• 제목/요약/키워드: Two-phase extraction

검색결과 275건 처리시간 0.128초

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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피페리딘계 이온성 액체와 포스페이트 염으로 구성된 수상이성분계를 이용한 숙신산의 추출 (Liquid Extraction of Succinic Acid by Aqueous Two-Phase Systems Composed of Piperidinium Ionic Liquids and Phosphate Salt)

  • 이우윤;홍연기
    • Korean Chemical Engineering Research
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    • 제54권1호
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    • pp.52-56
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    • 2016
  • 본 연구에서는 숙신산 추출을 위한 효과적인 분리공정으로서 피페리딘계 이온성 액체/$K_2HPO_4$에 의한 수상이성분계를 적용하여 수상이성분계 형성 특성 및 추출특성을 고찰하였다. 실험 결과 $K_2HPO_4$ 수용액에 적정량의 피페리딘계 이온성 액체를 첨가함에 따라 안정한 수상이성분계가 형성됨을 확인할 수 있었으며 이성분계 형성능력은 이온성 액체 내 양이온의 알킬 사슬길이에 따라 증가하였다. $K_2HPO_4$ 수용액을 기준으로 피페리딘계 이온성 액체의 수상이성분계 형성 능력은 이미다졸계나 피롤리딘계에 비해 우수하였다. 피페리딘계 이온성 액체를 이용한 수상이성분계 추출에 있어 숙신산 추출 효율은 75~95%의 범위를 가지며 이 값들은 이미다졸계나 피롤리딘계와 큰 차이가 없었다. 그러므로 피페리딘계 이온성 액체를 이용한 수상이성분계를 숙신산 추출에 적용할 경우 이미다졸계 및 피롤리딘계에 비해 적은 양의 이온성 액체를 사용하여 높은 추출효율을 얻을 수 있음을 확인할 수 있었다.

On-Channel Micro-Solid Phase Extraction Bed Based on 1-Dodecanethiol Self-Assembly on Gold-Deposited Colloidal Silica Packing on a Capillary Electrochromatographic Microchip

  • Park, Jongman;Kim, Shinseon
    • Bulletin of the Korean Chemical Society
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    • 제35권1호
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    • pp.45-50
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    • 2014
  • A fully packed capillary electrochromatographic (CEC) microchip with an on-column micro-solid phase extraction (SPE) bed for the preconcentration and separation of organic analytes was prepared. A linear microchannel with monodisperse colloidal silica packing was formed on a cyclic olefinic copolymer microchip with two reservoirs on both ends. Silver-cemented silica packing frit structure was formed at the entrance of the microchannel by electroless plating treatment as a base layer. A gold coating was formed on it by reducing $Au^{3+}$ to gold with hydroxylamine. Finally micro-SPE bed was formed by self-assembly adsorption of 1-dodecanethiol on it. Micro-SPE beds were about 100-150 ${\mu}m$ long. Approximately $10^3$ fold sensitivity enhancements for Sulforhodamine B, and Fluorescein in nM concentration levels were possible with 80 s preconcentration. Basic extraction characteristics were studied.

단계별 추출법을 이용한 망간각 구성 원소의 존재 형태 (Chemical Speciations of Elements in the Fe-Mn Crusts by Sequential Extraction)

  • 김종욱;문재운;지상범;고영탁;이현복
    • Ocean and Polar Research
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    • 제26권2호
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    • pp.231-243
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    • 2004
  • Sequential extraction was carried out on twenty two subsamples of three ferromanganese crusts from three seamounts (Lemkein, Lomilik, and Litakpooki) near the Marshall Islands in the western Pacific. The extraction was designed to fractionate Fe-Mn crust forming elements into low defined groups: (1) exchangeable and carbornate, (2) Mn-oxide, (3) Fe-oxyhyd.oxide, and (4) residual fraction. X-ray diffraction result shows that target material were well removed by each extraction step except for CFA in phosphatized crusts generation. According to chemical analysis of each leachate, most of elements in the Fe-Mn crusts are bound with two major phases. Mn, Ba, Co, Ni, Zn, (Fe, Sr, Cu, and V) are strongly bounded with Mn-oxide $({\delta}-MnO_2)$ phase, whereas Fe, Ti, Zr, Mo, Pb, Al, Cu,(V, P, and Zn) show chemical affinity with Fe-oxyhydroxide phase. This result indicates that significant amount of Al, Ti, and Zr can not be explained by detrital origin. Ca, Mg, K, and Sr mainly occur as exchangeable elements and/or carbonate phase. Outermost layer 1 and inner layer 2 which are both young crusts generations are similar in chemical speciations of elements. However, some of Fe-oxyhydroxide bounded elements (Pb, Y, Mo, Ba, Al, and V) in phosphatized innermost layer 3 are released during phosphatization and incorporated into phosphate (Pb, Y, Mo, and Ba) or Mn-oxide phase (Al and V). Our sequential extraction results reveal that chemical speciations of elements in the hydrogenetic crusts are more or less different from interelemental relationship calculated by statistical method based on bulk chemistry.

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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    • 제51권3호
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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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    • 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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발효배양액에서 Streptococcus parauberis Z49균주가 생산하는 Bacteriocin의 간편한 추출 (Simple and Rapid Extraction of a Bacteriocin Produced by Streptococcus parauberis Z49 from Fermented Cultures)

  • 박홍제;강용호
    • 미생물학회지
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    • 제46권3호
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    • pp.291-295
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    • 2010
  • Streptococcus parauberis Z49가 생산하는 새로운 bacteriocin의 특성을 조사하고 수성 이상계를 사용하여 발효배양액에서 효율적으로 추출하였다. Nisin과 유사한 S. parauberis Z49의 bacteriocin은 $121^{\circ}C$에서 15 min 열처리해도 활성이 있었으며, 넓은 범위의 pH (pH 2-12)에서 안정하며, M. luteus, Lactobacillus sp., L. fermentum, E. faecium, L. monocytogenes, and P. fluorescens의 생육을 억제하였다. S. parauberis Z49의 발효배양액에 있는 bacteriocin을 간편하게 분리하기 위한 PEG 600/$Na_2SO_4$ 수성 이상계의 최적조건은 PEG 600 15%, $Na_2SO_4$ 30%, NaCl 8% 이었으며, bacteriocin은 PEG층에서 농축이 되었다.

Optimization and validation of HPLC/DAD method for the determination of adenosine and cordycepin in cordyceps products

  • Sasikarn Panpraneecharoen;Tisorn Chatrakoon;Sompong Sansenya;Saowapa Chumanee
    • 분석과학
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    • 제36권4호
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    • pp.152-160
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    • 2023
  • Adenosine and cordycepin are bioactive compounds with health benefits. Therefore, both substances are often used to assess the quality of Cordyceps products. Optimization and validation of the HPLC/DAD method for determining two nucleosides were studied. The samples were prepared using an ultrasound-assisted extraction (ultrasonic bath). The result was optimal conditions for aqueous extraction, an extraction time of 35 min, and an extraction temperature of 40 ℃. The Chromatographic separation was achieved using a reverse phase column (InfinityLab Poroshell 120 EC-C18, 4.6 × 250 mm, 2.7 ㎛) at 30 ℃ with a mobile phase gradient elution of water and methanol at a flow rate of 0.7 mL/min. The eluents were monitored via a diode array detector at 260 nm. Two nucleosides were separated by less than 12 min after injection. The developed method was found to be excellent linear (r2 > 0.9999), accurate (% recovery 95.34-98.51), and precise (% relative standard deviation < 2.0). The limit of detection (LOD) and quantification (LOQ) were 0.45 and 1.38 mg/mL for adenosine and 0.47 and 1.43 mg/mL for cordycepin, respectively. This method was satisfactory for simultaneously quantitating two nucleoside contents, which were used to evaluate Cordyceps products.

Anticancer and Antioxidant Activity of Allergen-Removed Extract in Rhus verniciflua Stokes

  • Kim, Myong-Jo;Choi, Won-Cheol;Barshinikov, A. M.;Kobayashi, A.
    • 한국약용작물학회지
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    • 제10권4호
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    • pp.288-293
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    • 2002
  • Allergen-removed-extract was produced from Rush verniciflua by two phase methods. Phase one was high temperature treatment of Rush verniciflua tree to get allergen-removed-extract. Phase two was extraction of solution from phase one product using water or organic solvents. The solutions from above method show high antioxidant activity, anticancer activity, and improvement in lung function, but did not contain urushiol family compounds.

Poly(ethylene glycol)/인산염 용액 2상계를 이용한 Lactobacillus sporogenes가 생산하는 균체내 $\beta$-Galactosidase의 추출 분리에 관한 연구 (Purification of Intracellular $\beta$-Galactosidase from Lactobacillus sporogenes in an Aqueous Poly(ethylene glycol)- Potassium Phosphate Two-Phase System)

  • 이삼빈;김영만;이철호
    • 한국미생물·생명공학회지
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    • 제15권2호
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    • pp.84-88
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    • 1987
  • Poly(ethylene glycol)/salt가 형성하는 액상 2상계에서 균체내 효소인 $\beta$-galactosidase 분리를 최적화하는 실험을 수행하였다. L. sporogenes에 의해 생산된 $\beta$-galactosidase의 분획은 2상계를 형성하는 PEG 분자량이 작을수록 상층부로 이동하였으며, 상층부 PEG 농도가 증가할 때 salting-out 현상이 나타나서 단백질 뿐아니라 $\beta$-galactosidase는 상층부로 크게 이동하였다. 또한 균체내 효소 정제에서 문제가 되는 세포벽잔사(cell debris)들은 조성이 binodial line에 접근할수록 효소 fraction과 반대방향으로 이동되었다. PEG-salt를 이용한 $\beta$-galactosidase 분리는 조성을 binodial line레 접근시키면서 동시에 상층부의 부피를 줄이는 것이 효과적이었다. PEG1000, 300을 각각 단계적으로 이용한 2단계 추출(two-step extraction)로써 세포벽 잔사, bulk protein, 색소물질 및 핵산 등을 제거할 수 있었으며, 이때 효소의 회수율은 74%, 단백질 회수율 35%로서 정제도는 2 배 이상의 효과를 보였다.

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