• Title/Summary/Keyword: pH-dependent kinetic parameters

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The Cofactors Role on Chemical Mechanism of Recombinant Acetohydroxy Acid Synthase from Tobacco

  • Kim, Joung-Mok;Kim, Jung-Rim;Kim, Young-Tae;Choi, Jung-Do;Yoon, Moon-Young
    • Bulletin of the Korean Chemical Society
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    • v.25 no.5
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    • pp.721-725
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    • 2004
  • Acetohydroxy acid synthase (AHAS) is one of several enzymes that require thiamine diphosphate and a divalent cation as essential cofactors. The active site contains several conserved ionizable groups and all of these appear to be important as judged by the fact that mutation diminishes or abolishes catalytic activity. Recently, we have shown [Yoon, M.-Y., Hwang, J.-H., Choi, M.-K., Baek, D.-K., Kim, J., Kim, Y.-T., Choi, J.-D. FEBS Letters 555 (2003), 185-191] that the activity is pH-dependent due to changes in $V_{max}$ and V/$K_m$. Data were consistent with a mechanism in which substrate was selectively catalyzed by the enzyme with an unprotonated base having a pK 6.48, and a protonated group having a pK of 8.25 for catalysis. Here, we have in detail studied the pH dependence of the kinetic parameters of the cofactors (ThDP, FAD, $Mg^{2+}$) in order to obtain information about the chemical mechanism in the active site. The $V_{max}$ of kinetic parameters for all cofactors was pH-dependent on the basic side. The pK of ThDP, FAD and $Mg^{2+}$ was 9.5, 9.3 and 10.1, respectively. The V/$K_m$ of kinetic parameters for all cofactors was pH-dependent on the acidic and on the basic side. The pK of ThDP, FAD and $Mg^{2+}$ was 6.2-6.4 on the acidic side and 9.0-9.1 on the basic side. The well-conserved histidine mutant (H392) did not affect the pH-dependence of the kinetic parameters. The data are discussed in terms of the acid-base chemical mechanism.

A Parametric Study on the Sorption of U(VI) onto Granite (U(VI)의 화강암 수착에 대한 매개변수적 연구)

  • Min-Hoon Baik;Won-Jin Cho;Pil-Soo Hahn
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.2 no.2
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    • pp.135-143
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    • 2004
  • An experimental study on the sorption of U(VI) onto a Korean granite was performed as a function of the geochemical parameters such as contact time, pH, ionic strength, and carbonate concentration using a batch procedure. The distribution coefficient,$K_d$, was about 1-200 mL/g depending on the experimental conditions. The sorption of U(VI) onto granite particles was greatly dependent upon the contact time, pH, and carbonate concentration, but insignificantly dependent on the ionic strength. It was noticed that the sorption of U(VI) onto granite particles was highly correlated with the uranium speciation in the solution, which was dependent on the pH and carbonate concentrations. It was deduced from the kinetic sorption experiment that a two-step first-order kinetic behavior could dominate the kinetic sorption of U(VI) onto granite particles. In the alkaline range of a pH above 7, U(VI) sorption was greatly decreased and this might be due to the formation of anionic U(VI)-carbonate aqueous complexes as predicted by the speciation calculations.

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Adsorption Characteristics of As and Se Ions by HTMAB Modified Anthracite (HTMAB로 표면처리된 안트라사이트에 의한 비소 및 셀렌 이온의 흡착 특성)

  • Kim, Jeung-Bea
    • Journal of Environmental Science International
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    • v.27 no.3
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    • pp.167-177
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    • 2018
  • The removal characteristics of As and Se ions from aqueous solution by hexadecyl trimethyl ammonium bromide (HTMAB) modified anthracite (HTMAB-AT) were investigated under various conditions of contact time, pH and temperature. When the pH is 6, the zeta potential value of anthracite (AT) is -24 mV and on the other hand, the zeta potential value of the HTMAB-AT is +44 mV. It can be seen that the overall increase of about 60 mV. Increasing the (+) potential value indicates that the surface of the adsorbent had a stronger positive charge, so adsorption for the anion metal was increased. The isotherm data was well described by Langmuir and Temkin isotherm model. The maximum adsorption capacity was found to be 7.81 and 6.89 mg/g for As and Se ions from the Langmuir isotherm model at 298 K, respectively. The kinetic data was tested using pseudo first and pseudo second order models. The results indicated that adsorption fitted well with the pseudo second order kinetic model. The mechanism of the adsorption process showed that adsorption was dependent on intra particle diffusion model according to two step diffusion. The thermodynamic parameters(${\Delta}G^{\circ}$, ${\Delta}H^{\circ}$, and ${\Delta}S^{\circ}$) were also determined using the equilibrium constant value obtained at different temperatures. The thermodynamic parameters indicated that the adsorption process was physisorption, and also an endothermic and spontaneous process.

Evidence for two $Na^+$/$H^+$ Antiport Systems in Escherichia coli

  • Seo, Sung-Yum
    • Korean Journal of Microbiology
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    • v.30 no.4
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    • pp.269-277
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    • 1992
  • Several insertion mutants of Escherichia coli in the ant gene, coding for $Na^{+}$ $H^{+}$ antiport activity, showed littel, if any, reduction in the antiport activity. $Na^{+}$ dependent transport activity also remained at wild type level. These facts led to the idea that E. coli has evolved at least two distinct systems for extrusion of $Na^{+}$ The antiport activities were studied under various conditions to reveal different properties of these systems. For convenience these activities are referred to as major and minor activities. The distinguishing properties of the two systems include : kinetics (Km, Vm) at pH 7.8, competition pattern between $Na^{+}$ and Li$^{+}$ , pH profiles, pattern of the change in kinetic parameters as a function of pH, and sensitivity to protease, chemicals and heat.

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Adsorption properties of activated carbon prepared from pre-carbonized petroleum coke in the removal of organic pollutants from aqueous solution

  • Ahmed, S.A. Sayed;El-Enin, Reham M.M. Abo;El-Nabarawy, Th.
    • Carbon letters
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    • v.12 no.3
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    • pp.152-161
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    • 2011
  • Activated carbon was prepared from pre-carbonized petroleum coke. Textural properties were determined from studies of the adsorption of nitrogen at 77 K and the surface chemistry was obtained using the Fourier-transform infrared spectrometer technique and the Boehm titration process. The adsorption of three aromatic compounds, namely phenol (P), p-nitrophenol (PNP) and benzoic acid (BA) onto APC in aqueous solution was studied in a batch system with respect to contact time, pH, initial concentration of solutes and temperature. Active carbon APC obtained was found to possess a high surface area and a predominantly microporous structure; it also had an acidic surface character. The experimental data fitted the pseudo-second-order kinetic model well; also, the intraparticle diffusion was the only controlling process in determining the adsorption of the three pollutants investigated. The adsorption data fit well with the Langmuir and Freundlich models. The uptake of the three pollutants was found to be strongly dependent on the pH value and the temperature of the solution. Most of the experiments were conducted at pH 7; the $pH_{(PZC)}$ of the active carbon under study was 5.0; the surface of the active carbon was negatively charged. The thermodynamic parameters evaluated for APC revealed that the adsorption of P was spontaneous and exothermic in nature, while PNP and BA showed no-spontaneity of the adsorption process and that process was endothermic in nature.

Isolation and characterization of thioredoxin and NADPH-dependent thioredoxin reductase from tomato (Solanum lycopersicum)

  • Dai, Changbo;Wang, Myeong-Hyeon
    • BMB Reports
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    • v.44 no.10
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    • pp.692-697
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    • 2011
  • To investigate the pathways of oxidoreductases in plants, 2 key components in thioredox systems i.e. thioredoxin h (Trx h) and NADPH-dependent thioredoxin reductase (NTR) genes were first isolated from tomatoes (Solanum lycopersicum). Subsequently, the coding sequences of Trx h and NTR were inserted into pET expression vectors, and overexpressed in Escherichia coli. In the UV-Visible spectra of the purified proteins, tomato Trx h was shown to have a characteristic 'shoulder' at ~290 nm, while the NTR protein had the 3 typical peaks unique to flavoenzymes. The activities of both proteins were demonstrated by following insulin reduction, as well as DTNB reduction. Moreover, both NADPH and NADH could serve as substrates in the NTR reduction system, but the catalytic efficiency of NTR with NADPH was 2500-fold higher than with NADH. Additionally, our results reveal that the tomato Trx system might be involved in oxidative stress, but not in cold damage.

Kinetic Investigation on the Reaction between Cu(II) and Excess D-penicillamine in Aqueous Media

  • Lee, Yong-Hwan;Choi, Sung-Nak;Cho, Mi-Ae;Kim, Yong-Kyu
    • Bulletin of the Korean Chemical Society
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    • v.11 no.4
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    • pp.281-286
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    • 1990
  • The kinetics and mechanism of reduction of Cu(II) with an excess D-penicillamine have been examined at pH = 6.2 and 0.60M in ionic strength. The reaction at the initial stage is biphasic with a rapid complexation process to give "red" transient complex of $[Cu(II)(pen)_2]^2$- that is partially reduced to another transient "brown" intermediate. The "brown" intermediate is finally reduced to diamagnetic "yellow" complex, $[Cu(I)(Hpen)]_n$. The final reduction process is pseudo-first order in ["brown" transient] disappearance $with {\kappa} = {{\kappa}_{3a} + {\kappa}_{3b}[pen]^{2-}},$ where ${\kappa}_{3a} = (5.0{\pm}0.8){\times}10^{-3}sec^{-1}$ and ${\kappa} = (0.14{\pm}0.02) M^{-1}sec^{-1}$ at $25^{\circ}C$. The activation parameters for the $[H_2pen]$-independent and $[H_2pen]$-dependent paths are ${\Delta}H^{\neq} = (52{\pm}5)kJmol^{-1},$ and ${\Delta}S^{\neq} = ( - 27{\pm}3)JK^{-1}mo^{l-1},$ and ${\Delta}H^{\neq} = (56{\pm}2)kJmol^{-1}$ and ${\Delta} S^{\neq} = ( - 18{\pm}0.7)JK^{-1}mol^{-1}$ respectively. The nature of "brown" intermediate is not clearly identified, but this intermediate seems to be in the mixed-valence state, judging from the kinetic and spectroscopic informations.

Assessment of kinetics behavior of electrocoagulation process for the removal of suspended solids and metals from synthetic water

  • Singh, Hariraj;Mishra, Brijesh Kumar
    • Environmental Engineering Research
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    • v.22 no.2
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    • pp.141-148
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    • 2017
  • Globalization, industrialization, mining, and uncontrolled population growth have fostered a shortage of potable water. Therefore, it has become imperative to understand an effective and reasonable water purification technique. A renewed interest in electrocoagulation (EC) has been spurred by the search for reliable, cost-effective, water-treatment processes. This paper has elucidated a technical approach for getting rid of heavy metals and total suspended solids (TSS) from synthetic water using an aluminum electrode. The effect of operational parameters, such as current density, inter-electrode distance, operating time, and pH, were studied and evaluated for maximum efficiency. This study corroborates the correlation between current density and removal efficiency. Neutral pH and a low electrode gap have been found to aid the efficacy of the EC setup. The outcome indicates that a maximum TSS removal efficiency of 76.6% occurred at a current density of $5.3mA/cm^2$ during a contact time of 30 min. In the case of heavy metals remediation, 40 min of process time exhibited extremely reduced rates of 99%, 59.2%, and 82.1%, for Cu, Cr, and Zn, respectively. Moreover, kinetic study has also demonstrated that pollutants removal follows first-and second-order model with current density and EC time being dependent.

Equilibrium and kinetic studies of an electro-assisted lithium recovery system using lithium manganese oxide adsorbent material

  • Lee, Dong-Hee;Ryu, Taegong;Shin, Junho;Kim, Young Ho
    • Carbon letters
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    • v.28
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    • pp.87-95
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    • 2018
  • This study examined the influence of operating parameters on the electrosorptive recovery system of lithium ions from aqueous solutions using a spinel-type lithium manganese oxide adsorbent electrode and investigated the electrosorption kinetics and isotherms. The results revealed that the electrosorption data of lithium ions from the lithium containing aqueous solution were well-fitted to the Langmuir isotherm at electrical potentials lower than -0.4 V and to the Freundlich isotherm at electrical potentials higher than -0.4 V. This result may due to the formation of a thicker electrical double layer on the surface of the electrode at higher electrical potentials. The results showed that the electrosorption reached equilibrium within 200 min under an electrical potential of -1.0 V, and the pseudo-second-order kinetic model was correlated with the experimental data. Moreover, the adsorption of lithium ions was dependent on pH and temperature, and the results indicate that higher pH values and lower temperatures are more suitable for the electrosorptive adsorption of lithium ions from aqueous solutions. Thermodynamic results showed that the calculated activation energy of $22.61kJ\;mol^{-1}$ during the electrosorption of lithium ions onto the adsorbent electrode was primarily controlled by a physical adsorption process. The recovery of adsorbed lithium ions from the adsorbent electrode reached the desorption equilibrium within 200 min under reverse electrical potential of 3.5 V.

Inhibition and Chemical Mechanism of Protocatechuate 3,4-dioxygenase from Pseudomonas pseudoalcaligenes KF707 (Pseudomonas pseudoalcaligenes KF707에서 유래한 protocatechuate 3,4-dioxygenase 의 저해 및 화학적 메커니즘)

  • Kang, Taekyeong;Kim, Sang Ho;Jung, Mi Ja;Cho, Yong Kweon
    • 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.