• Title/Summary/Keyword: Monomeric form

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The Purification and Characterization of Bacillus subtilis Tripeptidase (PepT)

  • Park, Yong-Seek;Cha, Myung-Hoon;Yong, Whan-Mi;Kim, Hyo-Joon;Chung, Il-Yup;Lee, Young-Seek
    • BMB Reports
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    • v.32 no.3
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    • pp.239-246
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    • 1999
  • A tripeptidase (PepT) was purified to homogeneity from Bacillus subtilis through four sequential chromatographies including DEAE-Sepharose ion exchange, hydroxylapatite, mono-Q FPLC ion exchange, and Superose-12 FPLC gel filtration. The apparent molecular mass of the enzyme was 49,200 Da and 51,400 Da as determined by sodium dodecylsulfatepolyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration chromatography, respectively, and the enzyme exists in a monomeric form. The physicochemical properties of the enzyme were as follows: optimum pH at 7.5, optimum temperature at $60^{\circ}C$, and pI at 4.9. The $K_m$ and $V_{max}$ values of the enzyme were 4.3 mM and 2.5 mmol/min/mg, respectively, with MetAla-Ser as substrate. The B. subtilis PepT requires $Co^{2+}$ ion(s) for activation, while it is inactivated by EOTA and 1,10-phenanthroline, suggesting that it is a metalloprotein. The enzyme was not inhibited by any of serine protease, aspartic protease, or leucine aminopeptidase inhibitors. The enzyme showed comparable activities towards four different substrates including Met-Ala-Ser, Leu-Gly-Gly, Leu-Ser-Phe, and Leu-Leu-Tyr. The amino terminal sequence of PepT determined by Edman degradation was found to be MKEEIIERFTTYVXV and turned out to be identical to that of PepT deduced from a cloned B. subtilis pepT.

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Salen-Aluminum Complexes as Host Materials for Red Phosphorescent Organic Light-Emitting Diodes

  • Bae, Hye-Jin;Hwang, Kyu-Young;Lee, Min-Hyung;Do, Young-Kyu
    • Bulletin of the Korean Chemical Society
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    • v.32 no.9
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    • pp.3290-3294
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    • 2011
  • The properties of monomeric and dimeric salen-aluminum complexes, [salen(3,5-$^tBu)_2$Al(OR)], R = $OC_6H_4-p-C_6H_6$ (H1) and R = [salen(3,5-$^tBu$)AlOPh]C$(CH_3)_2$ (H2) (salen = N,N'-bis-(salicylidene)-ethylenediamine) as host layer materials in red phosphorescent organic light-emitting diodes (PhOLEDs) were investigated. H1 and H2 exhibit high thermal stability with decomposition temperature of 330 and $370^{\circ}C$. DSC analyses showed that the complexes form amorphous glasses upon cooling of melt samples with glass transition temperatures of 112 and $172^{\circ}C$. The HOMO (ca. -5.2~-5.3 eV) and LUMO (ca. -2.3~-2.4 eV) levels with a triplet energy of ca. 1.92 eV suggest that H1 and H2 are suitable for a host material for red emitters. The PhOLED devices based on H1 and H2 doped with a red emitter, $Ir(btp)_2$(acac) (btp = bis(2-(2'-benzothienyl)-pyridinato-N,$C^3$; acac = acetylacetonate) were fabricated by vacuum-deposition and solution process, respectively. The device based on vacuum-deposited H1 host displays high device performances in terms of brightness, luminous and quantum efficiencies comparable to those of the device based on a CBP (4,4'-bis(Ncarbazolyl) biphenyl) host while the solution-processed device with H2 host shows poor performance.

Synthesis and Oxidative Catalytic Property of Ruthenium-doped Titanate Nanosheets (루테늄이 도입된 티타네이트 나노시트의 합성 및 산화 촉매 활성 연구)

  • Lee, Yoonhee;Kwon, Ki-Young
    • Applied Chemistry for Engineering
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    • v.28 no.5
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    • pp.593-596
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    • 2017
  • Sodium titanate nanosheets were prepared by a hydrothermal synthesis method under basic conditions. Ruthenium was introduced on the surface of sodium titanate nanosheets through an UV irradiation in the aqueous $RuCl_3$ solution. The crystal phase and morphology of synthesized samples were analyzed by X-ray diffraction, transmission electron microscopy and energy dispersive spectroscopy. In addition, the content of Ru was evaluated by inductively coupled plasma. It was proposed that a monomeric form of ruthenium was incorporated on the surface of sodium titanate. Ruthenium incorporated sodium titanates were applied to alcohol oxidation using molecular oxygen as an oxidant. The sample with 7% ruthenium showed a catalytic activity with a turnover frequency value of $2.1h^{-1}$ in oxidizing benzyl alcohol to benzaldehyde without any other byproducts at $105^{\circ}C$ and 1 atmosphere.

Syntheses and Crystal Structures of Xylyl-Bridged NO2S2-Donor Macrocycles and Binuclear Mercury(II) Complex

  • Lee, Ji-Eun;Jin, Yong-ri;Seo, Joo-beom;Yoon, Il;Song, Mi-Ryoung;Lee, So-Young;Park, Ki-Min;Lee, Shim-Sung
    • Bulletin of the Korean Chemical Society
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    • v.27 no.2
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    • pp.203-207
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    • 2006
  • Isomeric series of dilinked $NO_2S_2$ macrocycles ($L^2$: para-, $L^3$: meta- and $L^4$: ortho-linked) capable of binuclear complexing ability were prepared from its monomeric analog $L^1$ in reasonable yields except ortho-type reaction, which led to mixture due to the formation of monomer-type macrocyclic quaternary ammonium bromide $L^5$. Moreover, L2 (as $2HNO_3$ form) and $L^5$ were confirmed by an X-ray crystallography. Reaction of $HgCl_2$ with $L^2$ yielded a binuclear complex $[Hg_2(L^2)Cl_4]$. In the complex, each mercury(II) has a distorted tetrahedral environment made up of S and N donors from an exodentate $L^2$ and two coordinated Cl atoms.

Solvent Mediated Hydrogen-bonded Supramolecular Network of a Cu(II) Complex Involving N2O Donor Ligand and Terephthalate (N2O 주개 리간드와 테레프탈레이트를 포함하는 구리(II) 착물의 용매를 매개로 한 수소결합형 초분자 네트워크)

  • Chakraborty, Jishnunil
    • Journal of the Korean Chemical Society
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    • v.55 no.2
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    • pp.199-203
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    • 2011
  • The title one-dimensional hydrogen-bonded coordination compound $[Cu^{II}(C_{13}H_{17}N_3OBr)(C_8H_5O_4)]{\cdot}2H_2O.CH_3OH$ has been synthesized and characterized by single crystal X-ray diffraction study. The monomeric unit contains a square-planar $Cu^{II}$ centre. The four coordination sites are occupied by a tridentate anionic Schiff base ligand (4-bromo-2-[(2-piperazin-1-yl-ethylimino)-methyl]-phenol) which furnishes an $N_2O$-donor set, with the fourth position being occupied by the oxygen atom of an adjacent terephthalate unit. Two adjacent neutral molecules are linked through intermolecular N-H---O and O-H---N hydrogen bonds and generate a dimeric pair. Each dimeric pair is connected with each other via discrete water and methanol molecules by hydrogen bonding to form a one-dimensional supramolecular network.

High Level of Soluble Expression in Escherichia coli and Characterisation of the Cloned Bacillus thuringiensis Cry4Ba Domain III Fragment

  • Chayaratanasin, Poramed;Moonsom, Seangdeun;Sakdee, Somsri;Chaisri, Urai;Katzenmeier, Gerd;Angsuthanasombat, Chanan
    • BMB Reports
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    • v.40 no.1
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    • pp.58-64
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    • 2007
  • Similar to the other known structures of Bacillus thuringiensis Cry $\delta$-endotoxins, the crystal structure of the 65-kDa activated Cry4Ba toxin comprises three domains which are, from the N- to C-terminus, a bundle of $\alpha$-helices, a three-$\beta$-sheet domain, and a $\beta$-sandwich. To investigate the properties of the C-terminal domain III in isolation from the rest of the toxin, the cloned Cry4Ba-domain III was over-expressed as a 21-kDa soluble protein in Escherichia coli, which cross-reacted with anti-Cry4Ba domain III monoclonal antibody. A highly-purified domain III was obtained in a monomeric form by ion-exchange and size-exclusion FPLC. Circular dichroism spectroscopy indicated that the isolated domain III fragment distinctly exists as a $\beta$-sheet structure, corresponding to the domain III structure embodied in the Cry4Ba crystal structure. In vitro binding analysis via immuno-histochemical assay revealed that the Cry4Ba-domain III protein was able to bind to the apical microvilli of the susceptible Stegomyia aegypti larval midguts, albeit at lower-binding activity when compared with the full-length active toxin. These results demonstrate for the first time that the C-terminal domain III of the Cry4Ba mosquito-larvicidal protein, which can be isolated as a native folded monomer, conceivably participates in toxin-receptor recognition.

Thermodynamics on the Mixed Micellar Formation of Dimethyldodecylamine Oxide in Water/n-Propanol (Dimethyldodecylamine Oxide 의 물/n-프로판올 용매에서 혼합미셸 형성에 관한 열역학적 연구)

  • Lee Byung-Hwan
    • Journal of the Korean Chemical Society
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    • v.37 no.6
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    • pp.562-569
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    • 1993
  • The pseudophase separation model is used to describe the effects of pH and n-propanol on the mixed micellar formation of protonated and unprotonated dimethyldodecylamine oxides. Dimethyl-dodecylamine oxide surfactant molecules may exist in aqueous solution in either nonionic (unprotonated) or cationic (protonated) form, and they can be modeled thermodynamically as a binary mixture of cationic and nonionic surfactants. The composition of the binary mixture is varied by adjusting the solution pH. And activities, micellar compositions, and monomeric compositions of two surfactant species can be calculated directly from the experimental titration data by applying pseudophase separation model to the micellar system of DDAO in water/n-propanol. The critical micellar concentrations and the p$K_a$ values of the binary mixture systems are dependent on the micellar composition of the protonated cationic surfactant (X); especially they show the minimum phenomena when they are plotted against the micellar composition of the protonated cationic surfactant (X). The critical micellar concentration of the binary mixed DDAO system is generally decreased when n-propanol is added to the binary mixture system, and the degree of decrease is dependent on the concentration of n-propanol.

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Characterization of Xylanase from Bacillus agaradhaerens DK-2386 Isolated from Korean Soil (토양으로부터 분리한 Bacillus agaradhaerens DK-2386 균주가 생산하는 Xylanase의 특성)

  • Choi, Ji-Hwi;Park, Young-Seo;Lee, Hyungjae;Bai, Dong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.43 no.4
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    • pp.330-335
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    • 2015
  • The optimum conditions for the production of xylanase from Bacillus agaradhaerens DK-2386 have been previously investigated. In this study xylanase was purified by ammonium sulfate precipitation and CM-sepharose ion exchange chromatography. The molecular mass of the xylanase as determined by SDS-PAGE was 23 kDa in a form of monomeric enzyme. The optimum pH and temperature for xylanase activity was 6.0 and $60^{\circ}C$, respectively. Xylanase activity was increased by the addition of EDTA and then stabilized at $40^{\circ}C$ for 24 h. The maximum xylanase activity was obtained when Birchwood xylan was used as a substrate and the $V_{max}$ and $K_m$ were $49,724{\mu}mol/min$ and 6.08 mg/ml, respectively.

The effect of Glomus intraradices on the physiological properties of Panax ginseng and on rhizospheric microbial diversity

  • Tian, Lei;Shi, Shaohua;Ma, Lina;Zhou, Xue;Luo, Shasha;Zhang, Jianfeng;Lu, Baohui;Tian, Chunjie
    • Journal of Ginseng Research
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    • v.43 no.1
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    • pp.77-85
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    • 2019
  • Background: Glomus intraradices is a species of arbuscular mycorrhizal fungi that, as an obligate endomycorrhiza, can form mutually beneficial associations with plants. Panax ginseng is a popular traditional Chinese medicine; however, problems associated with ginseng planting, such as pesticide residues, reduce the ginseng quality. Methods: In this experiment, we studied the effect of inoculating G. intraradices on several physiological properties and microbial communities of ginseng. UV-Visible Spectrum method was used to detect physical properties. Denaturing gradient gel electrophoresis method was used to analyze microbial communities. Results: The results indicated that inoculation with G. intraradices can improve the colonization rate of lateral ginseng roots, increase the levels of monomeric and total ginsenosides, and improve root activity as well as polyphenol oxidase and catalase activities. We also studied the bacterial and fungal communities in ginseng rhizospheric soil. In our study, G. intraradices inoculation improved the abundance and Shannon diversity of bacteria, whereas fungi showed a reciprocal effect. Furthermore, we found that G. intraradices inoculation might increase some beneficial bacterial species and decreased pathogenic fungi in rhizospheric soil of ginseng. Conclusion: Our results showed that G. intraradices can benefit ginseng planting which may have some instructive and practical significance for planting ginseng in farmland.

Spectroscopic Analysis of Silica Nanoparticles Modified with Silane Coupling Agent (실란 커플링제에 의해 표면이 개질된 실리카 나노입자의 분광학적 분석)

  • Song, Seong-Kyu;Kim, Jung-Hye;Hwang, Ki-Seob;Ha, Ki-Ryong
    • Korean Chemical Engineering Research
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    • v.49 no.2
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    • pp.181-186
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    • 2011
  • In this study, we used 3-(trimethoxysilyl)propylmethacrylate(MPS) silane coupling agent for surface modification of silica nanoparticles. We studied effects of reaction conditions such as solvent pH, MPS hydrolysis time, reaction time, and molar ratio of MPS to Si-OH groups on silica nanoparticle surfaces, on the surface modification reactions of silica nanoparticles. Fourier Transform Infrared Spectroscopy(FTIR), Elemental Analysis(EA) and solid state crosspolarization magic angle spinning(CP/MAS) Nuclear Magnetic Resonance Spectroscopy(NMR) techniques were used to determine the type and the degree of surface modification. We found MPS reacts preferentially with Si-OH groups of the silica nanoparticles as monomeric form at solvent pH = 4.5. But increasing hydrolysis time of MPS from 30 mins to 90 mins, and molar ratio of MPS to Si-OH groups on silica nanoparticle surfaces, we found that MPS reacts preferentially with Si-OH groups of the silica nanoparticles as oligomeric form.