• 제목/요약/키워드: Enzymatic characterization

검색결과 294건 처리시간 0.03초

High sensitivity biosensor for mycotoxin detection based on conducting polymer supported electrochemically polymerized biopolymers

  • Dhayal, Marshal;Park, Gye-Choon;Park, Kyung-Hee;Gu, Hal-Bon
    • 한국신재생에너지학회:학술대회논문집
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    • 한국신재생에너지학회 2010년도 춘계학술대회 초록집
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    • pp.243.1-243.1
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    • 2010
  • Devices based on nanomaterials platforms are emerging as a powerful tool for ultrasensitive sensors for the direct detection of biological and chemical species. In this talk, we will report the preparation and the full characterization of electrochemical polymerization of biopolymers platforms and nano-structure formation for electrochemical detection of enzymatic activity and toxic compound in electrolyte for biosensor applications. Formation of an electroactive polymer film of two different compounds has been quantified by observing new redox peak at higher potentials in cyclic voltammogram measurements. RCT value of at various biopolymer concentration based hybrid films has been obtained from electrochemical impedance spectroscopy analysis and possible mechanism for formation of complexes during electrochemical polymerization on conducting substrates has been investigated. Biosensors developed based on these hybrid biopolymers have very high sensitivity.

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Bacillus stearothermophilus KJ16이 생산하는 Cyclodextrin Glucanotransferase 의 정제와 효소특성 (Purification and Characterization of Cyclodextrin Glucanotransferase from Bacillus stearothermophilus KJ16)

  • 권현주;남수완;김광현;송승구;윤종원;김병우
    • 생명과학회지
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    • 제8권3호
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    • pp.326-332
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    • 1998
  • Cyclodextrin glucanotransferase from B. stearothermophilus KJ16 that can produce both cyclodextrin glucanotransferase and cyclodextrinase was purified by ammonium sulfate precipitation, DEAE-cellulose chromatography, Sephadex G-100 chromatography, and FPLC. The molecular weight of the purifice enzyme was about 65,000 dalton by SDS-PAGE. The optimal pH and temperature were 6.0 and $60^{\circ}C$, respectively. The enzyme was stable at $50^{\circ}C$ for 1 hr and in the pH range of 5.5 and 8.5. Mercaptoethanol and dithiothreitol inhibited the enzyme activity strongly. The enzyme produced 60% cyclodextrin(CD) from 5% soluble starch with the $^{\alpha}$, $^{\beta}$, $^{\gamma}$-CD ratio of 42:46:12. Amylopectin was the most suitable substrate with 67% conversion to CD.

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Characterization of Linear Polymer-Dendrimer Block Copolymer/Plasmid DNA Complexes: Formation of Core-shell Type Nanoparticles with DNA and Application to Gene Delivery in Vitro

  • Choi, Joon-Sig;Choi, Young-Hun;Park, Jong-Sang
    • Bulletin of the Korean Chemical Society
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    • 제25권7호
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    • pp.1025-1030
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    • 2004
  • A hybrid linear polymer-dendrimer block copolymer, poly(ethylene glycol)-block-poly(L-lysine) dendrimer, was synthesized and introduced to form polyionic complexes with DNA. The copolymer formed core-shell type nanoparticles with plasmid DNA. From dynamic light scattering experiments, the mean diameter of the polyplexes was observed to be 154.4 nm. The complex showed much increased water solubility compared to poly(L-lysine). The plasmid DNA in polyplexes was efficiently protected from the enzymatic digestion of DNase I. The cytotoxicity and transfection efficiency for 293 cells was measured in comparison with poly(Llysine).

A Cu, Zn superoxide dismutase (SOD1) from Cordyceps militaris: cDNA cloning, expression and characterization

  • Park, Nam-Sook;Lee, Sang-Mong;Sohn, Hung-Dae;Jin, Byung-Rae
    • 한국잠사학회:학술대회논문집
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    • 한국잠사학회 2003년도 International Symposium of Silkworm/Insect Biotechnology and Annual Meeting of Korea Society of Sericultural Science
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    • pp.66-70
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    • 2003
  • The first line of antioxidant defense against reactive oxygen species includes the enzymatic activity of the superoxide dismutase (SOD) that catalyzes the disproportionation of superoxide to hydrogen peroxide and water. The SOD mainly removes highly toxic $O_2$$^{[-10]}$ and also prevents $O_2$$^{[-10]}$ mediated reduction of iron and subsequent OH$^{[-10]}$ generation. Along with an interest in SOD as a first line of defense against damage mediated by the superoxide anion, the SOD1 enzyme has been subjected to investigation in the molecular and cellular level. (omitted)

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Acetobater sp.KM Alcohol Dehydrogenase의 분리 및 특성 (Purification and Characterization of Alcohol Dehydrogenase from Acetobacter sp. KM)

  • 전홍성;차영주
    • KSBB Journal
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    • 제10권1호
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    • pp.30-37
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    • 1995
  • Membrane-bound alcohol dehydrogenase(ADH) was purified to homogeneity from the acetic acid producing bacteria, Acetobacter sp. KM. The enzyme was solubilized and extracted with Triton X-100 and purified using the Mono-Q ion exchange chromatography and Superose 12 gel filtration chromatography. The enzyme was purified to 12-fold with a yield of 30%. The molecular weight of the purified enzyme was to be 335 KDa. SDS-PAGE of the enzyme showed two subunits with molecular weights of 79 KDa and 49 KDa. It indicated that the enzyme consisted of three subunits of the 79 KDa and two subunits of the 49 KDa. The purified .ADH preferentially oxidized straight chain aliphatic alcohol except methanol. Formaldehyde, acetaldehyde and glutaraldehyde were also oxidized. The apparent Km for ethanol was 1.04 mM and the optimum pH and temperature were 5.0∼6.0 and 32$^{\circ}C$, respectively. V2O5 and divalent cation such as ZnCl2 and NiCl2 inhibited enzymatic activity.

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Purification and Biochemical Properties of Glutathione S-Transferase from Lactuca sativa

  • Park, Hee-Joong;Cho, Hyun-Young;Kong, Kwang-Hoon
    • BMB Reports
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    • 제38권2호
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    • pp.232-237
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    • 2005
  • A glutathione S-transferase (GST) from Lactuca sativa was purified to electrophoretic homogeneity approximately 403-fold with a 9.6% activity yield by DEAE-Sephacel and glutathione (GSH)-Sepharose column chromatography. The molecular weight of the enzyme was determined to be approximately 23,000 by SDS-polyacrylamide gel electrophoresis and 48,000 by gel chromatography, indicating a homodimeric structure. The activity of the enzyme was significantly inhibited by S-hexylGSH and S-(2,4-dinitrophenyl) glutathione. The enzyme displayed activity towards 1-chloro-2,4-dinitrobenzene, a general GST substrate and high activities towards ethacrynic acid. It also exhibited glutathione peroxidase activity toward cumene hydroperoxide.

D-Amino Acid Aminotransferase 활성보유 고온성미생물의 탐색 및 분류학적 특성 연구 (Screening and Taxonomic Charactrization of D-Amino Acid Aminotransferase-producing Thermophiles)

  • 곽미선;이승구;정상철;서승현;이재흥;전영중;김영호;성문희
    • 한국미생물·생명공학회지
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    • 제27권3호
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    • pp.184-190
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    • 1999
  • To acquire an industrially useful biocatalyst for the enzymatic synthesis and production of various D-amino acid aminotransferase (D-AAT) activity. The enzyme activity was found from 110 strains of isolated thermophiles revealing its wide occurrence in thermophiles. Enzyme activity and thermal stability of the D-AAT producers were compared. Finally we have selected four thermophiles as producers of potent biocatalysts for the D-amino acid production; two thermophiles, Bacillus sp. Lk-1 and LK-2, having higher specific activity and two thermophiles, B. stearothermophilus KL-01 and Bacillus sp. KLS-01, having higher thermal stability than the D-AAT producers. Taxonomic and physiological characteristics of the four isolated thermophiles were described herein.

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Identification and Characterization of Rodent Germ Cells-Specific Hyaluronidases

  • Kim, Ekyune;Chang, Kyu-Tae
    • Reproductive and Developmental Biology
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    • 제36권3호
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    • pp.155-161
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    • 2012
  • Germ cell-specific hyaluronidases such as sperm adhesion molecule 1 (SPAM1) and hyaluronoglucosaminidase 5 (Hyal5) are in part responsible for dispersal of the cumulus cell mass, which is a critical step in establishing fertilization in mammals. In this study, we identified two testis-hyaluronidases, SPAM1 and Hyal5, in hamster and rat. These two genes were expressed specifically in the testis. At the protein level, hamster SPAM1 and Hyal5 display 78.7% and 75.4% identity with mouse SPAM1 and Hyal5. Further, the activity of the enzymes with respect to cumulus cell dispersion did not differ, although we observed that the enzymatic activity differed in pH range. These studies suggest that different sperm hyaluronidases are capable of dispersing the cumulus cell mass despite differences in enzyme activity.

Analytical Characterization of Aza-Indole Alkaloids in the Biosynthesis of Catharanthus Roseus

  • Lee, Hyang-Yeol
    • 한국응용과학기술학회지
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    • 제29권2호
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    • pp.248-256
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    • 2012
  • Aza-indoles are important pharmacophores that have similar size and biological properties of indole. Here we have synthesized 4- and 7-azaindole tryptamines and showed that they are successfully incorporated in the biosynthesis of monoterepene indole alkaloids (MIAs) to form novel azaindole alkaloids by enzymatic reactions of strictosidine synthase(STR) and strictosidine glucosidase(SDG) monitored by UPLC/MS. By using HPLC equipped with a HPLC photo diode array(PDA) detector, each of the UV spectra of azaindole alkaloids was obtained and characterized. When hydrophilicity of azaindole alkaloids was compared, 4-azaindole alkaloids were more hydrophilic than 7-azaindole alkaloids.