• Title/Summary/Keyword: Chemical Modification

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Critical Role of the Cysteine 323 Residue in the Catalytic Activity of Human Glutamate Dehydrogenase Isozymes

  • Yang, Seung-Ju;Cho, Eun Hee;Choi, Myung-Min;Lee, Hyun-Ju;Huh, Jae-Wan;Choi, Soo Young;Cho, Sung-Woo
    • Molecules and Cells
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    • v.19 no.1
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    • pp.97-103
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    • 2005
  • The role of residue C323 in catalysis by human glutamate dehydrogenase isozymes (hGDH1 and hGDH2) was examined by substituting Arg, Gly, Leu, Met, or Tyr at C323 by cassette mutagenesis using synthetic human GDH isozyme genes. As a result, the $K_m$ of the enzyme for NADH and ${\alpha}-ketoglutarate$ increased up to 1.6-fold and 1.1-fold, respectively. It seems likely that C323 is not responsible for substrate-binding or coenzyme-binding. The efficiency ($k_{cat}/K_m$) of the mutant enzymes was only 11-14% of that of the wild-type isozymes, mainly due to a decrease in $k_{cat}$ values. There was a linear relationship between incorporation of [$^{14}C$]p-chloromercuribenzoic acid and loss of enzyme activity that extrapolated to a stoichiometry of one mol of [$^{14}C$] incorporated per mol of monomer for wild type hGDHs. No incorporation of [$^{14}C$]p-chloromercuribenzoic acid was observed with the C323 mutants. ADP and GTP had no effect on the binding of p-chloromercuribenzoic acid, suggesting that C323 is not directly involved in allosteric regulation. There were no differences between the two hGDH isozymes in sensitivities to mutagenesis at C323. Our results suggest that C323 plays an important role in catalysis by human GDH isozymes.

Preparation of Higher Reinforced PVDF Hollow Fiber Microfiltration Membrane (고강도 PVDF 중공사 정밀여과막 제조 특성)

  • Choi, R.S.;Park, H.H.
    • Membrane Journal
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    • v.20 no.4
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    • pp.320-325
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    • 2010
  • This paper was carried out to study the preparation condition and the permeation flux of reinforced poly(vinylidene fluoride) (PVDF) hollow fiber microfiltration (MF) membrane with the solvent, additive, second miscible polymer, and preparation condition used poly(vinylidene fluoride) (PVDF) such as a material with the excellent chemical stability and the milder preparation condition. The performance of the reinforced MF membrane prepared obtained the average $0.3{\mu}m$ pore size, $42kg_f/cm^2$ tensile strength, and the high water flux of 600 LMH. The change of membrane performance with various additives was considerably affected on the water flux and rejection. For hydrophilic modification of hydrophobic PVDF MF membrane, the MF membrane might be prepared with a prefer water flux and rejection by addition of hydrophilic poly(methyl methacrylate) (PMMA).

Effect of DPBll Gene for the Transcriptional Induction by DNA Damage During Cell Cycle in Saccharomyces cerevisiae (출아효모의 세포주기동안 DNA 상해에 의한 발현 유도에 미치는 DPB11 유전자의 영향)

  • 선우양일;임선희;배호정;김중현;김은아;김승일;김수현;박정은;김재우
    • Korean Journal of Microbiology
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    • v.38 no.2
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    • pp.96-102
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    • 2002
  • The S-phase checkpoint mechanisms response to DNA damage or inhibition of DNA replication for maintenance of genetic stability in eukaryotic cells. These roles include cell cycle control arrest at S-phase and Iranscriptional induction of repair genes. To characterize the defects of dpbll mutant for both these responses, we examined the over-expression effect of DPBll gene, the sensitivity to HU, MMS, and the transcriptional pattern by DNA damage agent for RNRS mRNA. RNRS transcript is induced in response to a wide variety of agents that either damage D7A directly through chemical modification or induce stress by blocking DNA synthesis. As results, dpbll-1 cells are sensitive to DNA damage agents and the level of RNR3 mRNA is reduced approximately 40% than wild type cells. Moreover, we found the same results in dpb2-1 cells. Therefore, we propose that DPB2 and DPBll act as a sensor of replication that coordinates the transcriptional and cell cycle responses to replication blocks.

Synthesis, Characterization and Functionalization of the Coated Iron Oxide Nanostructures

  • Tursunkulov, Oybek;Allabergenov, Bunyod;Abidov, Amir;Jeong, Soon-Wook;Kim, Sungjin
    • Journal of Powder Materials
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    • v.20 no.3
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    • pp.180-185
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    • 2013
  • The iron oxides nanoparticles and iron oxide with other compounds are of importance in fields including biomedicine, clinical and bio-sensing applications, corrosion resistance, and magnetic properties of materials, catalyst, and geochemical processes etc. In this work we describe the preparation and investigation of the properties of coated magnetic nanoparticles consisting of the iron oxide core and organic modification of the residue. These fine iron oxide nanoparticles were prepared in air environment by the co-precipitation method using of $Fe^{2+}$: $Fe^{3+}$ where chemical precipitation was achieved by adding ammonia aqueous solution with vigorous stirring. During the synthesis of nanoparticles with a narrow size distribution, the techniques of separation and powdering of nanoparticles into rather monodisperse fractions are observed. This is done using controlled precipitation of particles from surfactant stabilized solutions in the form organic components. It is desirable to maintain the particle size within pH range, temperature, solution ratio wherein the particle growth is held at a minimum. The iron oxide nanoparticles can be well dispersed in an aqueous solution were prepared by the mentioned co-precipitation method. Besides the iron oxide nanowires were prepared by using similar method. These iron oxide nanoparticles and nanowires have controlled average size and the obtained products were investigated by X-ray diffraction, FESEM and other methods.

Assessment of organic matter biodegradation and physico-chemical parameters variation during co-composting of lignocellulosic wastes with Trametes trogii inoculation

  • Fersi, Mariem;Mbarki, Khadija;Gargouri, Kamel;Mechichi, Tahar;Hachicha, Ridha
    • Environmental Engineering Research
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    • v.24 no.4
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    • pp.670-679
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    • 2019
  • Lignin complexity molecule makes its biodegradation difficult during lignocellulosic wastes composting. So, the improvement of its biodegradation has usually been considered as an objective. This study aimed to determine the impact of Trametes trogii inoculation on organic matter and particularly on lignin and cellulose during green wastes co-composting with olive mill waste water sludge and coffee grounds. Three types of heaps (H1, H2 and H3) were investigated during 180 d. H3 and H2 were inoculated at the beginning of the process (t0) and 120 d later (t120), respectively while H1 was the control. Results showed the absence of pH stabilization in H3 during the first month. Also, in this period we observed a faster degradation of some easily available organic matter in H3 than in the other heaps. After 120 d, a better cellulose decomposition (25.28%) was noticed in H3 than in H1 and H2 (16%). Inoculation during the second fermentation phase induced supplementary lignin degradation in H2 with a percentage of 35% against 23 and 26% for H1 and H3, respectively. For all the runs, a Fourier Transform Infrared analysis showed aliphatic groups' decrease, OH groups' increase and lignin structural modification.

Study of surface modification and contact angle by electrospun PVdF-HFP membrane with DLC coating (DLC 코팅에 의한 PVdF-HFP 막의 표면변화 및 접촉각 연구)

  • Lee, Tae Dong;Cho, Hyun;Yoon, Su Jong;Kim, Tae Gyu
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.24 no.1
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    • pp.33-40
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    • 2014
  • Poly vinylidene fluoride-co-hexafluoropropylene (PVdF-HFP) membrane were prepared by the electrospinning technique. We had applied a DLC coating process and then the surface of the membrane and the contact angle change was investigated. Electrospun fibrous PVdF-HFP membrane surface became to wrinkled shape by Ar plasma treatment and treatment conditions. The wrinkled surface of PVdF-HFP membrane became super-hydrophilic. However, after DLC coating process, it became super-hydrophobic. The resulting surfaces were characterized by water contact angle measurement, X-ray photoelectron spectroscopy (XPS) and field emission scanning electron microscopy (FE-SEM). Resultantly it was recognized that the wettability characteristics of the membrane surfaces depended on the chemical composition and surface morphology.

Hydrophobic Self-assembled Monolayer(SAM) Coating for Enhanced Demolding Performance in Micromolding (마이크로몰딩의 이형성 향상을 위한 소수성 Self-assembled Monolayer(SAM) 코팅)

  • Park, Sang-Ha;Han, Seung-O;Park, Jong-Yeon;Mun, Seong-Uk;Park, Jeong-Ho
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.51 no.4
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    • pp.175-183
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    • 2002
  • In this paper, the surface modification effect of self-assembled monolayer(SAM) of 1-dodecanethiol [$CH_3$($CH_2$)$_{11}$SH] used as an anti-adhesive film in micromolding process was studied. Monolayers of 1-dodecanethiol[$CH_3$(CH$_2$)$_{11}$SH] were obtained by immersing a metal place in pure 1-dodecanethiol. SAM film on the nickel plate has been examined by using X-ray photoelectron spectroscopy(XPS). The focus has been placed on S-Ni bonding. From the XPS analysis, sulfur atoms were detected from the SAM film as a chemical composition of S-Ni. In order to measure an adhesion force of the SAM-coated nickel surface, atomic force microscopy(AFM) was used in force-distance mode, which whows the micro-adhesive force on solid surface. It was shown that adhesion forces measured from the SAM-coated nickel surface and the Ni surface without SAM coating were 3.52nN and 5.32nN, respectively. In order to investigate the effect of SAM coating on the surface foughness the replica in demolding process, hot embossing experiments were performed using a SAM-coated nickel master and a nickel master without SAM coating. Surface roughness of replica from the SAM-coated master showed 25nm and that of replica from master without SAM coating was 35nm. The smoother surface roughness of the replica from the SAM-coated, master is believed to result from reduction in the adhesion forces.ces.

Kinetic Properties of $\alpha$-Galactosidase from Aspergillus niger ATCC 16513 and Soybean(Glycine max. L) (Aspergillus niger ATCC 16513과 대두(Glycine max. L) $\alpha$-galactosidase의 kinetic 성질)

  • Geum, Jong-Hwa;Lee, Jong-Su;Sin, Cheol-Seung
    • The Journal of Natural Sciences
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    • v.5 no.1
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    • pp.53-57
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    • 1992
  • This experiment was carried out to elucidate some kinetic properties of the $\alpha$-galactosidase which produced and purified from Aspergillus niger ATCC 16513 and soybean(Glycine max. L). The Km value of Asp. niger and soybean $\alpha$-galactosidase were 37.0mM and 50.0mM for raffinose and55.5mM and 55.5mM for stachyose, respectively. The activity of Asp. niger and soybean $\alpha$-galactosidase were inhibited by galactose. Among the amino acids in active sites of both Asp. niger and soybean $\alpha$-galactosidase, histidine was identified by chemical modification of diethyl pyrocarbonate. Number of amino acids residues per mole of Asp. niger and soybean $\alpha$-galactosidase were 902 and 286, respectively.

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Potent Inhibition of Human Cytochrome P450 1 Enzymes by Dimethoxyphenylvinyl Thiophene

  • Lee, Sang-Kwang;Kim, Yongmo;Kim, Mie-Young;Kim, Sanghee;Chun, Young-Jin
    • Archives of Pharmacal Research
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    • v.27 no.2
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    • pp.199-205
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    • 2004
  • Cytochrome P450 (P450) 1 enzymes such as P450 1A1, 1A2, and 181 are known to be involved in the oxidative metabolism of various procarcinogens and are regarded as important target enzymes for cancer chemoprevention. Previously, several hydroxystilbene compounds were reported to inhibit P450 1 enzymes and were rated as candidate chemopreventive agents. In this study, we investigated the inhibitory effect of 2-[2-(3,5-dimethoxyphenyl)vinyl]-thiophene (DMPVT), produced from the chemical modification of oxyresveratrol, on the activities of P450 1 enzymes. The inhibitory potential by DMPVT on the P450 1 enzyme activity was evaluated with the Escherichia coli membranes of the recombinant human cytochrome P450 1A1, 1A2, or 1B1 coexpressed with human NADPH-P450 reductase. DMPVT significantly inhibited ethoxyresorufin O-deethylation (EROD) activities with $IC_{50}$ values of 61, 11, and 2 nM for 1A1, 1A2, and 1B1, respectively. The EROO activity in OMBA-treated rat lung microsomes was also significantly inhibited by OMPVT in a dose-dependent manner. The modes of inhibition by DMPVT were non-competitive for all three P450 enzymes. The inhibition of P450 1B1-mediated EROD activity by OMPVT did not show the irreversible mechanism-based effect. The loss of EROD activity in P450 1B1 with OMPVT incubation was not blocked by treatment with the trapping agents such as glutathione, N-acetylcysteine, or dithiothreitol. Taken together, the results suggested DMPVT to be a strong noncompetitive inhibitor of human P450 1 enzymes that should be considered as a good candidate for a cancer chemopreventive agent in humans.

Effect of Adding Crosslinked Particles on Rheological and Electrical Properties of Polystyrene/Carbon Nanotube Nanocomposites (가교 입자 첨가가 폴리스티렌/탄소나노튜브 나노복합재료의 유변물성 및 전기적 물성에 미치는 영향)

  • Yeom, Hyo Yeol;Na, Hyo Yeol;Lee, Seong Jae
    • Polymer(Korea)
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    • v.38 no.6
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    • pp.767-773
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    • 2014
  • Rheological and electrical properties of polystyrene (PS)/carbon nanotube (CNT) nanocomposites via coagulated precipitation were investigated. Chemical modification and surfactant wrapping of CNT to improve the dispersion of CNTs may reduce the intrinsic properties of pristine CNT. To avoid this problem, PS and CNTs were dissolved and dispersed in dimethylformamide and then PS/CNT nanocomposites were prepared by the coagulated precipitation of CNT-dispersed PS solution in water. The coagulated precipitation method was highly effective enhancing the electrical conductivity of nanocomposites. Furthermore, the effect of adding poly(styrene-co-divinylbenzene) crosslinked particles to PS matrix on the rheological and electrical properties was investigated. With the addition of the crosslinked particles, the electrical percolation threshold of CNT reduced to 0.25 wt% and electrical conductivity increased further. It is speculated that CNTs in the volume occupied by crosslinked particles helped electrical pathway formation.