• 제목/요약/키워드: 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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    • 제6권5호
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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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    • 제2권2호
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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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    • 제9권4호
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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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Poly(ethyleneglycol)과 인산염용액이 형성하는 2상계에서의 단백질 분획에 관한 연구 (Protein Partition in an Aqueous Poly (ethyleneglycol)-salt Two-phase System)

  • 이삼빈;이철호
    • 한국식품과학회지
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    • 제19권4호
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    • pp.285-289
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    • 1987
  • Poly(ethylene glycol)과 인산염 용액(PPB)이 형성 하는 액상 이상계에서 단백질의 분획 거동을 관찰하였다. PEG(12%)/PPB(12%) 액상 이상계에서 단백질의 표면 소수성이 다른 BSA, ${\beta}-lactoglobulin$, ovalbumin 모두 PPB-rich phase인 하층부로 몰려 그 분획계수는 대단히 낮았다. PPB의 농도를 15%로 증가 시키면 하층부의 전위는 +50mV 에서 0으로 낮아져 단백질의 분획계수가 다소 증가하는 경향을 나타내었다. PPB의 제2인산염과 제일인산염의 몰 비율을 1.43에서 9.55로 증가시키면 이상계의 상 하층부 부피비(Vt/Vb)가 다소 감소되면서 분획계수는 증가하였다. PPB농도를 $14{\sim}26%$로 증가시키면 이상계의 부피비는 1.5에서 0.39로 현저히 감소하면서 단백질의 분획계수는 ${\beta}-lactoglobalin$의 경우 74배, BSA 32배, ovalbumin 12배, lysozyme 5배로 크게 증가하였다.

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

  • 박병춘;임동준
    • 한국미생물·생명공학회지
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    • 제20권2호
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    • pp.190-195
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    • 1992
  • Polythylene glycol/dextran 수성 2단계에서 polyethylene glycol의분자량과 농도가 증가할수록 체적비는 증가하고 분배계수는 감소하였다. 또한 dextran의 분자량이 증가할 수록 체적비는 감소하고 분배계수는 증가하였다. 한편 dextran의 농도가 증가할수록 체적비와 분배계수는 감소하였다.

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

  • 이삼빈;이철호
    • 한국식품과학회지
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    • 제19권2호
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    • pp.140-145
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    • 1987
  • 폴리에틸렌 글라이콜-덱스트란 흔합수용액이 형성하는 수용액 2상계에서 표면 소수성의 크기를 알고 있는 표준 단백질들의 분획계수를 관찰하였다. 또한 폴리에틸렌 글라이콜(PEG)에 palmitate를 부착시킨PEG-P를 첨가함으로써 상층부의 소수성을 증가 시켰을때 단백질들의 분획계수 변화를 조사하였다. PEG/dextran 2상계에서 단백질의 분획계수는 딘백질의 유효 소수성에 관계없이 2상계의 용적비가 동일한 조건에서 PEG와 dextran의 농도가 함께 증가함에 따라 분획계도 증가하였다. 그러나 PEG에 78%친환도를 가지는 PEG-P를 소량 첨가하여 만든 2상계에서는 유효 소수법이 큰 bovine serum albumin과 $\beta-lactoglobulin$의 분획계수가 10배 이상 증가하였고. 유효 소수성이작은 ovalbumin은 큰 변화를 보이지 않았다. 이러한 분획계수의 급격한 중가는 PEG-P의 첨가량이 아주 낮은 2% 수준일 때 나타났으며 그 이상 침가시에는 모든 단백질이 공히 10% 첨가될때까지 완만히 증가하는 경향을 나타내었다.

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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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    • 제54권4호
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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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    • 제2권2호
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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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    • 제15권7호
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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.

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

  • 허병기;김진한목영일
    • KSBB Journal
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    • 제8권2호
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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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