• Title/Summary/Keyword: Catechol.

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Construction of Genetically Engineered Microorganisms for Overexpression of xylE Gene Encoding Catechol 2,3-dioxygenase and the Functional Stability of the Recombinant Plasmid pSW3a Containing xylE in Aquatic Environment

  • Han, Hyo-Yung;Kim, Chi-Kyung;Park, Yong-Keun;Ka, Jong-Ok;Lee, Byeong-Jae;Min, Kyung-Hee
    • Journal of Microbiology
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    • v.34 no.4
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    • pp.341-348
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    • 1996
  • The regulation of xylE gene expression was examined by using vector promoter and construction of genetically engineered microorganisms (GEMs) for application in microcosm. When the xylE gene wsa subcloned into pBluscript SK(+) under the control of lac promoter (pTY1) in E. coli, and the expression was induced by IPTG, the enzyme activity of catechol 2, 3-dioxygenase was increased 4.7 times more than that of the crude extracts from transformants harboring pTY1. We suggest that the xylE gene has its own promoter at the upstream portion, because it was able to be expressed even in the absence of IPTG. A recombinant plasmid, pSW3a harboring the xylE gene under the T7 promotor, showed the activity of 14.5 units/mg protein, higher than that of parental strain, E. coli PYT1. The xylE gene in recombinant plasmid pSW3a was used as reporter gene for the application in microcosm ecosystem, since it was used for detection of xylE-positive clones by catechol spray on the agar plates. The pSW3a in E. coli was introduced into Pseudomonas patida to construct GEM strain, and examined for the exxpression and functional stability in microcosms.

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Role of Two Sets of RND-Type Multidrug Efflux Pump Transporter Genes, mexAB-oprM and mexEF-oprN, in Virulence of Pseudomonas syringae pv. tabaci 6605

  • Ichinose, Yuki;Nishimura, Takafumi;Harada, Minori;Kashiwagi, Ryota;Yamamoto, Mikihiro;Noutoshi, Yoshiteru;Toyoda, Kazuhiro;Taguchi, Fumiko;Takemoto, Daigo;Matsui, Hidenori
    • The Plant Pathology Journal
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    • v.36 no.2
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    • pp.148-156
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    • 2020
  • Pseudomonas syringae pv. tabaci 6605 has two multidrug resistance (MDR) efflux pump transporters, MexAB-OprM and MexEF-OprN. To understand the role of these MDR efflux pumps in virulence, we generated deletion mutants, ΔmexB, ΔmexF, and ΔmexBΔmexF, and investigated their sensitivity to plant-derived antimicrobial compounds, antibiotics, and virulence. Growth inhibition assays with KB soft agar plate showed that growth of the wild-type (WT) was inhibited by 5 μl of 1 M catechol and 1 M coumarin but not by other plant-derived potential antimicrobial compounds tested including phytoalexins. The sensitivity to these compounds tended to increase in ΔmexB and ΔmexBΔmexF mutants. The ΔmexBΔmexF mutant was also sensitive to 2 M acetovanillone. The mexAB-oprM was constitutively expressed, and activated in the ΔmexF and ΔmexBΔmexF mutant strains. The swarming and swimming motilities were impaired in ΔmexF and ΔmexBΔmexF mutants. The flood inoculation test indicated that bacterial populations in all mutant strains were significantly lower than that of WT, although all mutants and WT caused similar disease symptoms. These results indicate that MexAB-OprM extrudes plant-derived catechol, acetovanillone, or coumarin, and contributes to bacterial virulence. Furthermore, MexAB-OprM and MexEF-OprN complemented each other's functions to some extent.

Changes of Polyphenols and Polyphenol Oxidase Active Bands during Apple Wine Fermentation (사과주(酒) 발효과정중(醱酵過程中) Polyphenol성(性) 물질(物質) 및 Polyphenol Oxidase 활성(活性) Band의 변화(變化))

  • Chung, Ki-Taek;Seo, Seung-Kyo;Song, Hyung-Ik
    • Korean Journal of Food Science and Technology
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    • v.16 no.4
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    • pp.413-417
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    • 1984
  • To examine enzymatic browning of apple wine, changes of active bands of polyphenol oxidase (EC 1.10.3.1) as well as polyphenol substances related to browning of apple wine were investigated during wine brewing. The decrease of total phenol was remarkably inhibited by the addition of sodium metabisulfite. In the meantime, auto-oxidation of catechol in a model system increased proportionally as the reaction pH and temperature increased. Catechol oxidation, however, was not detected at $4^{\circ}C$ below pH 5.0. Polyacrylamidegel electrophoretic patterns showed that the apple (Jonathan) indicated 4 bands with polyphenol oxidase activity, designated a, b, c and d whose Rm were 0.21, 0.30, 0.41 and 0.51, respectively. Among these, 2 bands, a and c remained until 5th day fermentation and only c band after 6th day fermentation. After pasteurization of apple wine at $60^{\circ}C$ for 30min, c band also remained.

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Bioassay-Guided Isolation and Identification of Compounds from Arecae Pericarpium with Anti-inflammatory, Anti-oxidative, and Melanogenesis Inhibition Activities

  • Indriana, Amelia;Lee, Kyoung Jin;Kim, Yeong Shik
    • Natural Product Sciences
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    • v.22 no.3
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    • pp.193-200
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    • 2016
  • This study describes the anti-inflammatory, anti-oxidant, and melanogenesis inhibition activities of methanol extract and various organic solvent fractions of Arecae Pericarpium. We examined the inhibition of lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 cells, 1,1-diphenyl-2-picrylhydrazine (DPPH) scavenging activity, mushroom tyrosinase inhibition activity and melanin contents. The study showed that, among all tested fractions, methylene chloride fraction showed the strongest inhibition of LPS-induced NO production in RAW 264.7 cells ($IC_{50}$ value $8.89{\mu}g/mL$) and DPPH radical scavenging activity ($EC_{50}$ value $21.39{\mu}g/mL$). Methylene chloride and ethyl acetate fractions similarly inhibited mushroom tyrosinase activity. Methanol extract exhibited strongest reduction of melanin content in B16F10 melanoma cells. Based on the bioactivity assay results, methylene chloride and ethyl acetate fractions were further separated. Eight phenolic compounds were isolated, which are dimeric syringol (1), catechol (2), 4-hydroxybenzaldehyde (3), vanillin (4), 4-hydroxyacetophenone (5), apocynin (6), protocatechuic acid (7) and 4-hydroxybenzoic acid (8). Among the isolated compounds tested, catechol showed the strongest inhibition of LPS-induced NO production in RAW 264.7 cells. Catechol also showed the concentration-dependent NF-${\kappa}B$ inhibition activity. Arecae Pericarpium might have potentials to be developed as anti-inflammatory agent or dermatological product for skin-whitening agent.

Purification and some properties of polyphenol oxidase from Spuriopimpinella bracycarpa (참나물로부터 추출한 polyphenol oxidase의 부분정제 및 성질)

  • Ham, Seung-Shi;Hong, Eun-Hee;Lee, Sang-Young;Park, Gwi-Gun;Omura, Hirohisa
    • Applied Biological Chemistry
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    • v.34 no.1
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    • pp.49-53
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    • 1991
  • Three polyphenol oxidase(polyphenol oxidase I, II and III ) were isolated from the crude extract of a Spuriopimpinella bracycarpa by $(NH_4)_2SO_4$ precipitation and subsequent Sephadex G-150 chromatography. The final preparation thus obtained showed three peaks of enzyme activity. Optimum pH and temperature for the activity of polyphenol oxidase were 7.5 and $30^{\circ}C$, respectively. The enzyme was completely inactivated when i4 was treated at$70^{\circ}C$ for 30min and at $80^{\circ}C$ for 5min at pH 6.5. The enzyme was partially inactivated by ascorbic acid, glutathione and potassium cyanide (0.1mM), and was completely inhibited by L-cysteine, ascorbic acid, glutathione and potassium cyanide(0.5 and 1.0mM). The enzyme has good activity on catechol and 3,4-dihydroxytoluene but was strongly inactivated on pyrogallol, dopamine and DL-dopa. The Michaelis cons4ant of the enzyme was 86.5mM with catechol as a substrate.

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Action Mechanism of LB10522, a New Catechol-Substituted Cephalosporin (카테콜 치환체를 가진 세파로스포린계 항생제 LB10522의 작용기전)

  • Kim, Mu-Yong;Oh, Jeong-In;Paek, Kyoung-Sook;Kim, In-Chull;Kwak, Jin-Hwan
    • YAKHAK HOEJI
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    • v.40 no.1
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    • pp.102-111
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    • 1996
  • LB10522 is a new parenteral broad spectrum cephalosporin with a catechol moiety at C-7 position of beta-lactam ring. This compound can utilize tonB-dependent iron transp ort system in addition to porin proteins to enter bacterial periplasmic space and access to penicillin-binding proteins (PBPs) which are the lethal targets of ${\beta}$-lactam antibiotics. The chelating activity of LB10522 to metal iron was measured by spectrophotometrically scanning the absorbance from 200 to 900nm. When $FeCl_3$ was added, optical density was increased between 450 and 800nm. LB10522 was more active against gram-negative strains in iron-depleted media than in iron-replete media. This is due to the increased expression of iron transport channels in iron-depleted condition. LB10522 showed a similar activity against E. coli DC2 (permeability mutant) and E. coli DCO (wild type strain) in both iron-depleted and iron-replete media, indicating a minimal permeaility barrier for LB10522 uptake. LB10522 had high affinities to PBP 3 and PBP 1A, 1B of E. coli. By blocking these proteins, LB10522 caused inhibition of cell division and the eventual death of cells. This result was correlated well with the morphological changes in E. coli exposed to LB10522. Although the in vitro MIC of LB10522 against P. aeruginosa 1912E mutant (tonB) was 8-times higher than that of the P. aeruginosa 1912E parent strain, LB10522 showed a similar in vivo protection efficacy against both strains in the mouse systemic infection model. This result suggested that tonB mutant, which requires a high level of iron for normal growth, might have a difficulty in surviving in their host with an iron-limited environment.

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A New Functional Model of Catechol Dioxygenases: Properties and Reactivity of [Fe(BLPA)DBC]$BPh_4$

  • Lim, Ji H.;Lee, Ho J.;이강봉;Jang, Ho G.
    • Bulletin of the Korean Chemical Society
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    • v.18 no.11
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    • pp.1166-1172
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    • 1997
  • [FeⅢ(BLPA)DBC]BPh4, a new functional model for the catechol dioxygenases, has been synthesized, where BLPA is bis((6-methyl-2-pyridyl)methyl)(2-pyridylmethyl)amine and DBC is 3,5-di-tert-butylcatecholate dianion. The BLPA complex has a structural feature that iron center has a six-coordinate geometry with N4O2 donor set. It exhibits EPR signals at g=5.5 and 8.0 which are typical values for the high-spin FeⅢ (S=5/2) complex with axial symmetry. The BLPA complex reacts with O2 within a few hours to afford intradiol cleavage (75%) and extradiol cleavage (15%) products which is very unique result of all [Fe(L)DBC] complexes studied. The iron-catecholate interaction of BLPA complex is significantly stronger, resulting in the enhanced covalency of the metal-catecholate bonds and low energy catecholate to FeⅢ charge transfer bands at 583 and 962 nm in CH3CN. The enhanced covalency is also reflected by the isotropic shifts exhibited by the DBC protons, which indicate increased semiquinone character. The greater semiquinone character in the BLPA complex correlates well with its high reactivity towards O2. Kinetic studies of the reaction of the BLPA complex with 1 atm O2 in CH3OH and CH2Cl2 under pseudo-first order conditions show that the BLPA complex reacts with O2 much slower than the TPA complex, where TPA is tris(2-pyridylmethyl)amine. It is presumably due to the steric effect of the methyl substituent on the pyridine ring. Nevertheless, both the high specificity and the fast kinetics can be rationalized on the basis of its low energy catecholate to FeⅢ charge transfer bands and large isotropic NMR shifts for the BLPA protons. These results provide insight into the nature of the oxygenation mechanism of the catechol dioxygenases.

The relationship between catechol-O-methyltransferase gene polymorphism and coronary artery abnormality in Kawasaki disease (가와사키병의 관상동맥 이상과 catechol-O-methyltransferase 유전자의 단일염기다형성)

  • Lee, Hyo Jin;Lee, Myung Sook;Kim, Ji Sook;Kim, Eun Ryoung;Kang, Sung Wook;Kim, Soo Kang;Chung, Joo Ho;Yoon, Kyung Lim;Han, Mi Young;Cha, Seong Ho
    • Clinical and Experimental Pediatrics
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    • v.52 no.1
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    • pp.87-92
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    • 2009
  • Purpose : Many gene polymorphisms are associated with coronary artery abnormalities in Kawasaki disease. Catechol-O- methyltransferase (COMT) plays an important role in the metabolism of catecholamines, catechol estrogen, and catechol drugs. Polymorphisms of the COMT gene are reported to be associated with myocardial infarction and coronary artery abnormalities. The aim of this study was to evaluate the relationship between COMT gene polymorphisms and coronary artery abnormalities in Kawasaki disease patients. Methods : One hundred and one Korean children with Kawasaki disease and 306 healthy Korean control subjects were enrolled in this study. The polymorphisms of the COMT gene were analyzed by direct sequencing. Results : There were no differences in the genotype and allelic frequency of the rs4680 and rs769224 polymorphic sites between Kawasaki disease and control subjects. Further, no significant difference was found in the rs4680 polymorphism between patients with coronary artery abnormalities and patients without coronary artery abnormalities (codominant P=0.32, dominant P=0.74, recessive P=0.13). However, the distribution of the rs769224 polymorphism was significantly different between patients with coronary artery abnormalities and patients without coronary artery abnormalities (codominant P= 0.0077, dominant P=0.0021, recessive P=0.16). Conclusion : Our results indicate that the polymorphisms of the rs769224 gene might be related to the development of coronary artery abnormalities in Kawasaki disease.

Volatile Substances and Physicochemical Characteristics of Pyroligneous Liquor (목초액의 휘발성 성분과 이화학적 특성)

  • Kim Jong-Soo;Park Seung-Woo;Choi Jung-Hwan;Lee Eun-Young;Lee Sang-Han;Chung Shin-Kyo
    • Food Science and Preservation
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    • v.12 no.6
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    • pp.656-661
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    • 2005
  • To develop food additives from pyroligneous liquor, we first examined the physicochemical characteristics of volatile substances. The pH, gravity, acidity, solution tar, and scorched remains in the pyroligneous liquor were 2.50, 1.020, $0.8\%$ $2.31\%$ and $0.008\%$ respectively. The main compounds of pyroligneous liquor were 2,6-dimethoxyphenol (syringol), 2-hydroxy-2-cyclopenten-l-one, 1,2-benzenediol (catechol), phenol, 2 (5H)-furanone, 2-methoxyphenol (guaiacol), 5-hydroxymethyl-2 furancarboxaldehyde were found in organic phase, whereas catechol, 1,4 : 3,6-dianhydro-${\alpha}d$-glucopyranose, cyclopropyl carbinol, 5-hydroxymethyl-2 furancarboxaldehyde in aqueous phase.

Desmutagenicity of the Enzymatic Browning Reaction Products Which Obtained from Prunus salicina (yellow) Enzyme and Polyphenol Compounds (재래종 황색자두효소 갈변반응 생성물의 돌연변이 억제작용)

  • Ham, Seung-Shi
    • Applied Biological Chemistry
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    • v.30 no.1
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    • pp.71-76
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    • 1987
  • The mutagenicity and desmutagenicity on enzymatic browning reaction products which obtained from prunes salicina (yellow) enzyme and polyphenol compounds were carried out. In the rec-assay on Bacillus subtilis strains H17 and M45, the enzymatic browning reaction products of pyrogallol, hydroxyhydroquinone, 3,4-dihydroxytoluene and catechol of $10^{-2}M$ did not showed mutagenicity. In the effects of various metal ions on the rec-assay, the enzymatic browning reaction products of pyrogallol showed mutagenic activity by $Fe^{3+},\;Mn^{2+},\;Zn^{2+},\;Ni^{2+}$ and $Al^{3+}$. In the enzymatic browning reaction products of hydroxyhydroquinone, $Cu^{2+},\;Mn^{2+}$ and $Pb^{2+}$ were effected in mutagenic action and the enzymatic browning reaction products of catechol was effected in mutagenic action by $Mn^{2+}$. In the DNA-breaking action of enzymatic browning reaction products of pyrogallol, hydroxyhydroquinone, 3,4-dihyroxytoluene and catechol did not show, DNA-breaking action. In the effects of various metal ions on the DNA-breaking action of enzymatic browning reaction products, $Cu^{2+}$ showed DNA-breaking action. In the mutagenicity test on Sal. typhimurium strains TA98 and TA 100 with S-9 mix, 4 kinds of browned substances did sot shove muragenicity, all the browned substances showed strong desmutagenic activity in the presence of benzo $({\alpha})-pyrene$ with S-9 mix.

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