• Title/Summary/Keyword: Catalyses

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Expression of Acetohydroxyacid Synthase from Bacillus anthracis and Its Potent Inhibitors

  • Choi, Kyoung-Jae;Pham, Chien Ngoc;Jung, Hoe-Il;Han, Sung-Hwan;Choi, Jung-Do;Kim, Jin-Heung;Yoon, Moon-Young
    • Bulletin of the Korean Chemical Society
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    • v.28 no.7
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    • pp.1109-1113
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    • 2007
  • Acetohydroxyacid synthase (AHAS, EC 2. 2. 1. 6) is the enzyme that catalyses the first step in the common pathway of the biosynthesis of the branched chain amino acids, valine, leucine and isoleucine. For the first time, the AHAS gene from Bacillus anthracis was cloned into the expression vector pET28a(+), and was expressed in the E. coli strain BL21(DE3). The purified enzyme was checked on 12% SDS-PAGE to be a single band with molecular weight of 65 kDa. The optimum pH and temperature for B. anthracis AHAS was at pH 7.5 and 37 oC, respectively. Kinetic parameters of B. anthracis were as follows: Km for pyruvate, K0.5 for ThDP and Mg2+ was 4.8, 0.28 and 1.16 mM respectively. AHAS from B. anthracis showed strong resistance to three classes of herbicides, Londax (a sulfonylurea), Cadre (an imidazolinone), and TP (a triazolopyrimidine). These results indicated that these herbicides could be used in the search for new anti-bacterial drugs.

Assay of the Bombyx mori Recombinant Protein Disulfide Isomerase (bPDI) Acivity

  • Goo, Tae-Won;Yun, Eun-Young;Kim, Sung-Wan;Park, Kang-Ho;Hwang, Jae-Sam;Kang, Seok-Woo;Park, Soo-Jung;Kwon, O-Yu
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.04a
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    • pp.45-45
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    • 2003
  • Protein disulfide isomerase (PDO) is an essential protein which is localized to the endoplasmic reticulum (ER) of eukaryotic cells. It catalyses the formation and isomerization of disulfide bonds during the folding of secretory proteins. We have isolarted a cDNA encoding PDI from Bombyx mori (bPDI), in which an open reading frame of 494 mino acid (55.6kDa) is shown. (omitted)

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The uniqueness of the plant mitochondrial potassium channel

  • Pastore, Donato;Soccio, Mario;Laus, Maura Nicoletta;Trono, Daniela
    • BMB Reports
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    • v.46 no.8
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    • pp.391-397
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    • 2013
  • The ATP-inhibited Plant Mitochondrial $K^+$ Channel ($PmitoK_{ATP}$) was discovered about fifteen years ago in Durum Wheat Mitochondria (DWM). $PmitoK_{ATP}$ catalyses the electrophoretic $K^+$ uniport through the inner mitochondrial membrane; moreover, the co-operation between $PmitoK_{ATP}$ and $K^+/H^+$ antiporter allows such a great operation of a $K^+$ cycle to collapse mitochondrial membrane potential (${\Delta}{\Psi}$) and ${\Delta}pH$, thus impairing protonmotive force (${\Delta}p$). A possible physiological role of such ${\Delta}{\Psi}$ control is the restriction of harmful reactive oxygen species (ROS) production under environmental/oxidative stress conditions. Interestingly, DWM lacking ${\Delta}p$ were found to be nevertheless fully coupled and able to regularly accomplish ATP synthesis; this unexpected behaviour makes necessary to recast in some way the classical chemiosmotic model. In the whole, $PmitoK_{ATP}$ may oppose to large scale ROS production by lowering ${\Delta}{\Psi}$ under environmental/oxidative stress, but, when stress is moderate, this occurs without impairing ATP synthesis in a crucial moment for cell and mitochondrial bioenergetics.

Molecular Cloning and Characterization of Catechol 2, 3-Dioxygenase Gene from Aniline-Degrading Psseudomonas acidovorans

  • Lee, Ji-Hyun;Bang, Sung-Ho;Park, Youn-Keun;Lee, Yung-Nok
    • Korean Journal of Microbiology
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    • v.30 no.4
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    • pp.316-321
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    • 1992
  • Catechol 2, 3-dioxygenase (C230) catalyses the oxidative ring cleavage of catechol to 2-hydroxymuconic semialdehyde. This is one of the key reactions in the metabolism of the widespresd pollutant aniline. We have cloned a gene encoding C230 from cells of the aniline degrading bacteria, Pseudomonas acidovorance KCTC2494 strain and expressed in E. coli, A 11.3-kilobase Sau3A partial digested DNA fragment from KCTC2494 was cloned into phagemid vector pBluescript and designated as pLP201. The C230 gene was mapped to a 2.8-kb region, and the derection of transcription was determined. The cloned C230 gene contains its own promoter which can be recognized and employed by E. coli transcriptional apparatus. C230 activities of subclones were identified by enzyme assay and activity staining. The T7 RNA promoter/polymerase system and maxicell analysis showed that a polypeptide with Mw of 35 kDa is the C230 gene product.

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Solandelactone G: A Novel Inhibitor of Farnesyl Protein Transferase (FPTase), Isolated from the Hydroid Solanderia secunda

  • Jongheon Shin;Youngwan Seo;Park, Jung-Rae;Kwon, Byoung-Mog;Nam, Ji-Youn;Jeong, Tae-Sook;Bok, Song-Hae
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.63-63
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    • 1995
  • Farnesyl Protein Transferase (FPTase) catalyses a post-translational modification of Ras that is obligatory for the cell transforming activity of this oncogene protein. The screening of natural products to identify inhibitors of this enzyme as a potential anticancer agents, has led to the isolation of a novel lactone, from the hydroid Solanderia secunda. Solandelactone G has been isolated from the hydroid Solanderia secunda collected along the offshore of Jaejudo and Keomunde. The structure of the compound has been determined as cyclopropane containing C$\_$22/ fatty acid lactone on the basis of the combined spectral and chemical methods

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F93-A: A Inhibitor of Farnesyl Protein Transferase from Aspergillus fumigatus KL93

  • Kwon, Byoung-Mog;Lee, Seung-Ho;Jeong, Tae-Sook;Kim, Sung-Uk;Son, Kwang-Hee;Park, Diol;Kim, Young-Kook;Nam, Ji-Youn;Bok, Song-Hae
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.66-66
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    • 1995
  • Mutated forms of the ras oncogenes are associated with about 30% of human tumors. The ras genes encode 21KDa proteins, called p21 or Ras, that are associated with the plasma membrane. FPTase is a dimeric enzyme that catalyses the transfer of the farnesyl group from farnesyl pyrophosphate onto cysteine 186 at the C-terminus of the Ras protein. This is mandatory process for triggering ras oncogene toward tumor formation. Therefore, selective inhibitors of FPTase have the potential to be used as antitumorgenic agents.

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Inter-Chain Interactions in Arrays of Metal-Organic Hybrid Chains on Ag(111)

  • Park, Ji-Hun;Jeong, Gyeong-Hun;Yun, Jong-Geon;Kim, Ho-Won;Gang, Se-Jong
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.02a
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    • pp.302-302
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    • 2011
  • Fabrications of metal-organic hybrid networks attracted much attention due to possible applications in gas storages, heterogeneous catalyses, information storages, and opto-electronic devices. One way to construct three-dimensional hybrid structures is to make the arrays of planar or linear metal-organic hybrid structures which are linked through electrostatic interactions. As a model study, we fabricated the arrays of one-dimensional hybrid chains and investigated inter-chain interactions between adjacent hybrid chains using scanning tunneling microscopy (STM) and spectroscopy (STS) on Ag(111). Brominated anthracene molecules were used to grow the arrays of hybrid chains on Ag(111). We proposed atomic models for the observed structures. Linear chains are made of repetition of Ag-anthracene units. Br atoms are attached to anthracene molecules through Br-H structures which mediate inter-chain interactions. Two different apparent heights were observed in anthracene molecules. Molecules having a Br-H connection look brighter than those with two connections due to electronic effect. When a chain is laterally manipulated with STM tip, Br atoms move together with the chain implying that Br-H inter-chain interactions are quite strong.

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Hydrogen Embrittlement and Surface Properties of Pd-coated Zr-based Amorphous Alloys (Pd 코팅된 Zr기 비정질 합금의 수소취성 및 표면특성)

  • Seok, Song;Lee, Dock-Young;Kim, Ki-Bae
    • Journal of Hydrogen and New Energy
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    • v.18 no.2
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    • pp.182-188
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    • 2007
  • [ $Zr_{50}-Ni_{27}-Nb_{18}-Co_5$ ] amorphous alloys ribbon was prepared by a single-roller melt-spinning technique. In order to improve the hydrogen kinetics Pd-coating were carried out on each side of the amorphous ribbon. Pd prevents oxidation of Zr and catalyses the dissociation of molecular hydrogen to atomic hydrogen. In this work, the hydrogen embrittlement and surface properties on Zr-based amorphous alloys were investigated. The Zr-based amorphous alloys were characterized by X-ray diffractometry(XRD) and differential scanning calorimetry(DSC). The morphology of surface and roughness was observed by using scanning electron microscopy(SEM) and atomic force microscopy (AFM). A lattice parameter of both Pd and Zr-based amorphous alloy was increased after hydrogen permeation at 473 K. After hydrogen permeation at 473 K, some cracks were observed on the surface of Pd, which was the cause for the hydrogen embrittlement. The crystallization temperature of Zr-based amorphous alloy was decreased due to the permeated hydrogen.

A Photocatalytic Degradation of Synthetic Detergent over $TiO_2$ Catalysts Prepared by Sol-Gel Method (졸-겔법으로 제조된 $TiO_2$촉매에 의한 합성세제의 광분해)

  • 양천회;홍필선
    • Journal of the Korean Society of Safety
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    • v.16 no.3
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    • pp.76-82
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    • 2001
  • An aqueous solution of a commercial liquid synthetic detergent for kitchen use was photodecomposed in the presence of titanium dioxides ponder under an atmosphere of air at room temperature. Titanium dioxides were prepared by sol-gel method from titanium iso-propoxide at different R R ratio($H_2O$/titanium iso-propoxide) and calcined at $500^{\circ}C$. All titanium dioxides m characterized by XRD, BET surface area analyzer and UV-VIS spectrometer. The surface mea of titanium dioxides prepared at R ratio=6 appeared higher volume about 20% than commercial $TiO_2$ catalyses. XRD patterns of titania particles were observed mixing phase together with rutile and anatase type. Titanium dioxides prepared by sol-gel method show higher activity about 6% than commercial $TiO_2$ catalysts on the Photocatalytic foundation of a commercial liquid synthetic detergent for kitchen. The concentration of the detergent decreased to about 90%, of its initial value at illumination times of 2 hour. illumination for 30 minutes decreased the concentration of oxygen to about one-fifth of the initial value.

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Site-directed Mutagenesis of Five Conserved Residues of Subunit I of the Cytochrome cbb3 Oxidase in Rhodobacter capsulatus

  • Ozturk, Mehmet;Gurel, Ekrem;Watmough, Nicholas J.;Mandaci, Sevnur
    • BMB Reports
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    • v.40 no.5
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    • pp.697-707
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    • 2007
  • Cytochrome $cbb_3$ oxidase is a member of the heme-copper oxidase superfamily that catalyses the reduction of molecular oxygen to the water and conserves the liberated energy in the form of a proton gradient. Comparison of the amino acid sequences of subunit I from different classes of heme-copper oxidases showed that transmembrane helix VIII and the loop between transmembrane helices IX and X contain five highly conserved polar residues; Ser333, Ser340, Thr350, Asn390 and Thr394. To determine the relationship between these conserved amino acids and the activity and assembly of the $cbb_3$ oxidase in Rhodobacter capsulatus, each of these five conserved amino acids was substituted for alanine by site-directed mutagenesis. The effects of these mutations on catalytic activity were determined using a NADI plate assay and by measurements of the rate of oxygen consumption. The consequence of these mutations for the structural integrity of the $cbb_3$ oxidase was determined by SDS-PAGE analysis of chromatophore membranes followed by TMBZ staining. The results indicate that the Asn390Ala mutation led to a complete loss of enzyme activity and that the Ser333Ala mutation decreased the activity significantly. The remaining mutants cause a partial loss of catalytic activity. All of the mutant enzymes, except Asn390Ala, were apparently correctly assembled and stable in the membrane of the R. capsulatus.