• Title/Summary/Keyword: Kinetic Constants

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Role of the Salt Bridge Between Arg176 and Glu126 in the Thermal Stability of the Bacillus amyloliquefaciens ${\alpha}$-Amylase (BAA)

  • Zonouzi, Roseata;Khajeh, Khosro;Monajjemi, Majid;Ghaemi, Naser
    • Journal of Microbiology and Biotechnology
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    • v.23 no.1
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    • pp.7-14
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    • 2013
  • In the Bacillus amyloliquefaciens ${\alpha}$-amylase (BAA), the loop (residues 176-185; region I) that is the part of the calcium-binding site (CaI, II) has two more amino acid residues than the ${\alpha}$-amylase from Bacillus licheniformis (BLA). Arg176 in this region makes an ionic interaction with Glu126 from region II (residues 118-130), but this interaction is lost in BLA owing to substitution of R176Q and E126V. The goal of the present work was to quantitatively estimate the effect of ionic interaction on the overall stability of the enzyme. To clarify the functional and structural significance of the corresponding salt bridge, Glu126 was deleted (${\Delta}$E126) and converted to Val (E126V), Asp (E126D), and Lys (E126K) by site-directed mutagenesis. Kinetic constants, thermodynamic parameters, and structural changes were examined for the wild-type and mutated forms using UV-visible, atomic absoption, and fluorescence emission spectroscopy. Wild-type exhibited higher $k_{cat}$ and $K_m$ but lower catalytic efficiency than the mutant enzymes. A decreased thermostability and an increased flexibility were also found in all of the mutant enzymes when compared with the wild-type. Additionally, the calcium content of the wild-type was more than ${\Delta}E126$. Thus, it may be suggested that ionic interaction could decrease the mobility of the discussed region, prevent the diffusion of cations, and improve the thermostability of the whole enzyme. Based on these observations, the contribution of loop destabilization may be compensated by the formation of a salt bridge that has been used as an evolutionary mechanism or structural adaptation by the mesophilic enzyme.

Saturated Hydraulic Conductivity of Surface Seals Estimated from Computed Tomography-Measured Porosity (고해상도 X-ray CT 를 이용한 토양표면 피막의 공극율 및 포화수리전도도 측정)

  • Lee, Sang-Soo;Gantzer, C.J.;Thompson, A.L.;Anderson, S.H.;Ketchum, R.A.;Ok, Yong-Sik
    • 한국환경농학회:학술대회논문집
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    • 2011.07a
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    • pp.207-222
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    • 2011
  • Relationships between soil saturated hydraulic conductivity ($K_s$) and porosity (${\phi}$) have been developed over many years; however, use of these relationships for evaluating rain-induced seals is limited mainly because of difficulties in estimating seal pore-size characteristics. The objectives of this study were to evaluate the $K_s$ of soil surface seals over a range of thicknesses, where seal thickness was determined using a High-Resolution-Computed-Tomography (HRCT) scanner, and to investigate relationships between $K_s$ and ${\phi}$ of developing seals in samples with equivalent diameters (e.d.) ${\geq}15\;{\mu}m$. A Mexico silt loam soil was packed to a bulk density (${\rho}_b$) of $1.1\;Mg\;m^{-3}$ in cylinders 160-mm i.d. by 160-mm long and subjected to $61-mm\;h^{-1}$ simulated rainfall having a kinetic energy (KE) of $25\;J\;m^{-2}\;min^{-1}$ for 7.5, 15, 30, and 60 min to create a range in seal development. Thicknesses of the seal layers were determined by analysis of HRCT images of seals. The $K_s$ values of the seals were estimated using an effective $K_s$ value ($K_{s-eff}$). The $K_s-{\phi}$ relationship was described by a Kozeny and Carmen equation, $K_s=B{\phi}^n$; where B and n are empirical constants and n = 31. This approach explained 86% of the variation between $K_s$ and ${\phi}$ within the soil seals. Knowledge of surface seal information and hydraulic conductivity can provide useful information to use in management of sites prone to sealing formation.

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The Kinetics of Transesterification between Dimethylterephthalate and 1,3-Propanediol (디메틸 테레프탈레이트와 1,3-프로판디올 사이의 에스테르교환반응에 관한 연구)

  • Na, Sang-Kuwon;Kong, Byeong-Gi;Choi, Chang-Yong;Kim, Jung-Gyu;Hong, Wan-Hae;Nah, Jae-Woon
    • Polymer(Korea)
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    • v.29 no.1
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    • pp.41-47
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    • 2005
  • The transesterification of dimethyl terephthalate (DMT) with 1,3-propanediol (PDO) was investigated in the presence of catalyst, titanium (IV) butoxide (TBO), at 175~190 $^{\circ}C$ . The degree of transesterification reaction was measured by the output of methanol which was distilled from the reactor. The amount of methanol increased as the reaction temperature, molar ratio and catalyst concentration increased. The observed overall rate of the transesterification was third order; first order with respect to DMT, PDO, and the concentration of catalyst, respectively. Using calculated rate constants, the activation energy for transesterification was 26.93 kcal/mole. The melting temperature of bis(2-hydroxytrimethyl) terephthalate (BHTMT) was 85.2$^{\circ}C$ and heat of fusion 141.3 J/g.

Removal of Geosmin and 2-MIB using Biological Activated Carbon Process (생물활성탄(BAC) 공정을 이용한 이취미물질(geosmin, 2-MIB)의 생분해 특성평가)

  • Son, Dong-Min;Son, Hee-Jong;Lee, Hwa-Ja;Kang, Lim-Seok
    • Journal of Korean Society of Water and Wastewater
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    • v.23 no.2
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    • pp.189-198
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    • 2009
  • Tastes and odor in water caused by geosmin and 2-MIB are the major customer complaints for water utilities. Therefore, control of geosmin and 2-MIB is a worldwide concern. In this study, the effects of biofilter media type (three different activated carbons and anthracite), empty bed contact time (EBCT) and temperature on the removal of geosmin and 2-MIB in BAC filters were investigated. Experiments were conducted at three different water temperatures (5, 15 and $25^{\circ}C$) and four different EBCTs (5, 10, 15, and 20 min). The experimental results indicated that the coal based BAC retained more bacterial biomass on the surface of the activated carbon than the other BACs, and increasing EBCT or increasing water temperature also increased the geosmin and 2-MIB removal in BAC filters. To achieve above 50% of removal efficiency for geosmin and 2-MIB in a BAC filter, above 10 min EBCT at $5^{\circ}C$ and 5 min EBCT at above $15^{\circ}C$ were required. The kinetic analysis for the biodegradation of geosmin and 2-MIB indicated a first-order reaction rate at various water temperatures. Data obtained from the BAC filters at various temperatures were also used to evaluate pseudo first-order rate constants for geosmin and 2-MIB. The half-lives evaluated at 5, 15, and $25^{\circ}C$ for geosmin and 2-MIB ranged from 2.39 to 10.31 min and 3.35 to 13.97 min, respectively, which can be used to assist water utilities in designing and operating BAC system.

Synthesis of Propylene Carbonate over Metal containing Ionic Liquid Catalysts (금속 함유 이온성 액체 촉매상에서의 프로필렌 카보네이트의 합성)

  • Moon, Ye-Ji;Ji, Dahye;Kim, Dong-Woo;Kim, Hyeon-Gook;Cho, Deug-Hee
    • Applied Chemistry for Engineering
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    • v.27 no.2
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    • pp.153-157
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    • 2016
  • In this study, three different metal-containing ionic liquid catalysts were prepared by metal insertion and characterized by various physicochemical analytic methods. The catalytic performance of the metal containing ionic liquids in the cycloaddition of $CO_2$ with propylene oxide (PO) to produce propylene carbonate (PC) was investigated under the solvent free condition. The order of approximate rate constants ($K_{app}$) for the metal containing ionic liquid catalysts was $(MeIm)_2ZnCl_2$, > $(MeIm)_2FeCl_2$ > $(MeIm)_2CuCl_2$. These results are in accord with the experimentally obtained activity order of the different metal containing ionic liquid catalysts.

Kinetics of Malonyl-CoA Synthetase from Rhizobium trifolii and Evidences for Malonyl-AMP Formation as a Reaction Intermediate

  • Kang, Sang-Won;Hong, Sung-Yu;Ryoo, Hyung-Don;Rhyu, Gyung-Ihm;Kim Yu-Sam
    • Bulletin of the Korean Chemical Society
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    • v.15 no.5
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    • pp.394-399
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    • 1994
  • The catalytic mechanism of malonyl-CoA synthetase from Rhizobium trifolii was investigated by the steady state kinetics and intermediate identification. Initial velocity studies and the product inhibition studies with AMP and PPi strongly suggested ordered Bi Uni Uni Bi Ping-Pong Ter Ter system as the most probable steady state kinetic mechanism of malonyl-CoA synthetase. Michaelis constants were $0.17{\pm}0.04 {\mu}M,\;0.24{\pm}0.18 {\mu}M\;and\;0.045{\pm}0.26 {\mu}$M for ATP, malonate and CoA, respectively. The TLC analysis of the $^{32}P-labelled$ products in reaction mixture containing $[{\gamma}-^{32}P]$ ATP in the absence of CoA showed that PPi was produced after the sequential addition of ATP and malonate. Formation of malonyl-AMP, suggested as an intermediate in the kinetically deduced mechanism, was confirmed by the analysis of $^{31}P-NMR$ spectra of AMP product isolated from the $^{18}O$ transfer experiment using $[^{18}O]$malonate. Two resonances were observed, corresponding to AMP labelled with zero and one atom of $^{18}O$, indicating that one atom of $^{18}O$ transferred from $[^{18}O]$malonate to AMP through the formation of malonyl-AMP. Formation of malonyl-AMP was also confirmed through the TLC analysis of reaction mixture containing $[{\alpha}-^{32}P]$ATP. These results strongly support the ordered Bi Uni Uni Bi Ping-Pong Ter Ter mechanism deduced from the initial velocity and product inhibition studies.

Synthesis of iron nanoparticles with poly(1-vinylpyrrolidone-co-vinyl acetate) and its application to nitrate reduction

  • Lee, Nara;Choi, Kyunghoon;Uthuppu, Basil;Jakobsen, Mogens H.;Hwang, Yuhoon;Broholm, Mette M.;Lee, Woojin
    • Advances in environmental research
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    • v.3 no.2
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    • pp.107-116
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    • 2014
  • This study aimed to synthesize dispersed and reactive nanoscale zero-valent iron (nZVI) with poly(1-vinylpyrrolidone-co-vinyl acetate) (PVP/VA), nontoxic and biodegradable stabilizer. The nZVI used for the experiments was prepared by reduction of ferric solution in the presence of PVP/VA with specific weight ratios to iron contents. Colloidal stability was investigated based on the rate of sedimentation, hydrodynamic radius and zeta potential measurement. The characteristic time, which demonstrated dispersivity of particles resisting aggregation, increased from 21.2 min (bare nZVI) to 97.8 min with increasing amount of PVP/VA (the ratios of 2). For the most stable nZVI coated by PVP/VA, its reactivity was examined by nitrate reduction in a closed batch system. The pseudo-first-order kinetic rate constants for the nitrate reduction by the nanoparticles with PVP/VA ratios of 0 and 2 were 0.1633 and $0.1395min^{-1}$ respectively. A nitrogen mass balance, established by quantitative analysis of aqueous nitrogen species, showed that the addition of PVP/VA to nZVI can change the reduction capacity of the nanoparticles.

Efficiency and Mechanism of Pb(II) Removal from Aqueous Solutions Using Cornus controversa and Quercus mongolica Biomass Waste (층층나무와 신갈나무 폐바이오매스를 활용한 수용액 중 납 제거 효율 및 기작)

  • Choi, Si Young;Jeong, Seok Soon;Yang, Jae E.;Kim, Hyuck Soo;Cho, Jun Hyung
    • Korean Journal of Environmental Agriculture
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    • v.40 no.4
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    • pp.239-247
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    • 2021
  • BACKGROUND: Enormous amounts of the wood biomass wastes have been produced through various wood processing. This study characterizes the surface characteristics of biomass powders of Cornus controversa (CC) and Quercus mongolica (QM) and investigates their removal efficiency and mechanism for Pb (II) in aqueous solution on which to base potential recycling alternative of the wood biomass. METHODS AND RESULTS: Batch experiments were conducted under different conditions of Pb concentrations, temperatures, time and solid/solution ratios. Adsorption isotherm of Pb by CC and QM biomass was explained significantly by the Langmuir model, indicating Pb was likely adsorbed on the monolayer of the surfaces. The adsorption kinetics were fitted significantly to the double first-order model consisting of rapid and slow steps. The respective rate constants (k1) of CC and QM for the rapid adsorption kinetic steps were 0.051 and 0.177 min-1, and most of the sorption reactions proceeded rapidly within 6-20 minutes. The maximum adsorption quantities (qmax) of Pb were 17.25 and 23.47 mg/g for CC and QM, respectively. Thermodynamic parameters revealed that adsorption of Pb on the biomass of CC and QM was a spontaneous endothermic reaction. CONCLUSION(S): Results demonstrate that biomass wastes of CC and QM can be used as Pb adsorbents judging from adsorption isotherm, kinetics, and thermodynamic parameters.

Removals of PAH-quinones Using Birnessite-Mediated Oxidative-Transformation Processes (망간산화물(Birnessite)을 매개로한 산화-변환반응을 이용한 PAH-퀴논화합물의 제거)

  • Choi, Chan-Kyu;Harn, Yoon-I;Kim, Seong-Uk;Shin, Hyun-Sang
    • Journal of Korean Society of Environmental Engineers
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    • v.33 no.6
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    • pp.396-404
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    • 2011
  • An investigation on the removals of PAH-quinone compounds, which are commonly produced from the biological and/or chemical treatments of PAH-contaminated soils, from the aqueous phase via birnessite (${\delta}-MnO_2$)-mediated oxidative transformation is described. It was demonstrated that acenaphthenequinone (APQ), p-PAH quinone can be removed via birnessite-mediated oxidative-coupling reactions, and anthraquinone (AQ) and 1,4-naphthoquinone (1,4-NPQ), o-PAH quinones were efficiently removed by birnessite-mediated cross-coupling reactions in the presence of catechol (CAT) as a reactive mediator. The removals of PAH-quinone compounds followed pseudo-first-order reactions, and the rate constant (k, $hr^{-1}$) for the removals of 1,4-NPQ under the experiment conditions (1,4-NPQ = 10 mg/L, CAT = 50 mg/L, ${\delta}-MnO_2$ = 1.0 g/L, pH 5, Reaction time = 6~96 hr) was 0.0426, which was about 4 times lower than that of APQ (0.173). With the observed pseudo-first order rate constants with respect to birnessite loadings under the same experimental conditions, the surface-normalized specific rate constant, $K_{surf}$, for 1,4-NPQ was determined to be $8.5{\times}10^{-4}L/m^2{\cdot}hr$. The analysis of the kinetic data with respect to birnessite loading indicated that the cross-coupling reactions of 1,4-NPQ consist of two different reaction steps over time and the results have also been discussed in terms of the reaction mechanisms.

Oxidation of Geosmin and 2-MIB in Water Using $O_3/H_2O_2$: Kinetic Evaluation (오존과 과산화수소를 이용한 Geosmin과 2-MIB 산화: 동력학적 평가)

  • Lee, Hwa-Ja;Son, Hee-Jong;Roh, Jae-Soon;Lee, Sang-Won;Ji, Ki-Won;Yoo, Pyung-Jong;Kang, Lim-Seok
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.7
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    • pp.826-832
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    • 2007
  • Unpleasant tastes and odors in drinking water cause same problems for water utilities across Korea. Even though tastes and odors do not create health problems, they are main concerns for consumers who determine the safety of their drinking water. In this study, two different odor producing compounds(geosmin 2-MIB) in the Nakdong river water and rapid sand filtered rater were treated by advanced oxidation of $O_3/H_2O_2$ process. The experimental results showed that the removal efficiency of geosmin with the use of 5 mg/L of $O_3$ and $H_2O_2$ was higher than efficiency with the use of $O_3$ alone for both the raw water and the sand filtered water. And in general, the removal efficiency of geosmin was higher than 2-MIB in the sand filtered water. Under the range of $O_3$ concentration $0.5\sim2.0$ mg/L, the removal rate constants(k) of geosmin for the raw and sand filtered waters, and the one of 2-MIB in the sand filtered water were increased rapidly as doses of $O_3$, and $H_2O_2$, increased. The removal rate constants(k) do not increase any more when $H_2O_2/O_3$ ratio increases above the optimum ratio. The optimum ratio of $H_2O_2/O_3$, dose was $1.0\sim2.0$ for both geosmin and 2-MIB. The removal rate constant(k) becomes lower when OH radical consuming materials are present in raw water. The half-life of geosmin decreased rapidly as the $O_3$ and $H_2O_2$ doses increase in the sand filtered water. The half life decreased about 8.5 times with the use of 2 mg/L of $O_3$ and 10 mg/L of $H_2O_2$ than with the use of 2 mg/L of $O_3$ alone for the sand filtered water.