• Title/Summary/Keyword: catalytic acid - sites

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Cloning and Characterization of Filamentous Fungal S-Nitrosoglutathione Reductase from Aspergillus nidulans

  • Zhou, Yao;Zhou, Shengmin;Yu, Haijun;Li, Jingyi;Xia, Yang;Li, Baoyi;Wang, Xiaoli;Wang, Ping
    • Journal of Microbiology and Biotechnology
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    • v.26 no.5
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    • pp.928-937
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    • 2016
  • S-Nitrosoglutathione reductase (GSNOR) metabolizes S-nitrosoglutathione (GSNO) and has been shown to play important roles in regulating cellular signaling and formulating host defense by modulating intracellular nitric oxide levels. The enzyme has been found in bacterial, yeast, mushroom, plant, and mammalian cells. However, to date, there is still no evidence of its occurrence in filamentous fungi. In this study, we cloned and investigated a GSNOR-like enzyme from the filamentous fungus Aspergillus nidulans. The enzyme occurred in native form as a homodimer and exhibited low thermal stability. GSNO was an ideal substrate for the enzyme. The apparent Km and kcat values were 0.55 mM and 34,100 min-1, respectively. Substrate binding sites and catalytic center amino acid residues based on those from known GSNORs were conserved in this enzyme, and the corresponding roles were verified using site-directed mutagenesis. Therefore, we demonstrated the presence of GSNOR in a filamentous fungus for the first time.

Directed Evolution of Beta-galactosidase from Escherichia coli into Beta-glucuronidase

  • Xiong, Ai-Sheng;Peng, Ri-He;Zhuang, Jing;Liu, Jin-Ge;Xu, Fang;Cai, Bin;Guo, Zhao-Kui;Qiao, Yu-Shan;Chen, Jian-Min;Zhang, Zhen;Yao, Quan-Hong
    • BMB Reports
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    • v.40 no.3
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    • pp.419-425
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    • 2007
  • In vitro directed evolution through DNA shuffling is a powerful molecular tool for creation of new biological phenotypes. E. coli $\beta$-galactosidase and $\beta$-glucuronidase are widely used, and their biological function, catalytic mechanism, and molecular structures are well characterized. We applied an in vitro directed evolution strategy through DNA shuffling and obtained five mutants named YG6764, YG6768, YG6769, YG6770 and YG6771 after two rounds of DNA shuffling and screening, which exhibited more $\beta$-glucuronidase activity than wild-type $\beta$-galactosidase. These variants had mutations at fourteen nucleic acid sites, resulting in changes in ten amino acids: S193N, T266A, Q267R, V411A, D448G, G466A, L527I, M543I, Q626R and Q951R. We expressed and purified those mutant proteins. Compared to the wild-type protein, five mutant proteins exhibited high $\beta$-glucuronidase activity. The comparison of molecular models of the mutated and wildtype enzymes revealed the relationship between protein function and structural modification.

Isolation and characterization of a cDNA encoding a mammalian cathepsin L-like cysteine proteinase from Acanthmoeba healui

  • Hong, Yeon-Chul;Hwang, Mi-Yul;Yun, Ho-Cheol;Yu, Hak-Sun;Kong, Hyun-Hee;Yong, Tai-Soon;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.40 no.1
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    • pp.17-24
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    • 2002
  • We have cloned a cDNA encoding a cysteine proteinase of the Acanthamoeba healui OC-3A strain isolated from the brain of a granulomatous amoebic encephalitis patient. A DNA probe for an A. healui cDNA library screening was amplified by PCR using degenerate oligonucleotide primers designed on the basis of conserved amino acids franking the active sites of cysteine and asparagine residues that are conserved in the eukaryotic cysteine proteinases. Cysteine proteinase gene of A. healui (AhCPI) was composed of 330 amino acids with signal sequence, a proposed pro-domain and a predicted active site made up of the catalytic residues, $Cys^{25},{\;}His^{159},{\;}and{\;}Asn^{175}$. Deduced amino acid sequence analysis indicates that AhCPI belong to ERFNIN subfamily of C 1 peptidases. By Northern blot analysis. no direct correlation was observed between AhCPI mRNA expression and virulence of Acanthamoeba, but the gene was expressed at higher level in amoebae isolated from soil than amoeba from clinical samples. These findings raise the possibility that AhCPI protein may play a role in protein metabolism and digestion of phagocytosed bacteria or host tissue debris rather than in invasion of amoebae into host tissue.

Phase Cooperation between Mo-V-O and SnO2 in Selective Oxidation of Acrolein -II. Supported Catalysts- (아크롤레인 선택 산화반응에서 Mo-V-O와 SnO2의 상간협동 - II. 담지촉매 -)

  • Park, Dae-Won;Na, Suk-Eun;Kim, Kyung-Hoon;Lee, Won-Ho;Chung, Jong Shik
    • Applied Chemistry for Engineering
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    • v.5 no.2
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    • pp.295-304
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    • 1994
  • $Mo-V-O/SnO_2$(VM/Sn) and $SnO_2/Mo-V-O$(Sn/VM) catalysts have been prepared and characterized by XRD, BET, SEM and TPD of ammonia. The catalytic reaction of acrolein oxidation with these catalysts, in a continuous-flow fixed-bed reactor, showed that they had higher conversion of acrolein and higher yield of acrylic acid than those of Mo-V-O itself. The origin of the observed synergy studied by TPD, TPR and TPO is explained by the cooperation of $SnO_2$ and Mo-V-O at their interfaces where electrons flow from Mo-V-O phase to $SnO_2$ and $SnO_2$ produces spill-over oxygens, which, by being transported onto the surface of Mo-V-O, reoxidize the partially reduced active sites.

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Dehydration of D-xylose over SAPO Catalysts Synthesized with Various Structure Directing Agents (다양한 구조 유도제로 합성된 SAPO촉매를 이용한 자일로오스의 탈수화반응)

  • Kim, Saet Byul;You, Su Jin;Kim, Yong Tae;Chae, Ho-Jeong;Jeong, Soon-Yong;Park, Eun Duck
    • Korean Chemical Engineering Research
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    • v.48 no.6
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    • pp.684-689
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    • 2010
  • We synthesized a variety of SAPO catalysts with various structure directing agents by the hydrothermal method and applied them to the D-xylose dehydration. Single or mixtures of organic amines, viz. tetraethylammonium hydroxide(TEAOH), dipropylamine(DPA), diethylamine(DEA), morpholine and diethanolamine(DEtA) were used as structure directing agents. The $N_2$-isotherm, $NH_3$-temperature programmed desorption(TPD) and temperature programmed oxidation(TPO) were conducted to characterize SAPO catalysts. Among tested SAPO catalysts, the SAPO-34 synthesized with morpholine showed the highest furfural yield. The external surface area as well as the surface concentration of acid sites appeared to affect the catalytic activity for the dehydration of xylose into furfural.

Platinum-Catalyzed and Ion-Selective Polystyrene Fibrous Membrane by Electrospinning and In-Situ Metallization Techniques

  • Hong, Seung-Hee;Lee, Sun-Ae;Nam, Jae-Do;Lee, Young-Kwan;Kim, Tae-Sung;Won, Sung-Ho
    • Macromolecular Research
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    • v.16 no.3
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    • pp.204-211
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    • 2008
  • A platinum-catalyzed polyelectrolyte porous membrane was prepared by solid-state compression of electrospun polystyrene (PS) fibers and in-situ metallization of counter-balanced ionic metal sources on the polymer surface. Using this ion-exchange metal-polymer composite system, fiber entangled pores were formed in the interstitial space of the fibers, which were surrounded by sulfonic acid sites ($SO_3^-$) to give a cation-selective polyelectrolyte porous bed with an ion exchange capacity ($I_{EC}$) of 3.0 meq/g and an ionic conductivity of 0.09 S/cm. The Pt loading was estimated to be 16.32 wt% from the $SO_3^-$ ions on the surface of the sulfonated PS fibers, which interact with the cationic platinum complex, $Pt(NH_3)_4^{2+}$, at a ratio of 3:1 based on steric hindrance and the arrangement of interacting ions. This is in good agreement with the Pt loading of 15.82 wt% measured by inductively coupled plasma-optical emission spectroscopy (ICP-OES). The Pt-loaded sulfonated PS media showed an ionic conductivity of 0.32 S/cm. The in-situ metallized platinum provided a nano-sized and strongly-bound catalyst in robust porous media, which highlights its potential use in various electrochemical and catalytic systems.

Ozone Oxidation of PAHs in the Presence of Soil (I): Ozonation of Soil Slurry Phase (Ozone에 의한 PAHs 오염토양 복원 연구(I): 토양슬러리상 오존 산화)

  • Lim, Hyung-Nam;Kim, Ji-Yeon;Choi, Heechul
    • Journal of Korean Society of Environmental Engineers
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    • v.22 no.5
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    • pp.869-877
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    • 2000
  • A mechanism of ozonation of simulated soil slurry contaminated by phenanthrene and benzo[a]pyrene has been studied under various conditions. The effects of soil media(BS, S, GB), radical scavenger, ozone input ratio(0.17~0.73mg/min), bicarbonate ion, and humic acid were investigated, BS showed the highest removal efficiency in media tested. The generation of OH-radical via the catalytic decomposition of ozone on active sites of the natural sand was confirmed by OH-radical scavenger experiments. The enhanced removal efficiency by OH-radical was indirectly quantified to be about 22%. As initial concentration of humic acid(as sodium salt) was increased, pseudo first-order rate constant ($k_o$) of phenanthrene was decreased from $1.37{\times}10^{-2}s^{-1}$ to $0.53{\times}10^{-2}s^{-1}$. The amount of ozone required to oxidize 80% of the initial mass of phenanthrene(10mg/kg) and benzo[a]pyrene(10mg/kg) was 67.2mg/kg-soil and 48.0mg/kg-soil, respectively.

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Benzyl Alcohol Oxidation over H5PMo10V2O40 Catalyst Chemically Immobilized on Sulfur-containing Mesoporous Carbon (황이 포함된 중형기공성 탄소에 화학적으로 고정화된 H5PMo10V2O40 촉매 상에서 Benzyl Alcohol 산화반응)

  • Gim, Min Yeong;Kang, Tae Hun;Choi, Jung Ho;Song, In Kyu
    • Korean Chemical Engineering Research
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    • v.54 no.3
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    • pp.419-424
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    • 2016
  • $H_5PMo_{10}V_2O_{40}$ ($PMo_{10}V_2$) catalyst chemically immobilized on sulfur-containing mesoporous carbon (S-MC) was prepared, and it was applied to the benzyl alcohol oxidation reaction. S-MC was synthesized by a templating method using SBA-15 and p-toluenesulfonic acid as a templating agent and a carbon precursor, respectively. S-MC was then modified to have a positive charge, and thus, to provide sites for the immobilization of $PMo_{10}V_2$. By taking advantage of the overall negative charge of $[PMo_{10}V_2O4_{40}]^{5-}$, $PMo_{10}V_2$ catalyst was immobilized on the S-MC support as a charge matching component. It was revealed that $PMo_{10}V_2$ species were finely and molecularly dispersed on the S-MC via chemical immobilization. In the vapor-phase oxidation of benzyl alcohol, $PMo_{10}V_2$/S-MC catalyst showed higher conversion of benzyl alcohol and higher yield for benzaldehyde and benzoic acid than unsupported $PMo_{10}V_2$ catalyst. The enhanced catalytic performance of $PMo_{10}V_2$/S-MC was due to fine dispersion of $PMo_{10}V_2$ species on the S-MC via chemical immobilization.

Identification of SNPs in Highly Variable Lysozyme Gene in Korean Native Chicken Populations (한국 재래닭의 고변이 Lysozyme 유전자의 SNP 확인)

  • Hoque, M.R.;Kang, B.S.;Lim, H.K.;Choi, K.D.;Lee, J.H.
    • Korean Journal of Agricultural Science
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    • v.37 no.3
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    • pp.399-404
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    • 2010
  • Single nucleotide polymorphisms (SNPs) in chicken lysozyme (LYZ) gene were investigated in this study. The identification of SNPs in both exon and intron in LYZ gene has led to understanding of evolution for the domestic chicken populations. A total of 24 samples from two Korean native commercial chicken populations (CCPs) were used for the initial identification of SNPs by mixing three DNA samples for sequencing experiments. By comparing with red jungle fowl (RJF), two commercial chicken populations have 18 common polymorphisms. Between two commercial chicken populations, 15 polymorphisms were identified. Of the 33 polymorphisms identified, two indels (21 and 4 bp) were found. Whereas, only one polymorphism in exon 2 at the bp position 1426 was a non-synonymous substitution (p.Ala49Val), indicating the amino acid changes. The identified non-synonymous substitution (p.Ala49Val) is located close to the catalytic sites of the enzyme, which might affect its activity. In our investigation, the polymorphisms in LYZ gene can provide broad ideas for the variation of Korean native chicken populations from the ancestor of chicken breeds as well as the some biological functions of the LYZ gene.

Effect of SO2 on the Simultaneous Removal of Mercury and NOx over CuCl2-loaded V2O5-WO3/TiO2 SCR Catalysts (CuCl2가 담지된 V2O5-WO3/TiO2 SCR 촉매에 의한 수은 및 NOx 동시 제거에서 SO2의 영향)

  • Ham, Sung-Won
    • Clean Technology
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    • v.28 no.1
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    • pp.38-45
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    • 2022
  • CuCl2-loaded V2O5-WO3/TiO2 catalyst showed excellent activity in the catalytic oxidation of elemental mercury to oxidized mercury even under SCR condition in the presence of NH3, which is well known to significantly inhibit the oxidation activity of elemental mercury by HCl. Moreover, it was confirmed that, when SO2 was present in the reaction gas together with HCl, excellent elemental mercury oxidation activity was maintained even though CuCl2 supported on the catalyst surface was converted to CuSO4. This is thought to be because not only HCl but also the SO4 component generated on the catalyst surface promotes the oxidation of elemental mercury. However, in the presence of SO2, the total mercury balance before and after the catalytic reaction was not matched, especially as the concentration of SO2 increased. In order to understand the cause of this, further studies are needed to investigate the effect of SO2 in the SnCl2 aqueous solution employed for mercury species analysis and the effect of sulfate ions generated on elemental mercury oxidation. It was confirmed that SO2 also promotes NOx removal activity, which is thought to be because the increase in acid sites by SO4 generated on the catalyst surface by SO2 facilitates NH3 adsorption. The composition change and structure of the components present on the catalyst surface under various reaction conditions were measured by XRD and XRF. These measurement results were presented as a rational explanation for the results that SO2 enhances the oxidation activity of elemental mercury and the NOx removal activity in this catalyst system.