• Title/Summary/Keyword: Enzyme electrode

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Purification and Characterization of Laccase from Wood-Degrading Fungus Trichophyton rubrum LKY-7

  • Hyunchae Jung;Park, Chongyawl;Feng Xu;Kaichang Li
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 2001.04a
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    • pp.18-25
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    • 2001
  • A new wood-degrading fungus Trichophyton rubrum LKY-7 secretes a high level of laccase in a glucose-peptone liquid medium. The production of laccase by the fungus was barely induced by 2,5-xylidine. The laccase has been purified to homogeneity through three chromatography steps in an overall yield of 40%. The molecular mass of the purified laccase was about 65 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified laccase had the distinct blue color and had basic spectroscopic features of a typical blue laccase: two absorption maxima at 278 and 610 nm and a shoulder at 338 nm. The N-terminus of the laccase has been sequenced, revealing high homology to laccases from wood-degrading white-rot fungi such as Ceriporiopsis subvermispora. The enzyme had a "low" redox potential (0.5 V vs normal hydrogen electrode), yet it was one of the most active laccases in oxidizing a series of representative substrates/mediators. Compared with other fungal laccases, the laccase has a very low Km value with ABTS [2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid] as a substrate and a very high Km value with violuric acid as a substrate. The laccase has the isoelectric point of 4.0. The laccase had very acidic optimal pH values (pH 3-4) while it was more stable at neutral pH than at acidic pH. The laccase oxidized hydroquinone faster than catechol and pyrogallol. The oxidation of tyrosine by the laccase was not detectable under the reaction conditions. The laccase was strongly inhibited by sodium azide and sodium fluoride. fluoride.

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Preparation of Enzyme Electrodes for Biofuel Cells Based on the Immobilization of Glucose Oxidase in Polyion Complex (폴리이온복합체를 이용하여 글루코스 산화효소를 고정화한 바이오전지용 효소전극 제조)

  • Nguyen, Linh Thi My;Li, Nan;Yoon, Hyon Hee
    • Applied Chemistry for Engineering
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    • v.24 no.1
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    • pp.99-103
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    • 2013
  • An emzymatic bioanode for a glucose/oxygen biofuel cell was prepared by the sequential coating of carbon nanotube (CNT), charge transfer complex (CTC) based on tetracyanoquinodimethane (TCNQ) and tetrathiafulvalene (TTF), glucose oxidase (GOx), and polyion complex (mixture of poly-L-lysine hydrobromide and poly (sodium 4-styrenesulfonate)) on a glassy carbon electrode. A biocathode was also prepared by the sequential coating of CNT, bilirubin oxidase (BOD), 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), and polyion complex. The effect of CNT and CTC on the electrochemical performance was investigated. The biofuel cell exhibited a promising performance with maximum power densities of 3.6, 10.1, and $46.5{\mu}W/cm^2$ at 5, 20, and 200 mM of glucose concentration, respectively. The result indicates that the biofuel cell architecture prepared in this study can be used in the development of biofuel cells and biosensors.

Non-Enzymatic Glucose Sensor Based on a Copper Oxide Nanoflowers Electrode Decorated with Pt Nanoparticles (백금 나노입자가 분산된 3차원 산화구리 나노구조체 기반의 글루코스 검출용 비효소적 전기화학 센서 개발)

  • Song, Min-Jung
    • Korean Chemical Engineering Research
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    • v.56 no.5
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    • pp.705-710
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    • 2018
  • An electrochemical glucose sensor with enzyme-free was fabricated using Pt nanoparticles (Pt NPs) decorated CuO nanoflowers (CuO NFs). 3-D CuO nanoflowers film was directly synthesized on Cu foil by a simple hydrothermal method and Pt NPs were dispersed on the petal surface of CuO NFs through electrochemical deposition. This prepared sample was noted to Pt NPs-CuO NF. Morphology of the Pt NPs-CuO NFs layer was analyzed using scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS). The electrochemical properties and sensing performances were investigated using cyclic voltammetry (CV) and chronoamperometry (CA) under alkaline condition. The sensor exhibited a high sensitivity, wide liner range and fast response time. Its excellent sensing performance was attributed to the synergistic effect of the Pt NPs and CuO nanostructure.

Study of Enzyme Immobilization on Composite of CTA and PCL Membrane for Biosensor (바이오센서용 CTA와 PCL 혼합막에의 효소고정화 기법의 개발)

  • 홍성현;김태진
    • KSBB Journal
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    • v.10 no.4
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    • pp.468-474
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    • 1995
  • The disposable glucose bio-sensor using composite of CTA and PCL membrane was developed for measurement of glucose. The most effective membrane was composed of CTA/PCL(80/20, w/w) and glutaraldehyde one-step immobilization method ($10{\mu}m$ thickness) for glucose sensor gave the best result among various methods, considering oxygen permeability and electronic sensitivity. A scanning electron micrograph of the cross-section of a typical asymmetric CTA/PCL composite membrane showed that the membrane fused with a dense layer covered with a GOD-glutaraldehyde. Glucose oxidase immoblilized on the membrane showed the linearity between difference of absolute amperometric values and glucose concentrations within 7mM when the GOD immobilized electrode was used. About 35% of activity was remained after 8 days when the tyrosinase was immobilized on CTA/PCL (80/20) membrane.

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Detection of Food-Grade Hydrogen Peroxide by HRP-Biocomposite Modified Biosensors

  • Chang, Seung-Cheol
    • Journal of Food Hygiene and Safety
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    • v.32 no.6
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    • pp.447-454
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    • 2017
  • A new amperometric biosensor has been developed for the detection of hydrogen peroxide ($H_2O_2$). The sensor was fabricated through the one-step deposition of a biocomposite layer onto a glassy carbon electrode at neutral pH. The biocomposite, as a $H_2O_2$ sensing element, was prepared by the electrochemical deposition of a homogeneous mixture of graphene oxide, aniline, and horseradish peroxidase. The experimental results clearly demonstrated of that the sensor possessed high electrocatalytic activity and responded to $H_2O_2$ with a stable and rapid manners. Scanning electron microscopy, cyclic voltammetry, and amperometry were performed to optimize the characteristics of the sensor and to evaluate its sensing chemistry. The sensor exhibited a linear response to $H_2O_2$ in the range of 10 to $500{\mu}M$ concentrations, and its detection limit was calculated to be $1.3{\mu}M$. The proposed sensing-chemistry strategy and the sensor format were simple, cost-effective, and feasible for analysis of "food-grade $H_2O_2$" in food samples.

Primer Evaluation for the Detection of Toxigenic Microcystis by PCR (독소 생성 Microcystis 검출을 위한 PCR primer의 평가)

  • 이현경;김준호;유순애;안태석;김치경;이동훈
    • Korean Journal of Microbiology
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    • v.39 no.3
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    • pp.166-174
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    • 2003
  • Microcystin produced by cyanobacteria in surface waters, such as eutrophic lake and river, is a kind of serious environmental problems due to its toxicity to human and wild animals. Microcystin is synthesized nonribosomally by the large modular multi-functional enzyme complex known as microcystin synthetase encoded by the mcy gene cluster. Amplification of mcy genes by PCR from cultures and environmental samples is a simple and efficient method to detect the toxigenic Microcystis. In order to evaluate primers designed to detect toxic microcystin-producing strains, 17 cyanobacterial strains and 20 environmental samples were examined by PCR with 7 pairs of primers. Some microcystin-producing cyanobacteria were not detected with FAA-RAA, TOX4F-TOX4R and FP-RP primers. The fragment of unexpected size was amplified with NSZW2-NSZW1 primers in Microcystis strains isolated from the lakes in Korea. TOX1P-TOX1F primers failed in amplification of toxin-producing strains. Only MSF-MSR and TOX2P- TOX2F primers amplified the fragments of mcy genes from 11 strains of microcystin-producing Microcystis. The water samples taken from 20 lakes in Korea were analyzed by PCR using each of the primers. In all the water samples, cyanobacteria capable of producing microcystin were detected by the PCR with TOX2P-TOX2F primers. These results indicate that TOX2P-TOX2F primers are better than the other primers for detection of microcystin-producing Microcystis strains in Korea. The nucleotide sequences of mcy gene in Microcystis aeruginosa NIER10010 suggest genetic diversity of Korean isolates.