• Title/Summary/Keyword: biosensor activity

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Strategies in Protein Immobilization on a Gold Surface

  • Park, Jeho;Kim, Moonil
    • Applied Science and Convergence Technology
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    • v.24 no.1
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    • pp.1-8
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    • 2015
  • Protein immobilization on a gold surface plays an important role in the usefulness of biosensors that utilize gold-coated surfaces such as surface plasmon resonance (SPR), quartz crystal microbalance (QCM), etc. For developing high performance biosensors, it is necessarily required that immobilized proteins must remain biologically active. Loss of protein activity and maintenance of its stability on transducer surfaces is directly associated with the choice of immobilization methods, affecting protein-protein interactions. During the past decade, a variety of strategies have been extensively developed for the effective immobilization of proteins in terms of the orientation, density, and stability of immobilized proteins on analytical devices operating on different principles. In this review, recent advances and novel strategies in protein immobilization technologies developed for biosensors are briefly discussed, thereby providing an useful information for the selection of appropriate immobilization approach.

Electrochemical Property of Immobilized Spinach Ferredoxin on HOPG Electrode

  • Nam Yun-Suk;Kim, You-Sung;Shin, Woon-Sup;Lee, Won-Hong;Choi, Jeong-Woo
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.1043-1046
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    • 2004
  • The stability and electrochemical properties of a self-assembled layer of spinach ferredoxin on a quartz substrate and on a highly oriented pyrolytic graphite electrode were investigated. To fabricate the ferredoxin self-assembly layer, dimyristoylphosphatidylcholine was first deposited onto a substrate for ferredoxin immobilization. Surface analysis of the ferredoxin layer was carried out by atomic force microscopy to verify the ferredoxin immobilization. To verify ferredoxin immobilization on the lipid layer and to confirm the maintenance of redox activity, absorption spectrum measurement was carried out. Finally, cyclic-voltammetry measurements were performed on the ferredoxin layers and the redox potentials were obtained. The redox potential of immobilized ferredoxin had a formal potential value of -540 mV. It is suggested that the redox-potential measurement of self-assembled ferredoxin molecules could be used to construct a biosensor and biodevice.

Electrochemical Biosensors based on Nanocomposites of Carbon-based Dots

  • Ngo, Yen-Linh Thi;Jana, Jayasmita;Chung, Jin Suk;Hur, Seung Hyun
    • Korean Chemical Engineering Research
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    • v.58 no.4
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    • pp.499-513
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    • 2020
  • Among the many studies of carbon-based nanomaterials, carbon-based dots (CDs) have attracted considerable interest owing to their large surface area, intrinsic low-toxicity, excellent biocompatibility, high solubility, and low-cost with environmentally friendly routes, as well as their ability for modification with other nanomaterials. CDs have several applications in biosensing, photocatalysis, bioimaging, and nanomedicine. In addition, the fascinating electrochemical properties of CDs, including high active surface area, excellent electrical conductivity, electrocatalytic activity, high porosity, and adsorption capability, make them potential candidates for electrochemical sensing materials. This paper reviews the recent developments and synthesis of CDs and their composites for the proposed electrochemical sensing platforms. The electrochemical principles and future perspective and challenges of electrochemical biosensors are also discussed based on CDs-nanocomposites.

Electrocatalytic Properties of Metal-dispersed Carbon Paste Electrodes for Reagentless L-lactate Biosensors (금속이 첨가된 탄소전극의 전기화학적 특성과 이를 이용한 L-lactate 바이오센서의 개발)

  • 윤현철;김학성
    • KSBB Journal
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    • v.11 no.4
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    • pp.489-496
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    • 1996
  • Metal dispersed carbon paste electrodes were fabricated, and their electrochemical properties were investigated. Among various metal dispersed carbons, platinum-dispersed carbon paste electrode showed most efficient electrocatalytic characteristics. The overpotential for the oxidation of NADH was significantly lowered in the platinum-dispersed carbon paste electrode, and catalytic current was also enhanced. Based on these electrocatalytic observations, L-lactate biosensor using L-lactate dehydrogenase was constructed to evaluate its performance in terms of sensitivity and stability.

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Enhancement in the Viability and Biosensing activity of Freeze-Dried Recombinant Bioluminescent Bacteria

  • Park, Sue-Hyung;Gu, Man-Bock
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.3
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    • pp.202-206
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    • 2000
  • The genetically-engineered Escherichia coli strain, DPD2540, which contains a fabA:::luxCDAbefusion gene, gives a bioluminescent output when membrane fatty acid synthesis is needed. For more pactical application of this strain in the filed as biosensor, freezedrying was adopted. A 12% surcrose solution with Luria-Bertani (LB) broth, as determined by the viability after freeze-drying, was found to be most most effective composition for lyophilization solution among various compositions testitons tested. Rapid freezing with liquid nitrogen also gave the best viability after freeze-drying as compared to samples frozen at-7$0^{\circ}C$ and -2$0^{\circ}C$. The biosensing activities of the cells showed a greater sensitivity when the cells from the expontial phase were freeze-dried. Finally, the optimum temperature for use of the freeze-dried cells in the biodencor field was determined.

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Expression of Plasma Membrane $H^ +-ATPase $ in the Roots of Plants Under Low Temperature (바이오센서에 의한 뿌리 원형질막에서의 $H^ +-ATPase $의 활성측정)

  • ;;;Etsuo Watanabe
    • KSBB Journal
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    • v.17 no.1
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    • pp.59-62
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    • 2002
  • The enzyme sensor for ATPase activity consisted of an immobilized membrane of two enzymes, purine nucloside phosphorylase (NP) and xanthine oxidase (XOD), and oxygen electrode. The $H^ +-ATPase $ rate of the plasma membranes increased by low root temperature. A cucumber and a pumpkin plasma membrane $H^ +-ATPase $$ activities measured by the proposed sensor system were in good agreement with the results obtained by a conventional UV spetrometer assay. One cycle of assay could be completed within 3 minutes.

Development of Cyanide Sensor Using S. cerevisiae (S. cerevisiae를 이용한 시안센서의 개발)

  • 김종민;이현우
    • KSBB Journal
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    • v.11 no.6
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    • pp.669-675
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    • 1996
  • Two types of cyanide sensor system were constructed for monitoring river water simply. One is membrane type flow system composed of S. cerevisiae coated membrane and oxygen electrode, the other is reactor type flow system composed of S. cevevisiae immobilized reactor and two oxygen electrode. S. cerevisiae and oxygen electrode were used for detecting cyanide compounds. These sensors are based on the inhibition of cyanides on the respiration activity of S. cerevisiae. Membrane type sensor system could detect cyanides ranging 0.10∼1.00ppm, and the respiration activity of S. cerevisiae continued for a day. Reactor type sensor system composed of immobilized chitopearl HP-5020 with S. cerevisiae could detect cyanides in the range of 0.10∼1.00ppm, and maintained 90% respiration activity for sixteen days. It was found that the optimum reactor size exist for the reactor type biosensor when respirating activity inhibition is used for detection of cyanides.

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Comparative Analysis of Src Activity in Plasma Membrane Subdomains via Genetically Encoded FRET Biosensors (유전적으로 암호화된 FRET 바이오센서를 통한 세포막 하위 도메인의 Src 활성 비교 분석)

  • Gyuho Choi;Yoon-Kwan Jang;Jung-Soo Suh;Heonsu Kim;Sanghyun Ahn;Tae-Jin Kim
    • Journal of Life Science
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    • v.33 no.2
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    • pp.191-198
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    • 2023
  • As a member of the focal adhesion complex of the plasma membrane, Src is a nonreceptor tyrosine kinase that controls cell adhesion and motility. However, how Src activity is regulated in the plasma membrane microdomain in response to components of the extracellular matrix (ECM) remains unclear. This study compared and investigated the activity of Src in response to three representative ECM proteins: collagen type 1, fibronectin, and laminin. Genetically encoded FRET-based Src biosensors for plasma membrane subdomains were used. FRET-based biosensors allow the real-time analysis of protein activity in living cells based on their high spatiotemporal resolution. The results showed that Src activity was maintained at a high level under all ECM conditions of the lipid raft, and there was no significant difference between the ECM conditions. In contrast, Src activity was maintained at a low level in the non-lipid raft membrane. In addition, the Src activity of lipid rafts remained significantly higher than that of non-lipid raft regions under the same ECM conditions. In conclusion, this study demonstrates that Src activity can be controlled differently by lipid rafts and non-lipid raft microdomains.

Enrichment of Electrochemically Active Bacteria Using a Three-Electrode Electrochemical Cell

  • Yoon, Seok-Min;Choi, Chang-Ho;Kim, Mi-A;Hyun, Moon-Sik;Shin, Sung-Hye;Yi, Dong-Heui;Kim, Hyung-Joon
    • Journal of Microbiology and Biotechnology
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    • v.17 no.1
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    • pp.110-115
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    • 2007
  • Electrochemically active bacteria were successfully enriched in an electrochemical cell using a positively poised working electrode. The positively poised working electrode (+0.7 V vs. Ag/AgCl) was used as an electron acceptor for enrichment and growth of electrochemically active bacteria. When activated sludge and synthetic wastewater were fed to the electrochemical cell, a gradual increase in amperometric current was observed. After a period of time in which the amperometric current was stabilized (generally 8 days), linear correlations between the amperometric signals from the electrochemical cell and added BOD (biochemical oxygen demand) concentrations were established. Cyclic voltammetry of the enriched electrode also showed prominent electrochemical activity. When the enriched electrodes were examined with electron microscopy and confocal scanning laser microscopy, a biofilm on the enriched electrode surface and bacterium-like particles were observed. These experimental results indicate that the electrochemical system in this study is a useful tool for the enrichment of an electrochemically active bacterial consortium and could be used as a novel microbial biosensor.

Real-time Fluorescence Assay of DNA Polymerase Using a Graphene Oxide Platform (산화 그래핀 플랫폼을 이용한 DNA 중합효소의 실시간 형광에세이)

  • Gang, Jongback
    • Microbiology and Biotechnology Letters
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    • v.41 no.4
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    • pp.456-461
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    • 2013
  • Using the different adsorption properties of ssDNA and dsDNA to GO, this study used a real time and efficient fluorescence assay to detect the enzymatic activity of the Klenow fragment with the adsorbed DNA to GO. Results showed that adsorption of fluorescein-tagged ssDNA to GO resulted in fluorescence quenching and DNA was released from GO by adding complementary DNA. In addition, fluorescence restoration was increased through a polymerization reaction by the Klenow fragment in the presence of a fluorescein-attached template, GO, and primer. Gel electrophoresis was conducted to confirm the hybridization and DNA polymerization reactions on GO.