• Journal of Microbiology and Biotechnology
• /
• v.19 no.7
• /
• pp.713-717
• /
• 2009

The dependence of the catalytic properties of horseradish peroxidase on the structural changes of ionic liquids was investigated with two water-miscible ionic liquids, N-butyl-3methypyridinium tetraftuoroborate ([$BMP_y$][$BF_4$]) and 1-butyl-3-methylimidazolium methylsulfate ([BMIM][$MeSO_4$]), each of which shares an anion ($BF_4^-$) or a cation ($BMIM^+$) with 1-butyl-3-methylimidazolium tetraftuoroborate ([BMIM][$BF_4$]), respectively. The oxidation of guaiacol (2-methoxyphenol) with $H_2O_2$was used as a model reaction. In order to minimize the effect of solution viscosity on the kinetic constants of the enzymatic catalysis, the enzymatic reactions for the kinetic study were performed in water-ionic liquid mixtures containing 25% (v/v) ionic liquid at maximum. Similarly to the previously reported results for [BMIM][$BF_4$], as the concentration of [$BMP_y$][$BF_4$] increased, the $K_m$value increased with a decrease in the $k_{cat}$value: the $K_m$value increased markedly from 2.8 mM in 100% water to 12.6 mM in 25% (v/v) ionic liquid, indicating that ionic liquid significantly weakens the binding affinity of guaiacol to the enzyme. On the contrary, [BMIM][$MeSO_4$] decreased the Km value to 1.4 mM in 25% (v/v) ionic liquid. [BMIM][$MeSO_4$] also decreased $k_{cat}$more than 3-folds [from 13.8 $s^{-1}$in 100% water to 4.1 $s^{-1}$in 25% (v/v) ionic liquid]. These results indicate that the ionic liquids interact with the enzyme at the molecular level as well as at a macroscopic thermodynamic scale. Specifically, the anionic component of the ionic liquids influenced the catalysis of horseradish peroxidase in different ways.

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

• Korean Journal of Microbiology
• /
• v.27 no.4
• /
• pp.348-356
• /
• 1989

An extracellular peroxidase found in culture broth of Pleurotus ostreatus was induced by syringic acid. This enzyme was fractionated by DEAE Sephadex A-50 ion exchange chromatography and gel filtration chromatogrphy on Sephadex G-150. The enzyme is a glycoprotein containing 35.7% carbohydrate. The results of SDS-linear polyacrylamide gradient gel electrophoresis and gel filtration indicate that the enzyme is a dimer consisted of identical subunits (Mr=72,400). The absorption spectrum of the enzyme indicates the presence of one mole of iron protoporphyrin IX per one mole of subunit. Isoelectric point of the enzyme is 4.26 and $K_m$ values for $H_2O_2$ is $7.2{\mu}M$. The enzyme showed its optimal activity at pH 3.5-4.0 and at $40^{\circ}C$. The Km values of this enzyme for ferulic acid and sinapic acid are 2.4 and 12.3 times higher than those of horseradish peroxidase, respectively.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.18 no.3
• /
• pp.874-879
• /
• 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.

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

• Proceedings of the Korean Environmental Sciences Society Conference
• /
• 2016.10a
• /
• pp.132-132
• /
• 2016

• Bulletin of the Korean Chemical Society
• /
• v.27 no.5
• /
• pp.672-678
• /
• 2006

A new approach to fabricate an enzyme electrode was described based on the immobilization of horseradish peroxidase (HRP) on dithiobis-N-succinimidyl propionate (DTSP) self-assembled monolayer (SAM) formed on gold-nanoparticles (Au-NPs) which were electrochemically deposited onto glassy carbon electrode (GCE) surface. The overall surface area and average size of Au-NPs could be controlled by varying deposition time and were examined by Field Emission-Scanning Electron Microscope (FE-SEM). The $O_2$ reduction capability of the surface demonstrated that Au-NPs were thermodynamically stable enough to stay on GCE surface. The immobilized HRP electrode based on Au-NPs/GCE presented faster, more stable and sensitive amperometric response in the reduction of hydrogen peroxide than a HRP immobilized on DTSP/gold plate electrode not containing Au-NPs. The effects of operating potential, mediator concentration, and pH of buffer electrolyte solution on the performance of the HRP biosensor were investigated. In the optimized experimental conditions, the HRP immobilized GCE incorporating smaller-sized Au-NPs showed higher electrocatalytic activity due to the high surface area to volume ratio of Au-NPs in the biosensor. The HRP electrode showed a linear response to $H_2O_2$ in the concentration range of 1.4 $\mu$M-3.1 mM. The apparent Michaelis-Menten constant ($K _M\; ^{app}$) determined for the immobilized HRP electrodes showed a trend to be decreased by decreasing size of Au-NPs electrodeposited onto GCE.

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

• Journal of the Korean Electrochemical Society
• /
• v.23 no.3
• /
• pp.73-80
• /
• 2020

Fenton oxidation method using hydrogen peroxide is an eco-friendly oxidation method used in water treatment and soil restoration. When removing pollutants by this method, it is quite important to properly regulate the concentration of hydrogen peroxide according to the concentration of the contaminants. In this study, electrochemical biosensors using HRP (horseradish peroxidase) enzymes were manufactured and studies were conducted on the activity of enzymes and the detection characteristics of hydrogen peroxide. HRP were electro deposited with chitosan and AuNP on the working electrode surface of the SPCE (Screen Printed Carbon Electrode). Then, the fixation of enzymes was confirmed using the cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The activity of HRP enzymes was also identified from chronoamperometry (CA) and UV spectroscopy. After immersing the biosensor in PBS solution the current generated from electrodes by titrating hydrogen peroxide was measured from CA analysis. The generated current increased linearly for the concentration of hydrogen peroxide, and a calibration curve was derived that could predict the concentration of hydrogen peroxide from the current.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.19 no.3
• /
• pp.224-228
• /
• 1990

Cabbage contained high peroxidase activity among tested plant sources. The cabbage peroxi-dase can replace horseradish peroxidase to assay glucose with glucose oxidase. The amount of glucose can be determined quantitatively by glucose oxidase-cabbage peroxidase. The opti-mum pH and temperature for enzymatic glucose determination by glucose oxidase-cabbage peroxidase were 6.0 and 35-45$^{\circ}C$ respectively. The glucose assay was inhibited by addition of various metal salts such as mercuric chloride lead acetate silver nitrate ammonium molyb-date sodium tunstate and cupric sulfate. The relationship between absorbance and amount of glucose was linear up to 8.33 mM glucose in the assay mixture under the assay conditions.