• Korean Journal of Microbiology
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• v.48 no.4
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• pp.293-297
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• 2012

Of the 169 strains of microorganisms stored in Polar and Alpine Microbial Collection of Korea Polar Research Institute, 27 strains were selected for their chitinase activity on ZoBell plates supplemented with 0.4% colloidal chitin. Among them, PAMC 21693 strain have shown the highest chitinolytic enzyme activity toward pNP-$(GlcNAc)_1$ at low temperature and the highest growth rate at $4^{\circ}C$. We cloned a full-length chitinase gene of 2,857 bp which contains an open reading frame of 2,169 bp encoding 872-amino acid polypeptide. Recombinant chitinase protein was expressed in E. coli and its molecular weight was confirmed 96 kDa. In this paper, we suggest the potential use of cold-active chitinase from polar microorganisms in the field of biotechnology.

• KSBB Journal
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• v.11 no.1
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• pp.8-14
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• 1996

Bacillus subtillis strains with transition state regulator mutations and a spore mutation were developed for the overexpression of apsE and for the enhancement of expression level. Among the many regulator genes, degU and hpr were chosen as a representative positive and negative regulator for the aprE, respectively. Spo II G was used for the construction of asporogeneous strains. All the mutants were constructed from two protease-deleted strain DB104 and the apsE gene was transformed with an integration vector pMK101. DB104(deg$U^h$(32) $his^+$)::pMK101(Cm) and DB104($\Delta$her(Em))::pMKl01(Cm) show 7-fold and about 2-fold increase in aprE expression level, respectively. But the effect of transition state regulator mutation on the aprE expression was diminished when the integrated aprE gene was amplified by the high concentration of chloramphenicol, i. e. 30 $\mu\textrm{g}$/ml. DB104($\Delta$spoIIG(Pm) degUh(32) his+)::pMK101(Cm) and DB104($\Delta$spoIIG(Pm) $\Delta$hpr(Em))::pMK101 double mutant show 10-fold and 3-fold increase in aprE expression level, respectively. The results suggest that sporulation mutation and transition state regulator mutation have independent and additive effect on the aprE expression, and the same gene dosage effect on the transition state regulator mutation was also identified.

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.5
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• pp.521-526
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• 2007

The effects of red ginseng powder on hepatotoxicity in $benzo(\alpha)pyrene\;[B(\alpha)P]$-treated mice were investigated. Male ICR mice were pretreated with red ginseng powder (50 or 100 mg/kg/day, for 5 days, intraperitoneally) before treatment with $B(\alpha)P$ (0.5 mg/kg, i.p, single dose). The ability of red ginseng powder to protect against oxidative damage to the mouse liver was examined by determining the level of lipid peroxide, glutathione, and the antioxidant enzyme activities. The glutathione content depleted by $B(\alpha)P$ were significantly increased by red ginseng powder, but elevation of lipid peroxide content induced by $B(\alpha)P$ was decreased by red ginseng powder. The increased activities of superoxide dismutase, catalase and glutathione peroxidase after $B(\alpha)P$-treatment were decreased by the treatment of red ginseng Powder; however, glutathione S-transferase activity depleted by $B(\alpha)P$ were significantly increased. These results suggest that red ginseng powder can protect against $B(\alpha)P$ intoxification through its antioxidant properties.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.45 no.6
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• pp.545-552
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• 2012

This study developed a natural seasoning (NS) and characterized its food components. Hydrolysate from Alaska Pollock Theragra chalcogramma heads and sea tangle Laminaria japonica byproduct were obtained by incubating them with Neutrase for 4 h. NS was prepared by mixing sorbitol 2%, salt 2%, ginger powder 0.04%, garlic powder 0.2%, onion powder 0.2% and inosine monophosphate (IMP) 0.1% based on concentrated hydrolysates from Alaska pollock head and sea tangle byproduct before vaccum filtering. The proximate composition of NS was 82.7% moisture, 9.0% crude protein, and 5.1% ash. It had a higher crude protein content than commercial anchovy sauce (CS), it was lower in moisture and ash. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and angiotensin-I converting enzyme (ACE) inhibiting activity of NS were 90.1% and 88.9%, respectively, which were superior to those of CS. The free amino acid content and total taste value of NS were 1,626.0 mg/100 mL and 165.86, respectively, which were higher than those of CS. According to the results of taste value, the major free amino acids were glutamic acid and aspartic acid. In the sensory evaluation, the color and taste of NS were superior to those of CS. No difference in fish odor between NS and CS was found.

• The Journal of Internal Korean Medicine
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• v.31 no.2
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• pp.254-263
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• 2010

Objectives : The neuroprotective effects of bee venom (BV) have been demonstrated in many studies, but bee venom has many side effects. So we used sweet bee venom (SBV), which has high molecular elements removed to reduce the side effects. I examined the neuroprotective effect of sweet bee venom in 1-methyl-4-phenylpyridine ($MPP^+$)-induced human neuroblastoma SH-SY5Y cells. Methods : To observe the possible toxicity of SBV itself, SH-SY5Y cells were treated with SBV in various concentrations for 3 h and $MPP^+$ in concentrations (1 and 5mM) for 24h. To investigate the protective effect of SBV against $MPP^+$ toxicity, SH-SY5Y cells were pretreated with vehicle or nontoxic concentrations of SBV for 3h and the cells were not washed, followed by incubation with respective concentrations of SBV and 1 mM $MPP^+$ for 24h. To investigate the protective effect of SBV against $MPP^+$ toxicity, SH-SY5Y cells were pretreated with vehicle or nontoxic concentrations of SBV for 3h and the cells were not washed, followed by incubation with respective of SBV(0.5%), 1 mM $MPP^+$, 5uM AKT inhibitor(LY984002) and 10uM ERK inhibitor(PD98059) for 24 h. The protective effect was measured by cell viability assay. To investigate the degree of apoptosis, caspase-3 enzyme activity was measured in control, $MPP^+$, SBV+$MPP^+$. Results : SBV (0.5%) pretreatment protected the SH-SY5Y cells against $MPP^+$-induced apoptotic cell death. The cell viability was higher in the SH-SY5Y cells that were pretreated with vehicle or nontoxic concentrations of SBV than those not pretreated. The caspase-3 activity was lower in the pretreated groups than these not pretreated. ERK and AKT enzymes have a role in the neuroprotective effects of the sweet bee venom. Conclusions : The results demonstrate that SBV has a protective effect on dopaminergic neurons against $MPP^+$ toxicity. This data suggest that SBV could be a potential therapeutic tool for neurodegenerative diseases such as Parkinson's disease(PD).

• The Korean Journal of Nuclear Medicine Technology
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• v.12 no.1
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• pp.62-65
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• 2008

Purpose: Vitamin $B_{12}$ and folic acid are type of water-soluble vitamin and those work as a secondary vital enzyme, but especially those are involved in the nuclear DNA synthesis. Those are mainly measured in order to diagnose megaloblastic anemia and to assess the storage of folic acid during pregnancy. It is generally known that the hemolyzed serum is useless for folic acid and vitamin $B_{12}$, but it is not easy to abide by this information because our lab (Samkwang Medical Laboratories) is reference laboratory. We tested how much the extent of hemolyzed serum had influence on the results. Materials and Method: We performed the tests of vitamin $B_{12}$, folic acid. For the hemolysis effect study, we used 40 cases. According to the level of A, B and C groups, each group had 10 samples which were mechanically hemolyzed serum. Results: Hemolysis did not affect the vitamin $B_{12}$ results. However in case of folic acid, the value increased according to the degree of hemolysis. And severe hemolyzed cases had the highest value (greater than 20 ng/ml). Conclusions: Preventing the hemolysis, it has to be informed that blood-collecting, separation and storage are performed more carefully. As you see from the above results, hemolyzed serum is not proper for folic acid test, and hemolysis does not affect the results of vitamin $B_{12}$. However, for the more accurate results, it is necessary to avoid hemolysis.

• Applied Biological Chemistry
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• v.34 no.2
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• pp.162-167
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• 1991

In an attempt to explore the mechanistic aspects of chilling injury in plants and their defensive measures against the low temperature stress, the time sequential measurements of pyruvate, superoxide radicals$(O_{\overline{2}})$ and antioxygenic enzymes during whole period of injury-inducing treatment were performed using mostly rice seedlings. Pyruvate was substantialy accumulated in leaf tissues during the exposure period to $5^{\circ}C$ of the seedlings ; the relative extent of the accumulation was increased with increasing time of the cold treatment. When the cold-treated plants were translocated to ambient temperature$({\sim}25^{\circ}C)$, the accumulation started to dissipate, concomitantly accompaning a remarkable increase in the $O_{\overline{2}}$ level of tissues. Superoxide dismutase(SOD) and catalase were also activated during post-chilling period, although they showed a considerable lag time for activation. In contrast, glutathione peroxidase, another antioxygenic enzyme in cells, was not activated at all by preceding cold treatment of plants. The uptake of exogenous $O_{\overline{2}}$ by the roots of rice seedlings resulted in increase in the activities of SOD and catalase in root tissues. The supply of $H_2O_2$ to plan st brought about the activation of catalase in situ, while failing to exert any effect on the activation state of glutathione peroxidase. The results obtained in this work suggest that pyruvate accumulation in cells is the direct cause of the overproduction of $O_{\overline{2}}$ and thereby other toxic activated oxygen species, and that SOD and catalase may play a crucial role in the protection of plant cells against active oxygen-mediated chilling injury.

• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.399-403
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• 2003

In order to understand the protection effects of antioxidant enzymes against oxidative stress caused by various environmental stresses, transgenic tobacco (Nicotiana tabacum cv, Xanthi) plants expressing both copper/zinc superoxide dismutase (CuZnSOD) and ascorbate peroxidase (APX) in chloroplasts (referred to as CA plants) were subjected to highlight (1,100$\mu$mol m$^{-2}$ sec$^{-1}$) and chilling at 4$^{\circ}C$. The protection effects of CA plants using leaf discs were compared with those of transgenic plants expressing either CuZnSOD or APX in chloroplasts (SOD plants or APX plants, respectively) and non-transgenic (NT) plants. CA plants showed about 15％ protection in the photosynthetic efficiency (Fv/Fm) of photosystem II relative to NT plants 1 hr after treatment of both highlight and chilling, whereas they showed about 23％ protection in the redox state of P700 in photosystem I at 3 hr after treatment. SOD plants or APX plants showed an intermediate protection effect between CA plants and NT plants. These results demonstrated that the coexpression of CuZnSOD and APX in chloroplasts importantly involves in the protection effects against oxidative stress caused by various environmental stresses.

• Journal of Environmental Impact Assessment
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• v.6 no.1
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• pp.105-110
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• 1997

Sanitary landfill is a general method as a final disposal of municipal solid waste(MSW), therefore leachate characteristics are very various as lime goes by because of highly concentrated organic acids are contained non biodegradable COD. So it is hard to abide by the mandatory standards of discharge eventhough applying the physicochemical and biological processes to treat the leachate. The process of treating leachate are determined by the degree of removal and components, but they are highly contained organic materials. It is a removal method to use jointly with the physicochemical process if the hard and fast rule is needed. The critical components of material are COD, ammonia, salts and heavy metals in the case of treating biologically. Biological process is to use metabolism of microorganism, therefore it is a desirable condition which heavy metals are not contained, because they acting as an inhibitor of enzyme. Of these are contained, organic decomposition and synthetic function of microorganisms decrease significantly. Consequently, this research paper lays emphasis on the concentration of heavy metals in leachate and for the purpose of forecasting the factors which are affecting the leaching of metalic waste in some degree, experimented the various reacting conditions. 1. When the concentration of heavy metals in leachate is in comparison with the level eluted after reaction, at pH 7.9 the result of reaction for PCB to CCL scrap showed that Zn, Mn, Cu was more eluted 11.6 times, 340.3 times, and 2,705.5 times respectively than the leachate undiluted solution. 2. At the condition of strong acid pH 4.7, the concentration of heavy metals in EM undiluted solution showed that Zn, Mn, Cu was more eluted 26.5 times, 147.3 times, and 3,656.3 times respectively than leachate undiluted solution. 3. When the ratio leachate to EM was 50 vs 50(V/V%), Mn was more eluted 198.7 times than leachate undiluted solution, but Zn and Cu do not show the meaningful results. 4. The color of landfill leachate was black-brown. And fulvic acid that is main ingredient of NBD COD contained, oxygen of 44~50%. For that reason, I estimated that the level of Zn, Mn, Cu was higher than the case of leachate. 5. COD of leachate from general landfill is difficult to remove. Because the solution of heavy metals is improved by the character of leachate(pH & ingredient of oxygen etc.) hence the Mn, Cu, Zn act as disturbing factor, the biochemical treatment is hard. Therefore the type of PCB & CCL scrap, iron, aluminum contained metals need to previously separate from general wastes as much as possible.

• Horticultural Science & Technology
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• v.34 no.6
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• pp.912-923
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• 2016

Watermelon production is often limited by powdery mildew in areas with a large daily temperature range. Development of resistant watermelon cultivars can protect against powdery mildew; however, little is known about the characteristics of its causal agents. Here, we identified the genus and race of a causal pathogen of powdery mildew in Ansung province of South Korea, and developed molecular markers for the generation of resistant watermelon cultivars. The causal pathogen was determined to be Podosphaera xanthii based on multiple sequence alignments of internal transcribed spacers (ITS) of rDNA. The physiological race was identified as 1W, and the Ansung isolate was named P. xanthii 1W-AN. Following inoculation with the identified P. xanthii 1W-AN, we found inheritance of the resistant gene fitting a single dominant Mendelian model in a segregated population ('SBA' ${\times}$ PI 254744). To develop molecular markers linked to fungus-resistant loci, random amplified polymorphic DNA (RAPD) was accomplished between DNA pooled from eight near-isogenic lines (NILs; $BC_4F_6$), originated from PI 254744 and susceptible 'SBB' watermelon. After sequencing bands from RAPD were identified in all eight NILs and PI254744, 42 sequence-characterized amplifiedregion (SCAR) markers were developed. Overall, 107 $F_2$ plants derived from $BC_4F_6$ NIL-1 ${\times}$ 'SBB' were tested, and one SCAR marker was selected. Sequence comparison between the SCAR marker and the reference watermelon genome identified three Nco I restriction enzyme sites harboring a single nucleotide polymorphism, and codominant cleavage-amplified polymorphic site markers were subsequently developed. A CAPS marker was converted to a high-resolution melt (HRM) marker, which can discriminate C/T SNP (254PMR-HRM3). The 254PMR-HRM3 marker was evaluated in 138 $F_{2:3}$ plants of a segregating population ('SBA' ${\times}$ PI254744) and was presumed to be 4.3 cM from the resistance locus. These results could ensure P. xanthii 1W-AN resistance in watermelon germplasm and aid watermelon cultivar development in marker-assist breeding programs.