• Analytical Science and Technology
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• v.12 no.1
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• pp.28-33
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• 1999

Glutathione(GSH) in biological samples was determined by high performance liquid chromatographic(HPLC) method with fluorescence detector(FLD) after monobromobimane(MBB) or 4-fluoro-7-sulfobenzofurazan(SBD-F) derivatization. The detection limit of $0.03{\mu}g/mL$ was obtained after MBB derivatization, derivative of MBB was about 200 times more sensitive than that of SBD-F. N-acetylcysteine was used as internal standard and tetrabutylammonium ion as counter ion for better separation. The determination by ion-pairing chromatography after MBB derivatization was characterized by linearity in the range between $0.08{\sim}8.33{\mu}g/mL$ with a good correlation coefficient of 0.998. By precision test appeared relative standard deviation at less than 5% at three different concentrations. This method can be used for the analysis of GSH in plasma and tissue.

• KSBB Journal
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• v.19 no.6 s.89
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• pp.462-466
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• 2004

Mycothiol (MSH), a low molecular antioxidant thiol compound, was purified and analyzed from Streptomyns coelicolor A[3]2 by the monobromobimane fluorescence detection method modified by this lab. Through HPLC chromatpgram, MSH fraction was obtained following the elution time of standard MSH (donated by Dr. Robert C. Fahey). That MSH showed the highest concentration among the thiol compounds contained in the cell indicated that MSH was the key thiol compound having antioxidant activity. To understand the role of gene of inositol monophosphatase (I-1-Pase) involved in the MSH biosynthesis, it was isolated from S. coelicolor A(3)2 and cloned and overexpressed in the Escherichia coli. The expressed I-1-Pase was purified through Ni-NTA column. The soluble protein consisted of 281 amino acids, and the molecular weight was 32 kDa. I-1-Pase of S. coelicolor A(3)2 had the sequence homology with those of human and E. coli by 24 and $25\%$, respectively, and had two conserved domains (mofif A and motif B) which were typical of I-1-Pase.

• Korean Journal of Environmental Agriculture
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• v.28 no.4
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• pp.409-411
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• 2009

Phytochelatins (PCs) are small-sized peptides synthesized by PC synthase (PCS) using glutathione (GSH) as a substrate, and they play an important role in the detoxification of toxic heavy metals in plants, fission yeast, and other living organisms. Recently, it has been suggested that PCS is also involved in degradation of some xenobiotics including monobromobimane. PCS cleaves the Gly residue from GSH-xenobiotics conjugates resulting in ${\gamma}$-Glu-Cys-xenobiotics, and this is to degraded further. Therefore, our research is focus on whether PCS is also involved in degradation of tolclofos-methyl, an important pesticide which has been used in ginseng cultivated areas. Heterologous expression of Arabidopsis PCS confers tolerance to tolclofos-methyl in yeast. Furthermore, PCS-deficient Cad1-3 Arabidopsis mutant showed high sensitivity to tolclofos-methyl compared with wild-type plants. These results imply that PCS is involved in degradation of tolclofos-methyl as other xenobiotics.

• Ocean Science Journal
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• v.43 no.1
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• pp.31-37
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• 2008

As a part of an ongoing search for antioxidants from marine sources, antioxidant activities of 24 kinds of seaweeds (4 green algae, 8 brown algae, and 12 red algae) were investigated. The seaweeds were extracted by acetone/dichloromethane and methanol, respectively. The antioxidant properties of both extracts were evaluated using four different activity tests, including degree of occurrence of intracellular reactive oxygen species (ROS), NO, lipid peroxidation, and GSH (glutathione) in mouse macrophage Raw 264.7 cells. The levels of intracellular reactive oxygen species (ROS) and GSH were measured using 2',7'-dichlorofluorescin diacetate (DCFDA) and monobromobimane as fluorescence probe, respectively. Moreover, the generation of NO and lipid peroxidation products were determined by each method based on the Griess reaction and TBARS assay. Solvent extracts from seaweeds such as Scytosiphon lomentaria, Prionitis cornea, Laruencia okamurae, Callophyllis japonica, Sargassum horneri, Dictyopteris divaricata, Lomentaria catenata, Corallina confuse, Ishige okamurae, and Ahnfeltiopsis flabelliformi exhibited high antioxidant activities in cellular oxidizing systems.