• Applied Biological Chemistry
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• v.38 no.2
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• pp.118-122
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• 1995

Escherichia Coli ornithine transcarbamylase is the enzyme which catalyzes the L-citrulline biosynthesis from L-ornithine and carbamyl phosphate. To facilitate the purification of enzyme which will be used for many biochemical studies such as structure and function relationships and catalytic mechanisms, the cloning and expression of E. coli argI gene for ornithine transcarbamylase was conducted. argI was amplified from genomic DNA of E. coli strain of $DH5{\alpha}$, by polymerization chain reaction (PCR) method. The amplified argI gene was ligated to the prokaryotic expression vector pKK223-3 and used for transformation of E. coli TB2 which was deficient of ornithine transcarbamylase. The over-produced enzyme by the tnansformant was purified by ammonium sulfate fractionation, heat denaturation and affinity chromatography. The result of SDS denaturation gel electrophoresis for the purified enzyme showed a single band of about 38 kDa of ornithine transcarbamylase. Kinetic data for the expressed enzyme gave almost the s?????? values as those of the wild type enzyme. The $k_{cat}$, of the enzyme was $1.0{\times}10^5min^{-1}$, and $K_ms$ for ornithine and carbamyl phosphate were 0.35 mM and 0.06 mM, respectively.

• Korean Journal of Microbiology
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• v.41 no.4
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• pp.300-305
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• 2005

A transcriptionally active fragment $(P_{19})$ isolated by utilizing the promoter-probe shuttle vector pSK1Cat was analyzed. By subcloning analysis, the 180 bp region $(P_{180})$ responsible for the activity was determined. Transcriptional fusion of the C. glutamicum metX gene to $P_{180}\;(P_{180}-metX)$ resulted in a 24-fold increase in MetX activity in a complex medium, while a 13-fold increase was observed with the $P_{tac}$ promoter. Additionally, the expression conferred by $P_{180}$ was not affected by methionine added to the growth medium, suggesting that the $P_{180}$ clone is useful for the deregulated expression of biosynthetic genes in C. glutamicum during amino acid fermentation. Introduction of $P_{180}-metX$ into a lysine-producing C. glutamicum resulted in the production of methionine to 0.8 g/l.

• Korean Journal of Human Ecology
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• v.15 no.2
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• pp.327-339
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• 2006

We have investigated the distribution of major local foods cultivated in Gwangju City and Jeollanam-Do in order to understand the food culture of Honam region. The local foods were divided into 4 classes: agricultural, aquatic, animal and other products. 124 species of local foods, including 51 agricultural products, 50 aquatic products, 6 animal products and 17 others, were identified from this region. In Jeollanam-Do area, the agricultural products have a variety of fruits and vegetables, and aquatic products have fishes. In particular, rice, cucumber, pear, anchovy, oyster, laver, oyster mushroom, shiitake mushroom, and honey were determined as major local foods in Jeollanam-Do area. Note that Mudeungsan watermelon is cultivated in Gwangju, and sweet smelt and cat fish caught only in Sumjin river and Tamjin river area. In recent years, pumpkin (Cucurbita maxima Duchesne), paprika, kiwi and melon were determined as major local foods in Jeollanam-Do. Furthermore, branding strategy of agricultural and animal products can be said to improve. Wild types of aquatic productions, mushrooms and wild edible greens have been decreasing, whereas cultivated production increasing. The kinds of special local foods in Gwangju City and Jeollanamdo area were influenced by geographical, religious and social factors.

• Journal of Environmental Impact Assessment
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• v.13 no.1
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• pp.21-31
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• 2004

Construction of highway would influence wildlife habitat being fragmented and deteriorated. It is inevitable to observe the death of animals that are killed by vehicles. Nonetheless the cause and status of animal roadkill have not been studied, and this study first attempted to analyze the roadkill in major highway in Korea. We collected 860 individuals of dead animals in Joongang Highway during 1996-2003. Among them Korean hare(Lepus sinensis) was totaled in 165 individuals(19.2%) with the highest figure; Korean racoon dog(Nyctereutes procyonoides) was 146(17.0%); Korean squirrel(Sciurus vulgaris) 56(6.5%); Korean roe deer(Capreolus capreolus) 26(3.0%). Domestic animals was totaled in 232 individuals(27.0%); domestic dog(Canis familiairis) 25, and domestic cat(Catus felis) 207. This study also categorized habitat as mountain-mountain, mountain-plain, mountain-river, plain-plain, plain-river, river-river in each side of the road. Habitat of mountain-plain was the heightest with 296(38.9%) individuals followed by mountain-mountain with 263(34.6%). This indicates that wild animals associated with mountain habitat was most influenced by the construction of roads. This study implied that we should mitigate the roadkill by designing artificial construction such as eco-bridge, fences along the sideway, and boxes under the highway, underdrain structure, etc. The monitoring and cause of habitat fragmentation with GIS approach should be followed to reduce the roadkill.

• BMB Reports
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• v.34 no.1
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• pp.51-58
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• 2001

Attempts using in vitro and in vivo selection procedures have been made to search for hammerhead ribozymes that have higher activities than the wild-type ribozyme and also to determine whether other sequences might be possible in the catalytic core of the hammerhead ribozyme. Active sequences selected in the past conformed broadly to the consensus core sequence except at A9, and no sequences were associated with higher activity than that of the hammerhead with the consensus core, an indication that the consensus sequence derived from viruses and virusoids is probably the optimal sequence [Vaish et al. (1997) Biochemistry 36, 6495-6501]. Recently, during construction of ribozyme expression vectors, we isolated a mutant hammerhead ribozyme, with an insertion of G between A9 and G10.1, that appeared to show significant activity [Kawasaki et al. (1996) Nucleic Acids Res. 24, 3010-3016; Kawasaki et al. (1998) Nature 393, 284-289]. We, therefore, characterized kinetic properties of the G-inserted mutant ribozymes in terms of the NUX rule. We demonstrate that the NUX rule is basically applicable to the G-inserted ribozymes and, more importantly, one type of G-inserted ribozyme was very active with $k_{cat}$, value of $6.4\;min^{-1}$ in 50 mM Tris-HCl (pH 8.0) and 10 mM $MgCl_2$ at $37^{\circ}C$.

• Journal of Microbiology and Biotechnology
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• v.20 no.7
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• pp.1069-1076
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• 2010

A novel laccase from Tricholoma mongolicum was purified by using a procedure that entailed ion-exchange chromatographies on DEAE-cellulose, CM-cellulose, and Q-Sepharose, and FPLC-gel filtration on Superdex 75. The purified enzyme was obtained with a specific activity of 1,480 U/mg-protein and a final yield of 15%. It was found to be a monomeric protein with a molecular mass of 66 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Its N-terminal amino acid sequence was GIGPVADLYVGNRIL, similar to some but also different to other mushroom laccases. The optimum pH and temperature for the purified enzyme were pH 2 to pH 3 and $30^{\circ}C$, respectively. It displayed a low $K_m$ toward 2,7-azinobis (3-ethylbenzothiazolone-6-sulfonic acid) diammonium salt (ABTS) and high $k_{cat}/K_m$ values. The purified laccase oxidized a wide range of lignin-related phenols, but exerted maximal activity on ABTS. It was significantly inhibited by $Hg^{2+}$ ions, and remarkably stimulated by $Cu^{2+}$ ions. It inhibited HIV-1 reverse transcriptase and proliferation of hepatoma HepG2 cells and breast cancer MCF7 cells with an $IC_{50}$ of 0.65 ${\mu}M$, 1.4 ${\mu}M$, and 4.2 ${\mu}M$, respectively, indicating that it is also an antipathogenic protein.

• Korean journal of food and cookery science
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• v.30 no.4
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• pp.369-377
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• 2014

Obesity increases oxidative stress, which could contribute to the development of insulin resistance and hyperglycemia. The purpose of this study was to investigate the hypoglycemic and antioxidant effect of sanchae-namul (SN) in mice with diet-induced obesity. Five-week-old male C57BL/6J mice were fed a basal or high-fat and high-sucrose (HFHS) diet with or without 3% freeze-dried SN powder composed of chamnamul, daraesoon, miyeokchwi, bangpung namul, and samnamul for 12 weeks after a 1-week adaptation. After sacrifice, serum glucose and insulin were measured and the homeostasis model assessment for insulin resistance (HOMA-IR) was determined as well. Hepatic lipid peroxidation, glutathione (GSH), and activities of the antioxidant enzymes were determined. SN given at 3% of the total diet did not significantly influence body weight and food intake in mice fed the HFHS diet. Serum glucose and insulin levels, as well as HOMA-IR values, were significantly lower in the SN group than those in the HFHS group. Thiobarbituric acid reactive substances (TBARS) levels in the liver were decreased significantly in the SN group compared with those in the HFHS group. SN significantly increased the GSH levels and the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) in the liver compared with those in the HFHS group. Overall, these findings suggest that SN may be useful in alleviating insulin resistance and hyperglycemia in mice fed HFHS diet; further, the improvement of insulin resistance could partly occur by reducing the oxidative stress.

• Journal of Korean Society of Environmental Engineers
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• v.36 no.11
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• pp.733-738
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• 2014

Within the scope that does not interfere with the performance of basic duty, the military has been actively supporting the national policy for the conservation of the natural environment in accordance with the "military environmental protection guidelines". Since there is a lack of information regarding inhabitation for wild fauna and flora, field investigations of the natural environment at Jinhae local unit were performed three times in 2003. As a result of the investigation, lycoris sanguinea as flora, marten, otter, and leopard cat as mammalia, and accipiter soloensis, accipiter gentilis schvedow, accipiter gularis, buteo hemilasius, falco tinnunculus and terpsiphone atrocaudata as bird were confirmed to inhabit. For the conservation of the natural ecosystem, the expansion of research for natural ecosystems within the military unit is required. Additionally, environmental pollution can be prevented through the proper maintenance of basic environmental facilities including sewage treatment facilities, soil pollution control facilities, and waste storage facilities. In cooperation with local governments and private organizations, the natural cleansing action and sustainable wildlife conservation should be expanded.

• The Korean Journal of Zoology
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• v.38 no.3
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• pp.324-329
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• 1995

The phylogenetic relationships between Cheju native horses and Tsushima native horses were studied by protein polymorphism analyses in 16 gene loci (Trypsin inhibitor: Ti, Chymotrypsin inhibitor: CTi, Albumin: Al, Esterase: Es, Transferrin: Tf, Hemoglobin: Hb, Catalase: Cat, Esterase D: EsD, Glutamate oxaloacetate transaminase: GOT, Glyoxalase I: GLO I, Acid phosphatase: AcP, Superoxide dismutase: SOD, Lactate dehydrogenase: LDH, Hexokinase: HK, Malate dehydrogenase: MDH, Malic enzyme: ME). All allelic patterns of the protein loci, except 5 loci (SOD, LDH, HK, MDH, ME), were polymorphic in both two populations. Gene frequencies of the polymorphic loci of the population of Cheju native horses were higher than those of Tsushima native horses. Average heterozygosity in Cheju native horses was 0.375, showing higher than that of Tsushima native horses (0.304). The Da distance and gene identity of two populations were 0.108 and 0.868, respectively. The phylogenetic tree constructed by these results and those previously reported in other horse populations, consisted of three clusters. From this phylogenetic tree, it could be suggested that Cheju native horses and Tsushima native horses had diverged from the Mongolian wild horse (Equus prsewolskii).

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.347-355
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• 2016

An exo-β-D-glucosaminidase (AorCsxA) from Aspergillus oryzae FL402 was heterologously expressed and purified. The deduced amino acid sequence indicated that AorCsxA belonged to glycoside hydrolase family 2. AorCsxA digested colloid chitosan into glucosamine but not into chitosan oligosaccharides, demonstrating exo-β-D-glucosaminidase (CsxA) activity. AorCsxA exhibited optimal activity at pH 5.5 and 50℃; however, the enzyme expressed in Pichia pastoris (PpAorCsxA) showed much stronger thermostability at 50℃ than that expressed in Escherichia coli (EcAorCsxA), which may be related to glycosylation. AorCsxA activity was inhibited by EDTA and most of the tested metal ions. A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of V_max and k_cat values with those of the wild-type enzyme. The three-dimensional structure suggested the tightened pocket at the active site of F769W enabled efficient substrate binding. The AorCsxA gene was heterologously expressed in P. pastoris, and one transformant was found to produce 222 U/ml activity during the high-cell-density fermentation. This AorCsxA-overexpressing P. pastoris strain is feasible for large-scale production of AorCsxA.