• Proceedings of the Botanical Society of Korea Conference
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• 1987.07a
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• pp.133-148
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• 1987

Gibberellin(GA) 3-$\beta$ hydroxylation is very important for the shoot elogation in the higher plants, since only 3$\beta$-hydryoxylated GAs promote shoot elogation in several plants. Fluctuation of 3$\beta$-hydryoxylase activity was examined during seed maturation using two cultivars of , P. vulgaris, Kentucky Wonder (normal) and Masterpiece (dwarf). Very immature seeds of both cultivars contain high level of 3$\beta$-hydroxylase activity (per mg protein). Both cultivars showed maximum of enzyme activity (per seed) in the middle of their maturation process. Gibberellin 3$\beta$-hydroxylase catalyzing the hydroxylation of GA20 to GA1 was purified 313-fold from very early immature seeds of P. vulgaris. Crude soluble enzyme extracts were purified by 15% methanol precipitation, hydrophobic interaction chromatogrphy, DEAE ion exchange column chromatography and gel filtration HPLC. The 3$\beta$-hydroxylase activity was unstable and lost much of its activity duting the purification. The molecular weight of purified enzyme was extimated to be 42, 000 by gel filtration HPLC and SDS-PAGE. The enzyme exhibited maximum activity at pH 7.7. The Km values for [2.3-3H] GA20 and [2.3-3H]GA9 were 0.29 $\mu$M and 0.33 $\mu$M, respectively. The enzyme requires 2-oxoglutarate as a cosubstrate; the Km value for 2-oxoglutarate was 250 $\mu$M using 3H GA20 as a substrate. Fe2+ and ascorbate significantly activated the enzyme at all purification steps, while catalase and BSA activated the purified enzyme only. The enzyme was inhibited by divalent cations Mn2+, Co2+, Ni2+, Cu2+, Zn2+, Cd2+ and Hg2+. Effects of several GAs and GA anaogues on the putrified 3$\beta$-hydroxylase were examined using [3H]GA9 and GA20 as a substrates. Among them, GA5, GA9, GA15, GA20 and GA44 inhibited the enzyme activity. [13C, 3H] GA20 was converted by the partially purified enzyme preparation to [13C, 3H]GA1, GA5 and GA6, which were identified by GC-MS, GA9 was converted only GA4, GA15 and GA44 were converted to GA37 and GA38, respectively. GA5 was epoxidized to GA6 by the preparation. This suggests that 3$\beta$-hydroxylation of GA20 and epoxidation of GA5 are catalyzed by the same enzyme in P, vulgaris.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.327-333
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• 2005

Dihydroxy-acid dehydratase (DHAD, 2,3-dihydroxy-acid hydrolyase, EC 4.2.1.9) is one of the key enzymes involved in the biosynthetic pathway of the branched chain amino acid isoleucine and valine. Although the enzyme have been purified and characterized in various mesophiles including bacteria and eukarya, the biochemical properties of DHAD has bee not yet reported from hyperthermophilic archaea. In this study, we cloned, expressed, and purified a DHAD homologue from the thermoacidophilic archaeon Sulfolobus solfataricus P2, which grows optimally at $80\;^{\circ}C$ and pH 3, in E. coli. Characterization of the recombinant S. solfataricus DHAD (rSso_DHAD) revealed that it is the dimeric protein with a subunit molecular weight of 64,000 Da in native structure. rDHAD showed the highest activity toward 2,3-dihydroxyisovaleric acid among 17 aldonic acid substrates Interestingly, this enzyme also displayed 50 % activities toward some pentonic acids and hexonic acids when compared with the activity of this enzyme to the natural substrate. Moreover, rSso_DHAD indicated relatively higher activity toward D-gluconate than any other hexonic acids tested in substrates. $K_m$ and $V_{max}$ values of rSso_DHAD were calculated as $0.54\;{\pm}\;0.04\;mM$ toward 2,3dihydroxyisovalerate and $2.42\;{\pm}\;0.19\;mM$ toward D-gluconate, and as $21.6\;{\pm}\;0.4\;U/mg$ toward 2,3-dihydroxyisovalerate and $13.8\;{\pm}\;0.4\;U/mg$ toward D-gluconate, respectively. In the study for biochemical properties, the enzyme shows maximal activity between $70^{\circ}C$ and $80^{\circ}C$, and the pH range of pH 7.5 to 8.5. The half life time at $80^{\circ}C$ was 30 min. A divalent metal ion, $Mn^{2+}$, was only powerful activators, whereas other metal ions made the enzyme activity reduced. $Hg^{2+}$, organic mercury, and EDTA also strongly inhibited enzyme activities. Particularly, the rSso_DHAD activity was very stable under aerobic condition although the counterparts reported from mesophiles had been deactivated by oxygen.

• Microbiology and Biotechnology Letters
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• v.31 no.3
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• pp.284-291
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• 2003

Bacillus amyloliquefaciens K-42, which produces strongly a fibrinolytic enzyme, Was isolated from Ganjang, a traditional Korean soy sauce. The fibrinolytic enzyme was purified to homogeneity by ammonium sulfate fractionation, ion-exchange chromatography on DEAE-Sephadex A-50, gel chromatography on Sephadex G-100, and gel chromatography on Sephadex G-75 of the culture filtrate of Bacillus amyloliquefaciens K42. The purified enzyme showed the specific activity of 59.4 units per milligram, which was increased by 17.1 fold over the culture broth. And the molecular weight of purified fibrinolytic enzyme was confirmed to be about 45,000 Dalton by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme activity was relatively stable at pH 4.0-10.0 and the optimum pH was 8.0. The activity of the purified enzyme was increased by $Mg^{2+}$ , Cu$^{2+}$ but the enzyme was totally inhibited by $Ba^{2+}$ $Hg^{2+}$ In addition, the enzyme activity was potently inhibited by EDTA, EGTA and CDTA. It was concluded that the purified enzyme was a metalloprotease. And Km value was 2.03 mg/ml to fibrin.

• Journal of Biomedical Engineering Research
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• v.28 no.3
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• pp.387-391
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• 2007

An oxygenator is a very important artificial organ and widely used for patients with lung failure or during open heart surgery. Although an oxygenator has been widely studied worldwide to enhance its efficiency, studies on oxygenators, in particular when using a pulsatile blood flow, are domestically limited. Therefore, a new oxygenator was developed in the lab and animal experimental results are described in the paper. The oxygenator is composed of polycarbonate housing and polypropylene hollow fibers. It has a total length of 400 mm and a surface area of $1.7 m^2$. The animal experiment lasted for 4 hours. The blood flow rate was set to 2 L/min and a pulsatile blood pump, T-PLS (Twin-Pulse Life Support), was used. Samples were drawn at the oxygenator's inlet and outlet. The total hemoglobin (Hb), saturation oxygen ($sO_2$), and partial oxygen pressure ($pO_2$), partial $CO_2$ pressure ($pCO_2$), and plasma bicarbonate ion concentration ($HCO_3^-$) were measured. The oxygen and carbon dioxide transfer rates were also calculated based on the experimental data in order to estimate the oxygenator's gas transfer efficiency. The oxygen and carbon dioxide transfer rates were $16.4{\pm}1.58$ and $165.7{\pm}10.96 mL/min$, respectively. The results showed a higher carbon dioxide transfer rate was achieved with the oxygenator. Also, the mean inlet and outlet blood pressures were 162.79 and 137.92 mmHg, respectively. The oxygenator has a low pressure drop between its inlet and outlet. The aim of own preliminary study was to make a new oxygenator and review its performance when applying a pulsatile blood pump thus, confirming the possibility of a new oxygenator suitable for pulsatile flow.

• The Korean Journal of Mycology
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• v.12 no.4
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• pp.133-140
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• 1984

Some properties of cellulolytic enzymes produced by Pleurotus sajor-caju JAFM 1017 during its growth in synthetic medium were investigated. The optimum pH of avicelase, CMCase, and ${\beta}-glucosidase$ was pH 5.5, pH 4.5 and pH 6.0, respectively. Avicelase and CMCase were stable within pH 5.0 to 6.0 and 4.0 to 6.0, respectively, and ,${\beta}-glucosidase$ was within pH 5.5 to 6.5. The optimum temperature of avicelase, CMCase and ${\beta}-glucosidase$ was the same of $40^{\circ}C$. The enzymes were stable below the optimum temperature, but the enzymes were unstable over the temperature of $50^{\circ}C$, and avicelase was losing about 91.7% of activity at $70^{\circ}C$ for 10 min. The enzyme activity of avicelase and CMCase was increased in proportion to the substrate concentration within 1% and 0.7%, respectively, and ${\beta}-glucosidase$ was within 0.1%. The Michaelis constants (Km) of avicelase and CMCase were 30.77mg avicel/ml and 14.64m Na-CMC/ml, respectively and ${\beta}-glucosidase$ was 5. 13mg salicin/ml. The reducing sugar production of avicelase was proportionaly increased until 120 min. and CMCase and ${\beta}-glucosidase$ were until 60min. The activity of three cellulolytic enzymes were increased by $Ca^{2+}$ at the concentration of $10^{-2}M$, but were inhibited by $Hg^{2+}$, $Ag^+$.

• Journal of the Korean Society of Food Science and Nutrition
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• v.19 no.6
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• pp.625-635
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• 1990

The purpose of this study was focused on investigation of biochemical properties of amylases in germinating corn(Zea mays L.) the amylase(I), (II) and (III) from germinating corn seeds were partially purified by ammonium sulfate precipitation, DEAE-Sephadex A-50 ion exchange column chromatography and Sephadex G-100 gel filtration. The last step was effective for separation of the corn amylases to a homogeneous slate. the purified amylase(I) was identified as a kind of $\alpha$-amylase from the fact that 5％ starch solution was hydrolysed into mainly maltose and maltotetrose by it, and amylase(II) and amylase(III) were enzymes producing maltotetrose as main product. The molecular weight and specific activity of the amylase(I), (II) and (III) were determined to be 54,000 and 70.47 unit/mg, 39,000 and 62.98 unit/mg, and 51,000 and 80.39 unit/mg, respectively. It showed a tendency to increase the amylases activities in presence of Ba, Ca, Co and Fe groups, but inhibits in that of Ag, Sn, Hg and Zn groups, and amylase(I), (II) and (III) remained stable at pH 5-6 and 2$0^{\circ}C$ for 40 days in containing of 1 mM CaCl$_2$. The optimum pH and optimum temperatures were pH 6, pH 5 and pH 6 and 35$^{\circ}C$, 55$^{\circ}C$ and 55$^{\circ}C$, respectively. These results suggest that the amylase(I), (II) and (III) were different amylases.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2004.09a
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• pp.119-120
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• 2004

Urban groundwater has a unique hydrologic system because of the complex surface and subsurface infrastructures such as deep foundation of many high buildings, subway systems, and sewers and public water supply systems. It generally has been considered that increased surface impermeability reduces the amount of groundwater recharge. On the other hand, leaks from sewers and public water supply systems may generate the large amounts of recharges. All of these urban facilities also may change the groundwater quality by the recharge of a myriad of contaminants. This study was performed to determine the factors controlling the recharge of deep groundwater in an urban area, based on the hydrogeochemical characteristics. The term ‘contamination’ in this study means any kind of inflow of shallow groundwater regardless of clean or contaminated. For this study, urban groundwater samples were collected from a total of 310 preexisting wells with the depth over 100 m. Random sampling method was used to select the wells for this study. Major cations together with Si, Al, Fe, Pb, Hg and Mn were analyzed by ICP-AES, and Cl, N $O_3$, N $H_4$, F, Br, S $O_4$and P $O_4$ were analyzed by IC. There are two groups of groundwater, based on hydrochemical characteristics. The first group is distributed broadly from Ca-HC $O_3$ type to Ca-C1+N $O_3$ type; the other group is the Na+K-HC $O_3$ type. The latter group is considered to represent the baseline quality of deep groundwater in the study area. Using the major ions data for the Na+K-HC $O_3$ type water, we evaluated the extent of groundwater contamination, assuming that if subtract the baseline composition from acquired data for a specific water, the remaining concentrations may indicate the degree of contamination. The remainder of each solute for each sample was simply averaged. The results showed that both Ca and HC $O_3$ represent the typical solutes which are quite enriched in urban groundwater. In particular, the P$CO_2$ values calculated using PHREEQC (version 2.8) showed a correlation with the concentrations of maior inorganic components (Na, Mg, Ca, N $O_3$, S $O_4$, etc.). The p$CO_2$ values for the first group waters widely ranged between about 10$^{-3.0}$ atm to 10$^{-1.0}$ atm and differed from those of the background water samples belonging to the Na+K-HC $O_3$ type (<10$^{-3.5}$ atm). Considering that the p$CO_2$ of soil water (near 10$^{-1.5}$ atm), this indicates that inflow of shallow water is very significant in deep groundwaters in the study area. Furthermore, the P$CO_2$ values can be used as an effective parameter to estimate the relative recharge of shallow water and thus the contamination susceptibility. The results of our present study suggest that down to considerable depth, urban groundwater in crystalline aquifer may be considerably affected by the recharge of shallow water (and pollutants) from an adjacent area. We also suggest that for such evaluation, careful examination of systematically collected hydrochemical data is requisite as an effective tool, in addition to hydrologic and hydrogeologic interpretation.ion.ion.

• Korean Journal of Mineralogy and Petrology
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• v.33 no.4
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• pp.391-405
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• 2020

Leaching chracteristics of Andong-dam sediment was conducted for heavy metal and nutrients. Five mixed sediment samples were prepared and leaching was conducted under aerobic and anaerobic condition for 60 days. Cd, Cu, Pb, Cr, Zn, Hg, As, Fe, Mn, phosphorus, and nitrogen were analyzed at each sampling time. The leaching rate of phosphorus was higher in anaerobic condition comparing with that of under aerobic condition. Some samples showed higher than the water-quality level IV. In case of As and Cd which showed highest contamination level in the sediment, leached concentration were 0.028 mg/L and 0.003 mg/L in maximum, respectively. The leached concentration is below than the lake water quality standard of Korea. Other heavy metals including Cu, Pb, and Cr also showed similar trend. Five step sequential extraction showed that easily extractable 1-2 step portion such as ion-exchangeable and adsorbed one was less than 10% and the most of the portion was residual. For As and Cd, the residual portion were 80% and 95% respectively indicating the risk by the heavy metal leaching into the lake for a short period was not high in comparing with the contamination levels.

• The Korean Journal of Physiology
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• v.2 no.2
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• pp.33-43
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• 1968

Histamine, 0.5 mg as histamine base in 4 ml of normal saline solution, was injected into rabbits anesthetized with nembutal and the mean blood pressure was kept in the range of $52{\sim}80\;mmHg$ for over one hour by supplemental additions. Following the injection of the test substances, 300 mg of urea and 200 mg of antipyrine intravenously, serial blood samples were obtained from the femoral artery and the internal jugular vein at $0.5{\sim}3$ minutes interval. The decreasing patterns in the concentrations of arterial and venous blood plasma samples were compared with each other. The ratio of the concentration of brain tissue to that of the final arterial plasma was also studied. By these measures the degrees of penetration of the test substances in the brain in the control and in the histamine treated rabbits were observed. The concentrations of antipyrine and urea in the arterial blood plasma were decreasing exponentially with respect to the time elapsed. The venous concentrations were anticipated to increase initially and to cross the arterial concentration curve in the point of equlibrium between the plasma and the tissue. On the contrary to the expectation venous concentration also revealed the decreasing tendency similar to that of arterial plasma. The similarity between these two curves, arterial and venous, would be atributable to the fact that the cerebral blood flow rate was large enough and the rising phase in the venous concentration curve was instantly over before serial blood samples were taken. Inspite of some similarity in the decreasing tedency in both concentration curves there were appreciable discrepancies between the arterial and venous plasma which would reflect the situation far from the equlibria among several compartments in the brain. Changes in plasma potassium levels caused by the injection of histamine or bleeding were observed, too. Using 8 rabbits as the control and 12 rabbits for the histamine treated group following results were obtained: 1. Both of the concentration curves, arterial and venous, declined rapidly at_first and slowly later on and approached same equilibrium concentration with the passage of time after a single injection. The time at which attained the same concentration was $2.0{\pm}0.54\;min.$ in the control and $4.3{\pm}1.92\;min.$ in the histamine treated group with respect to antipyrine. On the other hand in the case of urea they were $2.4{\pm}0.59\;min.$ in the control and $4.4{\pm}1.31\;min.$ in the histamine group, respectively. In the histamine treated group enlarged spaces for distribution of test substances were postulated. 2. The concentration of antipyrine in the brain tissue water revealed no significant differences between the control and experimental groups, showing $212{\pm}40.2\;mg/l$ in the control and $206{\pm}64.1\;mg/l$ in the histamine treated group. On the other hand urea revealed higher value in the histamine treated group than in the control, showing an enhanced penetration of urea into the tissue after injection of histamine. Urea concentration in the brain water was $32.3{\pm}3.36\;mg%$ in the control and $39.2{\pm}4.25\;mg%$ in the histamine treated group. 3. The distribution ratio of antipyrine in the brain tissue was very close to unity in the histamine treated animals as well as in the control. 4. The average of the distribution ratio of urea in the control animals was 0.77 and it showed the presence of blood-brain barrier with regard to urea. However in the histamine treated animals the distribution ratios climbed up to 0.86 and they were closer to unity than in the control animals. Out of 12 cases 5 were greater than 0.9 and 8 exceeded 0.85. It appeared that histamine enhanced the penetration of urea through the barrier. 5. Histamine injection and or hemorrhage caused an elevation of the concentration of potassium in plasma. In the event that histamine and hemorrhage were applied together the elevation of potassium exceed the elevation seen at the histamine alone. There was no evidence that the leakage of potassium from the brain tissue was dominant in comparison with the general leakage from the whole body.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.7
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• pp.1115-1124
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• 2013

The present study was conducted to ensure the diversity of domestic solar salt by analyzing the composition and microbiological characteristics of solar salt (from Docho island: DS) and the flower of salt produced in different Korean salt flats (Sinui island: SF, Bigum island: BF, and Docho island: DF). The analyses showed that the moisture content of the three types of flower of salt and solar salt ranged from 10.54~13.82% and NaCl content ranged from 78.81~84.61%. The mineral content of those salts ranged from 3.57~5.51%. The content of insoluble matter in these salts was $0.01{\pm}0.00{\sim}0.05{\pm}0.00%$. The sand content of these salts was $0.01{\pm}0.01{\sim}0.03{\pm}0.01%$. By Hunter's color value analysis, the color of the flower of salt was brighter and whiter than solar salt. The salinity of the flower of salt was a little higher than solar salt as well. The magnesium and potassium ion content of DF was $9,886.72{\pm}104.78mg/kg$ and $2,975.23{\pm}79.73mg/kg$, respectively, which was lower than the content in SF, BF, and DS. The heavy metal content of all salts was acceptable under the Korean Food Sanitation Law. The flower of salt was confirmed to be sweeter and preferable to solar salt. More than 80% of the solar salt crystals were 2~3 mm in size, whereas crystals from the flower of salt were 0.5~2 mm in size. The bacterial diversity of DF and DS were investigated by culture and denaturing gradient gel electrophoresis (DGGE) methods. The number of cultured bacteria in flower of salt was approximately three times more than solar salt. By DGGE analysis, major microbes of DF were Maritimibacter sp., Cupriavidus sp., and unculturable bacteria, and those of DS were Cupriavidus sp., Dunalidella salina and unculturable bacteria. The results of DGGE analysis showed that major microorganisms in solar salts were composed of unidentified and unculturable bacteria and only a few microorganisms were culturable.