• Biomolecules & Therapeutics
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• v.23 no.5
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• pp.486-492
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• 2015

Drug metabolism mostly occurs in the liver. Cytochrome P450 (CYP) is a drug-metabolizing enzyme that is responsible for many important drug metabolism reactions. Recently, the US FDA and EU EMA have suggested that CYP enzyme induction can be measured by both enzymatic activity and mRNA expression. However, these experiments are time-consuming and their interassay variability can lead to misinterpretations of the results. To resolve these problems and establish a more powerful method to measure CYP induction, we determined CYP induction by using luminescent assay. Luminescent CYP assays link CYP enzyme activity to firefly luciferase luminescence technology. In this study, we measured the induction of CYP isozymes (1A2, 2B6, 2C9, and 3A4) in cryopreserved human hepatocytes (HMC424, 478, and 493) using a luminometer. We then examined the potential induction abilities (unknown so far) of mesalazine, a drug for colitis, and mosapride citrate, which is used as an antispasmodic drug. The results showed that mesalazine promotes CYP2B6 and 3A4 activities, while mosapride citrate promotes CYP1A2, 2B6, and 3A4 activities. Luminescent CYP assays offer rapid and safe advantages over LC-MS/MS and qRT-PCR methods. Furthermore, luminescent CYP assays decrease the interference between the optical properties of the test compound and the CYP substrates. Therefore, luminescent CYP assays are less labor intensive, rapid, and can be used as robust tools for high-throughput CYP screening during early drug discovery.

• Korean Journal of Microbiology
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• v.51 no.2
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• pp.156-168
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• 2015

The objective of this study is to investigate the anti-Helicobacter pylori and anti-cancer activities of the live cells (LC), cell-free culture supernatants (CFCS), and bacteriocin solution (BS) obtained from Lactobacillus acidophilus BK13 and Lactobacillus paracasei BK57 strains. After incubation for 30 h in MRS broth, the concentration of lactic acid produced by L. paracasei BK57 ($155.9{\pm}10.2mM$) was higher than in MRS broth using L. acidophilus BK13 ($126.8{\pm}7.9mM$). Maximum bacteriocin activity (128 AU/ml) of BK13 strain was observed after 30 h of cultivation at $37^{\circ}C$, however its magnitude was significantly lower than that of BK57 strain (256 AU/ml). The LC of L. acidophilus BK13 and L. paracasei BK57 were able to inhibit the growth of H. pylori ATCC 43504 at different incubation times, depending on the initial inoculum of the LAB. These CFCS and BS obtained from BK13 and BK57 strains dramatically inhibited the growth, adhesive ability, and enzymatic activity of H. pylori. Meanwhile, the anti-cancer effect of the lactic acid from L. acidophilus BK13 and L. paracasei BK57 strains on AGS cells had significant differences with the control group. Therefore, these antagonistic substances-producing strains are potentially useful as new potential antimicrobial agents for the management and prevention of H. pylori infections.

• Development and Reproduction
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• v.3 no.1
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• pp.29-38
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• 1999

The pathogenesis of endometriosis is unknown, but retrograde menstruation is widely accepted as an etiology. Refluxed endometrium from endometriosis patients is more prone to implant and invade peritoneum possibly through the action of extracellular proteolysis. This proteolytic action may involve plasminogen activators and the collagenase system. Plasminogen activators (PAs) and matrix metalloproteinases (MMPs) play a critical role in the breakdown of extracellular matrix components and basement membrane in the processes of implantation and tumor invasion. PAs are inhibited by plasminogen activator inhibitor (PAI) and MMPs activity is inhibited by tissue inhibitor of metalloproteinase (TIMP). To test the hypothesis that lower expression of PAI-1 and TIMP-3 in endometrium from women with endometriosis, we investigated their PAI-1 and TIMP-3 expression by quantitative competitive RT PCR in endometrium from women with and without endometriosis. Endometrial tissues were obtained from 14 patients with severe endometriosis and 14 patients without endometriosis. Total RNA was extracted and reverse transcribed into cDNA, and quantitative competitive PCR (QC PCR) was performed to evaluate PAI-1 and TIMP-3 mRNA expression. Endometrium from patients with endometriosis showed decreased expression of PAI-1 and TIMP-3 mRNA compared to endometrium from control in luteal phase (p<0.05). Our results suggest that endometrium from women with endometriosis expresses lower levels of PAI-1 and TIMP-3 than endometrium from normal women. Endometrium from endometriosis patients may be more invasive and prone to peritoneal implantation than control because of higher PA and MMP enzymatic activity. Thus, increased proteolytic activity may be one of the reasons for the invasive properties of the endometrium resulting in the development of endometriosis.

• Korean Journal of Food Science and Technology
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• v.43 no.6
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• pp.735-741
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• 2011

Bealmijang is a short-term fermented regional product that is prepared with soybean and extra ingredients. In this study, starter strain candidates were screened from Bealmijang for fermented soybean paste products. Twenty one bacterial strains producing extracellular enzymes (amylase, cellulase, protease, xylanase and lipase) were isolated from Bealmijang, buckwheat sokseongjang. The isolates were assessed for fibrinolytic and antibacterial activities, and salt tolerance. Strain HJ18-4, identified as Bacillus subtilis (AB601598) by biochemical properties (89.6%) and 16S rDNA sequencing (100%), showed the highest enzymatic, fibrinolytic, and antibacterial activities among the isolates. Although the growth of HJ18-4 was inhibited by the increase of NaCl concentration, the growth still exceeded that of B. subtilis KACC 10114 at 5% and 10% NaCl. These results suggest that B. subtilis HJ18-4 is suitable as a starter for soybean paste manufacture.

• Journal of Life Science
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• v.20 no.11
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• pp.1589-1594
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• 2010

Glutaraldehyde was used to cross-link chitosan beads to immobilize the crude enzyme $\beta$-glucosidase from Exiguobacterium sp. DAU5. The conditions for preparing cross-linking chitosan beads and immobilization such as concentration of glutaradehyde, cross-linking time, immobilization pH and time were optimized. The chitosan beads were cross-linked with 1.5% glutaraldehyde for 1.5 hr. The immobilized $\beta$-glucosidase had an overall yield of 20% and specific activity of 5.22 U/g. The optimized pH and temperature were 9.0 and $55^{\circ}C$, respectively. More than 80% of its activity at pH 7.0-10.0, 80% at $40^{\circ}C$ for 2 hr and 48% at $50^{\circ}C$ for 1 hr, were retained. However, the immobilization product showed higher pH and thermal stabilities than free enzymes. It also showed high hydrolyzing activity on soybean isoflavone glycoside linkage. These results suggest the broad application prospects of immobilization enzymes.

• Korean Journal of Food Science and Technology
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• v.27 no.1
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• pp.86-91
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• 1995

In order to expand the utility of alginic acid in the food industry, we have investigated to prepare low viscous alginic acid using a method for degradation of alginic acid with the enzyme system of Vibrio sp. AL-145. The enzyme showed maximum activity at pH 8.0 and $37^{\circ}C$, and was stable in the pH range 7.5 to 8.5 and at temperature up to $30^{\circ}C$, and 0.5M NaCl needed for the enzyme activity. The viscosity of alginic acid decreased with the reaction time courses regardless of alginic acid and enzyme concentration, and 90% of viscosity decreased with 60 min of reaction time, but the changes of reducing sugar not exhibited. The soluble concentration of partially degradated alginic acid(PDA) in water was about 10%(w/v), and adsorption capacity of $Ca^{2+}$ ion increased with increasing the concentration of PDA. The alcohol concentration on precipitation of PDA was higher than Na-alginic acid.

• Korean Journal of Food Science and Technology
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• v.9 no.3
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• pp.183-189
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• 1977

Studies were made of the continuous production of Pyridoxal 5'-phosphate (pyridoxal-p) on simultaneously immobilized cell column. Whole-cell of Pseudomonas polycolor having high activity of pyridoxine 5'-phosphate (pyridoxine-p) oxidase and Kloeckera sp. No. 2201 having high activity of catalase were used as the enzyme materials. The enzyme sources were entrapped in a polyacrylamide gel. Enzymatic properties of the simultaneously immobilized cells were investigated, comparing with those of the mixed whole-cells of the microorganisms. The simultaneously immobilized cells had higher enzyme activity than singly immobilized cells of Pseudomonas polycolor. From this result, the simultaneously immobilized pyridoxine-p oxidase-catalase system could be available to exert a protective effect upon the pyridoxine-p oxidase by destroying $H_2O_2$ which is a by-product of pyridoxine-p oxidation. The optimum pH was 9.0 for the immobilized cells and the whole-cells. The optimum temperature was $45^{\circ}C$ for the immobilized cells and $40^{\circ}C$ for the whole-cells. The pyridoxine-p oxidase of the immobilized cells were activated by $Hg^{++}$ and some SH-compounds.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.6
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• pp.752-760
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• 2008

Scale-up production of low-trans fat containing conjugated linoleic acid (CLA-TFO) was performed through lipase-catalyzed synthesis. Blend of fully hydrogenated soybean oil, olive oil containing conjugated linoleic acid and palm oil with 1:2:7 ratio was interesterified through Lipozyme RM IM in the 1 L-batch type reactor at $65^{\circ}C$ for 12 hrs, and the physicochemical and melting properties of CLA-TFO were compared with conventional (high trans fat) or commercial low-trans fat shortening. The trans fatty acids content in the conventional shortening (48.8 area%) was much higher than that of low-trans shortening (0.4 area%) and CLA-TFO (0.3 area%+CLA; 7.6 area%). Acid, saponification and iodine values of CLA-TFO were 0.4, 173.9 and 59.0, respectively. Their ${\alpha}$-, ${\gamma}$-tocopherol contents showed 4.7, 1.0 mg/100 g. Differences were observed in the solid fat contents (SFC), melting point of the conventional or low-trans fat and CLA-TFO. Each SFC of conventional, low-trans fat and CLA-TFO was 32.0, 29.3 and 30.4% with melting point of 38.5, 43.0 and $39.5^{\circ}C$ at $35^{\circ}C$, respectively. In texture profile analysis, hardness of conventional, low-trans fat and CLA-TFO was 111.7, 75.2 and 63.8 g.

• Journal of Life Science
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• v.20 no.3
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• pp.326-335
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• 2010

The concentration of phenolics in Phellinus pini (CY001) extracts, expressed as mg of GAEs per g of P. pini fractions, and the EtOAc fraction (436.5 mg GAEs/g) of P. pini had a higher phenolic content than other fractions. Several biochemical assays were used to screen antioxidant properties such as reducing power, 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity, NBT/XO superoxide system and inhibition of DCF/AAPH peroxyl radicals. Among the six mushroom extracts, the EtOAc fraction from P. pini (CY001) showed the most potent DPPH radical, superoxide radical, and peroxyl radical scavenging activities, with $IC_{50}$ values of $11.49\;{\mu}g/ml$, $8.32\;{\mu}g/ml$, and $1.91\;{\mu}g/ml$, respectively. The EtOAc fraction of P. pini (CY001) significantly inhibited enzymatic lipid peroxidation and effectively attenuated LPS-induced NO production of RAW 264.7 cells without cytotoxicity. We also found that the EtOAc fraction had a significant hepato-protectant effect on tacrine-induced cytotoxicity in HepG2 cells. These findings suggest that P. pini (CY001) may have potential as a natural antioxidant, which contains compound(s) with radical scavenging activity.

• Applied Biological Chemistry
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• v.50 no.2
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• pp.87-94
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• 2007

A bacterium which has high enzymatic activities such as amylase, cellulase and protease was isolated from Korean traditional soybean food, doenjang. The isolated bacterium was identified to Bacillus subtilis HS25 by the test of morphological and biochemical properties according to Bergey's Manual of Systematic Bacteriology and API 50 CHL kit, and by the 16S rDNA sequence. The isolated B. subtilis HS25 had a potent antibacterial activity against food born causative or pathogenic bacteria. B. subtilis HS25 is endospore forming cell and contained flagella and abundant viscous material at the out layer of cell wall. It was rod type bacterium $(0.5{\sim}0.8{\times}3{\sim}5{\mu}m)$ having biochemical characteristics such as gram staining(+), catalase(+), oxidase(-) and hydrolysis of esculin(+). The optimal medium compositions for production of antibacterial substance in the B. subtilis HS25 were 1% of soluble starch, 0.5% of yeast extract, 0.5% of peptone and 0.05% of MgCl$_2{\cdot}6H_{2}O$. The optimum temperature and pH of the growth of the B. subtilis HS25 was 35$^{\circ}C$ and pH 7.5, respectively. The antibacterial activity was more high in neutral to a little alkaline pH (6.5-10.5) than in acidic pH. The optimal shaking speed to grow and to produce antibacterial substance of the B. subtilis HS25 was 160${\sim}$200 rpm. The optimal culture time for antibacterial activities of the bacterium were shown to be in the range of 12-36 hr.