• Journal of Life Science
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• v.21 no.1
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• pp.119-126
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• 2011

There is a growing recognition of the significance of $H_2S$ as a biological signaling molecule involved in vascular and nervous system functions. In mammals, two enzymes in the transsulfuration pathway, cystathionine ${\beta}$-synthase (CBS) and cystathionine ${\gamma}$-lyase (CGL), are believed to be chiefly responsible for $H_2S$ biogenesis. Genetic inborn error of CGL leads to human genetic disease, cystathioninuria, by accumulating cystathionine in the body. This disease is secondarily associated with a wide range of diseases including diabetes insipidus and Down's syndrome. Although the human CGL (hCGL) overexpression is essential for the investigation of its function, structure, reaction specificity, substrate specificity, and protein-protein interactions, there is no clear report concerning optimum overexpression conditions. In this study, we report a detailed analysis of the overexpression conditions of the hCGL using a bacterial system. Maximum overexpression was obtained in conditions of low culture temperature after inducer addition, performing low aeration during overexpression, and using a low concentration inducer (0.1 mM, IPTG) for induction. Expressed hCGL was purified by His-tag affinity column chromatography and confirmed by Western blot using hCGL antibody and enzyme activity analysis. We also report that the His tag with TEV site attached protein exhibits 76% activity for ${\alpha}-{\gamma}$ elimination reaction with L-cystathionine and 88% for ${\alpha}-{\beta}$ elimination reaction with L-cysteine compared to those of wild type hCGL, respectively. His tag with TEV site attached protein also exhibits a 420 nm absorption maximum, which is attributed to the binding cofactor, pyridoxal 5'-phosphate (PLP).

• Journal of Life Science
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• v.24 no.9
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• pp.935-945
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• 2014

Persimmon leaves were commonly consumed as beverages, but were also used as popular folk medicine in Asia. The purpose of this work was to assess the biological activities of Diospyros Lotus L. extracts (DLLE). Various solvent extracts, including n-Hexnae, $CHCl_3$, EtOAc, and n-BuOH fractions, were obtained from the methanol extract of Diospyros Lotus L. leaves. The increasing interest in the powerful biological activity of plant phenolics and flavonoids outlined the necessity for determining their content in medicinal herbs. In this study, the total polyphenol and flavonoid contents (TPC and TFC) in the EA fraction were higher than those of other fractions. The biological activities of DLLE were tested using the 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activity assay, as well as superoxide dismutase (SOD) activity as an anti-oxidant effect and ${\alpha}$-glucosidase inhibitory activity as an anti-diabetic effect. The EA fraction with high TPC and TFC values showed the highest anti-oxidant effect and high ${\alpha}$-glucosidase inhibition. The EA fractions were further purified into eight fractions using open column chromatography. Higher anti-oxidant and anti-${\alpha}$-glucosidase activity were observed in polar fractions. The content of the flavonoids, including quercein-3-O-rutinoside, kaempferol-3-O-glucoside, myricetin, luteolin, and kaempferol, were analyzed in effective fractions using high-performance liquid chromatography (HPLC). The results suggest that DLLE have anti-oxidative and anti-diabetic effects and thus, have the potential as anti-diabetic materials and as a source for natural health products.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.11
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• pp.1610-1616
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• 2016

This study aimed to establish an optimal extraction process and high performance liquid chromatography-ultraviolet (HPLC-UV) analytical method for determination of 3,5-dicaffeoylquinic acid (3,5-DCQA) as a part of materials standardization for the development of a xanthine oxidase inhibitor as a health functional food. The quantitative determination method of 3,5-DCQA as a marker compound was optimized by HPLC analysis using a Luna RP-18 column, and the correlation coefficient for the calibration curve showed good linearity of more than 0.9999 using a gradient eluent of water (1% acetic acid) and methanol as the mobile phase at a flow rate of 1.0 mL/min and a detection wavelength of 320 nm. The HPLC-UV method was applied successfully to quantification of the marker compound (3,5-DCQA) in Aster glehni extracts after validation of the method with linearity, accuracy, and precision. Ethanolic extracts of A. glehni (AGEs) were evaluated by reflux extraction at 70 and $80^{\circ}C$ with 30, 50, 70, and 80% ethanol for 3, 4, 5, and 6 h, respectively. Among AGEs, 70% AGE at $70^{\circ}C$ showed the highest content of 3,5-DCQA of $52.59{\pm}3.45mg/100g$ A. glehni. Furthermore, AGEs were analyzed for their inhibitory activities on uric acid production by the xanthine/xanthine oxidase system. The 70% AGE at $70^{\circ}C$ showed the most potent inhibitory activity with $IC_{50}$ values of $77.01{\pm}3.13{\sim}89.96{\pm}3.08{\mu}g/mL$. The results suggest that standardization of 3,5-DCQA in AGEs using HPLC-UV analysis would be an acceptable method for the development of health functional foods.

• Applied Biological Chemistry
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• v.5
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• pp.23-31
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• 1964

Six species of marine shell-fishes were subjects in this study. The content of RNA and DNA in mid-intestinal glands (liver) was determined and RNA was also extracted from above materials by phenol method and their nucleotide compositions were analysed by ion exchange column chromatography. C; citidylic acid, A; adenylic acid, G; guanylic acid, U; uridylic acid, Pu; purine nucleotide, Py; pyrimidine nucleotide 1) Their RNA and DNA content was summarized in the next table.(mg/100mg of lipid free powder) Materials RNA DNA RNA/DNA Meretrix meretrix Susoria (Gmelin) 6.82 0.82 6.5 Anadara(scapharea) inflata(Reeve) 4.62 0.70 6.6 Ostrea(crassostrea) gigas Thunberg 8.74 1.03 8.5 Turbo cornutus Solander 2.16 0.60 3.6 Haliotis gigantea Gmelin 7.02 0.22 36.0 2) Their RNA nucleatide compositions was summaries in following tabe. Materials/RNA nucleotide G+C/A+U G+U/A+C Pu/Py Meretrix meretrix Susoria(Gmelin) 1.12 1.36 1.08 Venerupis philippinarum (Adoms et Reeve) 1.15 1.31 1.13 Anadara(scapharea) inflata(Reeve) 0.97 1.26 1.02 Ostrea (crassostrea) gigas Thunberg 1.51 1.34 1.31 Turbo cornutus Solander 1.37 1.29 1.03 Haliotis gigantea Gmelin 1.33 1.46 1.22 In six species of marine fishes examined, Pu/Py and G+C/A+U ratios of RNA vary in respective wide ranges of 1.02-1.31, 0.97-1.51, while G+U/A+C ratio in the range of 1.26-1.46, not far from 1.3. This G+U/A+C ratio seems to be specific in this species.

• Journal of Food Hygiene and Safety
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• v.29 no.4
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• pp.327-333
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• 2014

A method for analysis of five artificial sweetners (sodium saccharin, aspartame, acesulfame-K, sucralose, cyclamate) in beverage samples was developed using high-performance liquid chromatography/triple quadrupole mass spectrometry (HPLC/MS/MS). The method uses a single-step dilution for sample preperation. Seperation was achieved on a $C_{18}$ column ($2.1{\times}150mm$, $3.5{\mu}m$) with A- 2% methanol (1 mM ammonium acetate), B-95% methanol (1 mM ammonium acetate) as mobile phase with gradient mode. The quantitation of target compounds was performed by external calibration in selected reaction monitorning (SRM) mode. The coefficient of determination of calibration curve for sodium saccharin, aspartame, acesulfame-K, sucralose and cyclamate were 0.9957, 0.9991, 0.9943, 0.9982 and 0.9948, respectively. The limits of detection (LODs) and limits of quantitation (LOQs) were in the range of 0.001~0.022 mg/L and 0.004~0.073 mg/L, repectively. Recoveries for beverage samples were in the range of 92.76~113.50% with RSD < 10.91%. The method has applied to the determination of the five sweetners in 102 beverage samples. Three artificial sweetners-aspartame, acesulfame-K, sucralose were detected from 42 samples. Sodium saccharin and cyclamate were not detected in all samples.

• Food Science of Animal Resources
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• v.27 no.4
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• pp.531-537
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• 2007

Small amounts of trans fatty acids exist naturally in beef and dairy foods. Also, they can be produced in the process of partial hydrogenation to manufacture shortning or margarine. They can provide a better palatability and shelf life. According to the recently studies, trans fatty acids can raise health risk such as heart diseases and coronary artery diseases. They can also increase low-density lipoprotein (LDL) cholesterol and decrease high-density lipoprotein (HDL) cholesterol in the blood plasma, therefore increasing the risk of atherosclerosis and diabetes. The aim of this study was to determine total lipids and trans fatty acids (TFAs) content in processed foods and meat products. The analysis of trans fatty acids was performed in 28 samples of donuts, 18 samples of bakeries, 4 samples of frozen doughs, 2 samples of popcorns, and 4 samples of meat products (ham, sausage, nuget, and bacon). Total lipids in processed foods and meat products were extracted by chloroform-Methanol method and acid digestion, respectively. They were analyzed by gas chromatography using a SP-2560 column and flame ionization detector. The amounts of TFAs per 100 g of foods were 0-3.3% (0.74% on average) in donuts, 0.2-5.8% (1.18% on average) in bakeries, 0.2-6.3% (1.93% on average) in frozen doughs, and 0-5.8% in popcorns. Meat products such as ham, sausage, and nuget analyzed 0.1% of TFAs, respectively and trans fatty acids in bacon were not detected. As a result, the distribution of TFAs in processed foods was widely ranged from O% to 6.3% according to manufacturers and types of products, whereas the content of TFAs in meat products ranged from 0% to 0.1%.

• Korean Journal of Food Science and Technology
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• v.33 no.5
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• pp.641-644
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• 2001

This study was performed for development of new analytical method of monascus pigments in foods. In this method, analysis of monascus pigment in foods has been carried out by detection of monascin and ankaflavin of the main color component of monascus pigment as indicator compounds. Monascin and ankaflavin were isolated and identified by TLC, HPLC, Prep. HPLC, $^{1}H-NMR$ and Mass spectrophotometer. The analysis of monascin and ankaflavin in foods such as massal, sausage, mixed press ham, mixed fish sausage, semi-dried sausage and syrup was performed by using reverse phase high performance liquid chromatograph with Capcell Pak C18 column at wave length 390 nm. The quantitative results of monascin were as follows : $0.01{\sim}3.31\;{\mu}g/g$ item in massal, $0.05{\sim}0.10\;{\mu}g/g$ in mixed fish sausage, and $0.34{\sim}0.35\;{\mu}g/g$ in semi-dried sausage. But the quantitative results of ankaflavin were as follows: $0.02{\sim}0.89\;{\mu}g/g$ in massal, ankaflavin were not founded in other samples.

• Korean Journal of Food Science and Technology
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• v.32 no.5
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• pp.1191-1197
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• 2000

In the preliminary study, we investigated the anti-complementary activities of 62 extracts from Korean edible seaweeds. Of those, Pachymeniopsis elliptica showed the highest anti-complementary activity. Therefore, it was purified as follows; i) PE-1 by ethanol precipitation, ii) PE-1-C by ultrafiltration, iii) PE-1-CIV by DEAE-Toyopearl 650C, and iv) PE-1-CIV-ii by Sepharose CL-6B. The purified compound, PE-1-CIV-ii, was the complexed homogeneous polysaccharide (molecular mass: 780 kDa) with 82.9% of anti-complementary activity. Also, it contained a significant amount of sulfate group (30.5%), which indicated it as a sulfated algal polysaccharide. Its structural monosaccharides were galactose (44.3%), 3,6-anhydrogalactose (34.0%), glucose (8.2%), fucose (5.4%), xylose (5.2%) and rhamnose (2.9%). After the treatment of periodate on a sample, a significant decrease in anti-complementary activity was found, which was a characteristic of bioactive polysaccharides. And-tumor activity of PE-1-A, B and C was tested in the sarcoma-180 solid tumor model. The PE-1-C with the largest molecular mass (more than 300 kDa) showed 81% of inhibition on the solid tumors, suggesting that the anti-complementary activity was, at least in part, related to anti-tumor activity. Based upon these results, the purified polysacchardes could be an immunopotentiator in vivo.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.1
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• pp.85-88
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• 2015

This study attempted to establish an HPLC analysis method for determination of marker compounds as a part of material standardization for the development of health functional food materials from Perilla frutescens Britton var. acuta Kudo. The quantitative determination method of rosmarinic acid as a marker compound of P. frutescens Britton var. acuta Kudo extract (PFE) was optimized by HPLC analysis using a C18 column ($4.6{\times}150mm$, $5{\mu}m$) with 0.1% acetic acid as the elution gradient and methanol as the mobile phase at a flow rate of 1 mL/min and detection wavelength of 280 nm. The HPLC/UV method was applied successfully to quantification of the marker compound in PFE after validation of the method with linearity, accuracy, and precision. The method showed high linearity in the calibration curve at a coefficient of correlation ($R^2$) of 0.9995, and the limit of detection and limit of quantitation were $0.36{\mu}g/mL$ and $1.2{\mu}g/mL$, respectively. Relative standard deviation (RSD) values of data from intra- and inter-day precision were less than 3.21% and 1.43%, respectively. Recovery rate test at rosmarinic acid concentrations of 12.5, 25 and $50{\mu}g/mL$ scored between 97.04~98.98% with RSD values from 0.25~1.97%. These results indicate that the established HPLC method is very useful for the determination of marker compound in PFE to develop a health functional material.

• The Journal of the Petrological Society of Korea
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• v.25 no.3
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• pp.195-209
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• 2016

The understanding of geometric complexity of strike-slip Fault system can be an important factor to control fault reactivation and surface rupture propagation under the regional stress regime. The Kumamoto earthquake was caused by dextral reactivation of the Futagawa-Hinagu Fault system under the E-W maximum horizontal principal stress. The earthquakes are a set of earthquakes, including a foreshock earthquake with a magnitude 6.2 at the northern tip of the Hinagu Fault on April 14, 2016 and a magnitude 7.0 mainshock which generated at the intersection of the two faults on April 16, 2016. The hypocenters of the main shock and aftershocks have moved toward NE direction along the Futagawa Fault and terminated at Mt. Aso area. The intersection of the two faults has a similar configuration of ${\lambda}$-fault. The geometries and kinematics, of these faults were comparable to the Yansan-Ulsan Fault system in SE Korea. But slip rate is little different. The results of age dating show that the Quaternary faults distributed along the northern segment of the Yangsan Fault and the Ulsan Fault are younger than those along the southern segment of the Yansan Fault. This result is well consistent with the previous study with Column stress model. Thus, the seismic activity along the middle and northern segment of the Yangsan Fault and the Ulsan Fault might be relatively active compared with that of the southern segment of the Yangsan Fault. Therefore, more detailed seismic hazard and paleoseismic studies should be carried out in this area.