• Analytical Science and Technology
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• v.8 no.3
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• pp.259-264
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• 1995

The separation efficiency of metal ions by using the poly-NTOE(1, 12-diaza-3, 4:9, 10-dibenzo-5, 8-dioxacyclopentadeca-1, 12-ylene-2, 7-dihydroxyoctamethylene) has been determined by column chromatography in aqueous solution. Binding constants and separation factors for several poly-NTOE interactions were measured in aqueous solution. The order of these binding constants and separation factors with metal ions were Co(II)Zn(II) for the transition metal ions and Cd(II)

• Journal of the Korean Chemical Society
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• v.41 no.11
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• pp.612-638
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• 1997

The methods of separation and recovery of rare earth elements in monazite sand have been studied by the ion exchange chromatography. Both of cation and anion exchange resin were used as ion exchange resins and the solutions of EDTA, DTPA, IMDA and Ln-EDTA were used as eluents. The H+, Zn2+, Fe3+, Al3+, Cu2+, and NH4+ forms of cation exchange resin were used as retaining ions. Ln-EDTA solution was loaded on the EDTA form of anion exchange resin and separated. The Ln-EDTA solution was also used as an eluent for a selective separation of one element from the rare earth mixture solution. The size effects of resin column, the elution mechanism for the various elution types and the separation of a large amount of rare earths were studied.

• Korean Journal of Food Science and Technology
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• v.29 no.5
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• pp.1067-1070
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• 1997

Imunoglobulins from bovine plasma proteins were isolated by IMAC which $Cu^{2+}$ was chelated on a chelating sepharose fast flow gel. Most plasma proteins were eluted by 1st (0.01 M $Na_2HPO_4$, 0.5 M NaCl, pH 4.0) and 2nd elution buffers (0.01 M imidazol). According to the reverse phase HPLC analysis, it was found that proteins which were eluted by 1st elution buffer were mainly composed of serum albumin, while most IgG and transferrin were eluted by 2nd elution buffer. When protein fractions obtained by 2nd elution buffer was applied to ultra filtration system (molecular weight cut off: 100 kD), IgG was further purified. These results indicate that IMAC is an excellent tool for isolating imunoglobulins from plasma proteins.

• The Korean Journal of Mycology
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• v.31 no.3
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• pp.168-174
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• 2003

In this study, Cordyceps militaris was grown in a liquid medium containing colloidal chitin. A chitinase was purified from the supernatant or cultured medium by ammonium sulfate fractionation, DEAE-Sephadex A-25 and Sephadex G-50 column chromatography. Optimum temperature and pH of this enzyme were $35^{\circ}C$ and 5.5, respectively. The molecular weight of the chitinase was estimated to be 48.5 kDa by SDS-PAGE and its Km value was 0.57 mM. The activity of this enzyme was inhibited by $Cu^{2+},\;Mn^{2+},\;Hg^{2+},\;Zn^{2+},\;CO_{3}^{2-},\;SO_4^{2-},\;CN^-,\;ion,\;and\;OCN^-$ maleic anhydride, acetic anhydride or N-bromo succinimide, especially strongly inhibited by sodium cyanate for 84.0 percentage. But its activity wag slightly stimulated by $Mg^{2+}\;and\;K^+$ ion, respectively. The products formed during hydrolysis of the hexa-N-acetylchitohexaose with this enzyme were N,N'-diacetylchitobiose and N,N',N'-triacetylchitotriose. These results imply that this purified enzyme may be an endo-chitinase.

• Analytical Science and Technology
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• v.13 no.1
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• pp.27-33
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• 2000

Simultaneous separation and analysis of Ni(II), Pd(II), Co(II), Cu(II) and Hg(II) in peperidinedithiocarbamate (PDTC) chelates were investigated by reversed phase liquid chromatography. The optimum conditions for the separation of PDTC metal chelates were examined with respect to the pH, extraction solvent, and mobile phase strength on Novapak $C_{18}$ column using methanol/water mixture as mobile phase. All metal PDTC chelates were eluted in an acceptable range of capacity factor value ($0{\leq}log\;k^{\prime}{\leq}1$). The linear calibration curves were obtained in the concentration range of $0{\sim}1.2{\mu}g/mL$ for five metal ions, and also good precision in the range of 1.96~3.41% RSD was obseved. Under the optimum conditions, trace metat ions in the composite water sample were successfully separated and determined with relative error of ${\pm}2.0%$.

• Resources Recycling
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• v.29 no.6
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• pp.41-47
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• 2020

This study was carried out to utilize the coal bottom ash generated in a circulating fluidized bed combustion boiler as a treatment agent for heavy metal ions, and experiments were conducted to remove heavy metal ions from the acid mine drainage. The batch experiments were conducted to investigate the influence of dosage of ash, initial concentration of solution on the removal capacity of heavy metal ions (Cu, Cd, Cr, Pb). The results of the experiment showed that the total removal capacity of heavy metals was 30.8 mg/L and 46.4 mg/g, respectively, under the condition that the concentration of coal ash was added as 15 g/L of heavy materials and 10 g/L of light materials. After that, a long-term column experiment was performed to determine the maximum removal capacity of heavy metal ions (Cu, Cd, Cr, Pb, As), and the removal capacity for each metal component was investigated. After approximately 60 days of operation, the maximum removal capacity of heavy metals was 23.6 mg/g at pH 9.25.

• Microbiology and Biotechnology Letters
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• v.37 no.3
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• pp.204-212
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• 2009

A strain producing ${\alpha}$-galactosidase and ${\beta}$-glucosidase was isolated from Kimchi. The isolated strain was identified as Weissella cibaria by 16S rDNA analysis and designated as Weissella cibaria K-M1-4. The enzyme activity of ${\alpha}$-galactosidase and ${\beta}$-glucosidase reached the maximum in the soy medium at $37^{\circ}C$ for 24 hr. The enzymes were purified by ethanol fractionation, DEAE sepharose fast flow, and sephacryl S-100HR column chromatography. ${\alpha}$-Galactosidase specific activity was shown by 576 Units/mg protein and the yield was 3.5% of the total activity of crude extracts. ${\beta}$-glucosidase specific activity was shown by 480 Units/mg protein and the yield was 2.9% of the total activity of crude extracts. The optimum temperature for ${\alpha}$-galactosidase was $60^{\circ}C$ and 43% of its original activity remained when it was treated at $80^{\circ}C$ for 30 min. For ${\alpha}$-galactosidase shows the optimum pH of 8.0 and is fairly stable between pH5.0 and pH9.0. The enzyme activity was increased in the presence of $Fe^{2+}$ and $Cu^{2+}$. The value of Km and Vmax for the enzyme were 0.98 mM and $1.81{\mu}$mole/min, respectively. The ${\beta}$-glucosidase has the optimum temperature of $50^{\circ}C$ and 46% of its original activity remained when it was treated at $80^{\circ}C$ for 30min. Its optimum pH of 7.0 and is fairly stable between pH5.0 and pH9.0. The enzyme activity was increased in the presence of $Fe^{2+},\;Co^{2+}$ and $Cu^{2+}$. The value of Km and Vmax for the enzyme were 1.24 mM and $6.81{\mu}$mole/min, respectively.

• Journal of Microbiology
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• v.44 no.6
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• pp.622-631
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• 2006

In this study we purified a fibrinolytic enzyme from Cordyceps militaris using a combination of ion-exchange chromatography on a DEAE Sephadex A-50 column, gel filtration chromatography on a Sephadex G-75 column, and FPLC on a HiLoad 16/60 Superdex 75 column. This purification protocol resulted in a 191.8-fold purification of the enzyme and a final yield of 12.9 %. The molecular mass of the purified enzyme was estimated to be 52 kDa by SDS-PAGE, fibrin-zymography, and gel filtration chromatography. The first 19 amino acid residues of the N-terminal sequence were ALTTQSNV THGLATISLRQ, which is similar to the subtilisin-like serine protease PR1J from Metarhizium anisopliae var. anisopliase. This enzyme is a neutral protease with an optimal reaction pH and temperature of 7.4 and $37^{\circ}C$, respectively. Results for the fibrinolysis pattern showed that the enzyme rapidly hydrolyzed the fibrin $\alpha$-chain followed by the $\gamma$-$\gamma$ chains. It also hydrolyzed the $\beta$-chain, but more slowly. The A$\alpha$, B$\beta$, and $\gamma$ chains of fibrinogen were also cleaved very rapidly. We found that enzyme activity was inhibited by $Cu^{2+}$ and $Co^{2+}$, but enhanced by the additions of $Ca^{2+}$ and $Mg^{2+}$ ions. Furthermore, fibrinolytic enzyme activity was potently inhibited by PMSF and APMSF. This enzyme exhibited a high specificity for the chymotrypsin substrate S-2586 indicating it's a chymotrypsin-like serine protease. The data we present suggest that the fibrinolytic enzyme derived from the edible and medicinal mushroom Cordyceps militaris has fibrin binding activity, which allows for the local activation of the fibrin degradation pathway.

• Current Research on Agriculture and Life Sciences
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• v.7
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• pp.153-163
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• 1989

The activity changes and biochemical properties of ${\beta}$-gal in tomato fruits during ripening were investigated. The total activity was increased during ripening and three isoenzymes (${\beta}$-gal I, II and III) were purified through DEAE Sephadex A-50 and Sephadex G-100 column chromatography. The activities of ${\beta}$gal isoenzymes (${\beta}$-gal I, II and III) during ripening were 69.8, 31.8 and 170.0 units in mature green phase, while those were 48.7, 88.4 and 136.8 units in Red phase, respectively. As the ripening proceeded the activities of ${\beta}$-gal I and III were some what decreased but the activity of ${\beta}$-gal II was incresed more than 2.8 fold. The optimum pH of ${\beta}$-gal I, II and III were 3.9, 4.2 and 3.9 and the optimum temperature of those were $60^{\circ}C$, $56^{\circ}C$ and $60^{\circ}C$, respectively. All isoenzymes were stable at pH 3.6~6.0 and lost their activity about 50% when it heated at $55^{\circ}C$ for 5 minute. $Mg^{{+}{+}}$-activated the three isoenzymes but $Ca^{{+}{+}}$ and SDS inhibited about 30~40%. $Hg^{{+}{+}}$ inhibited completely. The km value of ${\beta}$-gal I, II and III was 0.36mM, 0.63mM and 0.45mM, reaction rate was rapidly increased until the concentration of substrate was $6.0{\times}10^{-5}M$.

• Herbal Formula Science
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• v.21 no.2
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• pp.133-143
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• 2013

Objectives : We carry out the simultaneous quantification for quality control of four components in Bangpungtongseong-san (BPTSS) sample. In addition, we assessed the antioxidant effects of BPTSS sample. Methods : The used column for separation and analysis of four compounds was Luna C18 column and column oven temperature was maintained at $40^{\circ}C$. The mobile phase for simultaneous determination consisted of two solvent systems, 1.0% acetic acid in water and 1.0% acetic acid in acetonitrile. High performance liquid chromatography-photodiode array (HPLC-PDA) method for analysis was performed at a flow rate of 1.0 mL/min with PDA detection at 254 and 280 nm. The injection volume was 10 ${\mu}L$. The antioxidant activities of BPTSS were evaluated by measuring free radical scavenging activities on 2,2'-Azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) and 1-1-diphenyl-2-picrylhydrazyl (DPPH). The inhibitory effects on low-density lipoprotein (LDL) oxidation were evaluated by the formation of thiobarbituric acid relative substances (TBARS) and relative electrophoretic mobility (REM). Results : Calibration curves were acquired with $r^2{\geq}0.9999$. The values of limit of detection (LOD) and quantification (LOQ) were 0.06-0.29 ${\mu}g/mL$ and 0.20-0.98 ${\mu}g/mL$, respectively. The amounts of geniposide, liquiritin, baicalin, and glycyrrhizin in BPTSS were 5.06, 7.33, 27.56, and 7.81 mg/g, respectively. The BPTSS showed the radical scavenging activity in a dose-dependent manner. The concentration required for 50% reduction (RC50) against ABTS and DPPH radicals were 72.51 ${\mu}g/mL$ and 128.49 ${\mu}g/mL$. Furthermore, GMGHT reduced the oxidation properties of LDL induced by CuSO4. Conclusions : The established HPLC-PDA method will be helpful to improve quality control of BPTSS. In addition, BPTSS has potentials as therapeutic agent on anti-atherosclerosis.