Separation and partitioning of Green Fluorescent Protein from Escherichia coli homogenate in poly(ethylene glycol)/sodium-poly(acrylate) aqueous two-phase systems

The partitioning of Green Fluorescent Protein (GFP) in poly(ethylene glycol)/Na-poly(acrylate) aqueous two-phase systems (PEG/NaPA-ATPS) has been investigated. The aqueous two-phase systems are formed by mixing the polymers with a salt and a protein solution. The protein partitioning in the two-phase system was investigated at 25 degrees C. The concentration of the GFP was measured by fluorimetry. It was found that the partitioning of GFP depends on the salt type, pH and concentration of PEG. The data indicates that GFP partitions more strongly to the PEG phase in presence of Na2SO4 relative to NaCl. Furthermore, the GFP partitions more to the PEG phase at higher pH. The partition to the PEG phase is strongly favoured in systems with larger tie-line lengths (i.e. systems with higher polymer concentrations). The molecular weight of PEG is important since the partition coefficient (K) of GFP gradually decreases with increasing PEG size, from K ca. 300-400 for PEG 400 to K equal to 1.19 for PEG 8000. A separation process was developed where GFP was separated from a homogenate in two extraction steps: the GFP is first partitioned to the PEG phase in a PEG 3000/NaPA 8000 system containing 3 wt% Na2SO4, where the K value of GFP was 8. The GFP is then re-extracted to a salt phase formed by mixing the previous top-phase with a Na2SO4 solution. The K-value of GFP in this back-extraction was 0.22. The total recovery based on the start material was 74%. (c) 2008 Elsevier B.V. All rights reserved.

Plasmid DNA partitioning and separation using poly(ethylene glycol)/poly(acrylate)/salt aqueous two-phase systems

Phase diagrams of poly(ethylene glycol)/polyacrylate/Na2SO4 systems have been investigated with respect to polymer size and pH. Plasmid DNA from Escherichia coil can depending on pH and polymer molecular weight be directed to a poly(ethylene glycol) or to a polyacrylate-rich phase in an aqueous two-phase system formed by these polymers. Bovine serum albumin (BSA) and E. coil homogenate proteins can be directed opposite to the plasmid partitioning in these systems. Two bioseparation processes have been developed where in the final step the pDNA is partitioned to a salt-rich phase giving a total process yield of 60-70%. In one of them the pDNA is partitioned between the polyacrylate and PEG-phases in order to remove proteins. In a more simplified process the plasmid is partitioned to a PEG-phase and back-extracted into a Na2SO4-rich phase. The novel polyacrylate/PEG system allows a strong change of the partitioning between the phases with relatively small changes in composition or pH. (C) 2012 Elsevier B.V. All rights reserved.

Protein partitioning in poly(ethylene glycol)/sodium polyacrylate aqueous two-phase systems

The partition of hemoglobin, lysozyme and glucose-6-phospate dehydrogenase (G6PDH) in a novel inexpensive aqueous two-phase system (ATPS) composed by poly(ethylene glycol) (PEG) and sodium polyacrylate (NaPA) has been studied. The effect of NaCl and Na2SO4, pH and PEG molecular size on the partitioning has been studied. At high pH (above 9), hemoglobin partitions strongly to the PEG-phase. Although some precipitation of hemoglobin occurs, high recovery values are obtained particularly for lysozyme and G6PDH. The partitioning forces are dominated by the hydrophobic and electrochemical (salt) effects, since the positively charged lysozyme and negatively charged G6PDH partitions to the non-charged PEG and the strongly negatively charged polyacrylate enriched phase, respectively. (c) 2007 Elsevier B.V. All rights reserved.

Phase Diagrams of the Aqueous Two-Phase Systems of Poly(ethylene glycol)/Sodium Polyacrylate/Salts

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Harmonic identification using parallel neural networks in single-phase systems

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

An analysis of Generalized Symmetrical Components in non sinusoidal three phase systems

This paper presents the study of the so called Generalized Symmetrical Components, proposed by Tenti et. al. to the analysis of unbalanced periodic non sinusoidal three phase systems. As a result, it was possible to establish a proper relationship between such of generalized symmetrical components and Fortescue symmetrical components to the harmonic frequencies that compose a generic periodic non sinusoidal three phase system. © 2011 IEEE.

A new three-phase transformer modeling for three-phase harmonic analysis in distribution systems

This work presents a new three-phase transformer modeling suitable for simulations in Pspice environment, which until now represents the electrical characteristics of a real transformer. It is proposed the model comparison to a three-phase transformer modeling present in EMTP - ATP program, which includes the electrical and magnetic characteristics. In addition, a set including non-linear loads and a real three-phase transformer was prepared in order to compare and validate the results of this new proposed model. The three-phase Pspice transformer modeling, different from the conventional one using inductance coupling, is remarkable for its simplicity and ease in simulation process, since it uses available voltage and current sources present in Pspice program, enabling simulations of three-phase network system including the most common configuration, three wires in the primary side and four wires in the secondary side (three-phases and neutral). Finally, the proposed modeling becomes a powerful tool for three-phase network simulations due to its simplicity and accuracy, able to simulate and analyze harmonic flow in three-phase systems under balanced and unbalanced conditions.

Dextran sulfate/Triton X two-phase micellar systems as an alternative first purification step for clavulanic acid

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

ALGORITHM FOR PROBABILISTIC ANALYSIS of DISTRIBUTION SYSTEMS WITH DISTRIBUTED GENERATION

In this paper an efficient algorithm for probabilistic analysis of unbalanced three-phase weakly-meshed distribution systems is presented. This algorithm uses the technique of Two-Point Estimate Method for calculating the probabilistic behavior of the system random variables. Additionally, the deterministic analysis of the state variables is performed by means of a Compensation-Based Radial Load Flow (CBRLF). Such load flow efficiently exploits the topological characteristics of the network. To deal with distributed generation, a strategy to incorporate a simplified model of a generator in the CBRLF is proposed. Thus, depending on the type of control and generator operation conditions, the node with distributed generation can be modeled either as a PV or PQ node. To validate the efficiency of the proposed algorithm, the IEEE 37 bus test system is used. The probabilistic results are compared with those obtained using the Monte Carlo method.

Phase diagrams of a CTAB/organic solvent/buffer system applied to extraction of enzymes by reverse micelles

A partial pseudo-ternary phase diagram has been studied for the cethyltrimethylammonium bromide/isooctane:hexanol:butanol/potassium phosphate buffer system, where the two-phase diagram consisting of the reverse micelle phase (L-2) in equilibrium with the solvent is indicated. Based on these diagrams two-phase systems of reverse micelles were prepared with different compositions of the compounds and used for extraction and recovery of two enzymes, and the percentage of enzyme recovery yield monitored. The enzymes glucose-6-phosphate dehydrogenase (G6PD) and xylose redutase (XR) obtained from Candida guilliermondii yeast were used in the extraction procedures. The recovery yield data indicate that micelles having different composition give selective extraction of enzymes. The method can thus be used to optimize enzyme extraction processes. (c) 2007 Elsevier B.V. All rights reserved.

Eigenvalue analyses for non-transposed three-phase transmission line considering non-implicit ground wires

This paper presents a method for analyzing electromagnetic transients using real transformation matrices in three-phase systems considering the presence of ground wires. So, for the Z and Y matrices that represent the transmission line, the characteristics of ground wires are not implied in the values related to the phases. A first approach uses a real transformation matrix for the entire frequency range considered in this case. This transformation matrix is an approximation to the exact transformation matrix. For those elements related to the phases of the considered system, the transformation matrix is composed of the elements of Clarke's matrix. In part related to the ground wires, the elements of the transformation matrix must establish a relationship with the elements of the phases considering the establishment of a single homopolar reference in the mode domain. In the case of three-phase lines with the presence of two ground wires, it is unable to get the full diagonalization of the matrices Z and Y in the mode domain. This leads to the second proposal for the composition of real transformation matrix: obtain such transformation matrix from the multiplication of two real and constant matrices. In this case, the inclusion of a second matrix had the objective to minimize errors from the first proposal for the composition of the transformation matrix mentioned. © 2012 IEEE.

Using Universal Line Model (ULM) for Representing Electromagnetic Transients in Three-Phase Transmission Lines

The second-order differential equations that describe the polyphase transmission line are difficult to solve due to the mutual coupling among them and the fact that the parameters are distributed along their length. A method for the analysis of polyphase systems is the technique that decouples their phases. Thus, a system that has n phases coupled can be represented by n decoupled single-phase systems which are mathematically identical to the original system. Once obtained the n-phase circuit, it's possible to calculate the voltages and currents at any point on the line using computational methods. The Universal Line Model (ULM) transforms the differential equations in the time domain to algebraic equations in the frequency domain, solve them and obtain the solution in the frequency domain using the inverse Laplace transform. This work will analyze the method of modal decomposition in a three-phase transmission line for the evaluation of voltages and currents of the line during the energizing process.

5CN05 partitioning in an aqueous two-phase system: A new approach to the solubilization of hydrophobic drugs

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Instantaneous complex power applied to three-phase machines

The theory presented in this paper was primarily developed to give a physical interpretation for the instantaneous power flow on a three-phase induction machine, without a neutral conductor, on any operational state and may be extended to any three-phase load. It is a vectorial interpretation of the instantaneous reactive power theory presented by Akagi et al. Which, believe the authors, isn't enough developed and its physical meaning not yet completely understood. This vectorial interpretation is based on the instantaneous complex power concept defined by Torrens for single-phase, ac, steady-state circuits, and leads to a better understanding of the power phenomenon, particularly of the distortion power. This concept has been extended by the authors to three-phase systems, through the utilization of the instantaneous space vectors. The results of measurements of instantaneous complex power on a self-excited induction generator's terminals, during an over-load application transient, are presented for illustration. The compensation of reactive power proposed by Akagi is discussed and a new horizon for the theory application is opened.

Mitigation of symmetry condition in positive realness for adaptive control

Feasibility of nonlinear and adaptive control methodologies in multivariable linear time-invariant systems with state-space realization (A, B, C) is apparently limited by the standard strictly positive realness conditions that imply that the product CB must be positive definite symmetric. This paper expands the applicability of the strictly positive realness conditions used for the proofs of stability of adaptive control or control with uncertainty by showing that the not necessarily symmetric CB is only required to have a diagonal Jordan form and positive eigenvalues. The paper also shows that under the new condition any minimum-phase systems can be made strictly positive real via constant output feedback. The paper illustrates the usefulness of these extended properties with an adaptive control example. (C) 2006 Elsevier Ltd. All rights reserved.