Partition coefficient prediction of Baker's yeast invertase in aqueous two phase systems using hybrid group method data handling neural network

A hybrid GMDH neural network model has been developed in order to predict the partition coefficients of invertase from Baker's yeast. ATPS experiments were carried out changing the molar average mass of PEG(1500–6000 Da), p H(4.0–7.0), percentage of PEG(10.0–20.0 w/w), percentage of MgSO_4(8.0–16.0 w/w), percentage of the cell homogenate(10.0–20.0 w/w) and the percentage of MnSO_4(0–5.0 w/w) added as cosolute. The network evaluation was carried out comparing the partition coefficients obtained from the hybrid GMDH neural network with the experimental data using different statistical metrics. The hybrid GMDH neural network model showed better fitting(AARD = 32.752%) as well as good generalization capacity of the partition coefficients of the ATPS than the original GMDH network approach and a BPANN model. Therefore hybrid GMDH neural network model appears as a powerful tool for predicting partition coefficients during downstream processing of biomolecules.

MEASUREMENT AND CORRELATION OF PARTITION COEFFICIENTS OF UROKINASE IN AQUEOUS TWO-PHASE PEG-POTASSIUM PHOSPHATE SYSTEM

Partition coefficients of Urokinase(UK)were measured in aqueous two-phase systems con-taining polyethylene glycol and potassium phosphate at 273.2K.Based on the Diamond-Hsu model,a modified expression was obtained for the correlation of enzyme partitioning in theabove-mentioned systems.Utilizing a modified form,the partitioning data of UK and heteroproteinwere correlated.The results show that the modified model is simple gives good precision.and will fa-cilitate engineering scale-up of aqueous two-phase systems for certain proteins purification.

Effective extraction of elastase from Bacillus sp. fermentation broth using aqueous two-phase system

This paper presents the evaluation of an aqueous two-phase system (ATPS) for extracting elastase produced by Bacillus sp. EL31410. The elastase and cell partition behavior in polyethylene glycol (PEG)/salt systems was investigated. The suitable system for elastase extraction was PEG/KH2PO4-K2HPO4, in which elastase is mainly partitioned into the PEG-rich phase, while the cells remained in the other phase. The influence of defined system parameters (e.g. PEG molecular mass, pH, NaCl addition) on the partitioning behavior of elastase is described. The concentration of phase forming components, PEG and KH2PO4-K2HPO4, was optimized for elastase recovery by means of response surface methodology, and it was found that they greatly influenced extraction recovery. The optimal ATPS was 23.1% (w/w) PEG 2 000 and 11.7% (w/w) KH2PO4-K2HPO4. The predicted recovery was about 89.5%, so this process is suggested to be a rapid and convenient method for elastase extraction.

Measurement and Correlation of Equilibrium Data for Aqueous Two-phase System Ethanol+Water+ K_2HPO_4

The isothermal solubility data of aqueous two-phase system ethanol+water+K 2HPO 4 were determined with the turbidity titration method at 303.2 K. The binodal curves were described by using the Mistry equation very well. An experimental procedure for measuring the liquid-liquid equilibrium data of the aqueous two-phase system was proposed, in which the concentrations of the coexisting phases were determined with the corresponding densities of the solution. The tie lines were satisfactorily described by using the Othmer-Tobias and Bancroft equations.

Novel Vesicular Aqueous Two-Phase Systems

The interesting phenopmena of two aqueous phases coexisting in dilute aqueous solutions of sodium undecenoate-dodecyltrimethylammo brodride ndxture and sodium laurate-dodecyltrimethylammonium brondde mixture were investigated. Vesicles existing in both phases were shown by TEM images. The vesicles are dispersed in lower phase and flocculated in upper phase. Multilamellar structure of vesicles was found in the upper phase of sodium laurate-dodecyltrimethylammonium bromide system.

Measurement and Correlation of Partition Coefficients of Baicalin in EOPO/Salt Aqueous Two-Phase Systems

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 length, salt composition, molecular weight of EOPO, and solution pH, on the partition behavior was investigated in 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%.

Measurement of Mass Transfer Coefficients and Their Modelling of Continuous Counter-Current Aqueous Two-Phase System in a Packed Extraction Column

Overall dispersed side volumetric mare transfer coeffcieats for protein and amino acids were measured in continuous countercurreat PEG4000/KHP aqueous two-phase systems in a 57 mm I.D. packed extraction column. A model for overall dispersed side volumetric mass transfer coefficients was derived by describing the motion of the drops based upon Navier-Stokes equation combined with the relationship between mass transfer coefficients and the drop velocity. The model provides good predictions and can be successfully used in aqueous two-phase extraction. The awrage relative deviation between calculated values and experimental date ranges from 8% to 14%.

Partitioning and purification of extracellular β-1,3-1,4-glucanase in aqueous two-phase systems

The partition behaviors of β-1,3-1,4-glucanase, α-amylase and neutral proteases from clarified and whole fermenta-tion broths of Bacillus subtilis ZJF-1A5 were investigated. An aqueous two-phase system (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 enzyme partition and extraction. The MW and concentration of PEG were found to have significant effects on enzyme partition and extraction with low MW PEG showing the greatest benefit in the partition and extraction of β-glucanase with the PEG/MgSO4 system. MgSO4 concentration influenced the partition and extraction of β-glucanase significantly. pH had little effect on β-glucanase or proteases partition but affected α-amylase partition when pH was over 7.0. The addition of NaCl had little effect on the partition behavior of β-glucanase but had very significant effects on the partitioning of α-amylase and on the neutral proteases. The partition behaviors of β-glucanase, α-amylase and proteases in whole broth were also investigated and results were similar to those obtained with clarified fermentation broth. A two-step process for purifying β-glucanase was developed, which achieved β-glucanase recovery of 65.3% and specific activity of 14027 U/mg, 6.6 times improvement over the whole broth.

Theoretical Computation of Nonlinear System Equations of Heavier Pellets Movements for Two Phase Flow

This paper presents nonlinear ordinary differential equations (ODES) of the heavier pellets movement for two phase flow, which actually represent a system of equations. The usual methods of solution such as Runge -Kutta method and it's datum results are discussed. This paper solves ODES of general form using variable mesh-length, linearizing the nonlinear terms by finite analysis method, fuilding an iteration sequence, and amending the nonlinear terms by iteration . The conditions of convergent operation of iteration solution is checked. The movement orbit and velocity of the pellets are calculated. Analysis of research results and it's application examples are illustrated.

Copper(Ⅰ)-catalyzed Synthesis of Aryl, Heteroaryl and Vinyl Sulfides in Aqueous Two-phase System

A fast and convenient C―S bond formation reaction was achieved catalyzed by CuI/L-proline in aqueous two-phase system providing a simple method for synthesis of aryl sulfides in good yields in short time.

Enhancement of Shikonin Production in Cell Suspension Cultures of Arnebia Euchroma Employing Two-Liquid-Phase Systems

1 INTRODUCTIONThe production of secondary metabolites using plant cells has been the subject of much inter-est in recent years.Despite that tremendous research efforts had been made in this topic,notmany products have reached the commercial stage.It is generally acknowledged that the mainproblem in this field is the lack of basic knowledge of the biosynthetic routes,and the mechanisms found to bring about the production of such secondary metabolites.There are,however,some techniques that have beneficial effects on the production and ex-

Separation/Analysis Rhodamine B by Anion Surfactant/Ionic Liquid Aqueous Two-Phase Systems Coupled with Ultraviolet Spectrometry

A novel method for the separation/analysis rhodamine B has been described. The ionic liquid (1-tetradecyl-3-methylimidazolium bromide)/anion surfactant (sodium dodecyl sulfate)/NaCl two-phase systems (ATPSs) is presented as a simple, rapid and effective sample pretreatment technique coupled with ultraviolet spectrometry for analysis rhodamine B in soft drink. The effects of parameters on the ATPSs extraction of rhodamine B such as amount of surfactant, ionic liquid and salt, pH, temperature, stabilization and centrifugal time have been studied in details. Under the optimized conditions, the linear range of calibration curve for rhodamine B was 0.05 - 7.0 μg·mL-1 and the detection limit was 3.2 ng·mL-1. The phase equilibrium and the mechanism of phase separation for ATPSs have been discussed. This method has been applied to the determination of rhodamine B in soft drink.

Inversion of two-phase extensional basin systems during subduction of the Paleo-Pacific Plate in the SW Korean Peninsula:Implication for the Mesozoic “Laramide-style” orogeny along East Asian continental margin

During subduction, continental margins experience shortening along with inversion of extensional sedimentary basins. Here we explore a tectonic scenario for the inversion of two-phase extensional basin systems, where the Early-Middle Jurassic intra-arc volcano-sedimentary Oseosan Volcanic Complex was developed on top of the Late Triassic-Early Jurassic post-collisional sequences, namely the Chungnam Basin. The basin shortening was accommodated mostly by contractional faults and related folds. In the basement, regional high-angle reverse faults as well as low-angle thrusts accommodate the overall shortening, and are compatible with those preserved in the cover. This suggests that their spatial and temporal development is strongly dependent on the initial basin geometry and inherited structures.Changes in transport direction observed along the basement-sedimentary cover interface is a characteristic structural feature, reflecting sequential kinematic evolution during basin inversion. Propagation of basement faults also enhanced shortening of the overlying sedimentary cover sequences. We constrain timing of the Late Jurassic-Early Cretaceous(ca. 158-110 Ma) inversion from altered K-feldspar 40 Ar/39 Ar ages in stacked thrust sheets and K-Ar illite ages of fault gouges, along with previously reported geochronological data from the area. This "non-magmatic phase" of the Daebo Orogeny is contemporaneous with the timing of magmatic quiescence across the Korean Peninsula. We propose the role of flat/low-angle subduction of the Paleo-Pacific Plate for the development of the "Laramide-style" basement-involved orogenic event along East Asian continental margin.

Two-phase flow instability in a parallel multichannel system

The two-phase flow instabilities observed in through parallel multichannel can be classified into three types,of which only one is intrinsic to parallel multichannel systems.The intrinsic instabilities observed in parallel multichannel system have been studied experimentally.The stable boundary of the flow in such a parallel-channel system are sought,and the nature of inlet flow oscillation in the unstable region has been examined experimentally under various conditions of inlet velocity,heat flux,liquid temperature,cross section of channel and entrance throttling.The results show that parallel multichannel system possess a characteristic oscillation that is quite independent of the magnitude and duration of the initial disturbance,and the stable boundary is influenced by the characteristic frequency of the system as well as by the exit quality when this is low,and upon raising the exit quality and reducing the characteristic frequency,the system increases its instability,and entrance throttling effectively contributes to stabilization of the system.

The Pancharatnam Phase of a Three-Level Atom Coupled to Two Systems of N-Two Level Atoms

In this paper, we present the analytical solution for the model that describes the interaction between a three-level atom and two systems of N-two level atoms. The effects of the quantum numbers and the coupling parameters between spins on the Pancharatnam phase and the atomic inversion, for some special cases of the initial states, are investigated. The comparison between the two effects shows that the analytic results are well consistent.

NUMERICAL SIMULATION OF CHARGED GAS-LIQUID TWO PHASE JET FLOW IN ELECTROSTATIC SPRAYING

Multi-fluid k-e-kp, two phase turbulence model is used to simulate charged gas-liquid two phase coaxial jet, which is the transorting flow field in electrostatic spraying. Compared with the results of experiment, charged gas-liquid two-phase turbulence can be well predicted by this model.

Effect of grain size reduction on high temperature oxidation of binary two-phase alloys

The effect of grain size reduction on the high temperature oxidation of binary two phase alloys was discussed based on the recent research progress. The results show that for those two phase alloys with coarse grain prepared by the conventional methods, complex oxide scales are easily formed after oxidation under high oxygen pressure or under oxygen pressure below the stability limit of the less reactive component oxides. On the contrary, for the nano sized alloys, an exclusive external oxidation of the most reactive component usually occurs during oxidation in air or pure oxygen even for much lower content of the most reactive component. So the gain size reduction is not always beneficial to improve the oxidation resistance of the materials, but exhibits different effects depending mainly on the protective feature of the scales. The transition mechanisms between the different oxidation modes are discussed with respect to the thermodynamic and dynamic aspects.

Study on the Intermediate in the o-Phenylenediamine Oxidative Reaction Using Hemoglobin as A Mimetic Peroxidase in Aqueous-Organic Two Phase

Hemoglobin was used as a mimetic enzyme for peroxidase to catalyze the oxidative reaction of o-phenylenediamine with H2O2 which functioned as an oxidant. The relationship between physicochemical properties of the intermediate and enzymatic activity of hemoglobin was studied. Since the solubility of the intermediate in the reaction is higher in butanol phase than in water phase, the intermediate itself diffused from the aqueous phase to the butanol phase. The experimental results showed that the rate of product and the stability of intermediate were associated with the temperature and the pH value of the buffer. The formation rate of intermediate and half-life period reveal the maximal in pH7, nevertheless, the whole rate of the catalytic reaction is the greatest in pH5, which the ratio of the initial rate in final product formation compared to that intermediate formation is the greatest.