• Journal of Microbiology and Biotechnology
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• v.16 no.10
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• pp.1523-1528
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• 2006

The optimization of a batch-type quartz crystal microbalance (QCM)-precipitation sensor measuring acetylcholinesterase (AChE) activity was conducted. To covalently bind AChE onto the gold electrode of a QCM surface, glutaraldehyde cross-linking to a cystamine self-assembled monolayer was tried at different cystamine concentrations. At the optimum conditions of the QCM-precipitation sensor, 0.1 M potassium phosphate buffer (pH 8.0), containing 0.01% Tween 80, was used as the reaction buffer, with the enzyme amount of 5 units for immobilization and the substrate concentration of 50 mg/ml. The current biosensor might find a future applicability to the sum parameter detection on organophosphorus and carbamate pesticides.

• BMB Reports
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• v.33 no.5
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• pp.359-365
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• 2000

Ribozymes are catalytic RNAs that can cleave RNAs at specific sites, thus they have been employed to degrade a target mRNA in vivo. Development of allosterically controllable ribozymes is of great current interest, but it remained difficult to furnish such functions to ribozymes in cultured cells or in animals. Recently, we designed allosterically controllable ribozymes termed maxizymes, which have sensor arms that recognize target mRNA sequences and, in the presence of such target sequences only, they form a cavity that can capture catalytically indispensable $Mg^{2+}$ ions, cleaving the target. The maxizyme was applied to therapy for chronic myelogenous leukemia (CML). It cleaved specifically the chimeric BCR-ABL mRNA, which caused CML, without damaging the normal ABL or BCR mRNA in mammalian cells and also in mice, providing the first successful example for allosteric control of the activity of artificial ribozymes in vivo.

• Molecular & Cellular Toxicology
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• v.4 no.2
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• pp.100-105
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• 2008

Biochips are a powerful emerging technology for biomedical, environmental applications. Especially, making use of bioseonors in the evaluation of toxicity becomes increasingly important. For biosensor as a toxicity detection, biomolecules like antibodies or aptamers have been developed to specifically capture the toxic target molecules. In addition, the development of optimal chip materials capable of maintaining the activity of embedded biomolecules such as proteins or aptamers has proven challenging. Here, using sol-gel materials, new chip material, whose ability for immobilizing the embedded aptamers and maintaining the ability of embedded aptamers is optimal, was searched. We used sol-gel formulation screening methods previously developed and found the best formulation which shows high sensitive and specific interactions of aptamers. This study results will support the technological advancement for diagnosis and environmental sensor.

• Korean Journal of Chemical Engineering
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• v.35 no.12
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• pp.2421-2429
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• 2018

Thiol-based self-assembled anchor linked to glucose oxidase (GOx) and gold nanoparticle (GNP) cluster is suggested to enhance the performance of glucose biosensor. By the adoption of thiol-based anchors, the activity of biocatalyst consisting of GOx, GNP, polyethyleneimine (PEI) and carbon nanotube (CNT) is improved because they play a crucial role in preventing the leaching out of GOx. They also promote electron collection and transfer, and this is due to a strong hydrophobic interaction between the active site of GOx and the aromatic ring of anchor, while the effect is optimized with the use of thiophenol anchor due to its simple configuration. Based on that, it is quantified that by the adoption of thiophenol as anchor, the current density of flavin adenine dinucleotide (FAD) redox reaction increases about 42%, electron transfer rate constant ($k_s$) is $9.1{\pm}0.1s^{-1}$ and the value is 26% higher than that of catalyst that does not use the anchor structure.

• Analytical Science and Technology
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• v.23 no.2
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• pp.172-178
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• 2010

Glutathione S-transferase is known to play a crucial role in detoxification in many cases. To develop a herbicide detection biosensor, we in this study attempted to immobilize glutathione S-transferase enzyme on solid supports, polystyrene and agarose, and Na-alginate. These matrixes were attractive materials for the construction of biosensors and might also have utility for the production of immobilized enzyme bioreactors. We also compared the activities of glutathione-S-transferase immobilized OsGSTF3 and free OsGSTF3. The specific activity of the free enzyme in solution was 3.3 higher than the immobilized enzyme. These results suggest that 50% of the enzyme was bound with the catalytic site in polystyrene-alkylamine bead and immobilized enzymes showed 80% remaining activity until 3 times reuse.

• Journal of Life Science
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• v.32 no.2
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• pp.148-154
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• 2022

Focal adhesion kinase (FAK) is known to regulate cell adhesion, migration, and mechanotransduction in focal adhesions (FAs). However, studies on how FAK activity is regulated in the plasma membrane microdomains according to the composition of extracellular matrix (ECM) proteins are still lacking. A genetically encoded fluorescence resonance energy transfer (FRET)-based biosensor can provide useful information on the activity of intracellular signals with high spatiotemporal resolution. In this study, we analyzed the FAK activities in lipid raft (detergent-resistant membrane) and non-lipid raft (non-detergent-resistant membrane) microdomains using FRET-based membrane targeting FAK biosensors (FAK-Lyn and FAK-KRas biosensors) under four different ECM protein compositions: glass, type 1 collagen, fibronectin, and laminin. Interestingly, FAK activity in response to laminin in a lipid raft microdomain was lower than that in other ECM conditions. Cells subjected to fibronectin showed higher FAK activity in a lipid raft microdomain than that in a non-lipid raft microdomain. Therefore, this study demonstrates that the FAK activity can be distinctively regulated according to the ECM type and the environment of the plasma membrane microdomains.

• Applied Biological Chemistry
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• v.49 no.4
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• pp.315-321
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• 2006

Inhibitors of acetylcholinesterase(AchE), such as organophosphates and carbamates, interfere the action of AchE in nerve and may lead to a severe impairment of nerve functions or even death. Therefore, insect AchE is the biological target of predominant insecticides used in agriculture. Biosensors are sensitive and can be used as dispoisable sensors for environmental control. In recent years, the use of AchEs in biosensor technology has gained enormous attention, in particular with respect to insecticide detection. The principle of biosensors using AchE as a biological recognition element is based on the inhibition the catalytic activity by the agents to be detected. We here present a strip-type biosensor based on AchE inhibition. In this study, acetylcholinesterase and PVA-SbQ(polyvinyl alcohol functionalized with methyl pyridinium methyl sulfate) were co-immobilized on immobilone-P membranes. Immobilization of the enzymes showed a stability in 6 months without activity loss in $4^{\circ}C$ storage. Enzymes immobilized on surfaces of membrane responded to organophosphates and carbamate more sensitivitive than enzyme in solution. Organophosphates and carbamates concentrations could be detected by entrapped and surface immobilized enzymes, in 5 min. For chlorpyrifos, carbofuran, cabaryl, and methidathion, the detection limits of AChE-strip were similar to that of HPLC/GC method.

• Food Science and Biotechnology
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• v.17 no.2
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• pp.219-227
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• 2008

Bacillus thuringiensis was isolated as a pathogen of the sotto disease of silkmoth larvae about a hundred years ago. Since then, this bacterium has attracted attentions of not only insect pathologists but also many other scientists who are interested in its strong and specific insecticidal activity. This has led to the recent worldwide development of B. thuringiensis-based microbial insecticides and insect-resistant transgenic plants, as well as a landmark discovery of par asp orin, a cancer cell-specific cytotoxin produced by B. thuringiensis. In this review, we describe examination of interaction between inclusion proteins of B. thuringiensis and brush border membrane of insects using a surface plasmon resonance-based biosensor, identification and characterization of parasporin-4, the latest parasporin produced by the B. thuringiensis A1470 strain, and an effective method for preparing the parasporin-4 from inclusion bodies expressed in the recombinant Escherichia coli cells.

• Bulletin of the Korean Chemical Society
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• v.35 no.9
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• pp.2749-2752
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• 2014

Recent studies have shown that single-stranded DNA adsorbed onto graphene oxide is protected from DNase I cleavage. However, double-stranded DNA bound to graphene oxide and could be digested by DNase I. To elucidate whether single-stranded DNA is protect from DNase I in the presence of graphene oxide, this study conducted DNase I digestion using single-stranded DNA and single-stranded DNA containing the duplex region in the presence of graphene oxide. Addition of DNase I resulted in restoration of the fluorescence emission that had been quenched when DNA was adsorbed to graphene oxide. It indicates that DNase I cleaved the adsorbed single-stranded DNA onto graphene oxide, which was sufficient for the detection of DNase I activity.

• Journal of Pharmacopuncture
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• v.25 no.1
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• pp.37-45
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• 2022

Objectives: The quorum-sensing-inhibitory and anti-biofilm activities of the methanol extract of E. globulus leaves were determined against clinically isolated multidrug-resistant Pseudomonas aeruginosa. Methods: The preliminary anti-quorum-sensing (AQS) activity of eucalyptus was investigated against a biosensor strain Chromobacterium violaceum ATCC 12472 (CV12472) by using the agar well diffusion method. The effect of sub-minimum inhibitory concentrations (sub-MICs) of the methanol extract of eucalyptus on different quorum-sensing-regulated virulence factors, such as swarming motility, pyocyanin pigment, exopolysaccharide (EPS), and biofilm formation, against clinical isolates (CIs 2, 3, and 4) and reference PA01 of Pseudomonas aeruginosa were determined using the swarm diameter (mm)-measurement method, chloroform extraction method, phenol (5%)-sulphuric acid (concentrated) method, and the microtiter plate assay respectively, and the inhibition (%) in formation were calculated. Results: The preliminary AQS activity (violacein pigment inhibition) of eucalyptus was confirmed against Chromobacterium violaceum ATCC 12472 (CV12472). The eucalyptus extract also showed concentration-dependent inhibition (%) of swarming motility, pyocyanin pigment, EPS, and biofilm formation in different CIs and PA01 of P. aeruginosa. Conclusion: Our results revealed the effectiveness of the E. globulus extract for the regulation of quorum-sensing-dependent virulence factors and biofilm formation at a reduced dose (sub-MICs) and suggest that E. globulus may be a therapeutic agent for curing and controlling bacterial infection and thereby reducing the possibility of resistance development in pathogenic strains.