• Title/Summary/Keyword: 과산화효소

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A Study on the Electrochemical Characteristics of Biosensor with HRP Enzyme Immobilized on SPCE (SPCE에 HRP 효소가 고정화된 바이오센서의 전기화학적 특성에 관한 연구)

  • Han, Kyoung Ho;Lee, Dae Hyun;Yoon, Do Young;Choi, Sangil
    • Journal of the Korean Electrochemical Society
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    • v.23 no.3
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    • pp.73-80
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    • 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.

Electrochemical Kinetic Assessment of Rose Tissue Immobilized Biosensor for the Determination of Hydrogen Peroxide (과산화수소 정량을 위한 장미조직 함유 바이오센서의 전기화학 속도론적 고찰)

  • Rhyu, Keun-Bae
    • Applied Chemistry for Engineering
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    • v.25 no.1
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    • pp.107-112
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    • 2014
  • Using a chlorosulphonated polyethylene rubber solution for a binder of graphite powder and ferrocene for a mediator, a rose leaf tissue-embedded biosensor was built. Linearity on the Hanes-Woolf plot showed the reduction of the substrate was attained through the catalytic power of the rose peroxidase in the experimental range of electrode potential. Furthermore, 10 or more electrochemical parameters demonstrated that the electrode exerts its sensing ability quantitatively. The foregoing gave the full conviction that rose tissue can be used in place of the currently marketed enzyme for the practical use of enzyme electrode.

Stabilization of HRP Using Hsp90 in Water-miscible Organic Solvent (Hsp90을 이용한 유기용매에서의 과산화효소 안정화 연구)

  • Chung, Ja Hee;Choi, Yoo Seong;Song, Seung Hoon;Yoo, Young Je
    • Korean Chemical Engineering Research
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    • v.44 no.1
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    • pp.92-96
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    • 2006
  • Enzymes in organic media afford many advantages such as chiral synthesis and resolution, modification of fats and oils and production of biodegradable polymers. However, the nature of solvents influences the activity and stability of enzymes, and the presence of organic solvents always constitute a risk of enzyme inactivation. Heat-shock protein Hsp90, one of the molecular chaperone, was applied for understanding of enzyme inactivation and for increasing of enzyme stability in water-miscible organic solvent. Hsp90 showed stabilization effect on HRP in the 30% of DMSO, in the 30% and 50% of dioxane. Hsp90 also showed reactivation effect on the inactivated HRP by water-miscible organic solvent such as dioxane and DMSO. In addition, structural analysis using fluorescence spectrophotometry and circular dichroism showed that exposure of HRP in water-miscible organic solvent caused appreciable conformational changes and enzyme inactivation, and the unfolded HRP by water-miscible organic solvent was refolded by Hsp90.

Purification and characterization of versatile peroxidase from Pleurotus ostreatus produced in a rotary draft tube bioreactor (회전식 통풍관 생물반응기로부터 생산된 느타리균의 다목적 과산화효소(VP) 정제 및 특성)

  • Hyo-Cheol Ha
    • Journal of Mushroom
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    • v.21 no.4
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    • pp.209-214
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    • 2023
  • In this study, Pleurotus ostreatus No.42 was cultured in glucose-peptone-yeast-wheat bran medium using a previously reported novel rotary draft tube bioreactor. Versatile peroxidase (VP), a lignin-degrading enzyme, was isolated from a pellet-type mycelium culture grown in the medium for seven days. The VP was purified by sequentially applying ultra-filtration, DEAE-Sepharose CL-6B column, and Mono Q column. SDS-PAGE analysis revealed the molecular weight of VP to be 36.4 KDa with an isoelectric point of 3.65. The amino acid sequence was confirmed as VTCATGQTT. The purified VP was observed to possess the property of not only oxidizing Mn ions but also decomposing veratryl alcohol, a non-phenolic compound. The catalytic ability of VP is a subject for future research.

Purification and Pysiological Characterization of Isoperoxidase from Oat Root Treated with Alachlor (Alachlor 처리후(處理後) 귀리 근단(根端)에 존재(存在)하는 동위과산화효소(同位過酸化酵素) 정제(精製) 및 효소(酵素)의 생리적(生理的) 특성(特性))

  • Kwon, S.W.;Han, K.S.;Kim, J.C.
    • Korean Journal of Weed Science
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    • v.14 no.1
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    • pp.56-61
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    • 1994
  • The cationic isoperoxidases were isolated from oat root tips which had been grown in treatment with $1{\times}10^{-4}$ M alachlor and purified about 30-fold by treatment with ethylalchol and ion exchange chromatography on DEAF-celluose and CM-sephadex medium. The oat root was found to contain three isoperoxidase. The major activity peak (B) represented 65% of the total isoperoxidase activity. After purification, the major peak of isoperoxidase was purified about 37-fold from the oat root. Analysis of the major peroxidase peak(column fraction 58-78) by SDS revealed a single band which corresponed to a molecular mass of 42.5 kD. In vitro, isoperoxidase activies were inhibited by IAA. Isoperoxidase(50 unit) significantly inhibited 70.2% of cell division in oat root and 54.2% of cell elongation in oat coleoptile as compared with control.

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Antioxdative Effect of the Essential Oil form the Rhizomes of Acorus gramineus (석창포 정유성분의 항산화활성)

  • 구병수;이동웅
    • Journal of Life Science
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    • v.11 no.6
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    • pp.503-508
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    • 2001
  • The essential oil of Acorus gramineus(Aracease), which has been used as an antioconvulsant in Korean folk medicine, was evaluated for its effects on antioxidative system in vitro and in vivo. This mixture of terpenes showed inhibitory effects on xanthine oxidase activity with 13.3% at 10 $\mu\textrm{g}$/ml and on aldehyde oxidase activity with 5.0% at 1$\mu\textrm{g}$/ml. Lipid perosidation was inhibited by 49.4% at 1.0 mg/ml of the essential oil in vitro and by 16.7% after 7 days inhablation of an oil as compared to PTZ-treated control group. DPPH radical scavenging activity of this essential oil was ralatively werak.

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Effect of Polyacetylene Compounds from Korean Ginseng on Lipid Peroxidation (고려인삼의 폴리아세틸렌 성분이 과산화 지질 형성에 미치는 영향)

  • Kim Hyeyoung;Lee You Hui;Kim Shin Il;Jin Sung-Ha
    • Proceedings of the Ginseng society Conference
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    • 1988.08a
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    • pp.81-86
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    • 1988
  • The effect of three polyacetylene compounds. panaxydol. panaxynol and panaxytriol isolated from Korean ginseng on $CCI_4-induced$ lipid peroxidation in vitro and in vivo hepatic microsomal lipid peroxidation were investigated. Lipid peroxide levels both in serum and liver and serum enzyme (GOT. GPT. LDH) activities of normal or $CCI_4-treated$ mice and rats were also determined after administration of polyacetylenes. Hepatic microsomal cytochrome P-450 content and activities of aniline hydroxylase and aminopyrine demethylase were measured after treatment of polyacetylenes with or without carbon tetrachloride. As results. treatment with polyacetylenes to control mice did not influence the levels of lipid peroxides and serum enzyme activities while panaxynol did. Panaxynol itself inhibited liver lipid peroxidation in normal mice. Polyacetylene compounds protected hepatic lipid peroxidation and lowered serum lipid peroxide levels induced by $CCI_4$ Polyacetylenes prevented leakage of LDH to serum but elevated GOT and GPT levels caused by $CCI_4$ were not changed by polyacetylene pretreatment. $CCI_4$ caused losses in the content of cytochrome P-450 and activities of aniline hydroxylase and aminopyrine demethylase. When polyacetylenes were treated without $CCI_4$ panaxydol and panaxynol induced aniline hydroxylase and all three polyacetylenes induced aminopyrine demethylase. Cytochrome P-450 contents were not affected by polyacetylenes. In vitro hepatic microsomal lipid peroxidation was inhibited by polyacetylenes and $DL-{\alpha}-tocopherol$ in a concentration-dependent manner.

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Variation in the Pattern of Isoperoxidase Bands in the Four Parts of Lespedeza cyrtobotrya Miq. (참싸리 부위별(部位別) 과산화(過酸化) 동위효소형(同位酵素型)의 변이(變異))

  • Han, Young Chang
    • Journal of Korean Society of Forest Science
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    • v.30 no.1
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    • pp.42-49
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    • 1976
  • In order to study the variations of isoperoxidases of four parts of L. cyrtobotrya, leaves, secondary phloem, fibrous root, ovary were collected on september 29, 1975, respectively from 12 individuals which were planted in the compound of Institute of Forest Genetics, suwon, Korea. No variation of isoperoxidases appeared among the same parts which were collected from the same individual. There was a great variation in the pattern of isoperoxidase band among the 12 individuals in leaves, secondary phloem, fibrous root, and ovary. Regarding to the common occurrence band, the number was 7 in the leaves, secondary phloem, and fibrous root, while 35 bands were appeared in the ovary part. These was a great variation of occurrence band in four parts of Lespedeza. But the number of band in the parts of the Lespedeza was 4.50-5.16 on average, on the other hand there was no significant difference. No variation was observed in the activity of isoperoxidase in leaves. On the other hand, there was small varation in the secondary phloem, fibrous root and ovary.

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Antioxidative Effects of Silymarin and Silybin Purified from Silybum marianum on Lipid Peroxidation (엉겅퀴로부터 분리 정제한 Silymarin 및 Silybin의 지질 과산화에 대한 항산화 효과)

  • 이백천;박종옥;류병호
    • The Korean Journal of Food And Nutrition
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    • v.10 no.1
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    • pp.37-43
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    • 1997
  • This study was undertaken to evaluate as antioxidant activity against lipid peroxidation. Silymarin and silybin extracted from Silybum marianum were successively purified wit solvent fractionation by silica gel column chromatography. These isoflavonoid inhibited superoxide anion production in the xanthine oxidase system. In the rat liver microsomes, silymarin or silybin rapidly inhibited lipid peroxidation which was initiated enzymatically by reduced nicotinamide adenine dinucleotide phosphate(NADPH) or non-enzymatically by ascorbic acid or Fenton's reagent (H2O2+Fe2+). Mitochondrial lipid peroxidation was also inhibited by silymarin and silybin. silymarin and silybin inhibited on terminating radical chain reaction during lipid peroxidation in the enzymatic system of microsomes or in the linoleic acid hydroperoxide induced peroxidation system.

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