• Title/Summary/Keyword: Reverse ion-exchange

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Purification and Characterization of Cop, a Protein Involved in the Copy Number Control of Plasmid pE194

  • Kwak, Jin-Hwan;Kim, Jung-Ho;Kim, Mu-Yong;Choi, Eung-Chil
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.291-297
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    • 1998
  • Cop protein has been overexpressed in Escherichia coli using a T7 RNA polymerase system. Purification to apparent homogeneity was achieved by the sequential chromatography on ion exchange, affinity chromatography, and reverse phase high performance liquid chromatography system. The molecular weight of the purified Cop was estimated as 6.1 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). But the molecular mass of the native state Cop was shown to be 19 kDa by an analytical high performance size exclusion chromatography, suggesting a trimer-like structure in 50 mM Tris-HCI buffer (pH 7.5) containing 100 mM NaCl. Cop protein Was calculated to contain $39.1% {\alpha}-helix, 16.8% {\beta}-sheet$, 17.4% turn, and 26.8% random structure. The DNA binding property of Cop protein expressed in E. coli Was preserved during the expression and purification process. The isoelectric point of Cop was determined to be 9.0. The results of amino acid composition analysis and N-terminal amino acid sequencing of Cop showed that it has the same amino acid composition and N-terminal amino acid sequence as those deduced from its DNA sequence analysis, except for the partial removal of N-terminal methionine residue by methionyl-aminopeptidase in E. coli.

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Purification and Characterization of Novel Antimicrobial Peptide from the Skin of the Hagfish , Eptatretus burgeri

  • Hwang, Eun-Young;Seo, Jung-Kil;Kim, Chan-Hee;Go, Hye-Jin;Kim, Eun-jung;Chung, Joon-Ki;Rye, Hong-Soo;Park, Nam-Gyu
    • Preventive Nutrition and Food Science
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    • v.4 no.1
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    • pp.28-32
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    • 1999
  • A novel antimicrbial peptide , named HFS-I, was isolated and characterized from the skin of the hagfish, Eptatretus bugeri. The decapeptide with a molecular mass of 1279.5 Da was purified to homogeneity using a gel-filtration column, ion-exchange and C18 reverse-phase high performance liquid chromatograpy . The complete amino acid sequence of HFS-I, which was determined by a combination of an automated amino acid sequencing and FAB-MS, was F-P-W-W-L-S-G-K-Y-P-NH2. Comparison of the amino acid sequence with those of other known antimicrobial peptides revealed that HFS-I was a novel antimicrobial peptide. HFS-I showed a weak antimicrobial activity in vitro aganinst a broad spectrum of microorganism without hemolytic acitivity.

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Isolation of Angiotensin Converting Enzyme Inhibitors from Compositae Plants

  • Song, Kyung-Bin;Kim, Ji-youn;Jung, Hye-young
    • Preventive Nutrition and Food Science
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    • v.7 no.2
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    • pp.157-161
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    • 2002
  • Thirty plants of the Compositae family were screened for inhibitory activity of angiotensin converting enzyme (ACE). Among them, Chrysanthemum boreale Makino and Ixeris dentate Nakai were selected for further investigation since they had the highest inhibitory activity. Crude water extracts of the flowers of Chrysanthemum boreale Makino and the roots of Ixeris dentate Nakai were prepared by heating at 95$^{\circ}C$ and 6$0^{\circ}C$ for 2 hr, respectively, followed by centrifugation at 8000$\times$ g far 30 min. Crude extracts were then filtered using YM-3 and YM-1 membranes. The ACE inhibitors were isolated using consecutive chromatographic methods including: Sephadex G-15, ion exchange, FPLC, and reverse phase HPLC. The inhibitors were identified to have molecular masses of 204 and 283 daltons, respectively, by mass spectrometry. These results demonstrate that crude extracts of Compositae plants may be useful as functional food ingredients with anti-hypertensive properties.

Overexpression and purification of recombinant lysozyme from Agrius convolvuli expressed as inclusion body in Escherichia coli

  • Park, Soon-Ik;Yoe, Sung Moon
    • Animal cells and systems
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    • v.16 no.6
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    • pp.455-461
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    • 2012
  • Amongst the various antimicrobial peptides, lysozyme plays a central role in initiating and maintaining the antibacterial defense response of insect. Here we propose the biosynthesis and refolding of recombinant lysozyme in Escherichia coli expressed in inclusion body form. The Agrius lysozyme gene was amplified using gene specific primers and then ligated into the pGEX-4T-1 vector, which contained the glutathione S-transferase (GST) gene as a fusion partner. A recombinant lysozyme was expressed in E. coli Rosetta cells using a pGEX-4T-1 expression vector, and the fusion protein was induced by ioporpyl-${\beta}$-D-thiogalactopyranoside (IPTG). The recombinant protein produced as an inclusion body was resolubilized in solubilization buffer, and the resultant solution was dialyzed in refolding buffer. After thrombin cleavage, the recombinant lysozyme was purified by ion exchange chromatography and reverse phase chromatography. The recombinant lysozyme was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis and immunoreactivity against the anti-Agrius lysozyme was observed by western blot analysis of this protein. The recombinant lysozyme displayed antibacterial activity against Bacillus megaterium and Micrococcus luteus, which was confirmed by the inhibition zone assay.

Recombinant Expression, Isotope Labeling, and Purification of Cold shock Protein from Colwellia psychrerythraea for NMR Study

  • Moon, Chang-Hun;Jeong, Ki-Woong;Kim, Hak-Jun;Heo, Yong-Seok;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • v.30 no.11
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    • pp.2647-2650
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    • 2009
  • Cold shock proteins (Csps) are a subgroup of the cold-induced proteins on reduction of the growth temperature below the physiological temperature. They preferentially bind to single-stranded nucleic acids to translational regulation via RNA chaperoning. Csp plays important role in cold adaptations for the psychrophilic microorganism. Recently, Cold shock protein from psychrophilic bacteria, Colwellia psychrerythraea (CpCsp) has been identified. Three dimensional structures of a number of Csps from various microorganisms have been solved by NMR spectroscopy or X-ray crystallography, but structures of psychrophilic Csps were not studied yet. Therefore, cloning and purification protocols for further structural study of psychrophilic Csp have been optimized in this study. CpCsp was expressed in E. coli with pET-11a vector system and purified by ion exchange, size exclusion, and reverse phase chromatography. Expression and purification of CpCsp in M9 minimal media was carried out and $^{15}N$-labeled proteins with high purity over 90% was obtained. Further study will be carried out to investigate the tertiary structure and dynamics of CpCsp.

Synthesis of Low Concentration of NaOH Solution using $Na^+$ ion in the Concentrated Water from Membrane Separation Process (분리막 농축수에 포함된 Na를 이용한 저농도 NaOH 용액의 합성)

  • Lee, Yoon-Ji;Park, Youn-Jin;Choi, Jeong-Hak;Shin, Won-Sik;Choi, Sang-June;Chon, Uong
    • Korean Chemical Engineering Research
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    • v.49 no.6
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    • pp.810-815
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    • 2011
  • Concentrated water discharged from seawater desalination process contains a high concentration of $Na^+$ ion. Electrolysis was applied to synthesize NaOH solution from the highly concentrated NaCl solution. The effect of various operating parameters of composited laboratory-scale chlor-alkali (CA) membrane cell was investigated. The operating parameters such as membrane types (CIMS and Nafion membranes), pretreatment of the membrane, flow rate (73 mL/min~200 mL/min), initial $Na^+$ ion concentration (1.5 M, 3M and 5 M) and current (1.5A and 2A) were evaluated. It was observed that synthesis efficiency of NaOH solution with CIMS membrane was higher than that with Nafion membrane, but the durability of CIMS membrane on $Cl_2$ gas was poor. The synthesis efficiency of NaOH solution increased with increasing initial $Na^+$ ion concentration and current, while the efficiency decreased with increasing flow rate using Nafion membrane.

MXene Based Composite Membrane for Water Purification and Power Generation: A Review (정수 및 발전을 위한 맥신(MXene) 복합막에 관한 고찰)

  • Seohyun Kim;Rajkumar Patel
    • Membrane Journal
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    • v.33 no.4
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    • pp.181-190
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    • 2023
  • Wastewater purification is one of the most important techniques for controlling environmental pollution and fulfilling the demand for freshwater supply. Various technologies, such as different types of distillations and reverse osmosis processes, need higher energy input. Capacitive deionization (CDI) is an alternative method in which power consumption is deficient and works on the supercapacitor principle. Research is going on to improve the electrode materials to improve the efficiency of the process. A reverse electrodialysis (RED) is the most commonly used desalination technology and osmotic power generator. Among many studies conducted to enhance the efficiency of RED, MXene, as an ion exchange membrane (IEM) and 2D nanofluidic channels in IEM, is rising as a promising way to improve the physical and electrochemical properties of RED. It is used alone and other polymeric materials are mixed with MXene to enhance the performance of the membrane further. The maximum desalination performances of MXene with preconditioning, Ti3C2Tx, Nafion, and hetero-structures were respectively measured, proving the potential of MXene for a promising material in the desalination industry. In terms of osmotic power generating via RED, adopting MXene as asymmetric nanofluidic ion channels in IEM significantly improved the maximum osmotic output power density, most of them surpassing the commercialization benchmark, 5 Wm-2. By connecting the number of unit cells, the output voltage reaches the point where it can directly power the electronic devices without any intermediate aid. The studies around MXene have significantly increased in recent years, yet there is more to be revealed about the application of MXene in the membrane and osmotic power-generating industry. This review discusses the electrodialysis process based on MXene composite membrane.

Separation and Purification of Protease from Bacillus subtlils CCKS-111 in Korean Traditional Soy Sauce (한국재래간장으로 부터 분리한 Bacillus subtilis CCKS-111이 생성하는 Protease의 분리 및 정제)

  • Kim, Sung;Lim, Seong-Il;Lee, Hee-Duck;Lee, Seon-Ho;Son, Jun-Ho;Choi, Hee-Jin;Kim, Yeung-Hweal;Choi, Cheong
    • Applied Biological Chemistry
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    • v.40 no.3
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    • pp.178-183
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    • 1997
  • A protease was purified from Bacillus subtilis CCKS-111 by ammonium sulfate treatment, DEAE-cellulose ion-exchange chromatography, Sephadex G-100 gel filtration and high performance liquid chromatography (HPLC). The specific activity of the purified enzyme was 24.3 unit/mg protein and the purification fold of enzyme was 50.6. Molecular weight of the purified enzyme estimated about 28,000 by HPLC gel filtration. The amino acid residues of this enzyme were 251.3 except threonine, serine and glycine. This result was similar to Bacillus subtilis subtilisin DY. From the first N-terminal amino acid to the 32th amino acid, the amino acid sequence was estimated after RP-HPLC elution. N-terminal and the 32th amino acids were alanine and aspartic acid. Alanine, serine, glycine and arginine were four major acids in the enzyme.

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Development of Chemical and Biological Decontamination Technology for Radioactive Liquid Wastes and Feasibility Study for Application to Liquid Waste Management System in APR1400 (액체방사성폐기물에 대한 화학적, 생물학적 제염기술 개발 및 APR1400 액체폐기물관리계통 적용을 위한 타당성 연구)

  • Son, YoungJu;Lee, Seung Yeop;Jung, JaeYeon;Kim, Chang-Lak
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.17 no.1
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    • pp.59-73
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    • 2019
  • A decontamination technology for radioactive liquid wastes was newly developed and hypothetically applied to the liquid waste management system (LWMS) of the nuclear power plant (NPP) to evaluate its decontamination efficacy for the purpose of the fundamental reduction of spent resins. The basic principle of the developed technology is to convert major radionuclide ions in the liquid wastes into inorganic crystal minerals via chemical or biological techniques. In a laboratory batch experiment, the biological method selectively removed more than 80% of cesium within 24 hours, and the chemical method removed more than 95% of cesium. Other major nuclides (Co, Ni, Fe, Cr, Mn, Eu), which are commonly present in nuclear radioactive liquid wastes, were effectively scavenged by more than 99%. We have designed a module including the new technology that could be hypothetically installed between the reverse osmosis (R/O) package and the organic ion-exchange resin in the LWMS of the APR1400 reactor. From a technical evaluation for the virtual installation, we found that more than 90% of major radionuclides in the radioactive liquid wastes were selectively removed, resulting in a large volume reduction of spent resins. This means that if the new technology is commercialized in the future, it could possibly provide drastic cost reduction and significant extension of the life of resins in the management of spent resins, consequently leading to delay the saturation time of the Wolsong repository.

The Hardness Water Production By RO/NF/ED Linking Process From Deep Seawater (RO/NF/ED 연계 공정에 의한 고경도 담수 제조)

  • Moon, Deok-Soo;Kim, Kwang Soo;Gi, Ho;Choi, Mi Yeon;Jung, Hyun Ji;Kim, Hyun Ju
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.16 no.4
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    • pp.227-238
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    • 2013
  • The purpose of this study is to develop a process technology to produce high hardness drinking water which meet drinking water standard, remaining useful minerals like magnesium and calcium in the seawater desalination process while removing the sulfate ions and chloride ions. Seawater have been separated the concentrated seawater and desalted seawater by passing on Reverse Osmosis membrane (RO). Using Nano-filtration membrane (NF), We were prepared primary mineral concentrated water that sodium chloride were not removed. By the operation of electro-dialysis (ED) having ion exchange membrane, we were prepared concentrated mineral water (Mineral enriched desalted water) which the sodium chloride is removed. We have produced the high hardness water to meet the drinking water quality standards by diluting the mineral enriched desalted water with deionized water by RO. Reverse osmosis membranes (RO) can separate dissolved material and freshwater from seawater (deep seawater). The desalination water throughout the second reverse osmosis membrane was completely removed dissolved substances, which dissolved components was removed more than 99.9%, its the hardness concentration was 1 mg/L or less and its chloride concentration was 2.3 mg/L. Since the nano-filtration membrane pore size is $10^{-9}$ m, 50% of magnesium ions and calcium ions can not pass through the nano-filtration membrane, while more than 95% of sodium ions and chloride ions can pass through NF membrane. Nano-filtration membrane could be separated salt components like sodium ion and chloride ions and hardness ingredients like magnesium ions and calcium ions, but their separation was not perfect. Electric dialysis membrane system can be separated single charged ions (like sodium and chloride ions) and double charged ions (like magnesium and calcium ions) depending on its electrical conductivity. Above electrical conductivity 20mS/cm, hardness components (like magnesium and calcium ions) did not removed, on the other hand salt ingredients like sodium and chloride ions was removed continuously. Thus, we were able to concentrate hardness components (like magnesium and calcium ions) using nano-filtration membrane, also could be separated salts ingredients from the hardness concentration water using electrical dialysis membrane system. Finally, we were able to produce a highly concentrated mineral water removed chloride ions, which hardness concentration was 12,600 mg/L and chloride concentration was 2,446 mg/L. By diluting 10 times these high mineral water with secondary RO (Reverse Osmosis) desalination water, we could produce high mineral water suitable for drinking water standards, which chloride concentration was 244 mg/L at the same time hardness concentration 1,260 mg/L. Using the linked process with reverse osmosis (RO)/nano filteration (NF)/electric dialysis (ED), it could be concentrated hardness components like magnesium ions and calcium ions while at the same time removing salt ingredients like chloride ions and sodium ion without heating seawater. Thus, using only membrane as RO, NF and ED without heating seawater, it was possible to produce drinking water containing high hardness suitable for drinking water standard while reducing the energy required to evaporation.