moderate dye:dextran molar ratios (5–8) make the partitioning strongly salt-dependent. The dye-dextran can be directed to either the upper or the lower phase with partition coefficients from 0.02 to 28 by using salts. The dye-dextran in the two-phase system affects the partitioning of dye-binding enzymes (lactate dehydrogenase, glucose-6-phosphate dehydrogenase, 3-phosphoglycerate kinase) towards the dye-containing phase. Measurements of competition with nucleotide binding show an increased affinity of the dye for the enzyme with increasing ligand:dextran ratio. Theoretical considerations indicate that 1–2 dextran molecules are attached per enzyme molecule when affinity partitioning is fully developed.

Specifically increased solubility of enzymes in polyethyleneglycol solutions using polymer-bound triazine dyes

The enzymes glucose-6-phosphate dehydrogenase (EC 1.1.1.49) and 3-phosphoglycerate kinase (EC 2.7.2.3), present in an extract of Bakers' yeast, are largely kept insolution by minor amounts of polyethylene glycol-bound triazine dyes (Procion yellow HE-3G and Procion olive MX-3G) even when the solution contains such concentrations of polyethylene glycol (12.5% w w ) which normally precipitate the enzymes. The specific prevention from precipitation can be used for purification of enzyme, preferentially in dealing with crude extracts, which has been demonstrated in this work. A 3.4-fold purification of glucose-6-phosphate dehydrogenase has been achieved with good recovery (93%). Further purification has been possible by combining the recovered (enzyme-containing) supernatant liquid with a solution of dextran which generates an aqueous two-phase system. The lower, dextran-containing phase extracts part of the remaining bulk proteins leaving the target enzyme in the upper phase. The advantages of this method for enzyme purification in large scale are discussed.

Liquid-liquid extraction of lactate dehydrogenase from muscle using polymer-bound triazine dyes.

An extract from porcine muscle containing soluble enzymes has been partitioned between the two liquid phases of an aqueous, biphasic system consisting of dextran, polyethylene glycol, and water. The influence of polymer-bound triazine dyes (Cibacron blue F3G-A and Procion yellow HE-3G) on the partition of lactate dehydrogenase and total protein was studied. The effects of pH and concentrations of polymers and buffer on this so-called affinity partitioning were also determined. The two-phase systems were used in extraction procedures for purification of lactate dehydrogenase to a specific activity of 456-494 U (7.6-8.4 mukat) per mg protein. The use of these systems for extraction of enzymes in technical scale is discussed.

Effect of dextran- and poly(ethylene glycol)-bound procion yellow HE-3G on the partition of membranes from calf brain synaptosomes within an aqueous two-phase system

Membranes obtained by lysis and Yeda-press treatment of synaptosomes (nerve endings) from calf cortex have been partitioned within the aqueous phases (and the interfaces between them) of a Ficoll—dextran—poly(ethylene glycol)— water two-phase system. By introducing the dye Procion yello HE-3G in the upper phase, bound to poly(ethylene glycol), or in the lower phase, bound to dextran, the partition of the membranes could be strongly affected. The influence on the partition was more pronounced when the dye was bound to dextran. By using a number of two-phase systems in a counter-current distribution process, it was shown that the membrane preparation was inhomogeneous and that the fractions obtained differed in their contents of acethylcholinesterase, succinate dehydrogenase and ATPase. The affinity partitioning effect depended strongly on the concentration of polymer-bound dye. An optimum dye concentration was found when Procion yellow HE-3G was bound to poly(ethylene glycol). When the same dye was bound to dextran, the number of dye molecules per dextran molecule influence the effectiveness of the extraction.

Partial purification of d-glucose-6-phosphate dehydrogenase from bakers' yeast by affinity partitioning using polymer-bound triazine dyes

d-Glucose-6-phosphate dehydrogenase ( d-glucose-6-phosphate:NADP + 1-oxidoreductase EC 1.1.1.49) has been purified from bakers' yeast by liquid-liquid extraction using phase-restricted triazine dyes (Procion Yellow HE-3G, Procion Olive MX-3G, Procion Navy MX-RB and Cibacron Blue F3G-A). This method was combined with fractional precipitation with poly(ethylene) glycol) and batchwise treatment with DEAE-cellulose. This rapid procedure gave an enzyme preparation with a specific activity of 0.92 kat per kg protein within 5 h. The affinity extraction step can easily be scaled up and the good recovery of ligand-poly(ethylene glycol) should make the process useful for larger amounts of enzyme. The technical possibilities are discussed.

Counter-current distribution of yeast enzymes with polymer-bound triazine dye affinity ligands

A protein extract from bakers' yeast has been resolved into several of its enzyme components by counter-current distribution using aqueous polymeric two-phase systems comprising water, dextran and polyethylene glycol. The distribution of dehydrogenases and kinases has been effectively changed by binding triazine dyes (Cibacron Blue F3G-A, Procion Yellow HE-3G and Procion Olive MX-3G) to the polyethylene glycol, confined to the upper phase. The effect of the dye ligands on the enzymes decreased roughly in the order: phosphofructokinase, (EC 2.7.1.11), glucose 6-phosphate dehydrogenase, (EC 1.1.1.49), glyceraldehydephosphate dehydrogenase, (EC 1.2.1.12), alcohol dehydrogenase (EC 1.1.1.1), 3-phosphoglycerate kinase (EC 2.7.2.3), phosphoglycerate mutase (EC 2.7.5.3.), hexokinase (EC 2.7.1.1) and enolase (EC 4.2.1.11). However, the distribution of specific enzymes was strongly dependent also on the dye concentration. By appropriate selection of conditions, purifications of up to 10-fold could be achieved for phosphofructokinase and 100-fold for one of the isoenzymes of hexokinase. Furthermore, the two isoenzymes of hexokinase could be almost completely separated from each other. The counter-current distributions were performed in a newly developed apparatus where the separation of phases was enhanced by centrifugation. This allowed 55 transfers to be completed within about 8 h.