• 제목/요약/키워드: 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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    • 제25권5호
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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.

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

  • Min-Hoon Baik;Won-Jin Cho;Pil-Soo Hahn
    • 방사성폐기물학회지
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    • 제2권2호
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    • pp.135-143
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    • 2004
  • 국산화강암에 대한U(VI) 수착에 대한 실험적 연구를 지화학적 매개변수들인 접촉시간, pH, 이온강도, 탄산염 농도 등의 함수로 회분식으로 수행하였다. 국산 화강암에 대한 U(VI)의 수착의 분배계수 $K_{d}$ 는 실험 조건에 따라 약 1-200 mL/g의 값의 범위를 가지는 것으로 나타났다. 화강암 입자에 대한 U(VI)의 수착은 접촉시간, pH 및 탄산염 농도 등에는 크게 의존하였으나 이온강도에는 크게 의존하지 않는 것으로 나타났다. 화강암 입자들에 대한U(VI)의 수착은 용액에서 pH와 탄산염의 농도에 의존하는 우라늄 화학종과 밀집한 관계가 있음을 알 수 있었다. 또한 속도론적 수착 실험에 의하여 2 단계 일차식 속도론적 거동이 화강암 입자들에 대한 우라늄의 속도론적 수착을 지배할 수 있는 것으로 유추되었다. pH 7 이상의 알칼리 영역에서 화강암에 대한 우라늄의 수착이 크게 감소되었는데 이는 화학종 계산에 의해 예측된 바에 따라 음이온의 U(VI)-탄산염 복합체 형성에 기인하였을 것이다.

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

  • 김정배
    • 한국환경과학회지
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    • 제27권3호
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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
    • 미생물학회지
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    • 제30권4호
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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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    • 제12권3호
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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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    • 제44권10호
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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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    • 제11권4호
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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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    • 제22권2호
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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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    • 제28권
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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.

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

  • 강태경;김상호;정미자;조용권
    • 생명과학회지
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    • 제25권5호
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    • pp.487-495
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    • 2015
  • Pseudomonas pseudoalcaligenes KF707에서 정제한 protocatechuate 3,4-dioxygenase의 특징을 조사하기 위하여 pH안정성, 화학적 저해, 화학적 수식과 pH의존성 반응 상수에 대한 실험을 수행하였다. 이 효소는 pH 4.5~10.7에서 안정하였다. L-ascorbate와 glutathione은 Kis가 각각 0.17 mM과 0.86 mM인 경쟁적 저해제였으며, DL-dithiothreitol은 Kis 1.57 mM 및 Kii 8.08 mM의 비경쟁적 저해패턴을 나타내었다. Potassium cyanide, p-hydroxybenzoate 및 sodium azide는 Kis가 각각 55.7 mM, 0.22 mM 및15.64 mM이었으며, Kii는 각각94.1 mM, 8.08 mM, 및 662.64 mM인 비경쟁적 저해패턴을 나타내었다. $FeCl_{2}$는 Kis가 $29{\mu}M$로 가장 우수한 경쟁적 저해제였으며, $FeCl_{3}$, $MnCl_{2}$, $CoCl_{2}$, $HgCl_{2}$, $AlCl_{3}$도 각각 Kis가 1.21 mM, 0.85 mM, 3.98 mM, 0.17 mM 및 0.21 mM인 경쟁적 저해패턴을 보였다. 한편, 다른 금속이온들은 비경쟁적 저해패턴을 나타내었다. pH의존성 반응상수의 실험결과로부터 pK 6.2와 9.4의 촉매부위와 pK 5.5와 9.0의 결합부위가 존재함을 알 수 있었다. Lysine, cysteine, tyrosine, carboxyl과 histidine은 각각의 고유한 화학적 수식제에 의해 수식되었는데, 이는 이들 잔기들이 결합과 촉매에 관여한다는 것을 나타낸다. 위 결과를 토대로 화학적 메커니즘을 제시한다.