• 공업화학
• /
• 제34권2호
• /
• pp.131-136
• /
• 2023

가시광선 영역에서 빛을 흡수하는 고농도 시료의 농도를 색상 좌표 값을 이용해 측정하는 법을 개발하였다. 이성분계 고농도 시료와 색상 좌표 사이의 관계식을 실험식으로 구하고 Resazurin에서 Resorufin으로의 촉매 반응 속도를 평가하였다. 제시한 방법은 자외선가시광선분광법에서 비선형적으로 측정범위 한계를 넘어선 영역의 농도를 시료를 희석하지 않고 직접 측정할 수 있다.

• 공업화학
• /
• 제33권3호
• /
• pp.279-283
• /
• 2022

반응물과 생성물이 자외선 및 가시광선 영역에서 빛을 흡수하는 경우 시료의 농도 및 화학반응의 특성을 평가하는데 자외선가시광선분광법을 사용할 수 있다. 하지만 고농도와 높은 반응 온도 조건에서는 자외선가시광선분광법을 사용하는데 한계가 존재한다. 색을 가지고 있는 시료의 촉매 반응을 색상 분석으로 농도 및 조성을 외부에서 수집한 이미지를 분석하여 자외선가시광선분광법에서 동일한 결과를 얻을 수 있다. Resazurin은 촉매 및 환원제에 의해 resorufin으로 환원되며 농도에 따라 적색변광이 일어나며 카메라를 통해 수집하여 분석할 수 있다. 색상 분석을 위한 색공간은 CIE L^*a^*b^*를 사용하였고 소프트웨어를 통해 각각의 색상좌표 값을 추출하고 각 시료의 농도를 분석하였다. 시료의 농도와 촉매 반응에 대해 색공간을 이용한 분석법과 자외선가시광선분광법의 결과와 비교하여 제시된 방법의 유효성을 확인할 수 있다. 더욱이 색상 분석을 통한 농도 분석에서는 자외선가시광선분광법과 다르게 흡수파장이 중복이 있는 경우에도 디콘볼루션 없이 독립적으로 두 물질의 농도 측정이 가능하다.

• 센서학회지
• /
• 제33권2호
• /
• pp.70-77
• /
• 2024

In this study, a fluorescent ethanol sensor is developed to determine the ethanol concentration in the liquid phase. The sensor is developed using a complex of resazurin (RA)/resorufin (RO) and a hydrotalcite (HT) catalyst in a sol-gel matrix of methyltrimethoxysilane (MTMS) to produce a fluorescent ethanol-sensing membrane (RA/RO^*HT membrane). The operation mechanism of the RA/RO^*HT membrane is based on (i) the oxidation of ethanol to acetaldehyde and (ii) the reduction of RA to RO, through electron flows followed by EtOH ↔ HT ↔ RA/RO ↔ EtOH interactions. These possible redox reactions can lead to an increased fluorescence intensity of the RA/RO^*HT membrane as the ethanol concentration increases. The RA/RO^*HT membrane shows a linear detection range of 1-20 vol.% EtOH with limit of detection (LOD) of 0.178%. Additionally, the RA/RO^*HT membrane has high sensitivity and accuracy for determining the alcohol content in several Korean alcoholic beverages.

• 한국식품과학회지
• /
• 제53권2호
• /
• pp.204-212
• /
• 2021

본 연구에서는 살아있는 프로바이오틱스 균주에 의한 레자주린의 흡광 및 형광특성의 변화와 레소루핀과의 반응성을 분석하고, 균주별 레자주린에 대한 환원능을 비교하였다. LGG에 의해 레자주린은 흡광과 형광의 변화를 수반하며 레소루핀으로 환원되고 반응시간과 생균수의 증가에 따라 환원정도가 증가하였으며, 형광의 변화에서 더 정확하고 민감한 반응성을 보였다. 한편 LGG에 의한 레소루핀으로부터 다이하이드로레소루핀으로의 환원반응은 거의 유발되지 않았다. 프로바이오틱스 6개 균주 중 L. kimchicus의 생균에 의한 레자주린 환원력이 월등하게 높은 반면, 균체 파쇄 후의 ABTS 및 DPPH 라디칼 소거능은 다른 양상을 보이며 L. plantarum과 L. casei가 높은 활성을 나타냈다. 한편 생균의 MTT 환원능은 L. kimchicus가 LGG에 비해 현저히 높아 레자주린 환원능과 유사한 양상을 보였다. 본 연구결과는 레자주린이 단일 균주 프로바이오틱스의 생균수 측정에 유용하며, 프로바이오틱스의 선별 및 환원활성 측정에 활용될 수 있음을 시사한다.

• 한국양식학회지
• /
• 제16권2호
• /
• pp.99-103
• /
• 2003

Olive flounder (Paralichthys olivaceus) was exposed to water-borne phenanthrene for 4 weeks. After the exposure for 2-weeks, hepatic cytochrome P450 contents were significantly elevated. Induction of hepatic ethoxy resorufin-O-deethylase (PROD) activity was significantly increased in flounders treated with 1.0 and 2.0 $\mu$M phenanthrene, compared to untreated group and 0.5 $\mu$M treated group. However, there were no significant changes in pentoxyresorufin-O-deethylase (PROD) activity in hepatic microsome of all the phenanthrene-treated groups, compared to the untreated group. Phenanthrene has the potential to induce cytochrome P450 and EROD enzyme of the olive flounder.

• Food Science and Biotechnology
• /
• 제14권2호
• /
• pp.297-300
• /
• 2005

Effects of allyl sulfides on kinetic behavior of cytochrome P450 1 (CYP1)-catalyzed ethoxyresorufin O-deethylase (EROD) activity were studied using microsomes from benzo[a]pyrene-treated human hepatoma cells. Apparent $K_m$ and $V_{max}$ values were calculated as $2.8\;{\mu}M$ and $3.0\;{\mu}mol$ resorufin/min/mg protein based on Lineweaver-Burk plot of microsomal EROD activity, respectively. Diallyl disulfide (DADS) and diallyl trisulfide (DATS) affected $K_m$ and $V_{max}$ values of EROD activity and acted as mixed-type inhibitors for CYP1 isozymes. Apparent Ki values of DADS and DATS were calculated as 1.07 and 0.88 mM, respectively, by re-plotting slopes of Lineweaver-Burk plot and inhibitor concentrations.

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제10권4호
• /
• pp.367-371
• /
• 2005

About 3,000 bacterial colonies with esterase activities were isolated from soil samples by enrichment culture and halo-size on Luria broth-tributyrin (LT) plates. The colonies were assayed for esterase activity in microtiter plates using enantiomerically pure (R)- and (S)-2-phenylbutyric acid resorufin ester (2PB-O-res) as substrates. Two enantioselective strains (JH2 and JH13) were selected by the ratio of initial rate of hydrolysis of enantiomerically pure (R)- and (S)-2-PB-O-res. When cell pellets were used, both strains showed high apparent enantioselectivity ($E_{app}>100$) for (R)-2PB-O-res and were identified as Exiguobacterium acetylicum. The JH13 strain showed high esterase activity on p-nitrophenyl acetate (pNPA), but showed low lipase activity on p-nitrophenyl palmitate (pNPP). The esterase was located in the soluble fraction of the cell extract. The crude intracellular enzyme preparation was stable at a pH range from 6.0 to 11.0.

• Journal of Microbiology and Biotechnology
• /
• 제17권11호
• /
• pp.1826-1832
• /
• 2007

We describe the fabrication of poly(ethylene glycol) diacrylate (PEG-DA) hydrogel microstructures with a high aspect ratio and the use of hydrogel microstructures containing the enzyme ${\beta}$-galactosidase (${\beta}$-Gal) or glucose oxidase (GOx)/horseradish peroxidase (HRP) as biosensing components for the simultaneous detection of multiple analytes. The diameters of the hydrogel microstructures were almost the same at the top and at the bottom, indicating that no differential curing occurred through the thickness of the hydrogel microstructure. Using the hydrogel microstructures as microreactors, ${\beta}$-Gal or GOx/HRP was trapped in the hydrogel array, and the time-dependent fluorescence intensities of the hydrogel array were investigated to determine the dynamic uptake of substrates into the PEG-DA hydrogel. The time required to reach steady-state fluorescence by glucose diffusing into the hydrogel and its enzymatic reactions with GOx and HRP was half the time required for resorufin ${\beta}$-D-galactopyranoside (RGB) when used as the substrate for ${\beta}$-Gal. Spatially addressed hydrogel microarrays containing different enzymes were micropatterned for the simultaneous detection of multiple analytes, and glucose and RGB solutions were incubated as substrates. These results indicate that there was no cross-talk between the ${\beta}$-Gal-immobilizing hydrogel micropatches and the GOx/HRP-immobilizing micropatches.

• Biomolecules & Therapeutics
• /
• 제6권4호
• /
• pp.351-357
• /
• 1998

In order to understand the mechanism of the regulation of CYPIAI gene expression and ethoxy-resorufin deethylase (EROD) activity in ex vivo system, we have studied the action of TCDD and 3MC in theisolated perfused male rat liver. CYPIAI myNA level and EROD activity were measured in rat liver that wasisolated and perfused with va.ious chemicals such as 2,3,7,8-tet.achlorodibenzo-p-dioxin (TCDD), 3-methyl-cholanthrene (3MC), $17{\beta}$-est.adios ($E_2$), morin. TCDD or 3MC alone perfusion into male rat liver resulted in increase of CYPIAI mRNA level and the magnitude of stimulation was one and half times higher with TCDD treatment than 3MC treatment. However $E_2$ perfusion into male rat liver showed slight stimulation of CYPIAI mRNA level. When $10_{-8}$ M $E_2$ was perfused concomitantly with either $10_{-9}$ M TCDD or $10_{-9}$ M 3MC, stimulated CYPIAI mRNA by either TCDD or 3MC was inhibited. Morin was examined for its effects on CYPIAI mRNA level and result was similar to that was observed with estrogen except that morin alone did not change the level of CYPIAI mRNA. EROD activity was also stimulated with either TCDD or 3MC perfusion, and the magnitude of EROD stiumlation was similar to that of CYPIAI mRNA stimulation in response to TCDD or 3MC perfusion. This data is different from the data that we have obtained with female rat liver. Concomitant perfusion either $E_2$ or morin with TCDD or 3MC inhibited 3MC perFusion or TCDD perfusion stimulated EROD activity. These data confirm the hypothesis that TCDD and 3MC might act through the same mechanism of action on the regulation of CYPIAI gene expression in male rat liver.

• 약학회지
• /
• 제52권5호
• /
• pp.376-383
• /
• 2008

Chungkookjang (CKJ) is a fermented soybean product and one of favorite traditional foods in Korea. In this study, the alcoholic extract from Korean fermented soybean (CKJ) and its one of major flavonoids, genistein were evaluated for their protective effect against B(a)P induced cytotoxicity and DNA damage in HepG2 cells. CKJ extract and genistein decreased B(a)P-induced cell cytotoxicity. CKJ extract inhibited DNA single strand breaks evaluated by single cell gel electrophoresis. From RT-PCR study, it was revealed that CKJ extract decrease DNA damage induced in HepG2 cells expressing CYP1A1 and 1A2 by B(a)P. The metabolizing activities of CYP1A1 and CYP1A2, as measured by the 7-alkoxy resorufin O-deethylation (AROD) assay, showed that CKJ extract and genistein inhibited CYP1A1 and CYP1A2 activities. Genistein may contribute to these biological effects of CKJ extract at least in part. All these results indicate that CKJ extract and genistein may be useful for protection against B(a)P-induced cytotoxicity and DNA damage. Therefore, the alcoholic extract of Korean fermented soybean (CKJ) is suggested to be promising functional food which can prevent the cellular genotoxicity of dietary and lifestyle related carcinogens.