• Title/Summary/Keyword: two phase partition

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Salting-out Effects on the Partition of Proteins in Aqueous Two-phase Systems

  • KIM, CHAN-WHA;CHO KYUN RHA
    • Journal of Microbiology and Biotechnology
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    • v.6 no.5
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    • pp.352-357
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    • 1996
  • The partition of proteins in the salt-rich phase of polyethylene glycol (pEG)/salt aqueous two-phase systems is limited by the salting-out effects of salt. The logarithm of the concentration of proteins partitioned in the salt-rich phase decreases linearly with increases in the concentration of salt in the salt-rich phase (salting-out). Therefore, the partition of a given protein in the salt-rich phase of aqueous two-phase systems can be estimated from the salting-out constant. The slope of the solubility line (salting-out con-stant) for a given protein is determined by the type of salt in the two-phase systems.

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Production of Cyclodextrin Homologues Using Aqueous Two-Phase System

  • Chang, Woo-Jin;Koo, Yoon-Mo;Park, Sung-Sik
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.2 no.2
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    • pp.97-100
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    • 1997
  • Cyclodxtrin homologues(CDs), produced by cyclodextrin glycosyltransferase(CGTase), were simultaneously partitioned in aqueous two-phase system(ATPS). Partition coefficients of CDs were measured in PEG/dextran systems. Phosphate, citrate, sulfate were tested as salt. ATPS of PEG/salt and PEG/dextran had the partition coefficients of the CDs, larger than unity. However, PEG/dextran system was observed better than PEG/salt as CGTase activity decreased sharply with salt concentration. Enzymatic rection occurred mainly in PEG-rich bottom phase because of the low partition coefficient of CGTase. The resulting CDs transferred to the PEG-rich top phase, obeying the diffusional partition. In the ATPS of 7% PEG(M.W.40, 000), 7mg/ml of CDs were obtained in top phase at 4.5 hours.

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Large-Scale Purification of Protease Produced by Bacillus sp. from Meju by Consecutive Polyethylene Glycol/Potassium Phosphate Buffer Aqueous Two-Phase System

  • Cho, Seong-Jun;Kim, Chan-Hwa;Yim, Moo-Hyun;Lee, Cherl-Ho
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.498-503
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    • 1999
  • Protease produced from Bacillus sp. FSE-68 was isolated from Meju, a Korean fermented soybean starter, and was purified by a two-consecutive aqueous two-phase system. The change of partition coefficient (K) in the polyethylene glycol (PEG)/potassium phosphate buffer (PPB) aqueous two-phase system was measured at different pHs (6.0- 9.2), PPB concentrations (8-12%), and temperatures (4 and $20^{\circ}C$). As the PPB concentration in the aqueous two-phase system increased, the protease concentration in the top phase (PEG-rich phase) increased, thereby enhancing the partition coefficient. The minimum partition coefficient of the protease was achieved at pH 7.0, whereas that of the total protein was at pH 6.0. The biggest difference in partition coefficients of total protein and protease occurred at pH 6.0. It was interesting to note that the partition coefficient of protease decreased as the temperature increased. The optimum condition of the primary aqueous two-phase extraction of Bacillus sp. FSE-68 was pH 6.0, 14% (w/w) PPB, and 16% (w/w) PEG at $4^{\circ}C$, and the crude enzyme concentration in this system was 50% (w/w). The protease, which was concentrated in the top phase, was further mixed with 15% (w/w) PPB (pH 7.0) in the ratio of 1:1 at $20^{\circ}C$ to elute the bottom phase (PPB-rich phase). Using these steps, the purification fold achieved was 9.2 with a 44.7% yield.

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Protein Partition in an Aqueous Poly (ethyleneglycol)-salt Two-phase System (Poly(ethyleneglycol)과 인산염용액이 형성하는 2상계에서의 단백질 분획에 관한 연구)

  • Lee, Sam-Pin;Lee, Cherl-Ho
    • Korean Journal of Food Science and Technology
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    • v.19 no.4
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    • pp.285-289
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    • 1987
  • The partition behavior or proteins in an aqueous two-phase system of poly (ethyleneglycol)-potassium phosphate buffer (PEG/PPB) was investigated. The proteins of different surface hydrophobicity, i.e. Bovine serum albumin (BSA), ${\beta}-lactoglobulin$, ovalbumin. moved to the PPB-rich bottom phase in a PEG(12%)/PPB (12%) two-phase system resulting in very low partition coefficients. When the concentration of PPB increased to 15% level. the electric potential of bottom phase changed from +50 mV to zero and the partition coefficient tended to increase. The change In the molar ratio of $K_2HPO_4/KH_2PO_4$ in PPB from 1.43 to 9.55 caused the volume ratio of top to bottom phase $(V_t/V_b)$ to be decreased and protein partition coefficient increased. When the concentration of PPB was elevated from 14% to 26%, the $V_t/V_b$ decreased from 1.5 to 0.39 and the partition coefficient of proteins increased drastically; ${\beta}-lactoglobulin$ 74 fold. BSA 32 fold, ovalbumin 12 fold and lysozyme 5 fold.

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Formation of PEG/Dextran Aqueous Two-Phase System for Starch Hydrolysis Using $\alpha$-Amylase ($\alpha$-Amylase로 전분 가수분해를 위한 PEG/Dextran 수성 2상계 구성)

  • 박병춘;임동준
    • Microbiology and Biotechnology Letters
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    • v.20 no.2
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    • pp.190-195
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    • 1992
  • In the polyethylene glycol/dextran aqueous two-phase systems, volume ratio was increased and partition coefficient was decreased with the increase of potyethylene glycol molecular weight and concentration. However the volume ratio was decreased and the partition coefficient was increased with the increase of dextran molecular weight. On the other hand, the volume ratio and the partition coefficient were decreased with the increase of dextran concentration. Continuous enzymatic hydrolysis of soluble starch with $\alpha$-amylase which was produced by Bacillus amyloliquefaciens IF0 14141 was investigated in polyethylene glycol/dextran aqueous two-phase systems. Nonreacted soluble starch and $\alpha$-amylase were reused in these systems. $\alpha$-Amylase activity was maintained more than 100 hrs by recycling of $\alpha$-amylase from bottom of settler to reactor.

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Partition Coefficient of Proteins of Different Surface Hydrophobicity in Poly (ethylene glycol)-Dextran Aqueous Two Phase System (Poly(ethylene glycol)-Dextran 수용액 2상계에서 단백질들의 소수성에 따른 분획계수)

  • Lee, Sam-Pin;Lee, Cherl-Ho
    • Korean Journal of Food Science and Technology
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    • v.19 no.2
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    • pp.140-145
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    • 1987
  • The partition coefficient of the proteins of known effective hydrophobicity was determined in a poly (ethylene glycol)-dextran aqueous two-phase system. The changes in the partition coefficient was also determined when a fraction of PEG-palmitate (PEG-P) was added to the system. The partition coefficient of the proteins increased as the concentrations of PEG and dextran increased at a constant phase volume ration irrespective of the effective hydrophobicity of the proteins. When small amounts of PEG-P were added to the PEG phase, the partition coefficients of BSA and ${\beta}-lactoglobulin$, which had relative hydrophobicity (RI) of 700 and 120, respectively, increased more than ten-fold, whereas ovalbumin whose RI was 5 showed little change. The drastic increases m the partition coefficient were observed by the addition of PEG-P in 2% level to the PEG system. Addition of PEG-P over 5% level resulted in a slight further increase in the partition coefficient of all proteins tested.

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Partition method of wall friction and interfacial drag force model for horizontal two-phase flows

  • Hibiki, Takashi;Jeong, Jae Jun
    • Nuclear Engineering and Technology
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    • v.54 no.4
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    • pp.1495-1507
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    • 2022
  • The improvement of thermal-hydraulic analysis techniques is essential to ensure the safety and reliability of nuclear power plants. The one-dimensional two-fluid model has been adopted in state-of-the-art thermal-hydraulic system codes. Current constitutive equations used in the system codes reach a mature level. Some exceptions are the partition method of wall friction in the momentum equation of the two-fluid model and the interfacial drag force model for a horizontal two-phase flow. This study is focused on deriving the partition method of wall friction in the momentum equation of the two-fluid model and modeling the interfacial drag force model for a horizontal bubbly flow. The one-dimensional momentum equation in the two-fluid model is derived from the local momentum equation. The derived one-dimensional momentum equation demonstrates that total wall friction should be apportioned to gas and liquid phases based on the phasic volume fraction, which is the same as that used in the SPACE code. The constitutive equations for the interfacial drag force are also identified. Based on the assessments, the Rassame-Hibiki correlation, Hibiki-Ishii correlation, Ishii-Zuber correlation, and Rassame-Hibiki correlation are recommended for computing the distribution parameter, interfacial area concentration, drag coefficient, and relative velocity covariance of a horizontal bubbly flow, respectively.

Partitioning of Recombinant Human Interleukin-2 in a Poly(ethylene glycol)-Dextran Aqueous Two-Phase System

  • Lee, In-Young;Lee, Sun-Bok
    • Journal of Microbiology and Biotechnology
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    • v.2 no.2
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    • pp.135-140
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    • 1992
  • The partitioning of recombinant human interleukin-2(rhII-2) in PEG 8000-dextran 38800 aqueous two-phase system has been investigated using three different sources of rhIL-2. In the case of pure rhIL-2, the solubility in a PEG-dextran two-phase system was low and most of rhIL-2 was partitioned into the bottom phase. For the recovery of rhIL-2 from insoluble protein aggregates, the inclusion bodies of recombinant E. coli were solubilized by the treatment with sodium dodecyl sulfate (SDS). The addition of SDS significantly enhanced not only the solubility of rhIL-2 but also the partitioning of rhIL-2 to the top phase. When the ratio of SDS to rhIL-2 was 2.0, the partition coefficient(K) and the recovery yield(Y) at the top phase were 4.5 and 88%, respectively, at pH 6.8. In order to reduce the recovery steps further, SDS was directly added to the intact recombinant E. coli cells and then partitioned into the PEG/dextran aqueous two-phase system. The observed partition coefficient ($K{\cong{3.0$) and recovery yield ($Y{\geq}80%$ )of this method were comparable to the rhIL-2 recovery from insoluble protein aggregates. The results obtained in this work indicate that PEG-dextran two-phase partitioning might provide a simple way for the recovery and partial purification of recombinant proteins which are produced as inclusion bodies.

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Thermodynamic Model for Partition Coefficients in the Two Protein Systems

  • Jung, Chang-Min;Bae, Young-Chan;Kim, Jae-Jun
    • Macromolecular Research
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    • v.15 no.7
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    • pp.682-687
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    • 2007
  • The equation of state developed herein is predicated on a hard-sphere reference with perturbations introduced via a potential function to account for electrostatic forces and for attraction between protein particles. During this process, the generalized Lennard-Jones (GLJ) pair potential function is employed. The GLJ pair potential function is employed to represent the protein-protein interaction in two-protein systems. Via the use of the relation between the equation of state and the chemical potential, the phase behavior in the aqueous two-protein system can be estimated. The partition coefficients can be obtained via these processes. The calculated values of the coefficients agree fairly well with the experimental data in the given pH and ionic strength range, with no additional adjustable model parameters.

Process Development of Alcohol Production by Extractive Fermentation(III) -An Optimum Composition of PEG/Dx for Extractive Alcohol Fermentation- (추출발효에 의한 알코올.제조공정 개발(III) -추출 알코올 발효에 최적인 PEG/Dx의 조성-)

  • 허병기;김진한목영일
    • KSBB Journal
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    • v.8 no.2
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    • pp.178-184
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    • 1993
  • Extractive fermentations with the extract of Jerusalem artichoke in an aqueous-two-phase-system of polyethyleneglycol(PEG) and dextran(Dx) were investigated to obtain the effects of composition of PEG and Dx on both fermentation ,characteristics and partition ratio of alcohol. The specific growth rate of K. Fraglis CBS 1555 increased with a decrease of concentration of PEG and Dx. It augmented along with concentration of initial sugar up to 80g/l but decreased thereafter. The specific production rate of alcohol showed a rising tendency up to 100g/lof initial sugar, whereafter represented a decreasing trend. The partition ratio of alcohol between two phases augmented according to decrease of Dx comic. and increase of PEG cone. regardless of initial sugar concentrations. The ratio, however, decreased with Increment of initial sugar concentration at constant composition of PEG and Dx. The partition coefficient of alcohol had an ascending effect to the increase of PEG cone, but it had little effect on the changes of concentrations of Dx and initial sugar. The present study suggests that the optimum composition of PEG and Dx in the aqueous-two-phase-system by extractive fermentation were around 6.5%(w/v) of PEG and 3%(w/v) of Dx in considerations of emulsion state, sedimentation and separation of two phases, alcohol partition ratio, and specific growth rate.

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