• Journal of Applied Biological Chemistry
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• v.55 no.3
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• pp.173-178
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• 2012

Sucrose-glucan glucosyltransferase (Gtf) is an important enzyme involved in the cavity formation process where insoluble glucan is synthesized. In this study, we purified Gtf from Streptcoccus mutans Ingbritt through ammonium sulfate precipitation, Sephadex G-150, CM-Sephadex, and DEAE-Sephadex column chromatographies. A 13-fold of purification was achieved with a total yield of 6.3%. The apparent molecular mass of the enzyme was determined to be 66 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimal pH and temperature were established to be 6.0 and $40^{\circ}C$, respectively. The enzyme activity could be inhibited to 22-59% by 1 mM $Hg^{2+}$, $Cu^{2+}$ and $Al^{3+}$, and to 68% by 1 mM EDTA. It was also inhibited 40% by 2 mM xylitol and 35-45% by 0.05% soluble chitosan, glycol chitosan, and glycol chitin. This is the first report to reveal the inhibition effect of chitin derivatives on Gtf activity, which may be further applicable to develop gargles to overcome cavity.

• Journal of Microbiology and Biotechnology
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• v.19 no.3
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• pp.257-264
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• 2009

A $\beta$-agarase gene, agaB34, was functionally cloned from the genomic DNA of a marine bacterium, Agarivorans albus YKW-34. The open reading frame of agaB34 consisted of 1,362 bp encoding 453 amino acids. The deduced amino acid sequence, consisting of a typical N-terminal signal peptide followed by a catalytic domain of glycoside hydrolase family 16 (GH-16) and a carbohydrate-binding module (CBM), showed 37-86% identity to those of agarases belonging to family GH-16. The recombinant enzyme (rAgaB34) with a molecular mass of 49 kDa was produced extracellularly using Escherichia coli $DH5{\alpha}$ as a host. The purified rAgaB34 was a $\beta$-agarase yielding neoagarotetraose (NA4) as the main product. It acted on neoagarohexaose to produce NA4 and neoagarobiose, but it could not further degrade NA4. The maximal activity of rAgaB34 was observed at $30^{\circ}C$ and pH 7.0. It was stable over pH 5.0-9.0 and at temperatures up to $50^{\circ}C$. Its specific activity and $k_{cat}/K_m$ value for agarose were 242 U/mg and $1.7{\times}10^6/sM$, respectively. The activity of rAgaB34 was not affected by metal ions commonly existing in seawater. It was resistant to chelating reagents (EDTA, EGTA), reducing reagents (DTT, $\beta$-mercaptoethanol), and denaturing reagents (SDS and urea). The E. coli cell harboring the pUC18-derived agarase expression vector was able to efficiently excrete agarase into the culture medium. Hence, this expression system might be used to express secretory proteins.

• Journal of Microbiology and Biotechnology
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• v.22 no.1
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• pp.58-68
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• 2012

An investigation was conducted on the enhancement of production and purification of an oxidant and SDS-stable alkaline protease (BHAP) secreted by an alkalophilic Bacillus horikoshii, which was screened from the body fluid of a unique Korean polychaeta (Periserrula leucophryna) living in the tidal mud flats of Kwangwha Island in the Korean West Sea. A prominent effect on BHAP production was obtained by adding 2% maltose, 1% sodium citrate, 0.8% NaCl, and 0.6% sodium carbonate to the culturing medium. The optimal medium for BHAP production contained (g/l) SBM, 15; casein, 10; $K_2HPO_4$, 2; $KH_2PO_4$, 2; maltose, 20; sodium citrate, 10; $MgSO_4$, 0.06; NaCl, 8; and $Na_2CO_3$, 6. A protease yield of approximately 56,000 U/ml was achieved using the optimized medium, which is an increase of approximately 5.5-fold compared with the previous optimization (10,050 U/ml). The BHAP was homogenously purified 34-fold with an overall recovery of 34% and a specific activity of 223,090 U/mg protein using adsorption with Diaion HPA75, hydrophobic interaction chromatography (HIC) on Phenyl-Sepharose, and ion-exchange chromatography on a DEAE- and CM-Sepharose column. The purified BHAP was determined a homogeneous by SDS-PAGE, with an apparent molecular mass of 28 kDa, and it showed extreme stability towards organic solvents, SDS, and oxidizing agents. The $K_m$ and $k_{cat}$ values were 78.7 ${\mu}M$ and $217.4s^{-1}$ for N-succinyl-Ala-Ala-Pro-Phe-pNA at $37^{\circ}C$ and pH 9, respectively. The inhibition profile exhibited by PMSF suggested that the protease from B. horikoshii belongs to the family of serine proteases. The BHAP, which showed high stability against SDS and $H_2O_2$, has significance for industrial application, such as additives in detergent and feed industries.

• Korean Journal of Pharmacognosy
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• v.40 no.4
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• pp.408-414
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• 2009

This study was performed to investigate the anti-hyperglycemia effects of the Triticum aestivum wheat sprout (TAWS) water extracts in the diabetic mice. Diabetic experimental model was established by intraperitoneal injection of streptozotocin into male Balb/c mice. Mice were divided into five groups: normal (CON), diabetic control (DM), and three experimental groups (DM-100, diabetes with TAWS extracts 100mg/kg; DM-50, diabetes with TAWS extracts 50 mg/kg; DM-25, diabetes with TAWS extracts 25 mg/kg). TAWS extracts were administered orally in diabetic mice. Body weight, food intake, and blood glucose levels were recorded for 12 days and blood insulin levels were measured at the day 12. Oral administration of TAWS extracts reduced slightly food intake and induced a little body weight gain in DM-100 groups. The blood level of glucose was decreased in the dose-dependent manner; 55% in the DM-100 group and 39.7% in the DM-50 group. The blood level of insulin also was improved 10 folds in the DM-100 group and 3.6 folds in the DM-50 group compared to the DM group. The contents of total phenolic compounds and total flavonoids in 1 g dry mass of TAWS extracts were 6.6 mg of tannic acid equivalents and 1.0 mg of 8-hydroquinolline equivalents, respectively. In addition, the antioxidant and DPPH radical scavenging activity of TAWS extracts were 1.2 mM and 1.8 mM ascorbic acid equivalents, respectively. These results suggest that TAWS water extracts could contribute to attenuate clinical symptoms of diabetes mellitus.

• Korean Journal of Remote Sensing
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• v.21 no.1
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• pp.73-81
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• 2005

We construct improved geocentric digital elevation model (DEM), estimate tidal dynamics and ice stream velocity over Sulzberger Ice Shelf, West Antarctica employing differential interferograms from 12 ERS tandem mission Synthetic Aperture Radar (SAR) images acquired in austral fall of 1996. Ice, Cloud, and land Elevation Satellite (ICESat) laser altimetry profiles acquired in the same season as the SAR scenes in 2004 are used as ground control points (GCPs) for Interferometric SAR (InSAR) DEM generation. 20 additional ICESat profiles acquired in 2003-2004 are then used to assess the accuracy of the DEM. The vertical accuracy of the OEM is estimated by comparing elevations with laser altimetry data from ICESat. The mean height difference between all ICESat data and DEM is -0.57m with a standard deviation of 5.88m. We demonstrate that ICESat elevations can be successfully used as GCPs to improve the accuracy of an InSAR derived DEM. In addition, the magnitude and the direction of tidal changes estimated from interferogram are compared with those predicted tidal differences from four ocean tide models. Tidal deformation measured in InSAR is -16.7cm and it agrees well within 3cm with predicted ones from tide models. Lastly, ice surface velocity is estimated by combining speckle matching technique and InSAR line-of-sight measurement. This study shows that the maximum speed and mean speed are 509 m/yr and 131 m/yr, respectively. Our results can be useful for the mass balance study in this area and sea level change.

• Korean Journal of Food Science and Technology
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• v.35 no.2
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• pp.297-301
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• 2003

An inhibitor of squalene synthase, a key enzyme in the cholesterol biosynthetic pathways and a target for improved agents to lower plasma levels of low-density lipoprotein, was sequentially purified from Curcuma longa by acetone extraction, silica gel column chromatography, and sephadex LH-20 column chromatography. Active compound, YUF-01, was successfully purified and analyzed as $C_{20}H_{21}O_6$ by electron ionization mass spectrum. Through $^1H-NMR$ and $^{13}C-NMR$ analyses, YUF-01 was identified as curcumin, which showed strong inhibition of squalene synthase.

• Fisheries and Aquatic Sciences
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• v.9 no.4
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• pp.153-159
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• 2006

The ark shell (or 'blood clam') Scapharca broughtonii is a filter-feeding bivalve that has red blood and inhabits waters approximately 10m in depth off the southern coast of South Korea. This study was part of a larger research project investigating the causes of death and restoration of shellfish resources, which are important aquaculture products in South Korea. We examined physiological responses related to survival, respiration, excretion, and amino acid changes as a result of changes in salinity. The 9-day median lethal salinity ($LS_{50}$) was 16.5 psu with confidence limits of 14.9-18.1 psu. At $25^{\circ}C$, the oxygen consumption and ammonia-nitrogen excretion rates were increased with decreases in salinity. Although the osmolality of individuals was acclimated within 2 h at 26.4 psu and 12 h at 19.8 psu, it took more than 5 days at a salinity of 13.2 psu, whereas no individuals acclimated and all died at a salinity of 6.6 psu. Of the amino acids present in the blood, taurine and alanine increased in response to decreased salinity. Tissues of the gill and the mid-gut gland were affected by decreasing salinity. These data will provide important fundamental information for examining the causes of mass mortality of shellfish in the summer.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.45 no.3
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• pp.151-157
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• 2000

This study was carried out to isolate and identify the maysin and related flavonoid analogues in corn silks. Silks were covered with silk bag to prevent pollination and were sampled at 3-5 days after silking. The silks were filled with 100% MeOH and stored at $0^{\circ}C$ until analysis. The MeOH extracts of corn silks were filtered and concentrated at 35-4$0^{\circ}C$. The ${CH}_2$${Cl}_2$ was added on the concentrated aqueous solution to remove the chlorophyll and lipids. The Cis open column (25mm$\times$54 cm) was washed and activated with serial treatment of 500$m\ell$ of 100% MeOH(twice)longrightarrow75% MeOH longrightarrow50% MeOHlongrightarrow30% MeOHlongrightarrow100% $H_2$O(2 times). The concentrated aqueous solution was applied to the $C_{18}$ column and washed with $H_2O$ several times to remove the sugars and water soluble pigments. Neochlorogenic acid, chlorogenic acid and 4-caffeoylquinic acid were eluted with 10% MeOH, and rhamosyl isoorientin was eluted with 30% MeOH, but maysin was eluted with 50% MeOH from the $C_18$ open column. Collected fractions were analyzed with HPLC by using revers-phase Ultras-phere $C_{18}$ column (4.6$\times$250mm, 5$\mu\textrm{m}$) and $H_2$O (10% MeOH containing 0.1% $H_3$${PO}_4$)/MeOH (100% MeOH containing 0.1% H$_3$PO$_4$) linear gradient from 20% to 90% MeOH for 35 minutes, a flow rate of 1 $m\ell$/min and detection at 340nm. The selected fractions were concentrated and applied to the silicic acid column. Maysin was eluted with 500$m\ell$ of 100% ethyl acetate from the silicic acid column for the first purification, and the purity of collected fractions was about 75%, but the purity from the second purification with the Cis column (1/2 $\times$ 43") was greater than 95%. FAB-MS spectral data was obtained with VG7O-VSEQ VG analytical fast atom bombardment mass (UK). $^1$H-NMR and $^{13}$ C-NMR data were obtained with Bruker DPX 400 MHz NMR spectrometers (German) in DMSO-d$_{6}$ at 400 and 100 MHz, respectively.vely.

• Journal of Food Hygiene and Safety
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• v.34 no.5
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• pp.413-420
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• 2019

In this study, a sample preparation method and a simultaneous determination method by ultra-high performance liquid chromatography coupled with tandem mass spectrometry for 9 isomers of chlorogenic acid and arbutin in fruits were developed. The samples were extracted using 90% methanol (pH 3.0), with the solutions being shaken and then sonicated for 10 min each. After centrifugation at 4,000 rpm for 10 min, the extraction was concentrated under a vacuum at $40^{\circ}C$ using a vacuum evaporator. The residue was dissolved in 5 mL of 5% methanol and filtered through a $0.45{\mu}m$ membrane before UHPLC-MS/MS analysis. The separations were performed on a C18 column with gradient elution of water (containing 0.1% formic acid) and methanol (containing 0.1% formic acid). The specificity, linearity, limit of detection, limit of quantification, accuracy, and precision of the proposed methods were also evaluated.

• The Bulletin of The Korean Astronomical Society
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• v.40 no.2
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• pp.30.3-31
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• 2015

We present a kinematic analysis of 172 likely member galaxies of the Ursa Major Cluster. In order to understand the dynamical state of the cluster, we investigate the correlation of the cluster morphology with rotation, the velocity dispersion profile, and the rotation amplitude parallel to the global rotation direction. Both the minor axis and the rotation are very well-aligned with the global rotation axis in the outer region at half radius (> 0.5 $R_{max}$), but not in the inner region. The cluster exhibits low velocity dispersion and rotation amplitude profiles in the inner region, but higher in the outer. Both profiles exhibit outwardly increasing trends, suggesting an inside-out transfer of angular momentum of dark matter via violent relaxation, as revealed by a recent off-axis major-merging simulation. From Dressler-Schectman plots in the plane of galactic positions, and velocity versus position angle of galaxy, we are able to divide the Ursa Major Cluster into two substructures: Ursa Major South (UMS) and Ursa Major North (UMN). We derive a mass of $3.2{\times}10^{14}M_{\odot}$ for the cluster through the two-body analysis by the timing argument with the distance information (37 for UMN and 36 for UMS) and the spin parameter of ${\lambda}=0.049$. The two substructures appear to have passed each other 4.4 Gyr ago and are moving away to the maximum separation.