• Title/Summary/Keyword: Anion exchange

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Unsteady Mass Transfer Around Single Droplet Accompanied by Interfacial Extraction Reaction of Succinic Acid (숙신산 추출반응이 일어나는 단일 액적계에서의 비정상상태 물질 전달)

  • Jeon, Sangjun;Hong, Won Hi
    • Korean Chemical Engineering Research
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    • v.50 no.6
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    • pp.1021-1026
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    • 2012
  • The transient mass transfer in a single droplet system consisting of 1-octanol (continuous phase)/aqueous succinic acid solution (dispersed phase) was investigated in the presence of chemical reaction, which is acid/anion exchange reaction of succinic acid and tri-n-octylamine (TOA). This succinic acid extraction by TOA can be considered to occur at the interface between organic and aqueous phase, that is, heterogeneous reaction system. The basic properties of the system such as viscosity, density, distribution coefficient, terminal velocity of droplet, and diffusion coefficient were measured experimentally or calculated theoretically, and used for theoretical calculation of characteristic parameters of mass transfer later. The effects of succinic acid concentration on the terminal velocity was negligible in the existence of TOA, although the terminal velocity increases with succinic acid concentration in the absence of TOA. On the contrary, the terminal velocity decreases with TOA concentration. While droplets falls through organic phase, the trajectory of droplets is observed to oscillate around its vertical path. A mass trnasfer cell was prepared to monitor the mass transfer behavior in a single droplet and used to measure the mean concentration of succinic acid inside droplet. The results are expressed with dimensionless parameters. Under 50 g/L succinic acid condition, the system with 0.1 mol/kg TOA showed that the molar flux decreases in proportion to the decrease of concentration gradient, while in the case of 0.5 mol/kg TOA Sh increases rapidly with time indicating the molar flux of succinic acid decreases relatively slowly compared to the decrease in concentration gradient.

Development of Purification Process of Recombinant Human Vascular Endotherial Growth Factor (VEGF) using Fusion Protein (융합 단백질을 이용한 재조합 인간 혈관내피세포 성장인자의 정제공정 개발)

  • Sung, Keehyun;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.55 no.3
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    • pp.369-378
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    • 2017
  • Vascular endotherial growth factor (VEGF) is a potent mitogen that stimulates vascular permeability and angiogenesis and has a potential in therapeutic applications. An industrial production method that provides high yield as well as purity is needed. Researches for various factors of mild solubilization with combination of ubiquitin fusion protein to increase solubility were carried out as well as by changing pH and denaturant concentration. Usage of pET28-a bacteral expression vector in BL21 (DE3) host cell was capable of producing approximately 14 g/L VEGF fusion protein in 20L fermentor. A purification process consisting of four chromatography steps including refolding and digestion with UBP1 resulted in mild solublization under the conditions of 2M urea and pH 10.0 due to ubiquitin fusion tag protein that increases in solubility of target protein VEGF. High yield of refolding and dimerization could be obtained between two step Ni-affinity chromatography. Multimeric and misfolded proteins and endotoxin were removed by DEAE anion exchange chromatography. Final monomers were removed from dimers by gel filtration chromatography. Characterization analysis of purified dimeric VEGF was performed using SDS-PAGE and RP-HPLC with a purity of 97%.

Isolation of Mitogenic Glycophosphopeptides from Cheese Whey Protein Concentrate (유청 단백질에서 유도되는 생리활성 펩타이드에 관한 연구)

  • Yun, Sung-Seob
    • Journal of Dairy Science and Biotechnology
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    • v.15 no.1
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    • pp.33-44
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    • 1997
  • We investigated the immunological function of cheese whey protein concentrate (CWPC), which is a by-product of cheese production, using mitogenic activity in murine splenocytes as an index. A fraction isolated by gel filtration and anion exchange chromatography of CWPC showed high mitogenic activity, comparable to the activity of lipopolysaccharide (LPS). The fraction was detected as a single band on SDS-PAGE. It contained calcium, inorganic phosphorus, and carbo-hydrate, indicating the active component to be a glycophosphopeptide (GPP) Since pronase digestion of GPP did not reduce its mitogenic activity, carbohydrate rather than peptide may be important in the activity, When applied on an anti-${\beta}$-caseinophosphopeptide (${\beta}$-CPP ) antibody affinity column, the GPP was separated into two components, one with affinity to ${\beta}$-CPP and the other without such affinity. Both the components contained N-linked oligosaccharide chains and had the mitogenic activity. These results demonstrate that cheese whey contains a GPP having strong mitogenic activity

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Risk Assessment of Arsenic in Agricultural Products (농산물 중 비소 위해평가)

  • Choi, Hoon;Park, Sung-Kug;Kim, Dong-Sul;Kim, Mee-Hye
    • Korean Journal of Environmental Agriculture
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    • v.29 no.3
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    • pp.266-272
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    • 2010
  • The present study was carried out to assess exposure & risk for Korean by total and inorganic As intake through agricultural products. Total arsenic analysis was performed using microwave device and ICP-MS. 50% MeOH extraction and anion-exchange HPLC-ICP-MS method has been used to determine arsenic species. 329 samples covering 20 kinds of agricultural products were collected from various retail outlets and markets across Korea. The concentration of total As was in the range of 0.001~0.718 mg/kg, while inorganic and organic arsenic species in all samples was not determined. For risk assessment, probable daily intake was calculated and compared with provisional tolerable weekly intake (PTWI, 15 ${\mu}g$/kg b.w./week for inorganic arsenic) established by JECFA. The median daily exposure to total and inorganic As by intake of agricultural products except rice was ranged 0.0002~0.012, 0.0001~0.001 ${\mu}g$/kg b.w./day, corresponding to 0.01~0.5%, 0.002~0.1% of PTWI, respectively. The median level of total and inorganic As intake through rice was 0.603 and 0.041 ${\mu}g$/kg b.w./day, and 28.1% and 1.9% of PTWI, respectively. Therefore, the level of overall exposure to arsenic for Korean through agricultural products was below the recommended JECFA levels, indicating of least possibility of risk.

Electrochemical Properties of Kaolinite in Aqueous Suspension (수용액중(水溶液中)에서의 Kaolinite 입자(粒子)의 전기화학적(電氣化學的) 성질(性質))

  • Lim, Hyung-Sik;Baham, J.;Volk, V.V.
    • Korean Journal of Soil Science and Fertilizer
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    • v.16 no.4
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    • pp.318-324
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    • 1983
  • Electrochemical properties of Georgia kaolinite in aqueous suspension were studied by ion adsorption, potentiometric titration, and electrophoretic mobility measurements. Kaolinite in 0.001 M and 0.1 M NaCl solution showed qualitatively both pH independent and pH depender negative and positive charges through pH range 2.5-11.0 when dissolved aluminum ions from kaolinite were considered as well as $Na^+$ and $Cl^-$ as index ions. Electrophoretic mobilities (EM) of 0.02 wt. % kaolinite suspension in distilled water and 0.001 M NaCl solution were approximately constant against mobility measuring time consumed in the electrophoresis cell at different pH values, and isoelectric points(IEP) were around pH 4.7. EM values in 0.1 M NaCl solution were positive and constant against mobility measuring time below pH 4; but above pH 4, EM values were negative for the first 10 seconds followed by positive values which became approximately constant through stepped changes after 10 minutes. Hydrated cations may bind to the six- member oxygen ring sites having multiple partial negative charges on the exterior tetrahedral layer surface by both electrostatic and hydrogen bonding force while hydrated anions bind to the partially positively charged hydrogen atoms on the exterior octahedral layer surface. Parts of the aluminol groups on the exterior octahedral layer surface as well as edge faces may be involved in complex reactions and have both anion and cation exchange capacities in the electrolyte solution above pH 4.

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Purification and Characterization of Degradative Enzyme of Dental Plaque from Streptomyces sp. Y9343 (Streptomyces sp. Y9343이 生産하는 齒面細菌膜 分解酵素의 精製와 特性)

  • Kim, Seong-Joo;Han, Hong-Keun;Yoon, Jeong-Weon
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.9-18
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    • 1996
  • Streptococcus mutans has been implicated as primary causative agents of dental caries by insoluble glucan (IG) in human and experimental animals. An attempt was made to search for the ${\alpha}$-1,3 glucanase that degrades IG produced by S. mutans. ${\alpha}$-1,3 glucanase was detected in the culture supernatant of microorganisms, which are isolated from soils on agar medium containing IG as a sole carbon source. This Streptomyces sp. hydrolysed IG produced by immobilized S. mutans and was named as Y9373. This enzyme required ${\alpha}$-1,3 glucan (IG) as an inducer. The optimum conditions for enzyme production were studied. The enzyme was purified by 30~70% $(NH_4)_2SO_4$ precipitation, anion exchange chroma tography on DEAE-cellulose and gel filtration on Sepadex G-75. The purified enzyme has a specific activity of 7840.0 U/mg protein giving 32.1-fold purification and final yield of 0.53%. The molecular weight was estimated to be about 22.5 kDa by SDS-PAGE. The optimum pH and temperature for enzyme reaction were 6.5 and 37$^{\circ}C$, respectively and the enzyme was relatively stable at the temperature below 60$^{\circ}C$. The activity of purified enzyme was enhanced by adding $Co^{2+},\;Mn^{2+}\;and\;Mg^{2+}$ into the medium, whereas inhibited by adding $Hg^{2+},\;Zn^{2+}$ and SDS. The $K_m\;and\;V_{max}$ value of ${\alpha}$-1,3 glucanase for IG were estimated to be 2.50 mM and 0.0431 mM/min, respectively. The thin layer chromatographic analysis of hydrolysates from IG with ${\alpha}$-1,3 glucanase showed that glucose was the main product of reaction. This enzyme activity was about 14 times higher than marketing dextranase as preventive agent against artificial dental caries by S. mutans in TH medium including 5% sucrose after 30 minutes.

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Development of Enzyme-Linked Immunosorbent Assay for the Detection of Fumonisins (효소면역측정법을 이용한 Fumonisin의 검출법 개발)

  • Shon, Dong-Hwa;Hahn, Seong-Min;Lim, Sun-Hee;Lee, Yin-Won;Cho, Sun-Hee;Kang, Shin-Young;Lee, Kyung-Ae
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.119-125
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    • 1996
  • In order to develop enzyme-linked immunosorbent assay (ELISA) for fumonisins, production of specific antibodies, establishment of ELISA conditions, and quantitation of the toxin from spiked corns by ELISA were performed. Fumonisin $B_1(FB_1)$ conjugated to cholera toxin (CT) with or without Freund's adjuvant was subcutaneously injected into 2 groups of rabbits. When the titer of the antisera produced by each rabbit was tested, higher titer was observed in case of the immunization with the adjuvant. By use of the antiserum showing the highest titer (1:16,000) and its purified antibodies, competitive indirect and direct ELISA's (ciELISA and cdELISA) were established, respectively. When the cross-reactivity of the antibody against fumonisin analogs was investigated by the ciELISA, it was very low against $B_3$ (2%) but high against fumonisin $B_2$ (179%). The sensitivity of the ELISAs was also very high, because the detection limit for $FB_1$ was 0.03 ppb in ciELISA and 0.3 ppb in cdELISA. When the ELISA's were applied to the spiked corns after extraction with 75% methanol, the assay recovery of $FB_1$ was too unstable to assay. However, when cleanup by strong anion exchange (SAX) cartridge was introduced to remove interfering materials, the mean ELISA recovery of $FB_1$ from corns spiked to 3~10 ppm was found to be 34.0% and stable (mean of CV, 8.2%).

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Purification and Properties of Polygalacturonase from Ganoderma lucidum (Ganoderma lucidum이 생산하는 Polygalacturonase의 정제 및 특성)

  • Yoon, Sook;Kim, Myung-Kon;Hong, Jai-Sik;Kim, Myeong-Sook
    • The Korean Journal of Mycology
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    • v.22 no.4
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    • pp.298-308
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    • 1994
  • The properties of polygalacturonase by Ganoderma lucidum in liquid culture were investigated. The enzyme was composed of an endo- and an exo-polygalacturonase. The endo- and exo-polygalacturonase were purified approximately 56 and 9.2-fold, respectively, through ammonium sulfate fractionation, gel filtration on Biogel P-100, anion exchange chromatography on DEAE-cellulose, gel chromatography on Sephadex G-150 and re-gel chromatography on Sephadex G-150. The endo- and exo-polygalacturonase had higher affinity for apple pectin than for citrus pectin or pectic acid. The Km values of the endo- and exo-polygalacturonase for apple pectin, determined on the Lineweaver-Burk plot, were 1.44 and 10.6 mg $ml^{-1}$ for apple pectin, respectively. Purified endo-polygalacturonase was found to be homogeneous electrophoretically and had a molecular weight of 54,000 estimated on SDS polyacrylamide gel. The optimal pH for the activity of the enzymes was 4.0. The endo- and exo-polygalacturonase were stable in the pH range of 4.0 to 6.0 and 3.5 to 5.5, respectively. The optimal temperatures of the endo- and exo-polygalacturonase were 40 and $60^{\circ}C$, respectively. The exo-polygalacturonase was more resistant to heat than the endo-polygalacturonase, requiring heating for 40 min at $80^{\circ}C$ for complete inactivation. The activity of the endo-polygalacturonase was increased by $Ca^{++}$ and $Mn^{++}\;ions$, while that of the exo-polygalacturonase was increased by $Ca^{++}\;ion$ only, and was not affected by $Mn^{++}\;ion$.

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Copper Isotope Measurements Using a Neptune MC-ICP-MS (다검출기 유도결합 플라즈마 질량분석기를 이용한 구리 동위원소 분석법)

  • Park, Sanghee;Ryu, Jong-Sik;Shin, Hyung Seon;Kil, Youngwoo;Jo, Yunsoo
    • Journal of the Mineralogical Society of Korea
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    • v.29 no.4
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    • pp.221-227
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    • 2016
  • Copper is an essential transition metal involving in various biogeochemical processes. With the recent advances in analytical techniques and mass spectrometry, such as MC-ICP-MS, it is possible to measure Cu isotopes, which allows us to understand various biogeochemical processes in detail. Nonetheless, few studies have been performed in South Korea. In this study, we compared two purification methods previously reported using an anion exchange resin ($AG^{(R)}$ MP-1M), developed the best method in our lab environment, and then verified it by measuring Cu isotopic compositions in two USGS geological reference materials (BHVO-2 and BIR-1a). Although all matrix cations causing mass bias were effectively removed through both two methods with the yield of better than 95%, the method using the mixture of HCl and $H_2O_2$ only displays Cu isotopic compositions, in excellent agreement with reported values within the error. The method developed in this study is expected to be commonly applied to earth and environmental sciences.

Functional Characterization of the ${\alpha}$- and ${\beta}$-Subunits of a Group II Chaperonin from Aeropyrum pernix K1

  • Lee, Jin-Woo;Kim, Se Won;Kim, Jeong-Hwan;Jeon, Sung-Jong;Kwon, Hyun-Ju;Kim, Byung-Woo;Nam, Soo-Wan
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.818-825
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    • 2013
  • We isolated and functionally characterized the ${\alpha}$- and ${\beta}$-subunits (ApCpnA and ApCpnB) of a chaperonin from Aeropyrum pernix K1. The constructed vectors pET3d-ApCpnA and pET21a-ApCpnB were transformed into E. coli Rosetta (DE3), BL21 (DE3), or CodonPlus (DE3) cells. The expression of ApCpnA (60.7 kDa) and ApCpnB (61.2 kDa) was confirmed by SDS-PAGE analysis. Recombinant ApCpnA and ApCpnB were purified by heat-shock treatment and anion-exchange chromatography. ApCpnA and ApCpnB were able to hydrolyze not only ATP, but also CTP, GTP, and UTP, albeit with different efficacies. Purified ApCpnA and ApCpnB showed the highest ATPase, CTPase, UTPase, and GTPase activities at $80^{\circ}C$. Furthermore, the addition of ApCpnA and ApCpnB effectively protected citrate synthase (CS) and alcohol dehydrogenase (ADH) from thermal aggregation and inactivation at $43^{\circ}C$ and $50^{\circ}C$, respectively. In particular, the addition of ATP or CTP to ApCpnA and ApCpnB resulted in the most effective prevention of thermal aggregation and inactivation of CS and ADH. The ATPase activity of the two chaperonin subunits was dependent on the salt concentration. Among the ions we examined, potassium ions were the most effective at enhancing the ATP hydrolysis activity of ApCpnA and ApCpnB.