• Title/Summary/Keyword: Oxidation Kinetic

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Kinetic Evidence for the Interactive Inhibition of Laccase from Trametes versicolor by pH and Chloride

  • Raseda, Nasrin;Hong, Soonho;Kwon, O Yul;Ryu, Keungarp
    • Journal of Microbiology and Biotechnology
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    • v.24 no.12
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    • pp.1673-1678
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    • 2014
  • The interactive inhibitory effects of pH and chloride on the catalysis of laccase from Trametes versicolor were investigated by studying the alteration of inhibition characteristics of sodium chloride at different pHs for the oxidation of 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid). At pH 3.0, the addition of sodium chloride (50 mM) brought about a 40-fold increase in $K{_m}^{app}$ and a 4-fold decrease in $V_{max}{^{app}}$. As the pH increased to 7.0, the inhibitory effects of sodium chloride became significantly weakened. The mixed-inhibition mechanism was successfully used to quantitatively estimate the competitive and uncompetitive inhibition strengths by chloride at two different pHs (pH 3.0 and 6.0). At pH 3.0, the competitive inhibition constant, $K_i$, was 0.35 mM, whereas the uncompetitive inhibition constant, $K{_i}^{\prime}$, was 18.1 mM, indicating that the major cause of the laccase inhibition by chloride is due to the competitive inhibition step. At a higher pH of 6.0, where the inhibition of the laccase by hydroxide ions takes effect, the inhibition of the laccase by chloride diminished to a great extent, showing increased values of both the competitive inhibition constant ($K_i=23.7mM$) and uncompetitive inhibition constant ($K{_i}^{\prime}=324mM$). These kinetic results evidenced that the hydroxide anion and chloride share a common mechanism to inhibit the laccase activity.

Effects of Structural Difference of Ionic Liquids on the Catalysis of Horseradish Peroxidase

  • Hong, Eun-Sik;Park, Jung-Hee;Yoo, Ik-Keun;Ryu, Keun-Garp
    • Journal of Microbiology and Biotechnology
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    • v.19 no.7
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    • pp.713-717
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    • 2009
  • The dependence of the catalytic properties of horseradish peroxidase on the structural changes of ionic liquids was investigated with two water-miscible ionic liquids, N-butyl-3methypyridinium tetraftuoroborate ([$BMP_y$][$BF_4$]) and 1-butyl-3-methylimidazolium methylsulfate ([BMIM][$MeSO_4$]), each of which shares an anion ($BF_4^-$) or a cation ($BMIM^+$) with 1-butyl-3-methylimidazolium tetraftuoroborate ([BMIM][$BF_4$]), respectively. The oxidation of guaiacol (2-methoxyphenol) with $H_2O_2$was used as a model reaction. In order to minimize the effect of solution viscosity on the kinetic constants of the enzymatic catalysis, the enzymatic reactions for the kinetic study were performed in water-ionic liquid mixtures containing 25% (v/v) ionic liquid at maximum. Similarly to the previously reported results for [BMIM][$BF_4$], as the concentration of [$BMP_y$][$BF_4$] increased, the $K_m$value increased with a decrease in the $k_{cat}$value: the $K_m$value increased markedly from 2.8 mM in 100% water to 12.6 mM in 25% (v/v) ionic liquid, indicating that ionic liquid significantly weakens the binding affinity of guaiacol to the enzyme. On the contrary, [BMIM][$MeSO_4$] decreased the Km value to 1.4 mM in 25% (v/v) ionic liquid. [BMIM][$MeSO_4$] also decreased $k_{cat}$more than 3-folds [from 13.8 $s^{-1}$in 100% water to 4.1 $s^{-1}$in 25% (v/v) ionic liquid]. These results indicate that the ionic liquids interact with the enzyme at the molecular level as well as at a macroscopic thermodynamic scale. Specifically, the anionic component of the ionic liquids influenced the catalysis of horseradish peroxidase in different ways.

Photocatalytic Decomposition of Gaseous Ozone over $TiO_2$Thin Film

  • Cho, Ki-Chul;Hwang, Kyung-Chul;Yeo, Hyun-Gu;Taizo Sano;Koji Takeuchi;Sadao Matsuzawa
    • Journal of Korean Society for Atmospheric Environment
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    • v.19 no.E3
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    • pp.121-127
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    • 2003
  • The characteristics of heterogeneous photocatalytic decomposition were investigated at low concentration level of $O_3$on TiO$_2$for various operating parameters such as: loaded catalyst weight (0∼4 mg/$\textrm{cm}^2$), initial concentration of $O_3$(0.06∼10.0 ppm), gas flow rate (1.0 ∼ 2.5ι/min), and relative humidity (0∼80%). This study was conducted using a flow-type reactor at room temperature. Three kinds of pure TiO$_2$(P25, ST -01, and E- 23) were employed as photocatalyts. It was found that $O_3$removal ratio was identical, regardless of the loaded TiO$_2$weight in the range from 0.5 to 4.0 mg/$\textrm{cm}^2$. It was also found that higher initial ozone concentration results in greater oxidation rate of ozone and experimental data show kinetically a good agreement with Langmur-Hinshelwood kinetic model. We also observed that the removal ratio of $O_3$increases linearly with the increasing flow rate and also with the increasing relative humidity for each catalyst.

Stabilization Characteristics of the Pyrolyzed Oil from Waste Lubricating Oil (폐윤활유 열분해유의 안정화 특성 연구)

  • Kim, Seung-Soo;Kim, Young-Seok;Chun, Byung-Hee;Park, Chan Jin;Yoon, Wang Lai;Kim, Sung Hyun
    • Journal of Korean Society of Environmental Engineers
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    • v.22 no.6
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    • pp.1055-1061
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    • 2000
  • The kinetics of tar formation has been studied experimentally and modeled mathematically for waste lubricating oil after pyrolyzed at batch reactor. And stabilization of pyrolyzed oil has been studied. A combination of series and parallel reaction was assumed for the mechanism of tar formation. From the proposed kinetic model, pyrolyzed oil to tar was found to be rate limiting step for tar formation. It was found that the fly ash and coke had the ability to remove materials of tar formation and to protect oxidation of pyrolyzed oil.

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Metabolic Interactions of Cannabinoids with Steroid Hormones

  • Watanabe, Kazuhito
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2007.11a
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    • pp.57-64
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    • 2007
  • Metabolic interactions of the three major cannabinoids, ${\Delta}^9$-tetrahydrocannabinol (THC), cannabidiol (CBD), and cannabinol (CBN) with steroid hormones were investigated. These cannabioids concentration-dependently inhibited $3{\beta}$-hydroxysteroid dehydrogenase and $17{\alpha}$-hydroxylase in rat adrenal and testis microsomes. CBD and CBN were the most potent inhibitors of $3{\beta}$-phydroxysteroid dehydrogenase and progesterone $17{\alpha}$-hydroxylase, respectively, in rat testis microsomes. Three cannabinoids highly attenuated hCG-stimulated testosterone production in rat testicular interstitial cells. These cannabinoids also decreased in levels of mRNA and protein of StAR in the rat testis cells. These results indicate that the cannabinoids could interact with steroid hormones, and exert their modulatory effects on endocrine and testicular functions. Metabolic interaction of a THC metabolite, $7{\beta}$-hydroxy-${\Delta}^8$-THC with steroids is also investigated. Monkey liver microsomes catalyzed the stereoselective oxidation of $7{\beta}$-hydroxy-${\Delta}^8$-THC to 7-oxo-${\Delta}^8$-THC, so-called microsomal alcohol oxygenase (MALCO). The reaction is catalyzed by CYP3A8 in the monkey liver microsomes, and required NADH as well as NADPH as an efficient cofactor, and its activity is stimulated by some steroids such as testosterone and progesterone. Kinetic analyses revealed that MALCO-catalyze reaction showed positive cooperativity. In order to explain the metabolic interaction between the cannabinoid metabolite and testosterone, we propose a novel kinetic model involving at least three binding sites for mechanism of the metabolic interactions.

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Covalent Coupling of ${\beta}-Fructofuranosidase$ on Microbial Cells (미생물 세포에 공유결합으로 고정화시킨 ${\beta}-Fructofuranosidase$에 관한 연구)

  • Uhm, Tai-Boong;Byun, Si-Myung
    • Korean Journal of Food Science and Technology
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    • v.16 no.3
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    • pp.267-272
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    • 1984
  • ${\beta}-Fructofuranosidase$ was immobilized covalently on the oxidized microbial wall of a Penicillium spp. 'PS-8', which is totally different from the conventional whole cell immobilization in concept. The immobilization of ${\beta}-fructofuranosidase$ by a series of treatments; oxidation of microbial cells with sodium metaperiodate, enzyme loading on the oxidized cells, extrusion, and crosslinking induced by glutaradehyde, were carried out. The final product had a good mechanical strength and showed 26% of the applied enzyme activity. The specific activity was 750 units per g of the dry cell product. The immobilized enzyme showed the kinetic parameters as follows; optimum pH at 5, optimum temperature at $55^{\circ}C$, activation energy of 19 kJ $mol^{-1}$, and apparent Km of 55 mM.

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Functional Characterization of Drosophila melanogaster CYP6A8 Fatty Acid Hydroxylase

  • Sang-A Lee;Vitchan Kim;Byoungyun Choi;Hyein Lee;Young-Jin Chun;Kyoung Sang Cho;Donghak Kim
    • Biomolecules & Therapeutics
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    • v.31 no.1
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    • pp.82-88
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    • 2023
  • Genomic analysis indicated that the genome of Drosophila melanogaster contains more than 80 cytochrome P450 genes. To date, the enzymatic activity of these P450s has not been extensively studied. Here, the biochemical properties of CYP6A8 were characterized. CYP6A8 was cloned into the pCW vector, and its recombinant enzyme was expressed in Escherichia coli and purified using Ni2+-nitrilotriacetate affinity chromatography. Its expression level was approximately 130 nmol per liter of culture. Purified CYP6A8 exhibited a low-spin state in the absolute spectra of the ferric forms. Binding titration analysis indicated that lauric acid and capric acid produced type I spectral changes, with Kd values 28 ± 4 and 144 ± 20 µM, respectively. Ultra-performance liquid chromatography-mass spectrometry analysis showed that the oxidation reaction of lauric acid produced (ω-1)-hydroxylated lauric acid as a major product and ω-hydroxy-lauric acid as a minor product. Steady-state kinetic analysis of lauric acid hydroxylation yielded a kcat value of 0.038 ± 0.002 min-1 and a Km value of 10 ± 2 µM. In addition, capric acid hydroxylation of CYP6A8 yielded kinetic parameters with a kcat value of 0.135 ± 0.007 min-1 and a Km value of 21 ± 4 µM. Because of the importance of various lipids as carbon sources, the metabolic analysis of fatty acids using CYP6A8 in this study can provide an understanding of the biochemical roles of P450 enzymes in many insects, including Drosophila melanogaster.

Effects of High Dose Gamma Irradiation on Shelf Stability and Lipid Oxidation of Marinated and Precooked Pork Rib Steak (고선량 감마선 조사가 즉석 취식용 양념 돼지 갈비 구이의 저장성 및 지질 산화에 미치는 영향)

  • Lee, Ju-Woon;Park, Jae-Nam;Kim, Jae-Hun;Park, Jin-Gyu;Kim, Cheon-Jei;Kim, Kwan-Soo;Byun, Myung-Woo
    • Food Science of Animal Resources
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    • v.26 no.4
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    • pp.471-477
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    • 2006
  • This study was conducted to evaluate the application of high dose irradiation for ensuring shelf stability of marinated and precooked pork rib steak in the severe environments such as desert or space, etc. Marinated and precooked pork rib steak was manufactured, vacuum-packaged and gamma-irradiated with the absorbed doses of 10, 20, 30, 40 and 50 kGy, and used for the tests of the growth of microorganisms and lipid oxidation during storage at 35t of acceleration condition. Any growth of microorganisms was not observed in irradiated samples after irradiation immediately. However, the growths were observed in 10, 20 and 30 kGy samples at 4, 7 and 14 day storage, respectively. High dose (40 and 50 kGy) gamma irradiation retarded the growth of aerobic microorganisms by the analysis of kinetic parameter. The content of malondialdehyde increased in all samples during storage periods, and gamma irradiation accelerated the increase of lipid oxidation. Therefore, the application of combination of the various food processing technology should be considered for the sterilization of marinated and precooked pork rib steak without any deterioration of the quality occurred by high dose irradiation.

Synthetic, Characterization, Biological, Electrical and Catalytic Studies of Some Transition Metal Complexes of Unsymmetrical Quadridentate Schiff Base Ligand

  • Maldhure, A. K.;Pethe, G. B.;Yaul, A. R.;Aswar, A. S.
    • Journal of the Korean Chemical Society
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    • v.59 no.3
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    • pp.215-224
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    • 2015
  • Unsymmetrical tetradentate Schiff base N-(2-hydroxy-5-methylacetophenone)-N'-(2-hydroxy acetophenone) ethylene diamine (H2L) and its complexes with Cr(III), Mn(II), Fe(III), Co(II), Ni(II) and Cu(II) have been synthesized and characterized by elemental analyses, magnetic susceptibility measurements, IR, electronic spectra and thermogravimetric analyses. 1H, 13C-NMR and FAB Mass spectra of ligand clearly indicate the presence of OH and azomethine groups. Elemental analyses of the complexes indicate that the metal to ligand ratio is 1:1 in all complexes. Infrared spectra of complexes indicate a dibasic quadridentate nature of the ligand and its coordination to metal ions through phenolic oxygen and azomethine nitrogen atoms. The thermal behavior of these complexes showed the loss of lattice water in the first step followed by decomposition of the ligand in subsequent steps. The thermal data have also been analyzed for the kinetic parameters by using Horowitz-Metzger method. The dependence of the electrical conductivity on the temperature has been studied over the temperature range 313-403 K and the complexes are found to show semiconducting behavior. XRD and SEM images of some representative complexes have been recorded. The antimicrobial activity of the ligand and its complexes has been screened against various microorganisms and all of them were found to be active against the test organisms. The Fe(III) and Ni(II) complex have been tested for the catalytic oxidation of styrene.

Substrate Specificity of the Human Flavin-containing Monooxygenase for Organic Selenium Compounds (사람 Flavin-containing Monooxygenase의 셀레니움화합물에 대한 기질 특이성에 관한 연구)

  • Kim, Young-Mi
    • Environmental Analysis Health and Toxicology
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    • v.15 no.4
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    • pp.139-145
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    • 2000
  • The Flavin-containing monooxygenase (FMOs) (EC1.14.13.8) are NADPH-dependent flavoenzymes that catalyze oxidation of soft nucleophilic heteroatom centers in a range of structurally diverse compounds, including foods, drugs, pesticides, and other xenobiotics. In humans, FMO3 is quantitatively a major human liver monooxygenase. In the present study, the baculovirus expression vector system was used to overexpress human FMO3 in insect cells for catalytic studies. Six commercially available organic selenium compounds were examined for substrate activity with microsomes isolated from Spodoptera frugiperda (Sf)9 cells infected with human FMO3 recombinant baculovirus. While none of the aromatic heterocyclic selenides tested showed detectable activity, all dialkyl- and alkylaryl-selenides free from ionic groups catalyzed the NADPH- and O$_2$-dependent oxidation. Kinetic constants demonstrate that (based on Km) dialkyl-and alkylaryl- selenides are better substrates for human FMO3 than analogous nitrogen or sulfur compounds .

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