• Journal of Life Science
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• v.25 no.5
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• pp.487-495
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• 2015

We carried out pH stability, chemical inhibition, chemical modification, and pH-dependent kinetic parameter assessments to further characterize protocatechuate 3,4-dioxygenase from Pseudomonas pseudoalcaligenes KF707. Protocatechuate 3,4-dioxygenase was stable in the pH range of 4.5~10.5. L-ascorbate and glutathione were competitive inhibitors with $K_{is}$ values of 0.17 mM and 0.86 mM, respectively. DL-dithiothreitol was a noncompetitive inhibitor with a $K_{is}$ value of 1.57 mM and a $K_{ii}$ value of 8.08 mM. Potassium cyanide, p-hydroxybenzoate, and sodium azide showed a noncompetitive inhibition pattern with $K_{is}$ values of 55.7 mM, 0.22 mM, and 15.64 mM, and $K_{ii}$ values of 94.1 mM, 8.08 mM, and 662.64 mM, respectively. $FeCl_{2}$ was the best competitive inhibitor with a $K_{is}$ value of $29{\mu}M$. $FeCl_{3}$, $MnCl_{2}$, $CoCl_{2}$, and $AlCl_{3}$ were also competitive inhibitors with $K_{is}$ values of 1.21 mM, 0.85 mM, 3.98 mM, and 0.21 mM, respectively. Other metal ions showed noncompetitive inhibition patterns. The pH-dependent kinetic parameter data showed that there may be at least two catalytic groups with pK values of 6.2 and 9.4 and two binding groups with pK values of 5.5 and 9.0. Lysine, cysteine, tyrosine, carboxyl, and histidine were modified by their own specific chemical modifiers, indicating that they are involved in substrate binding and catalysis.

• 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.