• Bulletin of the Korean Chemical Society
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• 제25권8호
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• pp.1195-1201
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• 2004

Tyrosinase was covalently immobilized on platinum electrode according to the method we developed for laccase (Bull. Korean Chem. Soc. 2002, 23(7), 385) and p-chlorophenol, p-cresol, and phenol could be detected with sensitivities of 334, 139 and 122 nA/ ${\mu}M$ and the detection limits of 1.0, 2.0, and 2.5 ${\mu}M$, respectively. The response time ($t_{90\%}$) is 3 seconds for p-chlorophenol, and 5 seconds for p-cresol and phenol. The optimal pHs of the sensor are in the range of 5.0- 6.0. This sensor can tolerate at least 500 times repeated injections of p-chlorophenol with retaining 80% of initial activity. In case of tyrosinase and laccase co immobilized platinum electrode, the sensitivities are 560 nA/ ${\mu}M$ for p-phenylenediamine (PPD) and 195 nA/ ${\mu}M$ for p-chlorophenol, respectively. The sensitivity of the bi-enzyme sensor for PPD increases 70% compared to that of only laccase immobilized one, but the sensitivity for p-chlorophenol decreases 40% compared to that of only tyrosinase immobilized one. The sensitivity increase for the bi-enzyme sensor for PPD can be ascribed to the additional catalytic function of the co-immobilized tyrosinase. The sensitivity decrease for p-chlorophenol can be explained by the “blocking effect” of the co-immobilized laccase, which hinders the mass transport through the immobilized layer. If PPD was detected with the electrode that had been used for p-chlorophenol, the sensitivity decreased 20% compared to that of the electrode that had been used only for PPD. Similarly, if p-chlorophenol was detected with PPD detected electrode, the sensitivity also decreased 20%. The substrate-induced conformation changes of the enzymes in a confined layer may be responsible for the phenomena.

• Carbon letters
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• 제11권3호
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• pp.216-223
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• 2010

Two-types of ionically modified multi-walled carbon nanotube (MWNTs) based sensors were developed by radiationinduced graft polymerization using vinyl monomers such as 3-(butyl imidazol)-2-(hydroxyl)propyl methyl methacrylate and 1-[(4-ethenylphenyl)methyl]-3-buthyl-imidazolium chloride with ionic properties, in aqueous solution at room temperature. Subsequently, the tyrosinase-immobilized biosensor was fabricated by a hand-casting of the ionic property-modified MWNTs, tyrosinase, and chitosan solution as a binder onto ITO glass surface. The sensing ranges of the tyrosinase-biosensor for phenol in phosphate buffer solution was in the range of 0.005~0.2 mM. The total phenolic compounds mainly such as caffeine of the tyrosinase-immobilized biosensor for commercial coffee were also determined.

• 분석과학
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• 제10권1호
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• pp.66-74
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• 1997

지름 2.54cm, 길이 10cm인 유리관에 tyrosinase(EC. 1.14.18.1)를 입자의 크기 $550{\mu}m$인 탄소에 고정시켜 충진하고, 페놀과 산소를 기질로 사용하여 tyrosinase의 반응 특성을 조사하기 위해 axial dispersion 모델을 제안하였다. 본 논문에서 페놀의 농도는 55.5mM로 고정시키고 산소(2.7ppm, 5.4ppm, 그리고 9.5ppm)와 유속 (1~3mL/s)을 변화시키면서 탄소에 고정된 tyrosinase의 반응을 관찰하였다. 또한, Damkolher수를 계산하고 분산 특성과 식으로부터 효소반응 속도 및 분산의 영향을 예측하기 위해 수치적 해석을 하였다. 연구 결과 물질저항은 주로 외부 전달과 내부확산이었으며, 제안된 모델에서 Biot수는 64.25였다. 페놀은 1.0mL/s 정도의 느린 속도에서 산소의 농도가 높을수록 높은 전환율을 나타내었다. 한편, axial dispersion 모델과 plug flow 모델의 비교에서는 모두 같은 전환율을 나타내어 axial dispersion 모델이 반응속도와 무관함을 알 수 있었다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제7권4호
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• pp.212-215
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• 2002

Tyrosinase transcript In the blood Is known as the marker of malignant melanoma and it has been often determined by using reverse transcription-polymerase chain reaction (RT-PCA) . However, after the PCR process, the quantification of amplified CDMA by the gel electrophoresis is not reliable and time-consuming. for this reason, we tried to quantify the PCR product using a cuvette-type biosensor, where the oligonucleotide probe was immobilized on the cuvette surface and the single strand CDMA, the denatured PCH product, was then hybridized onto the immobilized probe to give a response signal. The response was Immediate and takes 15 min to obtain a stable signal. The biosensor was much more sensitive comparing to the gel electrophoresis method. The quantification of PCR product using a cuvette-type biosensor was feasible and rapid.

• Korean Chemical Engineering Research
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• 제54권6호
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• pp.817-821
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• 2016

Levodopa or L-3,4-dihydroxyphenylalanine (L-DOPA) is the direct precursor of the neurotransmitter dopamine. L-DOPA is a well-known neuroprotective agent for the treatment of Parkinson's disease symptoms. L-DOPA was synthesized using the enzyme, tyrosinase, as a biocatalyst for the conversion of L-tyrosine to L-DOPA and an electrochemical method for reducing L-DOPAquinone, the product resulting from enzymatic synthesis, to L-DOPA. In this study, three electrode systems were used: A glassy carbon electrode (GCE) as working electrode, a platinum, and a Ag/AgCl electrode as auxiliary and reference electrodes, respectively. GCE has been modified using electropolymerization of pyrrole to facilitate the electron transfer process and immobilize tyrosinase. Optimum conditions for the electropolymerization modified electrode were a temperature of $30^{\circ}C$ and a pH of 7 producing L-DOPA concentration 0.315 mM. After 40 days, the relative activity of an enzyme for electropolymerization remained 38.6%, respectively.

• 폴리머
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• 제36권4호
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• pp.470-477
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• 2012

다양한 아민 그룹으로 개질된 다중벽 탄소나노튜브(이하 MWNT) 지지체를 기반으로 하여 페놀 화합물을 검출하기 위한 티로시나아제가 고정된 바이오센서를 개발하였다. 방사선 중합법을 이용하여 MWNT에 글리시딜메타크릴레이트를 중합한 후 중합 사슬의 아미노화 반응을 통해 MWNT에 다양한 아민 그룹을 도입시켰다. 이렇게 제조된 물질의 물리적, 화학적 특성은 SEM, XPS 그리고 TGA에 의해 평가되었다. 그리고 제조된 물질을 기반으로 제작된 티로시나아제가 고정화된 바이오센서의 전기화학적 특성도 평가하였다. 본 효소 바이오센서는 0.1-0.9 mM의 페놀을 검출할 수 있다. 결합효과, pH, 온도 그리고 다양한 페놀화합물에 대한 반응과 같은 여러 가지 변수에 대하여도 최적화하였고 상용 레드와인에서의 페놀화합물 검출도 연구하였다.

• 분석과학
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• 제29권1호
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• pp.35-42
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• 2016

A new nano-composite carbon ink for the development of disposable dopamine (DA) biosensors based on screen-printed carbon electrodes (SPCEs) is introduced. The method developed uses SPCEs coupled with a tyrosinase modified nano-composite carbon ink. The ink was prepared by an “in-house” procedure with reduced graphene oxide (rGO), Pt nanoparticles (PtNP), and carbon materials such as carbon black and graphite. The rGO-PtNP carbon composite ink was used to print the working electrodes of the SPCEs and the reference counter electrodes were printed by using a commercial Ag/AgCl ink. After the construction of nano-composite SPCEs, tyrosinase was immobilized onto the working electrodes by using a biocompatible matrix, chitosan. The composite of nano-materials was characterized by X-ray photoelectron spectroscopy (XPS) and the performance characteristics of the sensors were evaluated by using voltammetric and amperometric techniques. The cyclic voltammetry results indicated that the sensors prepared with the rGO-PtNP-carbon composite ink revealed a significant improvement in electro-catalytic activity to DA compared with the results obtained from bare or only PtNP embedded carbon inks. Optimum experimental parameters such as pH and operating potential were evaluated and calibration curves for dopamine were constructed with the results obtained from a series of amperometric detections at −0.1 V vs. Ag/AgCl. The limit of detection was found to be 14 nM in a linear range of 10 nM to 100 µM of DA, and the sensor’s sensitivity was calculated to be 0.4 µAµM⁻¹cm⁻².

• 센서학회지
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• 제17권1호
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• pp.29-34
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• 2008

A new disposable amperometric tri-enzyme biosensor for the detection of paracetamol has been developed. The paracetamol sensors developed uses horseradish peroxidase modified screen-printed carbon electrodes (HRP-SPCEs) coupled with immobilized enzymes, tyrosinase and aryl acylamidase, prepared using a poly (vinyl alcohol) bearing styrylpyridinium groups (PVA-SbQ) matrix. Optimization of the experimental parameters has been performed and the paracetamol biosensor showed detection limit for paracetamol is as low as $100{\mu}M$ and the sensitivity of the sensor is $1.46nA{\mu}M^{-1}cm^{-2}$.

• KSBB Journal
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• 제10권4호
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• pp.468-474
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• 1995

Glucose biosensor용 glucose oxidase 고정화 막의 단일화 벚 일회용의 간편화를 위한 연구로서, glucose oxidase의 고정화 대상막으로는 CTA와 PCL의 촌합비율이 80/20인 혼합막을 사용한 고저화 방법 중에서는 glutaraldehyde one-steD 방법으로 효소고정화층의 건조 전 두께가 $10{\mu}m$ 인 효소고정화 막이 효과적이었다. 이 고정화 막을 천자현미쇠 으로 관찰해 본 결과, 고밀도의 CTA/PCL 막층 위에 GOD-glutaraldehyde 층이 건조 후 $3{\mu}m$ 정도로 덮여 있는 것을 알 수 있었다. Dissolved oxygen전극을 사용하여 glucose농도의 증가에 따른 전류세기의 차이 값을 측정해 본 결과, glucose 7mM의 안도범위 내에서 선형성을 나타냈으므로 이 고정화원이 glucose sensor용 효소고정화 방법으로 가장 적합하였다. 한편, tyrosmase는 CTA와PCL의 혼 합비율이 80/20인 혼합막에 direct CDI 방법으로 고정화한 것이 가장 효과적이었으며, 8일 후에도고 정화된 tyrosmase의 활성이 35% 이상 유지되었다.

• Bulletin of the Korean Chemical Society
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• 제24권8호
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• pp.1188-1192
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• 2003

In order to study the role of residue in the active site of glutathione S-transferase (GST), Tyr 108 residue in human GST P1-1 was replaced with alanine, phenylalanine and tryptophan by site-directed mutagenesis to obtain mutants Y108A, Y108F and Y108W. These three mutant enzymes were expressed in Escherichia coli and purified to electrophoretic homogeneity by affinity chromatography on immobilized GSH. The substitutions of Tyr108 significantly affected $K_m^{CDNB}$ and $K_m^{ETA}$, whereas scarcely affected $K_m^{GSH}$. The substitutions of Tyr108 also significantly affected $I_{50}$ of ETA, an electrophilic substrate-like compound. The effect of these substitutions on kinetic parameters and the response to inhibition suggests that tyrosine 108 in hGST P1-1 contributes to the binding of the electrophilic substrate and a major determinant in the binding of CDNB is the aromatic ring of Tyr108, not its hydroxyl group.