• Title/Summary/Keyword: Aqueous Phase

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Separation and Recovery of Cyclodextrin Glucanotransferase Using Aqueous Two-Phase Systems (수성2상계를 이용한 Cyclodextrin Glucanotransferase 분리 및 회수)

  • 김진현;홍승서;이현수
    • KSBB Journal
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    • v.15 no.6
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    • pp.556-559
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    • 2000
  • Cyclodextrin Glucanotransferase(EC 2.4.1.19 : 1,4-${\alpha}$-glucano) transferase, cyclizing; CGTase) can be separated and recovered in an aqueous two-phase system composed of poly(ethylene glycol)(PEG)/dextran and PEG/salt. In an aqueous two-phase system consisting of PEG 35000 (5%) and dextran T2000 (7%), all cell and debris were collected at the interphase. CGTase partitioned to the denser dextran phase at an yield of 83.4%. On the other hand, in an aqueous two-phase system consisting of PEG 35000 (10%) and sodium phosphate (15%), CGTase partitioned to the denser salt phase at an yield of 95.5%. In order to recover CGTase using an aqueous two-phase system, the PEG/salt system proved to be more efficient than the PEG/dextran system in terms of yield and cost.

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Extraction of Biomolcules by Ionic Liquids (이온성 액체를 이용한 생물분자의 추출)

  • Lee, Woo Yun;Lee, Yong Hwa;Lee, Jun;Hong, Yeon Ki
    • Journal of Institute of Convergence Technology
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    • v.4 no.2
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    • pp.23-26
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    • 2014
  • As an effective separation method for biomolecules, aqueous two-phase systems based on ionic liquids were suggested. Hydrophobic ionic liquids are more expensive and viscous in spite of their usage in the ionic liquid/water biphasic extraction compared with hydrophilic ionic liquids. In case of aqueous two-phase systems using hydrophilic ionic liquids, they can be diluted in aqueous phase. Experimental results show that aqueous two phase systems can be formed by adding appropriate amount of ionic liquids to aqueous salts solutions. The viscosity of ionic liquid aqueous phase is proportional to the cation chain length in ionic liquids. It is founded that the ionic liquid based aqueous two phase systems are effective for the separation of biomolecules such as acrylic acid.

Drop formation at submerged nozzles: Comparison of aqueous dispersed and organic dispersed cases for TBP-dodecane and nitric acid system

  • Roy, Amitava;Darekar, Mayur;Singh, K.K.;Shenoy, K.T.
    • Nuclear Engineering and Technology
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    • v.51 no.3
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    • pp.761-768
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    • 2019
  • Understanding the phenomena of formation of single drops is necessary to understand the hydrodynamics in solvent extraction equipment which are used for separation of nuclear materials. In this work, the phenomena of aqueous phase and organic phase drop formation at submerged nozzles are compared by conducting experiments with 30%TBP (v/v) in dodecane as the organic phase and nitric acid as the aqueous phase. Two different nozzles and three different nitric acid concentrations are used. For each nozzle and nitric acid concentration, velocity of the dispersed phase is varied. Drops of aqueous phase formed at downward oriented nozzles submerged in organic phase are observed to be smaller than the drops of organic phase formed at upward oriented nozzles submerged in aqueous phase. Correlations to estimate drop diameter are proposed.

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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Optimization of Conditions for Extractive Ethanol Fermentation in an Aqueous Two Phase System (수성이상계 에탄올 추출발효 조건의 최적화에 관한 연구)

  • 김진한;허병기;목영일
    • Microbiology and Biotechnology Letters
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    • v.22 no.5
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    • pp.531-537
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    • 1994
  • This study was undertaken with objective of optimizing the conditions of fermentation in an aqueous two-phase system which is composed of polyethylene glycol (PEG) 20000 and crude dextran (Dx). The data were obtained and analyzed using the Box-Wilson's experimental design protocol and the response surface methodology. To reach this end a multilinear polynomial regres- sion model was developed, which can be utilized for the purpose of optimizing the extractive fermentation. Optimum conditions for batch fermentation with aqueous two phase system were found to be at 4.2~5.4% PEG/3.2~4.2% Dx range. The composition of the center was 4.8% PEG/ 3.6% Dx. Optimum operating conditions for initial sugar concentration and fermentation time were approximately 160 g/l, and 21~22 hr, respectively. Fermentation in the aqueous two phase system composed of 5% PEG/4% Dx showed increase of 23% in ethanol concentration, of 9.5% in ethanol yield, and of 19% in ethanol productivity as compared to the case of fermentation of neat Jerusalem artichoke juice.

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Separation of Lactoferrin from Model Whey Protein Mixture by Reverse Micelles Formed by Cationic Surfactant

  • Noh, Kyung-Hyun;Rhee, Min-Suk;Imm, Jee-Young
    • Food Science and Biotechnology
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    • v.14 no.1
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    • pp.131-136
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    • 2005
  • The selective extraction behavior of lactoferrin (Lf) from whey protein mixture was examined using reverse micelles formed by the cationic surfactant, cetyldimethylammonium bromide (CDAB). The major whey proteins, including ${\beta}$-lactoglobulin, ${\alpha}$-lactalbumin and bovine serum albumin, were solubilized from aqueous phase to organic phase while Lf was recovered in the aqueous phase. The solubilization behaviors of the proteins were manipulated by the process parameters such as the pH and salt concentration of the aqueous phase and the surfactant concentration in the organic phase. Efficient forward extraction was achieved with sodium borate buffer (50 mM, pH 9) containing 50 mM KCl and organic phase containing 100 mM CDAB. Based on SDS-PAGE and densitometry, about 96% of the initial Lf remained in the aqueous phase after forward extraction. The dialyzed Lf fully maintained its bacteriostatic activity against E. coli O157:H7.

Partitioning of Lactobacillus helveticus Cells and Lactic Acid in Aqueous PEI/HEC Two-Phase Systems. (수용성 이상계에서의 젖산과 Lactobacillus helveticus세포의 분배특성)

  • 안한군;권윤중
    • Microbiology and Biotechnology Letters
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    • v.26 no.1
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    • pp.55-60
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    • 1998
  • For an ideal extractive bioconversion in aqueous two-phase systems, the product has to be preferentially partitioned into the phase opposite to the one in which the biocatalyst is located. Partitioning behaviors of Lactobacillus helveticus IAM 11090 and lactic acid in aqueous two-phase systems composed a polycation, poly(ethylenimine) (PEI), and an uncharged polymer (hydroxyethyl)cellulose (HEC) were investigated. L. helveticus cells were preferentially partitioned to the HEC-rich top phase while about 85% of lactic acid was partitioned to the PEI-rich bottom phase. These results indicate that extraction of charged, low molecular weight products in an aqueous two-phase systems can be promoted by using an oppositely charged polymer as one of the phase-forming polymer. By the ideal partitioning of the cells and lactic acid, an aqueous PEI/HEC two-phase system can be used as a potential system for the extractive lactic acid fermentation of cheese whey.

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Reverse Micellar Extraction of Fungal Glucoamylase Produced in Solid-State Fermentation Culture

  • Paraj, Aliakbar;Khanahmadi, Morteza;Karimi, Keikhosro;Taherzadeh, Mohammad J.
    • Journal of Microbiology and Biotechnology
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    • v.24 no.12
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    • pp.1690-1698
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    • 2014
  • Partial purification of glucoamylase from solid-state fermentation culture was, firstly, investigated by reverse micellar extraction (RME). To avoid back extraction problems, the glucoamylase was kept in the original aqueous phase, while the other undesired proteins/enzymes were moved to the reverse micellar organic phase. The individual and interaction effects of main factors (i.e., pH and NaCl concentration in the aqueous phase, and concentration of sodium bis-2-ethyl-hexyl-sulfosuccinate (AOT) in the organic phase) were studied using response surface methodology. The optimum conditions for the maximum recovery of the enzyme were pH 2.75, 100 mM NaCl, and 200 mM AOT. Furthermore, the optimum organic to aqueous volume ratio ($V_{org}/V_{aq}$) and appropriate number of sequential extraction stages were 2 and 3, respectively. Finally, 60% of the undesired enzymes including proteases and xylanases were removed from the aqueous phase, while 140% of glucoamylase activity was recovered in the aqueous phase and the purification factor of glucoamylase was found to be 3.0-fold.

Cultivation of Digitalis lanata Cell Suspension in an Aqueous Two-Phase System

  • Choi, Yeon-Sook;Lee, Sang-Yoon;Kim, Dong-Il
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.589-592
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    • 1999
  • Suspension cultures of Digitalis lanata were successfully performed in an aqueous two-phase system (ATPS) of 4.5% polyethylene glycol (PEG) 20,000 and 2.8% crude dextran. Cell growth in the medium containing an individual ATPS-forming polymer was inhibited due to the toxicity of PEG and a high viscosity of dextran. Formation of ATPS supported cell growth by showing a considerably decrease in viscosity and partitioning of cells into a PEG-lean dextran phase. It was found that an aqueous two-phase cultivation of plant cells in a stirred tank bioreactor could be successfully applied.

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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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