• KSBB Journal
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• v.13 no.2
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• pp.214-219
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• 1998

Polymerase chain reaction(PCR) was conducted to obtain the gene encoding glucose oxidase(GOD) from Aspergillus niger(ATCC 2110) and the DNA sequence determined was coincided with published GOD sequence from A. niger. Recombinant transforming vector containing GOD and hygromycin B(hyg.B) resistant gene(hph) was constructed and used for further transformation of A. niger ATCC 2110. Selectivity of hyg.B against A. niger differed depending on which media were used i.e., nutrient-rich media such as potato dextrose agar(PDA) and complete medium(CM) showed only 50% growth inhibition at 400 $\mu$m ml$^-1$ of hyg.B while the minimal media inhibited mycelial growth completely at 200 $\mu$m ml$^-1$ of hyg.B. Twenty to sixty putative transformants were isolated from the hyg.B-containing minimal top agar, transferred successively onto alternating selective and nonselective media for a mitotic stability of hyg.B resistance and, then, single-spored. Among the stable transformants, the transformant(GOD1-6) grown by flask culture showed the considerable increase of extracellular GOD activity, which was estimated to the degree of 50% - 100% comparing to that of wild type. Transformation of tGOD1-6 was resulted from integration of the vectors into heterologous as well as homologous regions of the A. niger genome. Southern blot analysis revealed that there were two independent integrations of vector into fungal genome and one into the GOD gene due to homologous recombination. In addition, GOD activity and sodium gluconate production when tGOD1-6 was fed-batch fermented were enhanced 11 fold and 2.25 fold, respectively, compared to that of the wild type.

• Journal of the Korean Society of Food Culture
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• v.19 no.1
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• pp.94-105
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• 2004

The biochemical components of Namcheondlggae, Miryangdlkkae 25, Boradlggae and Ipdlkkae 1 were measured. The samples were extracted with hot water, 60% acetone or 80% ethanol for screening physiological activity. The crude protein content (4.36%) was found in the Miryangdlkkae 25 and calcium content (497.5 mg%) was found in the Namcheondlggae among the tested 4 perilla loaves. Fructose was 30.86 mg% in the Namcheondlggae and free amino acids at all perilla leaves was detected seventeen. In Boradlggae, glutamic acid and alanine were 25.37 and 11.91 mg%. Totally nine non-volatile organic acids were also detected and the contents of malic acid and glutaric acid were 28.34 and 14.57 mg% in Boradlggae. The Miryangdlkkae 25 had the highest vitamin C amount which was 113.24 mg%. Angiotensin converting enzyme (ACE) inhibition activity of 60% acetone extract of Miryangdlkkae 25 was 39.20% when added as addition of 200 ppm level and xanthine oxidase inhibition activity of 80% ethanol extract of Boradlggae was 46.71%. Electron denoting activity of 60% acetone extract from Namcheohndlggae was the strongest inhibition activity as 98.19% when 200 ppm level of the sample extracts were added.

• The Korean Journal of Pharmacology
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• v.21 no.1
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• pp.1-11
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• 1985

The effects of feeding indole, indole-3-carbinol and benzofuran (all at 5 mmole/kg body wt./day) on various hepatic microsomal and cytosolic enzyme activities involved in xenobiotic metabolism have been compared. Benzofuran was found to elevate the activities of many enzymes both in microsomes (e.g., aniline hydroxylase, 7-ethoxycoumarin O-deethylase, p-nitrophenol UDPGA-transferase and epoxide hydrolase) and in cytosol (e.g., glutathione reductase, glutathione S-transferase, NADH:quinone reductase and UDP-glucose dehydrogenase). The structures of indole and indole-3-carbinol are similar to benzofuran except for the substitution of nitrogen with oxygen atom within the furan ring. Results showed that the activities of UDPGA-transferase and NADH:quinone reductase were not elevated by these indole compounds. While the chemical structure of these two indole compounds are identical except for the presence of the carbinol (methanol) group in indole-3-carbinol, there were marked differences in the types and activities of microsomal enzymes that were enhanced. Among the microsomal enzyme activities determined, indole elevated only the NADPH:cytochrome c reductase, while indole-3-carbinol increased several mixed function oxidase and particularly the epoxide hydrolase activities. Based on the chemical structures of tested compounds and the observed results, possible explanations for the mechanisms involved in elevating epoxide hydrolase activity by benzofuran and indole-3-carbinol are discussed.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.16-16
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• 2001

Apoptosis로 일컬어지는 예정된 세포사멸(programmed cell death)은 개별 세포의 입장에서는 곧바로 사멸을 의미하지만, 정상적인 고등 생물의 입장에서는 개체의 발생과 분화하는데 프로그램된 과정이다. 자발적 세포사멸은 다른 조직에 비해 생식 조직인 난소나 정소에서 복잡한 apoptosis 기작들을 가지리라 사료된다. 본 연구는 Bcl-2 family중 apoptotic protein인 Bax에 대해 suppression하는 유전자를 yeast system을 활용하여 돼지 정소와 난소로부터 각각 cDNA library를 구축한 후 탐색하였다. 탐색에 활용된 cDNA library는 돼지의 정소와 난소로부터 mRNA를 분리하여 yeast vector인 pAD-GAL4-2.1에 구축하였고, 마우스 bax 유전자는 gal 1 promoter의 조절 하에 glucose 배지에서는 유도되지 않고, galactose 배지에서만 선택적으로 Bax를 발현할 수 있는 효모 vector(pL19-bax)를 구축하였다. Bax에 의한 apoptosis suppressor를 탐색하기 위해 우선 효모 W303에 pL19-bax를 transform하여 glucose 배지에서 Bax의 발현을 억제하였다. pL19-bax를 가진 효모에 정소와 난소로부터 구축된 cDNA library를 transform 시키고, transform된 효모는 각각 Bax에 의한 toxicity를 저해하는 유전자를 찾기 위해 스크린되었다. 이러한 방법으로 정소 cDNA library 탐색에서는 5 $\times$ $10^{6}$ transformant중 39개, 난소cDNA library 탐색에서는 2 $\times$ $10^{6}$ transformant중 26개의 콜로니가 생존하였다. 이들 콜로니로부터 유전자를 분리하여 분석해 본 결과 여러 그룹으로 분류할 수 있었다. 각 그룹의 관련 유전자는 protein synthesis/degradation 12종, oxidation/reductation 5종, detoxin/ cell cycle promoter 3종, signal transduction/growth factor 5종, 그리고 알려지지 않은 유전자 9종이었다. 그 중, bax-toxicity inhibition에 강력한 survival phenotype을 가지는 유전자(pSEDL)를 동정하였다. 이것은 T3-4-1 콜로니로부터 분리하였는데 140개 아미노산으로 이루어진 인간 SEDL(GenBank, XM_013096) 유전자와 매우 유사한 homology를 가지며, bax와 관련된 기능은 밝혀져 있지 않다. 이외에도 분리된 유전자에는 NADH, thioreduction, 그리고 cytochrome oxidase와 같은 positive 유전자 군이 크로닝되어, Bax를 이용한 효모에서 apoptosis suppressor에 관련된 유전자를 손쉽게 스크린하는 것이 가능하고, 분리된 유전자의 기능을 예측할 수 있어 지금까지 보고된 유전자 크로닝법 보다는 강력한 수단으로 활용될 수 있다는 사실을 시사하였다. 그러나, ORF에 관계없이 Bax 발현에 저항하는 유전자군이 선발된다든지 하는 문제점은 금후 검토가 필요하리라 사료된다.

• Proceedings of the National Institute of Ecology of the Republic of Korea
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• v.4 no.3
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• pp.104-114
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• 2023

In this study, the distribution, bait preference, hosts, symbionts, habitat status, and cytochrome c oxidase subunit-I sequences of the ant Linepithema humile were analyzed. This species habitat comprised approximately 1.5 km along stone crevices, flower gardens, roadsides, and container yards in the vicinity of Busan station, Korea. Feeding activity of this species was confirmed in 9 species of 7 families of plants. Which was approximately 14% of total flora (63 species of 37 families) in the studied area. Particularly, it was observed that Dendranthema indicum and Camellia japonica were the most frequently visited. Further, we verified that they interacted with four species of aphids. We identified 22 species of ants (17 genera, 4 subfamilies) inhabiting the Busan station area, including L. humile, Tetramorium tsushimae, Nylanderia flavipes and the alien species Paratrechina longicornis. In areas where L. humile was the dominant species, the habitat of local native ants was reduced by approximately 30%. Bait preference was ranked in the following order: jelly, water with glucose (25%), and yogurt, with the highest preference for baits containing sugar and protein. The 630-bp cytochrome c oxidase subunit-I sequences of the local L. humile populations were 99% identical to those of L. humile in the National Center for Biotechnology Information database, and the Argentine ants that invaded Korea had a sequence identical to that of other invasive populations from China, Japan, and New Zealand. As L. humile rapidly increases due to domestic adaptation, continuous monitoring and control strategy for eradication are needed to protect domestic biodiversity.

• Toxicological Research
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• v.14 no.2
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• pp.171-176
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• 1998

The biological effects of the water extracts of Rhus Verniciflua Stokes (RVS) were evaluated by protection against hydroxyl radicals. Antioxidative activities were measured using both 1,1-diphenyl-2-picrylhydrazyl (DPPH) and thiocyanate method. Also we used the Glucose oxidase (GO) 20 mU/$\textrm{m}{\ell}$ hydroxyl radical generating system in mouse forebrain cell culture. Water was used for ex-traction from RVS as a solvent which has high polarity especially. In DPPH method, the antioxidative activities of the crude water extract were stronger than any other extracts of low polar-solvents. In the antioxidative effects of mouse forebrain culture using 20 mU/$\textrm{m}{\ell}$ GO, cell viabilities were evaluated 65.6%, 68.8% at 1 $\mu\textrm{g}$. 5 $\mu\textrm{g}$ addition of crude water extracts (30 mg/$\textrm{m}{\ell}$) respectively. 10 $\mu\textrm{g}$ addition of crude water extracts had more than 86.1% cell viabilities, P<0.0l significantly, compared with the group treated with GO alone. In comparison with the antioxidative activities of several commercial antioxidants (ascorbic acid, $\alpha$-tocopherol, catalase, serum), 273 $\mu\textrm{g}$/$\textrm{m}{\ell}$ addition of crude water extracts (300 $\mu\textrm{g}$/$\textrm{m}{\ell}$) showed equivalent antioxidative effect to 25 uM ascorbic acid.

• Preventive Nutrition and Food Science
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• v.17 no.2
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• pp.129-135
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• 2012

GPAR{elidium (G.) amansii is a red alga widely distributed in the shallow waters around East Asian countries. We investigated the effect of G. amansii on lipid accumulation and ROS (Reactive Oxygen Species) production in 3T3-L1 cells. G. amansii extracts dose-dependently inhibited lipid formation and ROS generation in cultured cells. Our results showed that anti-adipogenic effect of G. amansii was due to the reduction in mRNA expressions of PPAR${\gamma}$(peroxisome proliferator-activated receptor-${\gamma}$) and aP2 (adipocyte protein 2). G. amansii extracts significantly decreased mRNA levels of a ROS-generator, NOX4 (nicotinamide adenine dinucleotide phosphate hydrogen oxidase 4), and increased the protein levels of antioxidant enzymes including SOD1/2 (superoxide dismutases), Gpx (glutathione peroxidase), and GR (glutathione reductase), which can lead to the reduction of ROS in the cell. In addition, the G. amansii extract enhanced mRNA levels of adiponectin, one of the adipokines secreted from adipocytes, and GLUT4, glucose uptake protein. Taken together, our study shows that G. amansii extract inhibited lipid accumulation and ROS production by controlling adipogenic signals and ROS regulating genes.

• Biomedical Science Letters
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• v.2 no.2
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• pp.199-209
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• 1996

A new test strip to detect leukocytes using the myeloperoxidase in urine was developed. The reagent strip contains tetramethylbenzine, glucose and glucose oxidase. The detection limit was between 10 cells per 1$\mu$l urine(5 cells/hpf), showing greenish yellow color in the range of 10-25 cells/$\mu$l, green color in the range of 75-250 cells/$\mu$l, greenish blue color in the range of 500 cells/$\mu$l. The result can be obtained within two minute. The performance of the new method was evaluated by comparing the results of microscopic examination and other commercial products. Good correlations were shown between the values obtained by our urine strip and those by other commercial products with 172 urine samples. The results were proven that new methods were useful as primary screening reagents to detect leukocytes in urine.

• Journal of the Korean Society of Food Culture
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• v.34 no.4
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• pp.456-462
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• 2019

This study investigated the bread-making properties of rice bread supplemented with HPMC (hydroxypropyl methylcellulose) 1~3% and gluzyme (glucose oxidase), fungamyl (fungal ${\alpha}$-amylase) and pentopan (xylanase+hemicellulases) (0.0015~0.0090%). The viscoelastic properties of the dough with HPMC 1~3% were analyzed. When the rice flour was supplemented with HPMC 1~3%, the viscoelastic properties of the dough tended to increase as the amount of added HPMC was increased. The physicochemical characteristics of the rice bread with HPMC, gluzyme, fungamyl, and pentopan were analyzed. Supplementing the rice flour with HPMC, gluzyme, fungamyl, and pentopan had a significant effect on the volume (p<0.01) and specific volume (p<0.001) of the rice bread. Supplementing the rice flour with 3% HPMC and 0.0045% or 0.0090% pentopan had a significant effect on increasing the volume (p<0.01) and specific volume (p<0.001) of the rice bread. Supplementing the rice flour with 3% HPMC, 0.0023% gluzyme and 0.0015% fungamyl had a significant effect on increasing the volume (p<0.01) and specific volume (p<0.001) of the rice bread. These results suggest that supplementing the rice flour with HPMC, gluzyme, fungamyl and pentopan is effective for the production of rice bread.

• Advances in Traditional Medicine
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• v.1 no.2
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• pp.59-65
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• 2000

To clarify the toxic effect of oxidative stress, hydrogen peroxide $(H_{2}O_{2})-induced$ neurotoxicity was examined in cultured newborn mouse spinal motor neurons after spinal motor neurons were grown in the media containing various concentrations of glucose oxidase (GO). And also, the protective effect of Rhizoma gastrodiae extract against GO-induced neurotoxicity was evaluated. Cytotoxicity was expressed as a cell viability by 3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) assay. In this study, exposure of motor neurons to GO-induced cell death significantly, in a dose- and time-dependent manners in spinal motor neuron cultures. The decrease in cell viability of motor neurons damaged by GO was proventioned by Rhizoma gastrodiae extract. These results suggest that the neuroprotective effect of Rhizoma gastrodiae extract on GO-induced neurotoxicity may result from a attenuation of $H_{2}O_{2}-induced$ oxidative stress.