• Title/Summary/Keyword: Horseradish Peroxidase(HRP)

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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.

Peroxidase-mediated Formation of the Fungal Polyphenol 3,14'-Bihispidinyl

  • Lee, In-Kyoung;Yun, Bong-Sik
    • Journal of Microbiology and Biotechnology
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    • v.18 no.1
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    • pp.107-109
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    • 2008
  • Medicinal fungi, Phellinus linteus and Inonotus xeranticus, produce a cluster of yellow pigment in their fermentation broth that acts as an important element of biological activity. The pigment is composed of diverse polyphenols with a styrylpyrone moiety, mainly hispidin and its dimers, 3,14'-bihispidinyl, hypholomine B, and 1,1-distyrylpyrylethan. Although dimeric hispidins were proposed to be biosynthesized from two molecules of monomer via oxidative coupling by ligninolytic enzymes, laccase and peroxidase, the details of this process remain unknown. In this preliminary study, we attempted to achieve enzymatic synthesis of the hispidin dimer from hispidin by using commercially available horseradish peroxidase (HRP). Consequently, a hispidin dimer, 3,14'-bihispidinyl, was synthesized, whereas the other dimers, hypholomine B and 1,1-distyrylpyrylethan, were not produced. This result suggested that the oxidative coupling at the C-3 and C-14' positions of hispidins was dominant in the process of dimerization by HRP, and indicated that additional catalysts or substrates would be needed to synthesize other hispidin dimers present in the fungal metabolite.

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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Oxidative Coupling of Herbicide Propanil and Its Metabolite, DCA(3,4-dichloroaniline) to Humic Monomers (제초제 Propanil 및 그 분해산물인 DCA(3,4-dichloroaniline)와 Humic Monomer들과의 산화적 짝지움반응)

  • Kwon, Tae-Dong;Kim, Jang-Eok
    • Applied Biological Chemistry
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    • v.41 no.5
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    • pp.384-389
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    • 1998
  • The herbicide propanil and its metabolite, DCA were incubated with oxidative catalysts in the presence or absence of humic monomers to evaluate the incorporation of them into humic substances. Propanil and DCA underwent little or no transformation by oxidatve catalysts in the absence of humic monomers. In the presence of humic monomers, the most effective co-substrate for transformation of propanil was syringic acid by laccase and HRP, that of DCA was catechol by laccase and HRP, and protocatechuic acid by birnessite. The transformation of DCA was the highest when it was incubated with catechol at pH 8.0 during 24 hrs by laccase, and with catechol at pH 3.0 during 2 hrs by HRP, and with protocatechuic acid at pH 5.0 during 2 hrs by birnessite. The DCA transformation increased with increasing concentration of humic monomers. The transformation of DCA was increased with about 5 times when it was incubated with lactase and birnessite together than lactase alone, but that of it was not effected when it was incubated with HRP and birnessite together. When DCA was incubated with dissolved organic carbon in the presence of oxidative catalysts, the transformation of it was not increased by laccase and birnessite but increased by HRP.

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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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Peroxidase-Catalyzed Removal of Aromatic Pollutants (Peroxidase에 의한 방향성 유해물질의 제거)

  • 류근갑;박재홍임근호
    • KSBB Journal
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    • v.11 no.6
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    • pp.681-688
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    • 1996
  • In the removal of phenolic precipitates formed by horseradish peroxidase (HRP) and $H_2O_2$ from waste water, the effects of the concentrations of phenolic compounds and $H_2O_2$ on the removal efficiency of various phenols were studied. More than 90% of various phenolic compounds were removed from the aqueous solutions (pH 5-7) by HRP and H2O2. The removal efficiency of phenolic compounds by HRP was reduced to a great extent when the initial concentration of $H_2O_2$ was over 10mM. Furthermore, no phenolic compounds were removed when 50mM of $H_2O_2$ was used. The HRP's turnover number, which indicates the number of phenolic molecules removed per one molecule of HRP, was the largest as 18047 for p-ethoxyphenol while it was the smallest as 1244 for m-chlorophenol when the initial concentrations of phenolic compounds and H2O2 were the same at 1mM. HRP which was separated from the aqueous solution containing phenol and $H_2O_2$ after 24hr of reaction revealed structural changes and diminished activity. The Soret absorbance near 404nm of this HRP sample was decreased to 48% of that of fresh HRP. The values of kcat and kcat/Km of this HRP sample for the oxidation of guaiacol were also reduced to 41% and 51% of those of fresh HRP, respectively. The removals of nonphenolic aromatic compounds such as benzene, ethylbenzene, and toluene (BET) by HRP and $H_2O_2$ were enhanced when phenols were coexisting in the aqueous solutions of BET.

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Study on Antibody-enzyme Coupling and Enzyme Immunoassay Methods (효소-항체의 결합 및 효소면역측정 방법의 연구)

  • Jang Sean Il
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.3
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    • pp.874-879
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    • 2004
  • Alakaline phosphatase (ALP)- or horseradish peroxidase (HRP)-antibody conjugate was used frequently on the immunological detection methods such as enzyme-linked immunosobent assay (ELISA), immunobolt, immunohistochemistry. The classical enzyme-antibody coupling method by one-step (direction) injection of glutaraldehyde bring into being disadvantage such as low sensitivity of antigen detection because of homopolymers. This study was modified with the dialysis glutaraldehyde method to provide simple coupling through E-amino residues present in most protein. The dialysis glutaraldehyde coupling effects were better than the classical one-step glutaraldehyde injection in antigen detection of ELISA and immunobolt. Optimal dose of the dialysis glutaraldehyde solution was 0.10-0.25 %. This results suggest that the dialysis glutaraldehyde coupling method can readily applied to antigen detection of in vitro and in vivo.

Detection of luminol chemiluminescence using photodiode (화학 발광 검출을 위한 포토다이오드 제작)

  • Kim, Hyung-Il;Kang, Chul-Goo;Kang, Moon-Sik;Park, Jung-Ho;Min, Nam-Ki
    • Proceedings of the KIEE Conference
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    • 2002.07c
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    • pp.1552-1554
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    • 2002
  • 본 논문에서는 실리콘을 기초로 한 포토다이오드를 제작하여 현재 바이오센서에서 이용되고 있는 luminol 화학 발광을 전기적으로 검출하였다. 우선, 실리콘 웨이퍼에 이온주입을 통해 p-n 접합을 형성하고 Al 전극을 형성시켜 포토다이오드를 제작하였다. PDMS을 passivation 막으로 사용하여 horseradish peroxidase(HRP) 농도 변화에 따른 발광의 최적조건과 luminol, HRP가 섞인 용액에 서로 다른 양의 과산화수소를 넣어 발광이 최대로 발생하는 혼합비를 측정하였다. 최종적으로 2mM luminol, 25U HRP 조건 하에서 과산화수소 농도에 따른 화학 발광을 측정하였으며, $10{\mu}M$부터 $500{\mu}M$ 범위의 과산화수소 농도 변화에 따른 포토다이오드의 감도는 단위 면적당 23.429pA/ ${\mu}M$이다.

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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 %).