• Title/Summary/Keyword: horseradish peroxidase

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Electrochemical Properties of Tobacco Peroxidase Incorporated Enzyme Electrode Bound with CSM Rubber (CSM 고무로 결합된 담배 과산화효소 고정 효소전극의 전기화학적 특성)

  • Yoon, Kil Joong
    • Applied Chemistry for Engineering
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    • v.25 no.5
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    • pp.538-543
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    • 2014
  • In order to substitute for the marketed horseradish peroxidase, a hydrogen peroxide sensor embedded with tobacco leaf in carbon pastes was constructed and its sensing ability was electrochemically evaluated. Ten and more electrode parameters obtained implied that the enzyme electrode exerts its remarkable specificity quantitatively in the experimental range of potential. Especially the small symmetry factor (${\alpha}$, 0.21) showed that the electrode kinetics is very sensitive to the change of electrode potential. The experimental facts above suggested that our enzyme electrode functions as a hydrogen peroxide sensor normally and tobacco peroxidase can be used in the place of the marketed one as an alternative to marketed ones.

Ultrastructural Btudy on the Degranulation of Rat Peritoneal Mast Cells Induced by Horseradish Peroxidase (Horseradish Peroxidase에 의해 유도(誘導)된 백서(白鼠) 복강비만세포(腹腔肥滿細胞)의 탈과립(脫顆粒)에 관(關)한 미세구조적(微細構造的) 연구(硏究))

  • Cho, Byung-Pil;Kang, Ho-Suck;Kim, Woo-Kap
    • Applied Microscopy
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    • v.17 no.1
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    • pp.98-114
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    • 1987
  • Degranulation of the rat peritoneal mast cell induced by intraperitoneal injection of horseradish peroxidase(HRP) was studied using light and electron microscopes. 1. Rat peritoneal mast cells in the Tyrode's buffered salt solution injected control group did not show any particular morphological changes following the specified time course. 2. Under the light microscope, the majority of mast cells observed 10 minutes after HRP injection were nearly the same as those of the control group. However, after 30 minutes, granule densities or staining properties of certain cells began to decrease and these appearances increased gradually until 12 hours after injection, at which time small groups of granules being stained pale-red or pink with toluidine blue were easily identified in the cytoplasm of many cells, and numerous extruded granuleg were scattered around these cells. 3. In the mast cells representing the early stage of degranulation induced by HRP, the electron densities of certain granules decreased as the size enlarged, and perigranular cavities were formed by perigranular membrane expansion. As a result, a thin cytoplasmic septum was formed between the expanded perigranular membrane and the cytoplasmic membrane in the cell periphery, and fusion of the adjacent perigranular membranes was observed in the inner side of the cell. 4. In some mast cells, one or two changes in the peripheral cytoplasmic septum could be seen. One was a focal rupture of the peripheral septum and the other was the formation of a saccule containing one or more vesicles. This saccule was thought to be used for granule-extrusion site and/or material absorptive apparatus judging from the morphological characteristics. 5. As the degranulation proceeded, the granule was extruded from the cell after partial rupture of the peripheral cytoplasmic septum. This phenomenon proceeded to-ward the inner side of the cell through the fused perigranular cavities, and consequently several distinct cavities containing a few unextruded membrane-free granules were formed throughout the cytoplasm after 12 hours. As a rule, the granule-extrusion sites were relatively fewer while the cytoplasmic cavities resulting from degranulation were more numerously observed. Thus, it was thought that the granule-extrusion sites tended to be restricted in the HRP-induced degranulation.

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A Study on the Detection Characteristics in Glucose and Fabrication of Bi-Enzyme Electrode using Electrochemical Method (전기화학적 방법을 이용한 다중 효소 전극 제작 및 글루코스 검출 특성에 관한 연구)

  • Han, Kyoung Ho;Shin, In Seong;Yoon, Do-Young
    • Journal of the Korean Electrochemical Society
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    • v.23 no.3
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    • pp.66-72
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    • 2020
  • In this study, the development of biosensors capable of bi-enzyme reactions by including Horseradish peroxidase and glucose oxidase was carried out for detection of glucose. The sensors were manufactured using electro deposition method to reduce production time, and screen printed electrodes (SPE) were used to produce economical sensors. To check the bienzyme effect, the sensor was compared and analyzed with single enzyme biosensor. The characteristics of the sensor were evaluated using scanning electron microscopy(SEM), cyclic voltammetry(CV), electrochemical impedance spectroscopy(EIS), chronoamperometry(CA), and flow injection analysis(FIA). Analysis results from SEM, CV and EIS confirmed that the enzymes are well fixed to the electrode surface. In addition, it was confirmed that bi-enzyme biosensors manufactured from the CA method improved signal performance by 200% compared to single enzyme biosensors. From this results, we were able to explain that HRP and GOD react catalyzed to each other. And the results of FIA showed that the intensity of each current signal was constant when the same concentration of glucose was injected four times. In addition, by analyzing the intensity of current signals for glucose concentrations, the biosensors manufactured in this study showed excellent trends in signal sensitivity, reproducibility and stability.

Development of electrochemical biosensor for determination of galactose (4갈락토오즈 측정을 위한 전기화학적 바이오센서 개발)

  • Park, Kap Soo;Cho, Soon Sam;Quan, De;Lee, Jae Seon;Cha, Geun Sig;Nam, Hakhyun
    • Analytical Science and Technology
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    • v.20 no.5
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    • pp.393-399
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    • 2007
  • In principle, the blood galactose level may be determined conveniently with a strip-type biosensor similar to that for glucose. In this study, we describe the development of a disposable galactose biosensor strip for point-of-care testing. The sensor strip is constructed with screen-printed carbon paste electrode (SPCE) and sample amount (< $100{\mu}L$). The developed strip the galactose level in less than 90 s using bienzymatic system of galactose oxidase (GAO) and horseradish peroxidase (HRP). The effects of pH, mediator (1,1-ferrocenedimethanol) concentration, ratio of enzymes, and applied potential were determined preliminarily with glassy carbon electrodes, and optimized further with the strip-type electrodes. The sensor exhibits linear response in the range of $0{\sim}400{\mu}M$ ($r^2$ = 0.997, S/N = 3). Since a low working potential, in principle, the fabricated disposable galactose biosensor has -100 mV (vs. Ag/AgCl), it is applied for the detection of galactose, interfering responses from common interferents such as ascorbic acid, uric acid and acetaminophen could be minimized. The sensor has been used to determine the total galactose level in standard samples with satisfactory reproducibility (CV = 5 %).

Oxidative N-Debenzylation of N-Benzyl-N-substituted Benzylamines Catalyzed by Cytochrome P450

  • Kim, Sung-Soo;Lin, Gang;Yang, Ji-Won
    • Bulletin of the Korean Chemical Society
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    • v.25 no.2
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    • pp.249-252
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    • 2004
  • Cytochrome P450 (P450)/$O_2$/NADPH engender electron transfer reaction of N-benzyl-N-substituted benzylamines to yield corresponding radical cation 1 that is simultaneously converted into 2 and 3. Subsequently, expulsion of proton and hydroxylation yielding a-hydroxylamines are followed by formation of benzaldehydes and benzylamines.

Enzymatic Dyeing for Wool (효소를 이용한 wool의 염색)

  • Hoon Sik Shin;Byong Dae Jeon;Artur Cavaco-Paulo;Mitsuo Ueda
    • Proceedings of the Korean Fiber Society Conference
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    • 2001.10a
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    • pp.59-62
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    • 2001
  • Oxydoreductase enzymes such as laccases (benzenediol: oxygen oxidoreductase, EC 1.10.3.2) and horseradish peroxidase (donor: hydrogen peroxide oxidoreductase, HRP, EC 1.11.1.7) can provide novel ways for wool coloration in the face of actual state of the art of these enzymes. HRP has been reported as a very useful enzyme for the synthesis of phenolic polymers2). (omitted)

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Synthesis of Enzyme-Containing PEG Hydrogel Nanospheres for Optical Biosensors (광바이오센서용 효소를 함유한 PEG 수화젤 나노입자의 합성)

  • Kim, Bum-Sang
    • Polymer(Korea)
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    • v.29 no.6
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    • pp.613-616
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    • 2005
  • In this word as the first step to develop optical biosensors for a single cell level analysis, the preparation method of nano-scale polymer hydrogel spheres containing an enzyme was set up and the feasibility of the spheres as optical biosensors was investigated. The horseradish peroxidase (HRP) was encapsulated in the PEG hydrogel spheres by suspension photopolymerization, yielding spheres of the average size of 305 nm. After the polymerization, the incorporation and activity of HRP within the spheres were determined by the production of fluorescence resulted from the enzymatic reaction between HRP and $\H_{2}O_{2}$. The fluorescence emission response of the HRP-loaded PEG hydrogel spheres increased by nearly 300$\%$ as hydrogen peroxide concentration was changed from 0 to 11 nM in the presence of Amplex Red. The results suggest that the method to prepare the PEG hydrogel nanospheres containing an enzyme could be used for developing optical biosensors to measure various analytes in the very small samples like a single cell.

The Electrochemical Studies of Two Osmium Redox Polymer Films and Their Application for Multi-Detecting Biosensor (전기화학적인 방법을 이용한 두 개의 오스뮴 고분자 막의 고정화 및 다중 검출 바이오센서에 관한 연구)

  • Tae, Gun-Sik;Kim, Jin-Gu;Choi, Young-Bong;Kim, Hyug-Han
    • Journal of the Korean Electrochemical Society
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    • v.11 no.3
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    • pp.170-175
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    • 2008
  • Screen printed carbon electrodes (SPEs) modified with co-immobilized osmium-based redox polymers can be used to apply multi-detecting biosensors. In this study, we report our initial studies of multi-detecting biosensor concepts using two osmium-based redox polymers for horseradish peroxidase-mediated reduction of ${H_2}{O_2}$ coupled to glucose oxidase-mediated oxidation of glucose. We target to synthesize two osmium redox polymers of potentials use, a chloride-containing redox polymer ($E^{O'}$ + 0.520 vs. Ag/AgCl) and a methoxy-containing redox polymer $E^{O'}$ + 0.150 vs. Ag/AgCl). The former show good catalytic electrical signals with horseradish peroxidase and the latter's redox polymer is to be an effective redox mediator of glucose oxidation by glucose oxidase.

Isolation and Thermal Inactivation of Horseradish Peroxidase Isozymes (서양고추냉이에 있는 페르옥시다아제 이소짐의 분리(分離)와 열불활성화(熱不活性化))

  • Yoon, Jung-Ro;Park, Kwan-Hwa
    • Korean Journal of Food Science and Technology
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    • v.14 no.2
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    • pp.125-129
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    • 1982
  • Four peroxidase isozymes from horseradish roots (isozymes A, B, C and D) were isolated by chromatography and were thermally inactivated at $70{\sim}97^{\circ}C$ and pH 7.0. The four isozymes had different inactivation rates and the inactivation of each isozymes did not follow first order kinetics. D values of isozymes A, B, C, D and crude enzyme were 594s, 1850s, 2050s, 78s, 130s and z values were $24.0^{\circ}C$, $12.5^{\circ}C$, $18.0^{\circ}C$, $23.7^{\circ}C$ and $24.0^{\circ}C$, respectively. Sephadex gel chromatogram of the thermally treated isozyme C indicated that the shape and molecular weight of the native isozyme changed during inactivation.

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Phenol Removal Using Horseradish Peroxidase(HRP)-Mediated Polymerization Reaction in Saturated Porous Media (다공성 포화 매질에서 효소 중합반응을 이용한 페놀 제거)

  • Kim, Won-Gee;Lee, Seung-Mok
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.10
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    • pp.984-991
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    • 2008
  • This paper reports experimental results, demonstrating the feasibility of horseradish peroxidase(HRP) and H$_2$O$_2$ to reduce phenol transport in saturated porous media. A laboratory-scale packed column reactor(ID: 4.1 cm, sand-bed height 12 cm) column was utilized to simulate injection of HRP and H$_2$O$_2$ into an aquifer contaminated with phenol. Effluent concentrations of phenol and polymerization products were monitored before and after enzyme addition under various experimental conditions(enzyme dose: 0$\sim$2 AU/mL, [ionic strength]: 5$\sim$100 mM, pH: 5$\sim$9). The concentration of phenol in the column effluent was found to decrease by nearly 90% in the presence of HRP(2 AU/mL) and H$_2$O$_2$ in the continuous flow system at pH 7 and ionic strength 20 mM. The influent phenol was converted in the system to insoluble precipitate, which deposited in pore spaces. The remains were discharged as soluble oligomers. About 8% of total pore volume in column system was decreased by deposition of polymer produced.