• The Korean Journal of Pain
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• v.7 no.2
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• pp.238-241
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• 1994

Aim of this study was to discover the projection area of the first cervical spinal nerve. Subcutaneous injection of wheat germ agglutinin-horseradish peroxidase(WGA-HRP) was done at five points of young dogs scalp and face. After two days of survival time, animals were sacrificed by perfusion through the left ventricle of the heart. Trigeminal ganglion, first and second cervical dorsal root ganglion, superior cervical ganglion, middle cervical ganglion and stellate ganglion were removed. Projection area of wheat germ agglutinin-horseradish peroxidase in vestigated into above ganglions. Projection into the first cervical dorsal root ganglion and stellate ganglion was not found. This experiment is deemed valuable for the study of neuronal connection on the central nervous system.

• KSBB Journal
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• v.14 no.4
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• pp.465-469
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• 1999

Horseradish peroxidase(HRP) in organic solvent can be reactivated by evaporation. In order to measure the evaporation effect, the enzyme solutions were obtained by evaporation and dilution of organic solvent, respectively. Although two situations were thermodynamically identical, the activity from evaporation was higher than that from dilution. From the UV absorbance and the fluorescence intensity mesurements, it can be explained that reactivation of enzyme activity might be caused by reversible folding, and the enzyme obtained by evaporation was more refolded than that obtained by dilution.

• Journal of Korean Society of Environmental Engineers
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• v.32 no.11
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• pp.1054-1062
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• 2010

Oxidative coupling reactions of humic substances (HS) can be catalyzed by a variety of natural extracellular enzymes and metal oxides. In this study, property changes of HS induced by a natural enzyme, horseradish peroxidase (HRP), and the effect of it to microfiltration (MF) were investigated. PAHA was transformed by oxidative coupling reaction with HRP and hydrogen peroxide ($H_2O_2$), verifying the catalytic effects of the HRP. Size exclusion chromatography (SEC) revealed that weight-average molecular weight (MWw) of PAHA was proportionally increased with the dosages of HRP and $H_2O_2$, indicating the transform action of HS into larger and complex molecules. An increase in the conformational stability of HS was achieved through the promotion of intermolecular covalent bondings between heterogeneous humic molecules. Spectroscopic analysis (fluorescence and infrared spectroscopy) proved that functional groups were transformed by the reaction. Additionally, HS and transformed products were undergone microfiltration (MF) to examine the treatment potential of them in a water treatment facility. Original HS could not be removed by MF but larger molecules of transformed products could be removed. Meanwhile, transformed products caused more fouling on the filtration than original HS. This results proved that natural organic matter (NOM) can be removed by MF after its increase in molecular size by oxidative coupling reaction.

• Journal of Industrial and Engineering Chemistry
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• v.68
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• pp.364-371
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• 2018

Enzyme polymerization is a benign process exploiting the unique activity of enzymes. In this study, a peroxidase-mimetic catalyst is demonstrated as an alternative to horseradish peroxidase (HRP) for the polymerization of aniline. The mimetic catalyst successfully catalyzes the polymerization of aniline monomers to produce polyaniline (PANI) in an aqueous solution. The PANI produced is rich of para-structure that is generally observed when HRP is used as a catalyst. Compared to HRP, the peroxidase-mimetic catalyst shows a considerably higher catalytic activity at neutral and weak basic conditions (pH >6.5) and at temperatures over $45^{\circ}C$, at which HRP is denatured.

• Analytical Science and Technology
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• v.16 no.6
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• pp.504-508
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• 2003

A carbon paste electrode was constructed with peroxidase extracted from Horseradish and the variation of the response of the sensor with pH was investigated. Current profiles showed two highest sensitivities at two pH values respectively. In addition, two bands were observed in the electrophoretic expansion. A coincidence of the two experimental results added support to the possibility that the biosensor has two different isozymes. Assuming that current profiles are the sum of two gaussians, we deconvoluted them and determined the optimum pH of peroxidase isozymes.

• Journal of Microbiology and Biotechnology
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• v.27 no.2
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• pp.297-305
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• 2017

The use of peroxidase in the nitration of phenols is gaining interest as compared with traditional chemical reactions. We investigated the kinetic characteristics of phenol nitration catalyzed by horseradish peroxidase (HRP) in an aqueous-organic biphasic system using n-butanol as the organic solvent and ${NO_2}^-$ and $H_2O_2$ as substrates. The reaction rate was mainly controlled by the reaction kinetics in the aqueous phase when appropriate agitation was used to enhance mass transfer in the biphasic system. The initial velocity of the reaction increased with increasing HRP concentration. Additionally, an increase in the substrate concentrations of phenol (0-2 mM in organic phase) or $H_2O_2$ (0-0.1 mM in aqueous phase) enhanced the nitration efficiency catalyzed by HRP. In contrast, high concentrations of organic solvent decreased the kinetic parameter $V_{max}/K_m$. No inhibition of enzyme activity was observed when the concentrations of phenol and $H_2O_2$ were at or below 10 mM and 0.1 mM, respectively. On the basis of the peroxidase catalytic mechanism, a double-substrate ping-pong kinetic model was established. The kinetic parameters were ${K_m}^{H_2O_2}=1.09mM$, ${K_m}^{PhOH}=9.45mM$, and $V_{max}=0.196mM/min$. The proposed model was well fit to the data obtained from additional independent experiments under the suggested optimal synthesis conditions. The kinetic model developed in this paper lays a foundation for further comprehensive study of enzymatic nitration kinetics.

• Korean Journal of Environmental Agriculture
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• v.17 no.3
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• pp.239-245
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• 1998

To investigate the formation of bound residue with soil organic materials by oxidative coupling, nitroaromatics and their reduced metabolites, the insecticide parathion and the herbicide asulam were incubated with oxidoreductase, laccase or horseradish peroxidase, in the presence or absence of humic monomers. Most of aminotoluenes and amino-nitrophenols were completely transformed while most of nitrotoluenes and nitrophenols remained unchanged by a lactase or horseradish peroxidase in the presence or absence of humic monomers. Amino-nitrotoluenes were not transformed without humic monomers, but the addition of various humic monomers caused a considerable difference in the transformation of amino-nitrotoluenes by a lactase or horseradish peroxidase. Amino-nitrotoluenes were most transformed in the presence of catechol, syringaldehyde and protocatechuic acid. The insecticide parathion with nitro group and its metabolite were not mostly transformed in the presence or absence of humic monomers. The herbicide asulam with amino group remained unchanged without humic monomers as well, but the stimulating effect on the transformation of asulam was caused by the addition of catechol, syringaldehyde, protocatechuic acid or caffeic acid with a lactase.

• 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.

• Journal of Microbiology and Biotechnology
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• v.23 no.4
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• pp.527-533
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• 2013

Polyclonal antibodies labeled with a tracer have been commonly used as secondary antibodies in immunochemical assays to quantify the concentration of antibody-antigen complexes. The majority of these antibodies conjugated with a tracer are commercially available, with the exception of few untouched targets. This study focused on the production and application of mouse anti-quail IgY as an intermediate antibody to link between quail egg yolk IgY and goat anti-mouse IgG-HRP as primary and secondary antibodies, respectively. Subsequently, the produced mouse anti-quail IgY was labeled with horseradish peroxidase (HRP) and its efficiency on enzyme linked immunosorbent assay (ELISA) was compared with that of commercial rabbit anti-chicken IgY-HRP. As an intermediate antibody, mouse anti-quail IgY was successfully produced with good affinity and sensitivity (1:10,000) to the primary and secondary antibodies. Subsequently, mouse anti-quail IgY was effectively conjugated with HRP enzyme, resulting in a secondary antibody with good sensitivity (1:10,000) to quail anti-V. parahaemolyticus and V. vulnificus IgY. The detection limit was $10^5$ CFU/ml for both V. parahaemolyticus and V. vulnificus. The efficiency of the produced conjugate to detect quail IgY on ELISA was comparable to that of the commercial rabbit anti-chicken IgY-HRP, and hence the produced and labeled mouse anti-quail IgY-HRP can be used as a secondary antibody to detect any antibody produced in quail.

• Journal of Microbiology and Biotechnology
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• v.17 no.4
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• pp.600-603
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• 2007

The effect of a water-miscible ionic liquid, 1-butyl-3-methylimidazolium tetrafluoroborate $([BMIM][BF_4])$, on the horseradish peroxidase (HRP)-catalyzed oxidation of 2-methoxyphenol (guaiacol) with hydrogen peroxide $(H_2O_2)$ was investigated. HRP maintains its high activity in the aqueous mixtures containing various concentrations of the ionic liquid and even in 90% (v/v) ionic liquid. In order to minimize the effect of solution viscosity on the kinetic constants of HRP catalysis, the enzymatic reactions in the subsequent kinetic study were performed in water-ionic liquid mixtures containing 25% (v/v) ionic liquid at maximum. As the concentration of $[BMIM][BF_4]$ increased for the oxidation of guaiacol by HRP, the $K_m$ value increased with a slight decrease in the $K_{cat}$ value: The $K_m$ value increased from 2.8 mM in 100% (v/v) water to 22.5mM in 25% (v/v) ionic liquid, indicating that ionic liquid significantly weakens the binding affinity of guaiacol to HRP.