• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.5
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• pp.529-533
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• 2002

The peptides inhibiting angiotensin converting enzyme (ACE) were isolated from the hydrolysate of manila clam (Ruditapes philippinamm) proteins prepared with thermolysin. The thermolysin hydrolysate was pretreated with membrane filter (MW cut-off 10,000) to obtain the peptide fraction with ACE inhibition. The crude peptides were applied to a Sephadex LH-20 column and eluted with $30\%$ methanol. The three active fractions (A, B and C) were collected and concentrated, and then applied to a SP-Toyopearl 650S column equilibrated with distilled water and was eluted with a linear gradient of NaCl concentration (0 to 1 M). The four active fractions (A-1, A-2, B-1 and C-1) were collected and concentrated, and then applied to a SuperQ-Toyopearl 650S column equilibrated with distilled water and was eluted with a linear gradient of NaCl concentration (0 to 1 M). The maximum inhibitory activity was observed in the fraction B-1Q showed the IC_{50} values of 0.748 $\mu$g. The abundant amino acids obtained from active fraction B-1Q were leucine, isoleucine, alanine and threonine.

• Journal of the Korean Chemical Society
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• v.32 no.3
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• pp.211-226
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• 1988

Liquid chromatographic behavior of several metal ions in dithiocarbamate(DTC) chelates were investigated by reversed phase high performance liquid chromatography on Novapak $C_{18}$ and ${\mu}$-Bondapak $C_{18}$ columns. The optimum conditions for the separation of DTC-metal chelate were examined with respect to the pH, shaking time, flow rate, extraction solvent, and mobile phase strength. The metal ions in mixtures at trace level, chelated with some dithiocarbamate derivatives were separated successfully on Novapak $C_{18}$ column using acetonitrile/methanol/water or acetonitril/water mixtures as mobile phases. It was found that all DTC metal chelates studied were eluted in an acceptable range of capacity factor values ($0{\leqq}log\;k'{\leqq}1$). Although several foreign metal ions were coexisted, high recovery and good precision were attained ; 97.0-106.7 % for the recovery and 0.98-3.41% for the coefficient of variation. Under the optimum analytical conditions, trace metal ions in the composite water samples were determined sucessfully with in relative error of about {\pm}$6.7 %.

• Korean Journal of Microbiology
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• v.47 no.4
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• pp.302-307
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• 2011

Native chitin deacetylase of Aspergillus nidulans was purified to apparent homogeneity by a combination of phenyl-Sepharose and Q-Sepharose column chromatography. In order to analyze the amino acid residues involved in the enzyme activity, the enzyme was chemically modified with chemical agent, which selectively reacted with the specific amino acid residue on the protein. When the enzyme was chemically modified with diethylpyrocarbonate, which specifically reacted with histidine residues on the protein, the activity was eliminated. The chitin deacetylase, chemically modified with 100 ${\mu}M$ modifier at the residue of arginine or tyrosine, has shown to have decreased activities. It was shown that the modification at aspartic acid or glutamic acid did not affect the enzyme activity to a greater extent, which would not implicate that acid amino residues were directly involved in catalytic reaction and would affect on the global structures of the proteins. This results demonstrated that histidine and tyrosine residues of enzyme would participate in an important function of the chitin deacetylase activity.

• Journal of Life Science
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• v.25 no.9
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• pp.1021-1026
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• 2015

Invertase (β-D-fructosfuranosidase, EC 3.2.1.26) catalyzes the hydrolysis of sucrose into D-glucose and D-fructose. Three biochemical subgroups of invertases have been investigated in plants: vacuolar (soluble acid), cytoplasmic (soluble alkaline), and cell wall-bound (insoluble acid) invertases. An isoform of neutral invertase was purified from pea seedlings (Pisum sativum L.) and treated with gibberellic acid (GA) by sequential procedures consisting of ammonium sulfate precipitation, ion-exchange chromatography, absorption chromatography, and reactive green-19 affinity chromatography. The results of the overall insoluble invertase purification were a 430-fold increase. The purified neutral invertase was not glycosylated and had an optimum pH between neutral and alkaline (pH 6.8-7.5). It was inhibited by Tris, as well as by heavy metals, such as Hg²⁺ and Cu²⁺. Typical Michaelis–Menten kinetics were observed when the activity of the purified invertase was measured, with sucrose concentrations up to 100 mM. The K_m and V_max values were 12.95 mM and 2.98 U/min, respectively. The molecular mass was around 20 kDa. The sucrose-cleaving enzyme activity of this enzyme is similar to that of sucrose synthase and fructosyltransferase, but its biochemical characteristics are different from those of sucrose synthase and fructosyltransferase. Based on this biochemical characterization and existing knowledge, neutral INV is an invertase isoform in plants.

• Korean Journal of Microbiology
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• v.30 no.2
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• pp.101-107
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• 1992

Cytochrome c oxida5e from chemotrophically grown R p , geliitinosu was purified by cytochrome c affinity chromatography and DEAE-Sephacel ion exchange chromatography. The molecular weight of the cytochrome c oxidase was approximately 110.000 Da by sephacryl s-300 gel chromatography and approximately 52, 000 Da by SDS-gel electrophoresis, respectively. Therefore. cytochrolne c oxidase of Rps. gehtinosu seems to be dimer. The cytochrome c oxidasc was very sensitive to temperature. It's Km and Vmax were 20 pM and 44 unitlmg protein for horsc heart cytochrome c as a substrate. respectively, and its optimum pH and temperature were 6.4 and 25$^{\circ}$C. respectively. The absorption peaks of the reduced cytochrome c oxidase showed at 554 nm, 523 nm. and 422 nm. The activiiy of cytochrome c oxidase was inhibited by KCN, and NaN3, but not by CO, antimycir~ A. and myxothiazol. The cytochrome c-551 was produced either in phototrophically or chemotrophically grown Rps. gelaiinosci. The rcduced cytochrome c-551 was oxidized by b-type cytochrome c oxidase from Rp.v. gc.lrtino.sc~. Km and Vmax of cytochrome c oxidase was 26 pM and 31 unitlnlg protein For cytochrome c-551 as a substrate. respectively. Thercfore. thc electron transfer chain of chemotrophically grown Rps. glatinosa seems lo be ubiquinol cytochrome bc, complex -'cytochrome c-55lMb-type cytochrome c oxidase+02.

• Journal of Applied Biological Chemistry
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• v.58 no.2
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• pp.175-181
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• 2015

Compositions of essential oils extracted from mint herb such as Mentha piperita, Mentha spicata, and Mentha ${\times}$ piperita var. citrate produced in Jeju were analyzed using gas chromatography-mass spectrometry (GC-MS) and headspace-GC-MS (HS-GC-MS). By the GC-MS analysis, 13 compounds were tentatively identified in Mentha piperita, Mentha spicata, and Mentha ${\times}$ piperita var. citrate, respectively. Peperitenone oxide, carvone, and linalool were detected as major compounds in Mentha piperita, in Mentha spicata, in Mentha ${\times}$ piperita var. citrate, respectively, based on the ratio of peak intensity in the total ion chromatogram. The greater number of compounds, including volatile alcohols and acetates were identified by HS-GC-MsS than by GC-MS in these all three essential oils. Similar patterns of composition were detected in both Mentha spicata and Mentha ${\times}$ piperita var. citrate by either one of GC-MS methods. However, in case of Mentha piperita, $\small{L}$-(-)-menthol, which was identified as the major compound by HS-GC-MS was detected in dramatically reduced quantity by GC-MS. Interestingly, we found that both linalyl acetate and linalool were identified as the dominant compounds in the essential oil of Mentha ${\times}$ piperita var. citrate.

• Analytical Science and Technology
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• v.18 no.1
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• pp.13-22
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• 2005

The elution patterns of seventeen 2,3,7,8-substituted PCDDs/Fs and 12 dioxin-like PCBs were studied by both manual open columns and automatic parallel LC columns in cleanup procedure. PCDDs/Fs and non-ortho-PCBs from other mono-ortho-PCBs were separated on automatic LC column, whereas they were not separated on an open manual column. The elution study on two cleanup methods was carried out using the PAR solution of unlabeled congeners and checked the recovery of each congener. Total recoveries of cleanup fractionation were ranged between 61.9 ~ 96.0% for PCDDs/Fs and 70.4 ~ 79.0% for PCBs by manual open columns and 71.8 ~ 104.5% for PCDDs/Fs and 61.3-120.3% for PCBs by automatic parallel LC columns, respectively. Unfortunately, #169-HxCB and 1,2,3,7,8-PeCDD were not separated on DB-5MS capillary column. The ions of 1,2,3,7,8-PeCDD were selected at M/M+2 instead of M+2/M+4 suggested by EPA method 1613. It is possible to discriminate 1,2,3,7,8-PeCDD and PCB #169 in HRGC/HRMS analysis.

• Analytical Science and Technology
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• v.7 no.3
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• pp.379-386
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• 1994

The elution behaviors in reversed-phase liquid chromatography were investigated thermodynamically for 2-hydroxy-arylazopyrazolone chelates with Ni(II), Cu(II), Co(III), Cr(III) on Novapak $C_{18}$ column. There was a good linear dependence of the capacity factor(k') on the variations of column temperature in van't Hoff plot. From this result, it was confirmed that the retention mechanism of these chelates in the reversed phase liquid chromatography system was invariant under the condition of various temperatures. For the most cases of the chelates studied, the dependence of capacity factor(1n k') on enthalpy$(-{\Delta}H)^{\circ}$, calculated by van't Hoff plot showed a good linearity(r=0.980~0.999) except [Pm(2-OH_(5-Cl)PaPz](r=0.787) and also the compensation temperatures(${\beta}$) showed constant values. The range of compensation temperature values calculated from the slope of $-{\Delta}H^{\circ}$ vs 1n k' plots was 374.3~806.9K. It was suggested that the retention of metal-2-hydroxy-arylazopyrazolone chelates in the reversed phase liquid chromatography system was largely affected by the hydrophobic effect.

• Korean Chemical Engineering Research
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• v.49 no.2
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• pp.129-136
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• 2011

Simulated Moving Bed(SMB) process consists of multiple chromatographic columns, which are usually partitioned into four zones. Such a process characteristic allows a continuous binary separations those are impracticable in conventional batch chromatographic processes. Compared with batch chromatography, SMB has advantages of continuity, high purity and productivity. Various researches have been reported for the integration of reaction and recovery during process operation on the purpose of economics and effectiveness. Simulated Moving Bed Reactor(SMBR) is introduced to combine SMB as a continuous separation process and reactor. Several cases of SMBR have been reported for diverse reactions with catalytic, enzymatic and chemical reaction on ion exchange resin as main streams. With an early type of fixed bed using catalyst, SMBR has been developed as SMB using fluidized enzyme, SMB with immobilized enzyme and SMB with discrete reaction region. For simple modeling and optimization of SMBR, a method considering convection only is possible. A complex method considering axial dispersion and mass transfer resistance is needed to explain the real behavior of solutes in SMBR. By combining reaction and separation, SMBR has benefits of lower installation cost by minimizing equipment use, higher purity and yield by avoiding the equilibrium restriction in case of reversible reaction.

• Analytical Science and Technology
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• v.31 no.1
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• pp.23-30
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• 2018

Twenty-two allergy-induced aromatics in children were analyzed using a gas chromatography flame ionization detector (GC-FID) and gas chromatography mass spectrometer (GC-MSD). Analytes were extracted using an automatic Soxhlet extractor and centrifuged for 10 min in a fast freezing centrifuge, and the supernatant was transferred into a 2 mL vial and injected in split mode. Under the established conditions, the calibration curve showed linearity with a correlation coefficient of 0.996 or more. A wide range of sensitivity of 6.7 to 1,859,839 depending on the device characteristics and detector used was shown. The detection limit of the device was 0.0032 to $0.0335{\mu}g/mL$, and the maximum detection limit was less than $0.1{\mu}g/mL$. The detection limit of the method ranged from 0.0033 to $0.1161{\mu}g/mL$. In addition, the limit of quantification ranged from 0.0100 to $0.5422{\mu}g/mL$, with a level of precision ranging from 0.21 % to 4.89 % and a degree of accuracy ranging from 89 % to 111 %. The analytical method developed in this study was applied to commercial products.