• Title/Summary/Keyword: Aqueous Phase

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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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Optimization of Catechol Production Using Immobilized Resting Cells of Pseudomonas putida in Aqueous/organic Two-phase System

  • Chae, Hee-Jeong;Yoo, Young-Je
    • Journal of Microbiology and Biotechnology
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    • v.7 no.5
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    • pp.345-351
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    • 1997
  • An aqueous/organic two-phase reaction system was applied to the production of catechol using immobilized resting cells of Pseudomonas putida CY 400. Water/ethyl ether system was used because of high partition coefficient of catechol and thus to reduce the product inhibition and degradation. Among the tested immobilization carriers, polyacrylamide gel gave the highest catechol productivity. The immobilization seemed to protect the cells against solvent toxicity. From the simulation of reaction conditions based on two-phase models, it was found that there was an optimum acetate concentration at fixed benzoate and cell concentrations for the catechol productivity. A lower phase volume ratio (lower fraction of organic phase) gave a higher productivity. However, the substrate conversion was low at low phase volume ratio.

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Mass Transfer of Lysozyme Extraction Using Reversed Micelles (역미셀을 이용한 Lysozyme 추출에 대한 물질 전달)

  • 전병수;김석규;윤성옥;송승구
    • KSBB Journal
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    • v.16 no.3
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    • pp.241-245
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    • 2001
  • Mass transfer rates have been measured for the extraction of enzyme from aqueous solution into a reverse-micelle phase at $25^{\circ}C$. The 420 mL vessel was carefully designed to maintain a planar interface between the aqueous and solvent phases, so allowing precise measurement of interfacial area, has been investigated. Sodium di-2-ethylhexyl sulfosuccinate(AOT) was the surfactant used. Factors varied included: agitator speed, pH, ionic strength and surfactant concentration. Samples were taken from the solvent phase at 15min intervals, and the amount of enzyme extracted was measured by UV absorption at 280 nm. The observed Sherwood numbers for the aqueous phase $Sh_1$were correlated interms of the aqueous phase Reynolds number $Re_1$, and modified Schmidt number $Sc_1$. $Sh_1=0.664Re_1^{0.5}Sc_1^{0.33}$

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Effect of Sophora Subprostrata Fractions on Focal Ischemic Brain Damage Induced by Middle Cerebral Artery Occlusion in Rats(I) (광두근(廣豆根) 분획물이 중대뇌동맥폐쇄(中大腦動脈閉鎖)에 의한 뇌허혈손상에 미치는 효과(I) - 행동평가를 기준으로)

  • Choi, Moon-Seok;Kim, Youn-Sub
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.3
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    • pp.760-764
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    • 2005
  • This research was performed to investigate protective effect of Sophora Subprostrata fractions against focal ischemic brain damage after middle cerebral artery(MCA) occlusion. Rats were divided into six groups: MCA-occluded group(Control); each administered groups with Sophora Subprostrata total phase(Total), Sophora Subprostrata Aqueous phase (Aqueous), Sophora Subprostrata BuOH phase(BuOH), and Sophora Subprostrata Alkaloid phase(Alkaloid) after MCA-occlusion; sham-operated group(Sham). The right MCA was occluded by A poly-L-lysine coated 4-0 nylon suture thread through the internal carotid artery permanently. Sophora Subprostrata and fractions were administered orally(5mg/ml) for 7 days after MCA-occlusion. The behavior of ischemic rats were examined at 24 hours, 3, 5 and 7 days after MCA-occlusion from the views of 4 different aspects: posture & balance tests(4 subtests), reflex tests(6 subtests), muscle-tone tests(3 subtests), and foot-fault test. The results showed that 1) in muscle tone test, Sophora Subprostrata total phase only increased reduced muscle tone function from 3 to 7 days, 2) in reflex test, Sophora Subprostrata total and Aqueous phase increased fast recovery from 24 hours and 3 days, 3) in posture & balance test, Sophora Subprostrata total and Aqueous phase increased fast recovery from 24 hours, and Sophora Subprostrata BuOH and Alkaloid phase increased posture & balance function from 3 days, but 4) in motor function test, Sophora Subprostrata did not show effective recovery compared with control group. In conclusion, Sophora Subprostrata has protective effects against brain damage at the early stage of focal cerebral ischemia. Sophora Subprostrata total and Aqueous phase produced more pronounced protective effect against focal ischemic brain damage.

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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Influence of Xanthan, Emulsification Temperature, and Environmental Stresses on the Preparation of Water-in-Corn Oil Emulsions Droplets Coated by Polyglycerol Polyricinoleate

  • Surh, Jeong-Hee
    • Food Science and Biotechnology
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    • v.18 no.2
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    • pp.299-306
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    • 2009
  • The purpose of this study was to prepare stable water-in-corn oil (W/O) emulsion droplets coated by polyglycerol polyricinoleate (PGPR). W/O emulsions (20 wt% aqueous phase, 80 wt% oil phase containing 8 wt% PGPR) were produced by high pressure homogenization (Emulsions 1), however, appreciable amount of relatively large water droplets (d>$10{\mu}m$) were found. To facilitate droplet disruption, viscosity of each phase was adjusted: (i) increased the viscosity of aqueous phase by adding 0.1 wt% xanthan (Emulsions 2); (ii) decreased the viscosity of oil phase and aqueous phase by heating them separately at $50^{\circ}C$ for 1 hr immediately before emulsification (Emulsions 3). Homogenizing at the elevated temperature clearly led to a smaller water droplet size, whereas xanthan neither improved nor adversely affected on the microstructures of the emulsions. In addition, the Emulsions 3 had good stability to droplet aggregation under shearing stress, thermal processing, and long term storage.

계면활성제/1-Hoxanol/물 혼합물의 상태도와 전기 전도도에 관한 연구

  • 오성근;김종득
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.12 no.1
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    • pp.34-61
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    • 1986
  • The microstructural transitions of aqueous micellar solutions of cetyltrime-thylammonium bromide and sodiumdodecyl sulfate by adding 1-hexanol were investigated, measuring the concentrations of equilibrated phases and the electrical conductivities at the low concentrations of surfactants, where the solobilities of 1-hexanol varied significantly, at 3$0^{\circ}C$ and 45$^{\circ}C$. Ternary phase diagrams of multiphase regions, constructed by liquid chromatography analysis and by counting the number of phase of samples, consisted of one three-phase region and three two-phase regions. One of the two-phase regions was found to equilibrate an aqueous micellar solution and a liquid crystal, and had a critical point between them. Near this region, the solubility curve varied abruptly, and the isotropic solution turned birefrigent. The conductivities of the single phase regions above the critical point increased up to a certain point as 1-hexanol added, and then decreased, representing the microstructural transition at the supercritical region. Further, the solubility of 1-hexanol in aqueous micellar solution was found to increase as temperature and the number of hydrophilelipophile balance of surfactants increase.

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Purification and Characterization of Crystalins by Aqueous Two-Phase Extraction

  • Bermudez, Ondrea;Forciniti, Daniel
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.6
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    • pp.395-401
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    • 2001
  • Crystallins are a family of water-soluble proteins that constitute up to 90% of the wa-ter-soluble proteins in mammalian eye lenses, We present in this paper an alternative purification method for these proteins using polyethylene glycol/dextran aqueous two-phase extraction. Un-der the appropriate conditions, we were able to recover the γ-crystallin fraction essentially free of the remaining proteins. High concentrations of salt at a neutral pH maximize the recovery of γ-crystallins in the top phase and minimize the contamination by the other proteins present in the lenses. The proposed protocol decreases the separation time by about 50%. The complex partition behavior observed for these proteins reflects a delicate balance between protein/phase-forming species(various polymers and salts) and protein interactions. This is evidenced, in part, by the role played by the largest proteins in this group as a "pseudo"phase-forming species.

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X-Ray Diffraction Studies of Uranyl Hydrolysis Precipitates Synthesized in Neutral to Alkaline Aqueous Solutions

  • 박용준;표형렬;김원호;전관식
    • Bulletin of the Korean Chemical Society
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    • v.17 no.10
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    • pp.925-929
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    • 1996
  • Uranyl hydrolysis precipitates were obtained by increasing pH value of aqueous uranyl solution in the range of neutral to alkaline pH value and their phase transformation during the solubility experiment under various conditions has been examined. The precipitates formed in the hydrolysis reaction of uranyl ion had a layered structure such as a meta-schoepite phase, a schoepite structure, or a mixed phase of meta-schoepite and schoepite. Phase transformation between them was strongly dependent on the pH value at which the precipitate was formed. The distance between the layers in meta-schoepite or schoepite phase was ∼7.35 Å, and it was increased with the pH value at which the precipitate was synthesized as well as the pH values of the aqueous solution. The phase transformation from a meta-schoepite to schoepite was fast for the precipitates formed at low pH values, however, it was not the case for the precipitates formed at high pH values. A small difference of pH value in aqueous solution gave a great change on its solubilities near pH 9.7, because a layered structure of the precipitates became amorphous above that pH value. Greater solubility for the precipitate formed at higher pH value can be explained from the fact that the precipitates formed at low pH value had a better crystallinity and also that the precipitates formed at higher pH value has a slower rate of crystallization.

Separation of Immunoglobulin G from Colostrum by Reverse Micelles of Cationic Surfactant (양이온 계면활성제로 형성된 역미셀을 이용한 초유 Immunoglobulin G의 분리)

  • 노경현;임지영
    • Food Science of Animal Resources
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    • v.24 no.1
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    • pp.80-85
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    • 2004
  • This study was carried out to separate immunoglobulin G(IgG) from colostrum using reverse micellar extraction of cationic surfactant and to suggest suitable extraction conditions. The reconstituted colostrum powder was solubilized into a reverse micellar phase containing CDAB(cetyldimethylethyl ammonium bromide) by mixing equal volume of the aqueous and organic phase with constant stirring. The solubilization of proteins from the aqueous to the organic phase was manipulated by pH and ionic strength of the aqueous phase and concentration of surfactant in the organic phase. Based on the SDS-PAGE and densitometry, about more than 90% of initial IgG was remained in the aqueous phase after reverse micellar extraction. Although the aqueous phase contained lactoferrin and bovine serum albumin as minor components, about 93% of the total protein was IgG. The efficient extraction was achieved by the reaction of sodium phosphate buffer(pH 8) containing 50 mM KCl and organic phase containing 100 mM CDAB. The separation of IgG using reverse micellar extraction was simple, highly efficient and easy to be scaled up.