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

• Journal of Electrochemical Science and Technology
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• v.9 no.1
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• pp.28-36
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• 2018

Metal-organic frameworks have recently been considered very promising modifiers in electrochemical analysis due to their unique characteristics among which tunable pore sizes, crystalline ordered structures, large surface areas and chemical tenability are worth noting. In the present research, $Cu(btec)_{0.5}DMF$ was electrodeposited on the surface of glassy carbon electrode at room temperature under cathodic potential and was initially used as the active materials for the detection of $H_2O_2$. The cyclic voltammogram of $Cu(btec)_{0.5}DMF$ modified GC electrode shows distinct redox peaks potentials at +0.002 and +0.212 V in 0.1 M phosphate buffer solution (pH 6.5) corresponding to $Cu^{(II)}/Cu^{(I)}$ in $Cu(btec)_{0.5}DMF$. Acting as the electrode materials of a non-enzymatic $H_2O_2$ biosensor, the $Cu(btec)_{0.5}DMF$ brings about a promising electrocatalytic performance. The high electrocatalytic activity of the $Cu(btec)_{0.5}DMF$ modified GC electrode is demonstrated by the amperometric response towards $H_2O_2$ reduction with a wide linear range from $5{\mu}M$ to $8000{\mu}M$, a low detection limit of $0.865{\mu}M$, good stability and high selectivity at an applied potential of -0.2 V, which was higher than some $H_2O_2$ biosensors.

• 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.

• Journal of Microbiology and Biotechnology
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• v.32 no.11
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• pp.1471-1478
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• 2022

2'-Fucosyllactose (2'-FL), the most abundant fucosylated oligosaccharide in human milk, has multiple beneficial effects on human health. However, its biosynthesis by metabolically engineered Escherichia coli is often hampered owing to the insolubility and instability of α-1,2-fucosyltransferase (the rate-limiting enzyme). In this study, we aimed to enhance 2'-FL production by increasing the expression of soluble α-1,2-fucosyltransferase from Helicobacter pylori (FucT2). Because structural information regarding FucT2 has not been unveiled, we decided to improve the expression of soluble FucT2 in E. coli via directed evolution using a protein solubility biosensor that links protein solubility to antimicrobial resistance. For such a system to be viable, the activity of kanamycin resistance protein (Kan^R) should be dependent on FucT2 solubility. Kan^R was fused to the C-terminus of mutant libraries of FucT2, which were generated using a combination of error-prone PCR and DNA shuffling. Notably, one round of the directed evolution process, which consisted of mutant library generation and selection based on kanamycin resistance, resulted in a significant increase in the expression level of soluble FucT2. As a result, a batch fermentation with the ΔL M15 pBCGW strain, expressing the FucT2 mutant (F#1-5) isolated from the first round of the directed evolution process, resulted in the production of 0.31 g/l 2'-FL with a yield of 0.22 g 2'-FL/g lactose, showing 1.72- and 1.51-fold increase in the titer and yield, respectively, compared to those of the control strain. The simple and powerful method developed in this study could be applied to enhance the solubility of other unstable enzymes.

• Journal of Sensor Science and Technology
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• v.3 no.2
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• pp.50-56
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• 1994

Fiber-optic biosensor for the detection of organophosphorus compounds in a contaminated water was developed, which was the component of pesticides and agricultural agent. The detection principle of designed sensor was the pH variance induced by a reaction of acetylcholinesterase enzyme inhibited by organophosphorus compounds. The pH variance was detected by the optical system to measure the organophosphorus compounds. Litmus was selected as the pH-sensitive dye suitable to the enzyme reaction and a light source to be detected by the optical system. The enzyme entrapped in Ca-alginate gel was immobilized at the inner wall to maintain the high activity of enzyme and to be reused for a long period. The optical fiber was used to miniaturize and control remotely the sensor system. The He-Ne laser with 632 nm was selected as the light source to prevent light intensity fluctuation by the product. Cheap plastic optical fibers were used as the transmission part of the light and the phototransistor was used as the reception part of light based on the wavelength of He-Ne laser. The proposed fiber-optic biosensor has the linear analytical range of 0 ppm-1.5 ppm with response time of 5 minutes.

• Membrane and Water Treatment
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• v.6 no.3
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• pp.189-203
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• 2015

Membrane biofouling impedes wide application of membrane bioreactor (MBR) for wastewater treatment. Recently, quorum sensing (QS) mechanisms are accounted for one of major mechanisms in biofouling of MBRs. In this study, vanillin was applied to investigate reduction of biofouling in MBRs. MBR sludge was analyzed to contain QS signal molecules by cross-feeding biosensor assay and HPLC. In addition, the inhibitory activity of vanillin against bacterial quorum sensing was verified using an indicator strain CV026. The vanillin doses greater than 125 mg/L to 100 mL of MBR sludge showed 25% reduction of biofilm formed on the membrane surfaces. Two MBRs, i.e., a typical MBR as a control and an MBR with vanillin, were operated. The TMP increases of the control MBR were more rapid compared to those of the MBR with the vanillin dose of 250 mg/L. The treatment efficiencies of the two MBRs on organic removal and MLSS were maintained relatively constant. Extracellular polymeric substance concentrations measured at the end of the MBR operation were 173 mg/g biocake for the control MBR and 119 mg/g biocake for the MBR with vanillin. Vanillin shows great potential as an anti-biofouling agent for MBRs without any interference on microbial activity for wastewater treatment.

• Journal of Electrochemical Science and Technology
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• v.4 no.1
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• pp.41-45
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• 2013

Platinum is a well known element which shows a significant electrocatalytic activity in many important applications. In glucose sensor, because of the poisoning effect of reaction intermediates and the low surface area, the electrocatalytic activity towards the glucose oxidation is low which cause the low sensitivity. So, we fabricate a nanoporous PtZn alloy electrode by deposition-dissolution method. It provides a high active surface and a large enzyme encapsulating space per unit area when it used for an enzymatic glucose sensor. Glucose oxidase was immobilized on the electrode surface by capping with PEDOT composite and PPDA. The composite and PPDA also can exclude the interference ion such as ascorbic acid and uric acid to improve the selectivity. The surface area was determined by cyclic voltametry method and the surface structure and the element were analyzed by Scanning Electron Microscope (SEM) and Energy Dispersive X-ray spectroscopy (EDX), respectively. The sensitivity is $13.5{\mu}A/mM\;cm^2$. It is a remarkable value with such simply prepared senor has high selectivity.

• Food Science and Biotechnology
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• v.17 no.1
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• pp.15-19
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• 2008

This research was conducted to develop a quick and sensitive method of detecting carbamate residues using the immobilization of antibody-antigen interactions with surface plasmon resonance (SPR). We have used commercialized surface plasmon resonance equipment (Biacore 3000). The antibody used for the immunoassay was specific for glutathione-s-transferase (GST) and the antigens included several carbamate pesticides (carbofuran, carbaryl, and benfuracarb). When antigens were applied to the protein GST, the detection limit was 2 ng/mL of carbamate pesticide. The fabricated protein GST maintained its activity for over 200 measurements. Thus we determined that the SPR biosensors could detect the specific reversible binding of a reactant in solution to a binding partner immobilized on the surface of the sensor and allow real-time detection and monitoring.

• KSBB Journal
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• v.17 no.3
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• pp.213-227
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• 2002

To date, the majority of biosensor technologies use binding components such as enzymes antibodies, nucleic acids and protein ligands. In contrast, the goal underlying the use of cells and tissues of animals and plants for a sensor system is to obtain systems capable of extracting information based on the biological activity, mechanisms of action and consequences of exposure to a chemical or biological agent of interest. These systems enable the interrogation of more complex biological response and offer the potential to gather higher information content from measuring physiologic and metabolic response. In these articles, same of the recent trends and applications of microbial biosensors in environmental monitoring and for use in food and fermentations have been reviewed. This endeavor presents many technological challenges to fabricate new microbial biosensors for other scientific field.